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Edward's Blog
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The dissolution of sulfide minerals such as pyrite (FeS2), arsenopyrite (FeAsS), chalcopyrite (CuFeS2), sphalerite (ZnS), and marcasite (FeS2) yields hot, sulfuric acid-rich solutions that contain high concentrations of toxic metals. In locations where access of oxidants to sulfide mineral surfaces is increased by mining, the resulting acid mine drainage (AMD) may contaminate surrounding ecosystems. Communities of autotrophic and heterotrophic archaea and bacteria catalyze iron and sulfur oxidation, thus may ultimately determine the rate of release of metals and sulfur to the environment. AMD communities contain fewer prokaryotic lineages than many other environments. However, it is notable that at least two archaeal and eight bacterial divisions have representatives able to thrive under the extreme conditions typical of AMD. AMD communities are characterized by a very limited number of distinct species, probably due to the small number of metabolically beneficial reactions available. The metabolisms that underpin these communities include organoheterotrophy and autotrophic iron and sulfur oxidation. Other metabolic activity is based on anaerobic sulfur oxidation and ferric iron reduction. Evidence for physiological synergy in iron, sulfur, and carbon flow in these communities is reviewed. The microbial and geochemical simplicity of these systems makes them ideal targets for quantitative, genomic-based analyses of microbial ecology and evolution and community function.
Acidic, metal-rich fluids are formed by chemical weathering of metal sulfide-rich rocks. These acid rock drainage (ARD) solutions are hot because metal sulfide oxidation reactions are highly exothermic. The predominant metal sulfide mineral in most rocks is pyrite (FeS2). Pyrite-rich deposits are often mined for metals such as Au, Ag, Cu, Zn, and Pb, which are typically present as impurities in pyrite or occur in sulfide minerals such as chalcopyrite (CuFeS2), sphalerite (ZnS), and galena (PbS). Mining increases the surface area of sulfide ores exposed to air and water, thus, increases rates of acid generation. Regions where rocks have low buffering capacity generate highly acidic toxic solutions that are referred to as acid mine drainage (AMD).
Despite the extreme acidity, heat, and high concentrations of sulfate and toxic metals, a diverse range of microorganisms populate AMD environments. These organisms can form a chemoautotrophically-based biosphere in the subsurface, ultimately sustained by electron donors derived from sulfide minerals, CO2, O2, and N2 derived from air, and phosphate liberated by water–rock interaction. Microbial activity increases the rate of AMD formation and may be responsible for the bulk of AMD generated [1].
Microbe–mineral interactions are of importance because AMD is a very widespread environmental problem. The organisms can be used in ore processing and are a source of novel biomolecules (especially enzymes) for industrial processes.
DNA-based studies of organisms populating mining environments have provided insights into the diversity of acidophilic, metal-tolerant species. Here, we review the importance of archaeal and bacterial lineages, and integrate microbiological, geochemical, mineralogical, and molecular information necessary for quantitative descriptions of the ecology of AMD. Eukaryotes (protists, fungi, and yeasts) are abundant and important in some parts of acid systems. However, due to the paucity of data on eukaryotes in AMD, our review focuses primarily on the prokaryotic components of these communities. We show that the prokaryotic richness of acidophilic communities is low compared to other extremophile and non-extremophile assemblages, yet the species are broadly distributed across the tree of life. Because of their biological and geochemical simplicity, AMD environments have potential as model systems for analysis of biogeochemical interactions and feedbacks and microbial community structure and function.
Dissolution of sulfide minerals
Many factors impact AMD generation. In part, rates of dissolution reactions are determined by fluid chemistry and flow, mineral and rock type, and temperature. The rate of supply of oxidant to the mineral surface influences the rate at which pyrite dissolves. The typical oxidants are oxygen and ferric iron. The concentration of oxygen in groundwater is very small compared to the large requirement for O2 in the overall reaction:2].
Geochemical studies have established that oxygen is a less effective sulfide oxidant than ferric iron. Thus, the dominant pathway for pyrite dissolution involves oxidation of ferrous iron by oxygen:Eqs. 2 and 3, required to describe the sustainable process, yields the reaction in Eq. 1. Ferrous iron oxidation by O2 at low pH is slow, thus the rate of the reaction in Eq. 2 may limit the rate of AMD generation. However, iron-oxidizing prokaryotes catalyze ferrous iron oxidation, thus can determine the rate of pyrite dissolution [1,3]. The feedback between metabolic activity and mineral dissolution can drive the pH down to values <2, thus selecting for community members optimized for life in acid.
Physiological experiments have shown that AMD microorganisms can impact rates of sulfur oxidation during dissolution of pyrite, arsenopyrite, chalcopyrite, marcasite, and sphalerite [4,5]. Oxidation of sulfide ions to sulfate occurs via a series of intermediate sulfur-bearing compounds. Because protons are generated in the subset of reactions that add oxygen to the reduced sulfur species, microbial utilization of sulfide and intermediate sulfur compounds can significantly affect acidification, as well as pyrite dissolution rates.
There is debate about the details of the mechanisms by which microbe–sulfide mineral interactions occur. Pyrite oxidation has been proposed to proceed via indirect and direct pathways [1]. Increase in the rate of sulfide mineral dissolution due to oxidation of aqueous ferric iron is typically referred to as an 'indirect' mechanism. Oxidation of intermediate sulfur species can occur on the mineral surface or in solution. In contrast, 'direct' catalysis is used to describe the possibility of a direct enzymatic interaction with ions bound to the pyrite surface [6].
Enzymatic oxidation via a direct mechanism requires that the cells are either in close proximity or attached to the solid surface. Larsson et al. [7] showed that close proximity of the archaeon Acidianus brierleyi to pyrite was necessary for optimal growth and oxidation rates. Cell-sized pits, often observed on pyrite surface after reaction with Acidithiobacillus ferrooxidans[8], have contributed to speculation about a 'direct' enzymatic pathway.
Experiments were conducted by Edwards et al. [9] to determine whether cell attachment was necessary to generate cell-sized pits. High-resolution scanning electron microscopy (SEM) was used to characterize dissolution patterns on sulfides by Ferroplasma acidarmanus (at 37°C) and Acidithiobacillus ferrooxidans (at 25°C). Elliptical pits (1–2 μm) formed on pyrite surfaces in the presence of these species. However, they also formed in abiotic experiments with just Fe3+, indicating that the presence of cells was not required for pit formation. Cell-sized pits were not observed on marcasite or arsenopyrite reacted with A. ferrooxidans, or on marcasite surfaces reacted with F. acidarmanus. However, rounded F. acidarmanus cells were found within round, deep pits (<0.5 μm) in arsenopyrite surfaces, clearly indicating enhanced dissolution in proximity to cells.
Fowler et al. [10] used constant ferric and ferrous ion concentrations in continuous flow reactors, with and without Acidithiobacillus ferrooxidans, to determine if that organism enhances leaching of pyrite by a mechanism other than increasing Fe3+ in the reactor. Dissolution rates with cells were faster, implying the presence of leaching enzyme(s) or a localized accumulation of Fe3+ at the cell surface.
Edwards et al. [11] conducted surface colonization experiments with pyrite in situ at Iron Mountain, CA, USA and in the laboratory using enrichments from the site. They found that bacteria tended to orient parallel pyrite surface steps in {110} and {100} orientations. Edwards et al. [5] also found that attachment of Acidithiobacillus caldus to pyrite, marcasite, and arsenopyrite was non-random. No cells attached to quartz inclusions within pyrite, indicating that attachment is mineral specific [12]. These observations suggest that cells are not simply using Fe2+ in solution, but exhibit chemotaxis, probably to gradients in [Fe2+]aq outward from dissolving surfaces.
In addition to 'indirect' and 'direct' approaches to sulfide mineral dissolution, cells may make soluble organic molecules to shuttle electrons from insoluble elemental sulfur on pyrite surfaces to cell-associated enzymes [13]. It is also possible that oxygen and hydroxyl radical species are important in some sulfur transformations [2]. Alternatively, Sand et al. [6] proposed that cells attach to the mineral surface and accumulate Fe3+ in their outer membranes, causing local enhanced reactivity via a non-enzymatic direct mechanism.
Microorganisms accelerate iron oxidation rates at low pH by five orders of magnitude [14], thus can increase rates of pyrite dissolution by the reaction in Eq. 3. Numerous microbial pyrite dissolution rates have been reported. Typical experimental values range between 10−5 and 10−7 mol m−2 s−1 for microbial and ferric iron experiments compared to 10−6 to 10−9 mol m−2 s−1 for inorganic oxygen-mediated rates [11].
The degree to which microorganisms enhance sulfide mineral dissolution is determined by the number of iron-oxidizing cells present and the level of activity of the cells. These parameters are not incorporated in most reported microbial rates. Edwards et al. [11] determined the increase in release of iron from pyrite in two enrichments containing Leptospirillum ferriphilum to be 5×10−18 mol Fe cell−1 s−1, comparable to the Thiobacillus ferrooxidans iron oxidation rate of 6×10−18 mol Fe cell−1 s−1[15]. Using cell-normalized rates, it is possible to estimate the magnitude of the microbial impact in a natural environment if information about numbers of iron-oxidizing microorganisms is available. Using this approach, Edwards et al. [5] suggested that microbial activity accounts for about 75% of the AMD produced.
It is essential to define the microbial population in order to analyze an ecological system. Currently, the primary method of determining which organisms comprise a microbial community involves determining the sequence of 16S ribosomal RNA genes (16S rRNA) from environmental samples. This approach provides information about species richness as well as the evolutionary relationships between lineages. Microbial species composition can then be correlated with environmental data to determine how communities are shaped by geochemical factors.
Any AMD system has many microbial niches. Variations in temperature, ionic strength, and pH yield communities characterized by different mixtures of organisms, but all habitats are restricted in membership to a few species (Table 1) [16,17]. This can likely be attributed to the limited number of energy-deriving reactions available in AMD environments. The number of substrates multiplied by the number of geochemically distinct habitats might be loosely related to the total number of AMD-adapted species. This would predict that the total number of AMD lineages would be small, given that AMD solution chemistry is strongly controlled by pyrite dissolution (see details below). Microbes from 11 putative divisions have been detected in AMD environments (Fig. 1).
|Number of putative divisions
|Number of genera and novel lineagesa
|Arid southwestern US soil
|Wisconsin agricultural soil
|Grass pasture rhizospheres
|Contaminated aquifer
|Marine bacterioplanktonic community
|High-temperature petroleum reservoirs
|Mid-Atlantic Hydrothermal Vent
|Yellowstone N.P. hot spring
|Antarctic sea ice and water
|Deep subsurface paleosol
|Iron Mountain Mine, CA 1998
|Total of all studied AMD sites
|compiled here
|Number of putative divisions
|Number of genera and novel lineagesa
|Arid southwestern US soil
|Wisconsin agricultural soil
|Grass pasture rhizospheres
|Contaminated aquifer
|Marine bacterioplanktonic community
|High-temperature petroleum reservoirs
|Mid-Atlantic Hydrothermal Vent
|Yellowstone N.P. hot spring
|Antarctic sea ice and water
|Deep subsurface paleosol
|Iron Mountain Mine, CA 1998
|Total of all studied AMD sites
|compiled here
'+' indicates that clones of unresolved phylogenetic positions were recovered.
aNovel lineages were counted subjectively by phylogeny provided in paper, each deeply branched cluster was counted.
The AMD system most intensively studied by culture-independent molecular methods is within the Richmond Mine at Iron Mountain in northern California. The methods used include 16S rDNA clone library analyses and cell imaging using multiple fluorescent probes designed to bind to the ribosomal RNA with varying levels of specificity. Pyrite-associated communities, subaqueous slime streamers, and subaerial biofilms in pH 0.5–1.4, 27–50°C solutions have been monitored and community responses to seasonal variations have been determined. Results confirm that a handful of prokaryotic taxa (often less than five groups distinct at the genera level) make up the communities in any specific microenvironment [17,28]. Low diversity has also been noted using cultivation-based approaches [29,30].
Among the bacterial lines of descent are divisions within the proteobacteria, nitrospira, firmicutes, and acidobacteria. The most extensively studied group (but possibly the least relevant under AMD-generating conditions, see below) are the γ-proteobacteria, specifically Acidithiobacillus spp. (formerly T. ferrooxidans, Thiobacillus caldus[31]) and Thiobacillus spp.
Two β-proteobacterial groups have been detected to date. Among this group are Thiomonas sp. (strains Ynys1 and Ynys3 [32]) and subsequently an isolate designated NO-16 from a Norwegian mine [30].
There are six species of heterotrophic α-proteobacteria of the genus Acidiphilum in pure culture. Acidiphilum cryptum (strain JF-5) was originally isolated from acidic (pH 3.0) iron-rich sediment from a lake associated with a coal mine in eastern Germany [33]. Another group of α-proteobacteria have been detected in clone libraries constructed from pH<1.0 Iron Mountain AMD environments. However, these live in near neutral pH habitats inside acidophilic protists [34].
Recently, a novel group of δ-proteobacteria clones were recovered from Iron Mountain (Fig. 1) [16,17]. However, because this group has defined cultivation, their metabolic characteristics remain uncertain.
'Leptospirillum ferrooxidans'-group organisms are commonly detected in AMD and bioleaching systems. To date, Leptospirillum isolates and environmentally-derived clones cluster within one of three phylogenetically distinct groups (I, II, and III), as designated by Bond et al. [16]. Two species have been named: L. ferrooxidans[35] (group I) and L. ferriphilum (group II) [36]. To date, group III Leptospirillum has only been detected via clone library analysis of Iron Mountain microbial communities [16,17].
Three distinct groups of AMD organisms fall within the firmicutes division. These include an Actinobacteria group containing Acidimicrobium ferrooxidans and Ferromicrobium acidophilus (Fig. 1) and the polyphyletic (two lineages) low G+C Gram-positive Sulfobacillus. Sulfobacillus thermosulfidooxidans VKLM and Sulfobacillus disulfidooxidans SD-11 belong to the original Sulfobacillus spp. group, which is related to the Alicyclobacillus lineage (Fig. 1). Another large bacterial group named Sulfobacillus, represented by some S. thermosulfidooxidans and Sulfobacillus acidophilus, is distinct from the group represented by 'Sulfobacillus' and separate from the Alicyclobacillus lineage. We suggest that in the future organisms within the Alicyclobacillus (including the type strain VKLM) be formally renamed to Alicyclobacillus based on phylogeny (Fig. 1).
The first isolate of the Acidobacteria division, Acidobacterium capsulatum, was recovered from an AMD site by Kishimoto et al. [37]. Closely related environmental clones have been reported from Iron Mountain (Fig. 1) [12,17].
Archaeal lineages reported from AMD environments are restricted to the Thermoplasmatales and the Sulfolobales. A number of clones phylogenetically divergent from Thermoplasma spp. have been detected in clone libraries (Fig. 2) [16,17]. Further surveys and isolation efforts are needed to resolve the tentative groupings within this lineage, currently referred to as the 'alphabet plasmas' (Fig. 2).
One member of the Sulfolobales, Metallosphaera prunae, has been detected in AMD environments. Two other Sulfolobales genera, Acidianus and Sulfolobus, have only been found in geothermal acidic environments, thus are not considered further here.
There have been a few reports of eukaryotes in AMD environments [38–41]. Ciliates belonging to Cinetochilium genus and an amoeba related to Vahlkampfia sp. within the lineage Heterolobosea, and three flagellates (Eutreptial spp.) were isolated from an AMD site and shown to be able to graze on mineral-oxidizing acidophilic bacteria [42]. Lutz et al. [38] recovered clones related to Vahlkampfia sp. from Iron Mountain. At Iron Mountain fungal filaments provide structure to subaqueous biofilms termed 'slime streamers' (Figs. 3 and 4). Recently, Amaral Zettler et al. [43] reported a diverse community eukarya (including alga) present in the pH 2 Rio Tinto River, Spain. In this instance, eukaryotes comprised at least 60% of the total biomass. It remains to be seen if subsurface non-phototrophic protistan communities growing in more acidic environments (such as encountered at Iron Mountain) are as diverse. Metabolically active protists are associated with Richmond Mine biofilms (pH 0.5–2.0; Fig. 3).
Evolution of acidophilic metal-tolerant organisms
It is widely believed that early Earth environments were relatively anoxic prior to the appearance of bacterial oxygenic photosynthesis. Given this, and the low abundance of sulfates in the early Earth record, it is likely that ARD environments were rare in the Archaean. Once the concentration of O2 increased, new metabolic options, such as oxidation of iron and sulfur coupled to reduction of O2, may have stimulated diversification within existing lineages, explaining the broad distribution of acidophiles over the tree of life (Figs. 1 and 2).
ARD and, more recently, AMD environments exist as isolated point sources that are exposed then removed via erosion within a few thousand to a few millions of years. Lateral gene transfer (LGT) is a mechanism by which some AMD survival genes could be introduced to create new acid- and metal-tolerant lineages. Recent analyses of LGT provide evidence for exchange of genes between extremely acidophilic trading partners [45]. However, there is no evidence to suggest that AMD organisms evolve from non-extremophiles when local acidic environments appear. In fact, organisms that colonize AMD environments possess habitat-specific genes (e.g., involved in metal resistance) [44] whose phylogeny matches the 16S rRNA gene phylogeny and modern inhabitants of AMD environments have very close relatives that are widely geographically distributed. Organisms are apparently introduced to newly created extremely acidophilic habitats from distant sites via as yet poorly understood dispersal mechanisms. The survival of these lineages over geologic timescales depends upon this.
Metabolic options and pathways
In regions of AMD systems exposed to sunlight, photosynthesis is an important source of energy and fixed carbon [46]. However, in subsurface ARD and AMD systems, inputs of externally-derived fixed carbon and nitrogen are minimal. The primary metabolic groups detected in AMD-generating regions are lithoautotrophs that oxidize Fe2+ and S− released by pyrite dissolution, organoheterotrophs that utilize carbon produced by lithoautotrophs, lithoheterotrophic Fe and S oxidizers, and anaerobes that couple oxidation of sulfur or organic carbon to Fe3+ reduction. A subset of these organisms must produce all fixed carbon and nitrogen required by the community.
In addition to S−, intermediate sulfur and sulfoxy oxidation products can serve as electron donors. Electrons from these compounds are passed either to O2 or Fe3+ (generated by oxidation of Fe2+ by O2) because these are typically the only suitable electron acceptors available in significant concentrations.
Utilization of Fe3+, and possibly sulfate, as electron acceptors is possible under microaerophilic and anoxic conditions. O2 consumption by microbes and pyrite dissolution can induce some typically aerobic species such as A. cryptum, Sulfobacillus, and possibly F. acidarmanus and related 'alphabet plasmas' to grow anaerobically. To date, no obligate anaerobes have been cultivated from an AMD habitat. However, uncharacterized d-proteobacteria phylogenetically closely related to anaerobic species have been detected [16,17].
The acidophilic lineages documented to date have diverse evolutionary histories. If, as suggested above, most arose after the divergence of the three domains, the enzymes involved in the key transformations may be diverse. Our ability to test this hypothesis is limited due to the restricted amount of biochemical and genome sequence data currently available. Work of Blake and coworkers [47] suggests that the critical process of iron oxidation involves unrelated enzymes in most lineages examined to date.
Iron oxidation
The autotrophic, facultative anaerobic Fe oxidizer Acidithiobacillus ferrooxidans (formerly T. ferrooxidans) is readily cultivated and has been the focus of extensive genetic, genomic, and physiological studies [4,10,65]. Consequently, it is the only organism for which there are biochemical models of the electron transport chain for iron oxidation.
All characterized L. ferrooxidans and L. ferriphilum isolates are iron oxidizers [48,49,36,50,29]. Although no members of Leptospirillum group III (Fig. 1) have been isolated yet, it is likely that these are also iron oxidizers. S. acidophilus, S. thermosulfidooxidans, and Acidimicrobium ferrooxidans strains TH3 and ICP are also iron oxidizers [51–53].
A few archaea associated with AMD have been shown to oxidize iron. Ferroplasma acidiphilum is an obligate autotroph able to oxidize ferrous iron as a sole energy source [54]. F. acidarmanus can oxidize ferrous iron and can grow heterotrophically [55]. A crenarchaeota, Metallosphaera sedula, can oxidize iron and sulfur at high temperatures (optimal growth at 74°C and pH 2.0). Thus, it is a candidate for high through-put industrial bioleaching [56].
Iron reduction
AMD solutions are iron rich because ferric and ferrous iron are very soluble at low pH (<2.5). In some cases, Fe3+ concentrations in AMD may exceed oxygen concentrations by several orders of magnitude [17]. Thus, Fe3+ may be widely used as an electron acceptor in microbial metabolism.
Johnson and McGinness [57] showed that the ability to reduce soluble Fe3+ is widespread among heterotrophic acidophiles. Moderately thermophilic species S. thermosulfidooxidans (strain TH1), S. acidophilus (strains ALV, THWX, and YTF1), and Acidimicrobium ferrooxidans (TH3) use ferric iron as the sole electron acceptor when grown heterotrophically under oxygen-limited conditions (with yeast extract and glycerol [57]). Thiobacillus acidophilus and a number of pure cultures of mesophilic heterotrophs are capable of mixotrophic Fe3+ reduction [58]. A. cryptum can couple Fe3+ reduction to oxidation of a variety of substrates including H2 and glucose. For example, Kusel et al. [59] demonstrated that A. cryptum strain JF-5 couples oxidation of glucose to reduction of either O2 or soluble Fe3+.
Some microorganisms can utilize Fe3+ even if it is not in solution. S. acidophilus is capable of anaerobic dissolution of iron hydroxide, jarosite, and goethite [51]. Bridge and Johnson [57] demonstrated that A. cryptum strain SJH is capable of reductive dissolution of a wide range of ferric iron minerals (akageneite, goethite, jarosite, natrojarosite, magnetite, and amorphous ferric hydroxide). They reported that contact was necessary for dissolution to occur and that rates were faster at lower pH (2.0 vs. 2.8–3.0).
Anoxic conditions are not required for Fe3+ reduction by mixed cultures of acidophiles [58]. Recently, Kusel et al. [59] demonstrated that under oxic conditions JF-5 reduced soluble Fe3+ and schwertmannite (a Fe3+-sulfate mineral) in sediment microcosms at pH 3.
In addition to coupling Fe3+ reduction to oxidation of organic carbon or hydrogen, some organisms can use Fe3+ to oxidize sulfur compounds. Bridge and Johnson [57] showed that S. acidophilus strains THWX, ALV, and TH1 use ferric iron to oxidize tetrathionate anaerobically.
Sulfur oxidation
Most members of AMD communities that oxidize sulfur also can fix CO2. An exception is A. cryptum, which may utilize sulfur oxidation to some extent but is normally considered to be an organoheterotroph [60]. T. acidophilus can oxidize sulfur, iron, and organic carbon and is capable of autotrophic growth [61]. Autotrophic sulfur oxidation by pure cultures of Acidithiobacillus ferrooxidans[62], Thiobacillus thiooxidans[63], Sulfobacillus (more correctly Alicyclobacillus) disulfidooxidans SD-11 [64], T. albertis[65], S. acidophilus[66] has been demonstrated. A. ferrooxidans also can grow under anoxic conditions using ferric iron as the electron acceptor and S° the electron donor [62].
Several autotrophic sulfur oxidizers can also oxidize organic carbon and/or ferrous iron. For example, S. disulfidooxidans is capable of mixotrophic S oxidation [64], but does not oxidize iron. However, SB37, a recent S. disulfidooxidans isolate of close phylogenetic association, is able to oxidize Fe2+ (Baker, unpublished data). Fourteen strains of Gram-positive sulfide mineral-oxidizing bacteria, including S. acidophilus (one being ALV) and S. thermosulfidooxidans (including TH1), can grow autotrophically on ferrous iron and S°, mixotrophically in the presence of yeast extract, and heterotrophically on just yeast extract [66].
A variety of sulfur compounds with oxidation states intermediate between 2− (e.g., sulfide in sphalerite) and 6+ (sulfate) form during metal sulfide oxidation (e.g., polysulfide: Sn2−, elemental sulfur: S°, thiosulfate: S2O32−). Some of these can be utilized by AMD microorganisms. Elemental sulfur and tetrathionate (S4O62−) are thought to be especially biologically important because they are relatively stable in low pH solutions. T. acidophilus grows autotrophically on trithionate (S3O62−), elemental sulfur, and tetrathionate [66,67]. As noted above, some strains of S. acidophilus couple autotrophic oxidation of tetrathionate to reduction of Fe3+[51,66]. It is interesting to note that none of the strains described as Thiobacilli that were adapted to higher temperatures (∼50°C) was capable of tetrathionate oxidation [66]. This is likely due to a decrease in bioavailability because of faster reaction rates.
Other metabolic requirements
Most subsurface AMD sites receive relatively minimal inputs of fixed carbon and nitrogen from external sources. The Thiobacilli (including Acidithiobacillus ferrooxidans) are probably the dominant group responsible for CO2 fixation in lower temperature (<30°C), higher pH (>2) environments. In lower pH, higher temperature communities, autotrophic taxa include Leptospirillum spp., Ferroplasma spp., Sulfobacillus spp., Ferromicrobium spp., and Acidimicrobium spp. At high temperatures (∼65–95°C) archaea such as Metallosphaera spp. may be the key sources of fixed carbon for the AMD biome. Although it is known that the Acidithiobacillus ferrooxidans pathway involves a bc1 and an NADH-Q oxidoreductase complex functioning in reverse cycle for CO2 fixation [68], little is known about the pathways used by other acidophilic autotrophs.
Fixation of N2 in largely aerobic and microaerophilic AMD environments is potentially problematic due to inhibition of nitrogenase by O2. Acidithiobacillus ferrooxidans, a moderate acidophile, may overcome this problem by using tetrathionate as an electron donor and ferric iron (rather than O2) as an electron acceptor when fixing nitrogen [69]. The fixation of N2 in very low pH environments is enigmatic, because no one has observed it directly. L. ferriphilum has been shown to possess nifH genes [70]. However, attempts to demonstrate active nitrogenase in pure cultures of L. ferriphilum were unsuccessful. Determination of which organisms are responsible for N2 fixation in the full range of AMD environments is an important goal for further work.
Abundance, community structure, and physical/chemical regimes
Studies over the last decade indicate microbial communities in acidic environments are dominated by the Thiobacilli. It now appears this is an artifact of the culturing and sampling methods used, at least in some cases. Schrenk et al. [71] and Edwards et al. [72] used fluorescent in situ hybridization (FISH) probes to quantify the relative abundances of Acidithiobacillus ferrooxidans within the Richmond Mine at Iron Mountain (pH 0.3–0.7, 30–50°C). They reported that A. ferrooxidans were nearly undetectable. Subsequent studies demonstrated that A. ferrooxidans occur in relatively low numbers in pyrite-dominated bioreactor systems [73]. The failure of A. ferrooxidans to thrive in more extreme AMD environments is attributed to their mesophilic growth optima (26°C) and moderately acidophilic nature (pH 1.3–4.5). Druschel et al. [17] did not obtain any A. ferrooxidans clones in six libraries constructed from six pH<1.0 AMD communities associated with the Richmond Mine. However, they demonstrated that an A. ferrooxidans strain was abundant in an oxidized, pH 1.4 pool.
Schrenk et al. [71] used FISH probes to show that A. ferrooxidans cells are abundant in environments with pH>1.3 and temperatures <30°C at the Richmond Mine. In this case, the higher pH, lower temperature environments occur outside the ore body, thus away from sites of active pyrite dissolution. However, pH 2–4, <30°C environments are encountered in many AMD systems early in weathering, when rocks are less enriched in pyrite, or when the buffering capacity is higher.
Leptospirillum group isolates have been obtained from AMD environments characterized by a wide range of temperatures and pHs. They are as abundant, or more abundant, in bioleaching systems than A. ferrooxidans over much of the A. ferrooxidans growth range. Sand et al. [74] note that the growth rate of A. ferrooxidans only outcompetes that of L. ferrooxidans (group I) below 14°C. They also note that in their bioreactors high ratios of Fe3+ to Fe2+ appear to be less inhibitory to Leptospirillum than to A. ferrooxidans.
L. ferrooxidans DMZ2705 (group I) is reported to grow in the pH range of 1.3–4.0, with an optimal pH range of 1.6–2.0 (compared to 1.4–1.8 for L. ferriphilum) [36], and an upper temperature limit around 55°C [75,35]. Goebel and Stackerbrandt [29] isolated strain Lf30-A (group II, thus L. ferriphilum) from a continuous bioreactor (pH 1–2 and 35–40°C) and showed that the isolate was capable of growth at 28, 37, and 45°C. Coram and Rawlings [36] noted that L. ferriphilum-dominated commercial biooxidation tanks operated at 40°C, and that some strains were able to grow at 45°C (no L. ferrooxidans strains were capable of growth at 45°C). Golovacheva et al. [50] characterized an isolate from an iron-containing hydrothermal spring (pH 2.0, 45°C) and named it 'Leptospirillum thermoferrooxidans' (also see [35]), based on greater G+C content and higher growth temperature (45–60°C) than previously described 'L. ferrooxidans' strains. The status of isolates described as 'L. thermoferrooxidans' is uncertain because these strains are not available for molecular characterization.
Oligonucleotide probe-based studies within the Richmond deposit indicate that Leptospirillum strains often dominate microbial communities growing at temperatures up to 50°C in solutions with pH values as low as 0.5 [16,17,71,72]. Leptospirillum (group III) were shown to comprise the majority of bacteria in subaerial slimes on the surface of a 'slump' of fine-grained pyrite ore [16] and, at times, to dominate subaqueous environments otherwise populated by L. ferriphilum[17]. L. ferriphilum and Leptospirillum group III primarily reside in lower pH microenvironments in the mine, while L. ferrooxidans occurs in higher pH environments (>1) [17].
There are several other groups of bacteria whose abundance and distribution have been studied in AMD. F. acidophilus has a pH range of 1.3–4.8 and is adapted to temperatures from <20 to 40°C [75]. Environmental clones closely related to F. acidophilus were recovered from the Richmond Mine AMD system (43°C) [16,17]. Ferromicrobium spp. were shown to be minor community members via probe-based studies [76]. Acidimicrobium ferrooxidans has been cultivated at temperatures between 34 and 57°C [53] and has been reported from a surprisingly diverse range of environments [77].
Sulfobacillus spp. have a broad range of physical growth regimes. Some isolates are capable of growth up to 65°C [78]. Recently, Yahya et al. [79] described Sulfobacillus-like strains (L-15 and Riv-14) isolated from geothermal sites. The strains are effective pyrite oxidizers at pH<1, suggesting that thermophilic species may be encountered in AMD systems. The FISH image in Fig. 3 shows Sulfobacilli rods within a Richmond Mine biofilm growing at pH∼0.7. Sulfobacillus clones have been recovered from pH 0.7–0.9, 35–43°C [16] and have been shown to comprise up to ∼6–8% of microbial communities (in pH 0.8–1.4, 27–32°C environments [17]).
Six members of the genus Acidiphilum are adapted to temperatures ranging from 17 to 45°C and pH values from 1.5 to 6.0 [75]. Strain SJH has a pH optimum of ∼3.0 [57]. One isolate was retrieved from a continuous bioreactor [29] and a clone was identified from the pH<1.0 solutions at Richmond Mine [12], though later studies at this site have not detected this group [16,17]. Peccia et al. [80] used a 16S rRNA probe to determine the abundance of Acidiphilum in sediments from an active AMD site in Colorado and in laboratory reactors. They found that the Acidiphilum outnumbered Acidithiobacillus ferrooxidans in their mixed culture bioreactors and that Acidiphilum and A. ferrooxidans populations were approximately equal in the sediment samples. In general, Acidiphilum sp. tends to occupy lower temperature, higher pH microenvironments. Under anoxic conditions they may contribute to iron cycling by redissolving ferric iron-based minerals precipitated when the pH is increased by mixing with groundwater and in streams.
The metabolic role of the δ-proteobacteria has not been elucidated because no members have been isolated. As δ-proteobacteria are often anaerobic sulfate or metal reducers, Bond et al. [16] suggested that anaerobic microenvironments may exist within the slime samples from which the clones were recovered. Attempts to amplify the highly conserved dissimilatory sulfite reductase (Dsr) gene were unsuccessful, and therefore it has not been possible to confirm these bacteria as sulfate reducers (Baker and Banfield, unpublished data).
Acidobacteria spp. populate relatively moderate AMD environments (20–37°C and pH 3.0–6.0). Recent analyses suggest that this group is limited to the higher pH environments (∼pH 1.4) at the Richmond Mine site [17].
Application of culture-independent approaches led to recognition that archaea are important and abundant in AMD environments. Iron-oxidizing archaea of the genus Ferroplasma have been isolated from a bioleaching plant [54] and the Richmond Mine [55]. Ferroplasma lack a cell wall and are bounded only by a cytoplasmic membrane. However, like other Thermoplasma and Sulfolobus spp., cells may possess an S-layer (Fig. 4) [81,82,76]. F. acidarmanus, isolated from the Richmond Mine, was shown to be the dominant organism in some habitats (see Fig. 3), comprising as much as 85% of the population in pH∼0.5, ∼40°C, high ionic strength habitats [55]. F. acidarmanus has a lower pH growth range than F. acidiphilum (0–2.5 vs. 1.3–2.2).
Clones within the Thermoplasmales (e.g., Thermoplasma acidiphilum and Thermoplasma volcanium), but distinct from Ferroplasma spp., have been detected in clone libraries created from samples collected from many very acidic geothermal hot spring environments [83]. T. acidophilum and T. volcanium have moderately thermophilic (45–67°C) growth ranges [81,82] and are adapted to pH ranges typical of AMD. T. acidophilum has been isolated from a coal refuse pile [81].
The metabolic role of the 'alphabet plasmas' within the order 'Thermoplasmatales' (Fig. 2) [16,17] is yet to be determined. Their environmental distribution indicates adaptation to high biomass, metal-rich, pH 0.5–1.4, 30–50°C habitats close to the air–biofilm interface [17]. Within the Richmond Mine system, all archaea are low abundance or absent from in pH 1.3–2.4 habitats [17,71]. Metallosphaera spp. (Sulfolobales order) can impact pyrite dissolution via catalysis of ferrous iron oxidation [84] and has been isolated from an acidic uranium mine [85] and a bioleaching reactor [56].
Synergistic interactions involving iron, sulfur, and carbon oxidizers
As indicated by the above discussion, community membership varies with pH and temperature, mineralogy, as well as with metal concentrations. However, in most environments (particularly in the subsurface) some species, or combination of species, within communities probably carry out iron oxidation, sulfur oxidation, organic carbon oxidation, carbon fixation, nitrogen fixation, extracellular polymeric slime production, as well as iron and sulfur reduction. It is also apparent from the above review that microbial communities in AMD systems tend to contain few distinct taxa (though these taxa may be remarkably phylogenetically diverse). Fig. 5 illustrates a simplified AMD system and provides examples of organisms that may control flow of iron, sulfur, nitrogen, carbon, and energy in different temperature regimes.
Interactions between members of microbial consortia are probably critical in optimization of AMD microbial community activity. For example, an important symbiosis exists between heterotrophic and certain autotrophic species: autotrophs may depend on coexisting heterotrophs to remove organic compounds that are toxic to them. Heterotrophic acidophiles are able to utilize organic materials produced by acidophilic autotrophs [86,56]. The culture filtrate from the autotroph Acidithiobacillus ferrooxidans contained sufficient organic matter to support heterotrophic growth of S. thermosulfidooxidans (strain TH1) [87]. Clark and Norris [53] conducted mixed culture experiments with Acidimicrobium ferrooxidans and both S. acidophilus and S. thermosulfidooxidans. They found iron oxidation by mixed cultures of Sulfobacillus spp. and Acidithiobacillus ferrooxidans was more extensive than in pure cultures containing any isolate. They attributed this greater leaching in mixed cultures to mixotrophic growth of the Sulfobacillus removing organic byproducts of A. ferrooxidans. This may explain the close correlation in populations of A. ferrooxidans/T. thiooxidans and Acidiphilum in environments studied by Peccia et al. [80].
Raman spectroscopy has been used to show that abundant S° forms on the surfaces of arsenopyrite [88] and galena [89] and pyrite [90] during oxidative dissolution. It has been hypothesized that microbial removal of elemental sulfur (S°) and other compounds is critical for continued access of the Fe3+ oxidant to pyrite surface sulfur-bearing groups. Dopson and Lindstrom [91] examined the effect of defined mixed cultures of the sulfur oxidizers A. caldus and S. thermosulfidooxidans (capable of Fe and S oxidation) on arsenopyrite dissolution. By following concentrations of released iron, tetrathionate, and sulfur in the cultures they found that A. caldus grows on surface sulfur compounds, resulting in an increase in the arsenopyrite dissolution rate. They also suggested that A. caldus produces surface-active agents to mobilize the extremely hydrophobic S°. Subsequently, Hu et al. [90] determined that elemental sulfur accumulations on galena significantly decrease dissolution rates. Because removal of S° from mineral surfaces is insignificant in abiotic controls, microbial oxidation of sulfur is likely to be a key determinant of acid generation rates (because all protons are released in conversion of sulfur to sulfate).
Perhaps the most exciting aspect of AMD communities is that they provide the opportunity to study largely self-contained biomes (independent of sunlight and most other life). Culture-independent methods have provided a detailed understanding of the full diversity and phylogeny of organisms populating AMD systems. Progress in understanding the biochemistry of acidophiles has been accelerated due to availability of genome sequences for a few isolates (e.g., F. acidarmanus and Acidithiobacillus ferrooxidans). However, we know essentially nothing about the subset of organisms that have not been isolated.
An important approach for analysis of uncultivated organisms is the construction and sequencing of large insert genomic libraries from environmental DNA [92]. Cultivation-independent sampling of genomes of acidophiles will reveal enzymes potentially involved in the major metabolic pathways and provide insights into pH homeostasis and metal resistance mechanisms. The low species richness of AMD communities may permit us to largely reconstruct genomes of populations from whole community libraries. Related gene expression studies should enable development of AMD ecosystem models that resolve the contributions of individual species at the molecular (pathway) level, describe and quantify flows of materials and energy, and identify symbioses and competitive strategies. Thus, through integration of geochemical and biological information, a comprehensive model for AMD production may be feasible. Studies of AMD systems could have broader significance. The remarkable simplicity of these habitats may permit a more fundamental understanding of how microbial communities work than is possible through study of more complex ecosystems.
We are grateful to Jenn Macalady and Philip Hugenholtz for critically reviewing this manuscript and discussion, and Ian Lo for preparation of FISH slides. We also thank Joe Coggliati (Stouffer Corporation), Rudy Carver (IT Corporation), Don Dodds (North Pacific Research), Ted Arman (Iron Mountain Mines), Richard Sugarek (EPA), and others for access to the field site and assistance with field work. This work was funded by the NSF LExEn and EGB programs and the US Department of Energy Microbial Genomics Program.


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Red Roof Inn Greensboro Airport places you within a few miles (5 km) of All-A-Flutter Butterfly Farm and Piedmont Triad Farmers Market. This 112-room motel welcomes guests with conveniences like free breakfast, free in-room WiFi, and free self parking. Greensboro Coliseum is also within a 15-minute drive of this Greensboro motel.
Free continental breakfast is included with your stay and awaits you each morning from 6 AM to 10 AM.
All of Red Roof Inn Greensboro Airport's 112 air-conditioned rooms provide free local calls, safes, and ironing boards. Guests can expect to find free WiFi and flat-screen TVs with cable channels. Bathrooms have hair dryers and free toiletries. Desks are also included.
Property features
Red Roof Inn Greensboro Airport features free WiFi in public areas, coffee in a common area, and a computer station. If you drive, take advantage of free parking. The 24-hour front desk has staff standing by to help secure valuables, and answer any questions about the accommodations. Additional amenities include express check-out and free newspapers.
- Free continental breakfast
- Free WiFi in rooms and public areas
- Free parking
- 24-hour front desk
Red Roof Inn Greensboro Airport
Hotel Amenities
Hotel Amenities
The motel offers a snack bar/deli. Guests can enjoy a complimentary breakfast each morning. A computer station is located on site and high-speed wireless Internet access is complimentary. This Greensboro motel also offers coffee/tea in a common area, complimentary newspapers in the lobby, and express check-out. Onsite self parking is complimentary.
Red Roof Inn Greensboro Airport has designated areas for smoking.
- Free RV, bus, truck parking
- Number of buildings/towers - 3
- Express check-out
- 24-hour front desk
- Number of floors - 2
- Free newspapers in lobby
- Free breakfast
- Designated smoking areas
- Computer station
- Total number of rooms - 112
- Snack bar/deli
- Free self parking
- Coffee/tea in common areas
- Free WiFi
- Safe-deposit box at front desk
- Year Built 1986
Family Friendly Amenities
- Family Hotel
- Free Wi-Fi
- Snack bar/deli
- Cribs/infant beds (complimentary)
Available in all rooms: Free WiFi
Available in some public areas: Free WiFi
Free self parking, Free RV, bus, truck parking
Room Amenities
Red Roof Inn Greensboro Airport offers 112 air-conditioned accommodations, which are accessible via exterior corridors and feature safes and hair dryers. Flat-screen televisions come with premium cable channels. Bathrooms include shower/tub combinations and complimentary toiletries.
Guests can surf the web using the complimentary wireless high-speed Internet access. Business-friendly amenities include desks and phones; free local calls are provided (restrictions may apply). Additionally, rooms include irons/ironing boards and blackout drapes/curtains. Housekeeping is provided daily.
- Connecting/adjoining rooms available
- Flat-panel TV
- Air conditioning
- Free local calls
- Premium TV channels
- Daily housekeeping
- Free toiletries
- Hair dryer
- Iron/ironing board
- In-room safe
- Access via exterior corridors
- In-room climate control (air conditioning)
- Phone
- Cable TV service
- Private bathroom
- Shower/tub combination
- Free cribs/infant beds
- Desk
- Free WiFi
- Blackout drapes/curtains
Where to Eat
A complimentary continental breakfast is served each morning between 6 AM and 10 AM.
Start your day with a free continental breakfast, available daily from 6 AM to 10 AM.
If you have requests for specific accessibility needs, please note them at check-out when you book your room.
- Accessible bathroom
- Roll-in shower
- In-room accessibility
Hotel Policies
Check-in time starts at 2 PM
Minimum check-in age is 18
Check-out time is noon
Payment types
Children and extra beds
- Children are welcome.
- Kids stay free! Children 17 years old and younger stay free when using existing bedding. Children may not be eligible for complimentary breakfast.
- Free cribs (infant beds)!
- Pets stay free
- 1 per room
- Specific rooms only, restrictions apply
- Pet-friendly rooms can be requested by contacting the property at the number on the booking confirmation
You need to know
Extra-person charges may apply and vary depending on property policy.
Government-issued photo identification and a credit card or cash deposit are required at check-in for incidental charges.
Special requests are subject to availability upon check-in and may incur additional charges. Special requests cannot be guaranteed.
Pets allowed (no fees)
No rollaway/extra beds available
No elevators
Hotel Name
- Red Roof Inn Greensboro Airport
- Red Roof Inn Hotel Greensboro Airport
- Red Roof Inn Greensboro Coliseum Hotel Greensboro
- Red Roof Inn Greensboro Airport Motel
- Red Roof Greensboro Airport Hotel Greensboro
- Red Roof Inn Greensboro
- Greensboro Red Roof Inn
We should mention
Children 17 years old and younger stay free when occupying the parent or guardian's room, using existing bedding. Children may not be eligible for complimentary breakfast. The property has connecting/adjoining rooms, which are subject to availability and can be requested by contacting the property using the number on the booking confirmation. The property allows pets in specific rooms only and has other pet restrictions. Guests can arrange to bring pets by contacting the property directly, using the contact information on the booking confirmation.


text: These Male Ants Are Copulating With a Dead Queen While a Spider Eats HerAlan Smithee3/23/12 4:33pmFiled to: WTFridayBugkkakeAntsSpiderCopulatingVideoClipsshowcase81EditPromoteShare to KinjaToggle Conversation toolsGo to permalink These winged ants are copulating with their queen. Apparently, they didn't notice the big spider who is eating it from the top. Ants are really really really stupid or really really really perverted. Advertisement In any case, it's a macabre orgy. One that I can't stop looking at, even if it grosses the hell out of me. [LiveScience]Reply81 repliesLeave a reply


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- 1[countable, usually singular] a sudden fall The jockey took a nasty tumble at the third fence. Share prices took a sharp tumble following news of the merger. see also rough and tumble
- 2[singular] tumble (of something) an untidy group of things a tumble of blond curls Word Origin Middle English (as a verb, also in the sense 'dance with contortions'): from Middle Low German tummelen; compare with Old English tumbian 'to dance'. The sense was probably influenced by Old French tomber
BrE BrE//ˈtʌmbl//; NAmE NAmE//ˈtʌmbl//
Check pronunciation: tumble


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#423 PayPal
- Market Cap As of May 2016
- $48.9 Billion
- Industry
- Consumer Financial Services
- Country
- United States
- Daniel Schulman
- Employees
- 16,800
- Sales
- Headquarters
- San Jose, California
PayPal on Forbes Lists
- #423 Global 2000
- #988 in Sales
- #476 in Profit
- #824 in Assets
- #187 in Market value
- #205 America's Best Employers
PayPal Holdings, Inc. operates as a holding company of PayPal, Inc. It engages in the digital and mobile payments on behalf of consumers and merchants worldwide. The company accepts payments from merchant websites, mobile devices, and applications; and at offline retail locations through payment solutions across its payments platform, including PayPal, PayPal Credit, Venmo and Braintree products. The company was founded in January 2015 and is headquartered in San Jose, CA.
More on PayPal
More on Global 2000
The World's Largest Companies 2016Steve Schaefer, Forbes Staff
China's Largest Companies 2016Shreya Agarwal, Forbes Staff


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At least 28% of schoolgirls in South Africa are HIV positive because 'sugar daddies' are taking advantage of them, the country's health minister warned today.
In contrast, Aaron Motsoaledi highlighted the fact that only 4% of schoolboys were HIV positive, The Sowetan newspaper reported.
Mr Motsoaledi released the figures at a public meeting at Carolina in South Africa's Mpumalanga province.
He said: "It is clear that it is not young boys who are sleeping with these girls. It is old men. We must take a stand against sugar daddies because they are destroying our children."
He said 94,000 schoolgirls fell pregnant in South Africa in 2011, with around 77 000 having abortions at public facilities.
"We can no longer live like that. We want to put an end to it, " he declared.
Some of the girls who became pregnant and tested positive for HIV were reportedly between the ages of 10 and 14.
Mr Motsoaledi has been widely praised for his efforts to help stop the spread of HIV, with the country backing the world's largest anti-retroviral programme since Jacob Zuma made him health minister in 2009.
Around 10% of South Africa's population is now living with HIV, official figures show.Reuse content


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A critical part of a long-tail keyword strategy is tracking your keywords and content to get them to show first in Google search engine rankings.
[Members] The Ultimate SEO Strategy Template for 2016 (Excel Template)
Designed to make your SEO efforts future-proof, this step-by-step Google SEO template uses an approach that has only improved in ranking over time – a cheat sheet for the post Hummingbird SEO. This comprehensive Excel template includes: SEO benchmarking for any website Keyword Strategy for 3 different types of conversion stages (click, lead


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Chatham Lodging Acquires Residence Inn in Bellevue, WA
Friday November 1st, 2013 - 11:16AM
These are shortcuts to your favorite social networking and bookmark sites. Add this story to your Facebook page, del.icio.us, DiggIt, and many others!
PALM BEACH, FL—Chatham Lodging Trust has acquired the 231-room Residence Inn by Marriott Seattle Bellevue/Downtown for a net cash purchase price of $71.8 million, plus customary, pro-rated amounts and closing costs.
The purchase price represents a forward 12 month capitalization rate of approximately 7.8% on the hotel's projected net operating income. Year to date through September 30, 2013, RevPAR at the Bellevue hotel is up 13% to $131.
Jeffrey H. Fisher, Chatham's CEO, said in a statement, "This hotel aligns perfectly with our strategy of acquiring coastal area hotels where demand is driven primarily by corporate travelers focused in the ever-growing technology, energy or medical sectors."
The hotel opened in 2008 and is situated along Interstate 405, providing access to businesses such as Microsoft, Amazon, Expedia, T-Mobile and AT&T.
The hotel occupies an approximately seven-acre site and features a lobby with contemporary décor and custom furniture and artwork. Facilities include 3,076 sq. ft. of meeting space in three rooms, an indoor, heated swimming pool, a fitness center, three levels of covered and secured parking, Starbuck's Café and a Microsoft game room.
The Residence Inn by Marriott Seattle Bellevue/Downtown is managed by Island Hospitality Management (IHM), which is 90% owned by Fisher.
Chatham funded the purchase with available cash and borrowings on its secured revolving credit facility of $59 million.
Tags: Chatham Lodging Trust • Marriott • Residence Inn • Bellevue • Seattle • Hospitality • Acquisitions • Ownership •
As the year rolls on, Hotel Business will continue to roll out initiatives to create a more interactive experience, to create more dialogue. Whether through our social media channels or new video programming, we're going to continue to implement initiatives to keep you engaged with us, us engaged with you—and you engaged with each other. We're on this journey together.
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meh 12/14/2010
This review is from: Kingston 4GB microSDHC Flash Card with Adapters & USB Reader Model MBLYG2/4GB
the micro flash works, as well as the sd adapter
the usb adapter stopped working on the first use. would get unbearably hot which was it's ultimate downfall,
Other Thoughts:
of coarse that was the purpose for getting the adapter pack with many choices for transferring data to a micro sd
Did you find this review helpful? Yes / No Thanks for the valuable feedback you provided!


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The increasingly popular New Balance H710 hiker sneaker hybrid factors in prominently once again this year with a July release of the following pack. Three different styles include two more understated versions not without bold contrast colors as well as a hard to miss red version. Look for the sneaker at select retailers including mita sneakers.


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Designed for fashion-conscious women, this yellow coloured kurta from Abhishti is a must-have in your ethnic wear collection. Made from Chanderi cotton, this kurta will surely get you noticed wherever you go. Team this sleeveless kurta with a contrast coloured churidar and Kolhapuris to look your ethnic best.
- FabricChanderi Cotton
- LengthThigh Length
- SleevesSleeveless
- NeckBoat Neck
- FitRegular
- Wash CareDry Clean Only
- ColorYellow
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- REVIEWSREVIEWS FOR Yellow Chanderi Cotton Zari Work Kurta
Be the first to write a review about this productRicha45Nice kurta
Looks really pretty.Sindhu55Amazing one!
Hi,This is a beautiful kurta in terms of quality, color, style and it also comes with a lining giving a long durability..overall the best kurta added to my wardrobe!!!priyanka singh45nice product
i bought it from jabing n the kurti is very nice n elegant material is also nicei m sastisfied after seeing this productShweta55elegant look
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Lovely kurta. Is exactly like it looks in the picture. Good quality and fit :)Almost tempted to buy the same one in pink :PVer nice55Awsome collection!
It is just awesome.Naina45excellent!
unique design not available elsewhere. Delivered 6 days ahead, in time for Diwali.very happy!Shreyatama55Very elegant
Absolutely unique and elegant design. Looks great on me. Loved it!


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Skip to comments.Shooting reported near Brookfield Square Mall (Wisconsin "mass casualty" situation)
Posted on 10/21/2012 10:36:02 AM PDT by kristinn
WISN 12 News has learned that a mass shooting has taken place near the Brookfield Square Mall.
WISN 12 News has been told that this is a "mass casualty" situation and that the scene is not secured.
Link to live coverage.
(Excerpt) Read more at wisn.com ...
"Bald man shoots up hair salon"
Just wanted to get that out there.
You asked, and here's your answer Pappy smear.
This text was located down about 15 deep, give or take a few.
In 2005, a man killed seven people at a church service at a hotel near Brookfield Square.
by Amy Smith/Staff Writer 11:08 AM
Thanks TornadoAlley3
"Authorities have retracted an earlier statement, saying that the suspected shooter is not African-American, but is "ethnic with dark skin."
If it is a Muzzie, that won't bode well for the Obama part of the Obama/Romney debate Monday on international relations.
The scanner just said the suspect is a balding black man in camo jacket again.
what's with the differences in descriptions.
one is grey sweater, black pants, black and white back-pack.
another cream shirt and dark brown pants.
just heard another with camo-jacket
If Ubama had a son...
scanner repeating black male, may be wearing camo jacket.
There is now a small map that shows the location of the hotel where the church service was attacked in 2005.
Checked the online reviews... It's a regular beauty salon, the kind our mothers and sisters go to. Nothing sordid about it. If CNN just suggested this was a gay bath house (which is what a "gay salon" is) to a national audience, this salon should own a dying cable news channel in short order.
Thanks. Sure wish they had this guy in custody by now.
Just heard that the police apparently know who the suspect is and that he has "fled the scene".
More than one perp?
They just talked about checking the cemetery, that sounds like a disgruntled husband/boyfriend type thing.
Asymmetrical warfare "soft targets"? That IS already here.
Well, at least they know who it was (hopefully).
So far no people have been killed (to my knowledge), and I'm hoping it stays that way.
(If fatalities were mentioned up thread and I missed it, I apologize.)
Yeah, CNN, "If only we can make homosexuals the victims here..."
The employee who spoke to Patch said the woman's estranged husband was out of work and his wife was working every day to bring in money to pay the bills. However, she recently decided to leave him, and that's when the problems started.
The two have two daughters, ages 12 and 21, the Azana employee told Patch.
"Authorities have retracted an earlier statement, saying that the suspected shooter is not African-American, but is "ethnic with dark skin."
The suspected shooter is also more than 200 lbs, but may not be as heavy-set as authorities said earlier."
If they already know who it is, that to me leans towards a husband/boyfriend of an employee. Or an ex-employee...
Bald man shoots up hair salon"
Envy killing.
I hope the loser didn't hurt his daughters. I mean, any worse than they will be if their papa went on a rampage aginst his wife.
David Gosh, of West Allis, told the Milwaukee Journal Sentinel he was driving past when they saw a woman coming from the salon and the shooter chasing her before the shooter ran into the building or behind the building.
"She ran right out into the street and was pounding on cars," Gosh told the Milwaukee newspaper.
You aren't kidding. Even in our relatively decent-size city where not that much happens, we've had a string of events lately we normally don't see - long-distance chases with one still at large and another resulting in the perp being shot dead, people burned in their homes or apartments, murders...last night, a bunch of Asians got into a fight and some stabbing happened with one dead as a result. Our LEO have been kept busy.
It's not usually blacks doing these killings because most blacks aren't STUPID ENOUGH to go on these psych drugs, or put their kids on them...or don't have the health benefits to have someone pay for them.
Same reason black suicides are much lower than whites.
a divorce...
good info at that link
2:12 p.m.
TODAY'S TMJ4 is reporting that 45-year-old Radcliffe Haughton from Brown Deer is sought for the shooting in Brookfield.
Different information from across the pond...
I wonder if it is Paula on the Facebook—somebody asks if Paula is back yet.
she is shown getting her hair done
Which will be identified by CNN as an AK-47 before the day's over.
Too bad we don't have another town hall debate coming up. We need more AK-47 questions from randomly selected suburban housewives.
1:01 p.m.
- Seven people have been transported from the scene of the shooting to area hospitals including Froedtert Hospital.
Four people were said to have non life-threatening injuries.
The other three people are in unknown condition.
12:37 p.m.
Newsradio 620 WTMJ's Jodi Becker is on scene.
- She reports that authorities are searching for a 6'1", 270 lb. African-American with a bald head and brown eyes wearing an Omama For America t-shirt.
He was reportedly driving a black 2003 Mazda.
Authorities believe he may be from Brown Deer.
She says Flight for Life has also gone to the scene to assist the injured.
12:32 p.m.
Here's what we have so far:
- Brookfield Police say they are looking for a suspect after shootings at a hair salon near Brookfield Square Mall Sunday morning.
The incident occurred at around 11:00.
- Multiple witnesses tell him and our news team that the shootings happened either at or near the Azana Salon and Spa, across Moorland Road from the mall.
- A number of stores have gone on lockdown inside Brookfield Square.
TODAY'S TMJ4's Todd Hicks was first on scene.
He reports that ambulances have gone to the scene while SWAT Team members were searching for a shooter.
Check back for more details on this breaking story as they come in to us, and catch TODAY'S TMJ4 for the latest.
They weren't/aren't sure if he is black or not. So he's been described as African-American, dark-skinned, and ethnic. So he could be anything.
scanner- sounds like they believe shooter is in spa.
Well, we're lucky a lot of people haven't been killed.
Thanks for the update.
I just don't let people get under my skin enough to want to do this sort of thing. Disliking someone doesn't have to mean you want to end their life unless they're threatening yours at the moment.
To many people out there are being convinced that others have really screwed them, and, "It's not your fault."
Something's sure up...
This could never be barry's son. He gives his kids Russian names, not high society English names.
Wisconsin court records show domestic restraining order against Radcliffe Haughton
Why can't you believe they published the description? How else are they going to find him/warn people?
2:12 p.m.
TODAY'S TMJ4 is reporting that 45-year-old Radcliffe Haughton from Brown Deer is sought for the shooting in Brookfield. Authorities are looking for a 2003 Mazda Protege with license plate: 171-KZD.
I would say he has definitely violated the restraining order.
He has a history of charges for disorderly, but they were dismissed.
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Colonial House Motel
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Grandson's 6th birthday today.
Day 18 – Tue 24 March – Kerikeri
It's Michael's 6th birthday today so we'll Skype them tonight. It's overcast with showers this morning so hope the washing will get dry later. The rain set in as we went off to the laundromat – hope it clears after lunch. Fat chance – it rained all day!
The oven decided to go on the blink today just as the buns were due to go ...
Cruise to Hole in the Rock
Lois, Olive and I took a cruise to the hole in the rock and also were blessed to see two pods of dolphins.
In the evening we took the ferry across the water to Russel and Greg took "Hi 3 ladies" to dinner at the Duke of Marlborough...very posh!!!
No drama sheep farmer
... a month). Not having to sleep on a bunk bed and having an on suite bathroom I only have had to share with three people – what luxury!! I'm loving it!
We had an incredibly long day on the bus as we made our way to the top of the North Island of New Zealand. We went to Paihia the Bay of Islands. It was beautiful! I have made many mentions of how spoiled I was by Hawaii, and how it is hard ...
Peace and Quiet - New Years in Paihia
... didn't clear for the rest of the evening. We decided rather watch the fireworks in the rain and jostle with the other punters for a good spot, we would instead stay in, play Scrabble and watch Monty Python. I don't think we even stayed up till midnight, boring gits that we were were passed out by 11pm in front of the telly like a pair of wallies.
Happy New Year 2015!
Well, there's next year! Who knows where I'll be by then? Watch this ...
... of spots where dolphins are regularly viewed. We were in luck and saw firstly Bottle Nosed Dolphins and then Common dolphins. We also saw Australasian gannets diving for fish, very difficult to photo though. The Hole in the Rock is in Piercy island which was named after Captain Cook's First Lord of the Admiralty at the time. When we got to the hole we actually sailed through which was pretty tight as you will see in ...


text:
Wood flies with eagle to first European title
DOHA, Qatar Chris Wood eagled the 18th hole to win the Qatar Masters for his first European Tour victory Saturday, overtaking Sergio Garcia and George Coetzee, who finished a shot back.
The 142nd-ranked Englishman shot 200 yards to the green with a 6-iron and watched the ball roll to within 12 feet. Wood calmly sank the putt to finish the final round at 3-under 69 for an overall 18-under 270. Garcia and Coetzee had looked set for at least a two-man playoff after both birdied the last hole.
"Winning on the European Tour is not easy," said Wood, who earned a victory in his 115th tour event. "It was a nice way to finish. It feels good to pick up the trophy. I've been waiting patiently for my turn and fortunately it came today,"
But Wood almost threw the victory away early, after he missed a birdie putt on No. 2 and then double-bogeyed the par-3 third.
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Featuring a main body made from 100% New Zealand Merino wool with water resistant nanotechnology, the Icebreaker Men's Sierra + Hood is a warm and windproof outer layer that's ready for adventures in cooler, wet conditions.
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Best Use Hiking, Casual Jacket Back Length Hip-Length Warranty Lifetime Type Water-Resistant Fleece/Wool Jacket Shell Material 260g Brushed Fleece, Merino Wool w/ Water-Resistant nanotechnology Other Media Cord Port Number of Pockets 3 Insulation Material n/a Hood Yes; Adjustable Fit Regular Fit
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Each day the word gets louder. Tiger Woods is to make his comeback in two weeks' time. So if the word proves true, will this be a time to rejoice, to welcome back the returning hero like the prodigal son? Or a time to scream: "You what? After all that 'I've changed... I'm so sincere... I've got such a long way to go' baloney a fortnight ago?" You decide.
What surely is unarguable is the perception Woods gave off to the world in his televised statement. He said: "I do plan to return to golf one day, I just don't know when that day will be. I don't rule out that it will be this year." Strictly by those statements it was impossible to foresee anything other than a lengthy absence as a broken man sought to piece back a shattered life.
Does a month represent a lengthy absence? Is it possible to piece back a shattered life in the time it takes to stage four tournaments? Who knows? Woods might think so. But then again, he may since have decided to do the repair job on the move. That's his choice, his right. Just as it would be everyone else's right to look back on that 14-minute soliloquy and wonder about an individual who looked more inclined to don a strait-jacket than a green jacket. If Woods is considering entering the Arnold Palmer Invitational before its deadline on Friday week then he should also be considering the flak he would receive. And he would be wise to line up a few more apologies.
First off he should say sorry to all those who took his "mea culpa" to be genuine. If he is teeing it up in Florida then the saddest aspect to me will be the scoffing of the cynics. You know, that smug band who saw the purposeful, look-into-the-camera delivery of Woods's first "I am truly sorry", who then watched the hand going across his heart and who then assured us that this was about as real as those counterfeit clubs sold on eBay. It's all about sponsorship, about the brand, about the reconstruction of the image, not the person. You'll see, they laughed.
Well, many of us in the gullible gallery would finally see. Most of his fans, certainly his British fans, didn't feel it necessary for him to apologise. Just turn up and play again. We always admired him as a golfer. Nothing else. But no, he chose to confess, and so his motives for doing so will inevitably be analysed. By the fans who believed his contriteness more than anyone.
Tiger should also apologise to his fellow players. Regardless of the timing of his reintroduction to their ranks, his fellow pros already have ample reason to be angry. When Ernie Els came out and called Woods "selfish" for taking over the airwaves on the third day of the Accenture Match Play, he was instructed by Woods's agent, Mark Steinberg, "to get your information right before commenting". The players were then informed Woods had no choice but to address the globe that particular morning as he was rechecking into rehab the next day. By and large the players accepted that explanation. The patient would be back under lock and key awhile.
It turned out that "awhile" lasted barely a week. Woods was back at his Orlando home the following Saturday. Question: why couldn't he wait until, say, last Monday to speak? What difference would it have made? Never mind whether it deflected any attention from the Match Play, it undeniably had the potential to distract the players in Tucson. Before they went out for their Friday matches the pros were badgered for their opinions on Tiger's "performance". It was the last thing they needed.
At this point, Woods's apologists will doubtless argue that he owes the rest of the Tour nothing. In fact, they owe him. Without Woods there would be half the interest, half the money. But none of them asked for Tiger to come along. None of them lay in their boyhood bed whispering, "I'm going to grab me some of them there Tiger millions". The majority are too old even to have heard of Tiger when they set off on the long fairway to the big time. They would have done so regardless of his rise and regardless of any associated riches. Tell me, why should somebody like Els feel indebted to Woods one iota? Right now, the South African and his peers have every right to feel fury. Woods should thank them for not being as vitriolic as they are justified to be.
There is one other section of the circus Woods should thank (other than the PGA Tour, of course, who have been quite pathetic in bending over backwards for their No 1 at every conceivable opportunity). Woods should thank the media. Without the media it must be suspected he would still be engaging in his extra-marital activities and in his own words not "have this opportunity to become a better person".
Woods no doubt detests the mudrakers and the mudslingers; but if he is true to his statement, he cannot deny they were ultimately a positive force in his "rebirth". Unless, of course, he was merely sorry about being caught. Return so early and that damnable taunt will undoubtedly receive airtime.
But perhaps all this hypothesising is unfair. Maybe the rumours of an imminent return aren't correct. If so, we should extend our best wishes for a speedy recovery. Speedy, that is. Not downright miraculous. We should then also be thankful as sports fans. A Masters minus the returning Tiger would be a Masters minus ridiculous hype. It would be what it set out to be: a golf tournament. Not some excruciating one-man stake-out.
If the professional game has learned anything from a saga which somehow still threatens to become yet more unsavoury, it is surely never again to rely too heavily on one player and, more pertinently, on one myth. Webs by their very nature are fragile, and deceit spins the most fragile webs of all. Woods and his advisers might be wise to remember that in the next few weeks.
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The AgN-GLEE workshop for Asia was held on 19th - 21st March 2013, in Bangkok.
The purpose of this workshop was to strengthen current and new USAID Feed the Future investments to achieve nutrition outcomes in the Feed the Future target countries.
Presentation made at the workshop: Aquaculture - Nutrition Linkages in FtF Aqua, Bangladesh: a case study
Presenter: Shakuntala Haraksingh Thilsted, Senior Nutrition Scientist, WorldFish.


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Sunset Drugs
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With Instructables you can share what you make with the world, and tap into an ever-growing community of creative experts.
Layout the gun
Look for the release button on the side of the gun
Push the button
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Get the right size bb's
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The Letarte Sisters' Labor Day Playlist
Say goodbye to summer with these tropical tunes.
Advertisement - Continue Reading Below
Lisa Letarte Cabrinha and Michele Letarte Ross share their favorite summer songs with T&C (a soundtrack that's a perfect match for their bohemian island chic swimwear line Letarte). With Michele in the NYC-area and Lisa in Maui, the two gave us a great collaboration of songs with an east coast/west coast/mid-Pacific vibe.
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Spitzer wins bid to return to O.C. Board of Supervisors
Todd Spitzer will return to the Orange County Board of Supervisors, winning an unofficial 68 percent of the vote over fellow Republican Deborah Pauly on Tuesday.
Spitzer previously represented the county's 3rd District – covering Orange, Tustin, Villa Park, Yorba Linda and portions of Anaheim and Irvine – on the board from 1997 to 2002. His return would represent a bit of electoral musical chairs, as he would replace termed-out Supervisor Bill Campbell, who in 2003 was elected to replace Spitzer after Spitzer succeeded Campbell in the state Assembly.
In the other supervisorial race, incumbent Janet Nguyen trounced her eccentric opponent, Steve Rocco, a perennial candidate for local offices who made no apparent effort to campaign. Final unofficial returns gave Nguyen 74 percent of the vote.
The seat represents the 1st District, which covers Santa Ana, Garden Grove, Westminster and part of Fountain Valley.
Neither race was expected to be close. Although Pauly ran a vigorous campaign against Spitzer, she was badly outspent by a candidate known for his prodigious fundraising abilities.
On Tuesday night, at a victory party at a Wahoo's Fish Taco restaurant in Orange, Spitzer joked that he had campaigned against the advice of his mentors.
"In fact, the advice given to me was to disappear for three months and then come back tonight," Spitzer said, as a few dozen supporters laughed.
In an interview Friday, Pauly said of Spitzer: "He's got all the money, all the name ID, all the establishment support. ... Those are all the reasons I ran against him."
Pauly succeeded in making a campaign issue of some of Spitzer's previous legislative votes, including his support – while a county supervisor – for a retroactive boost in pensions for deputy sheriffs known as "3 percent at 50." That 2001 decision has contributed to a growing unfunded pension liability for the county, which now totals more than $4 billion.
Pauly campaigned as a fiscal conservative, but she failed to win key endorsements. The central committee of the Orange County Republican Party voted last month to remove her from the position of first vice chairman, although she remains a voting member of the committee.
Spitzer last week denounced Pauly as "a political pyromaniac," a reference to some of her past public statements. For example, after the U.S. Congress passed the health care reform bill in 2010, Pauly wrote on her Facebook page: "Applauding this Health Care Bill is like applauding a mugging or a rape. Do you feel sodomized?"
"If I had said things like 'Oh this is just horrible' about 'Obamacare,' would that have brought any attention to my extreme concern about what happened?" Pauly asked Friday. "I do intend to be attention-getting."
Acknowledging that some aspects of health care need to be changed, Pauly said, "It would have been better to do something more incremental, (then) assess the results of that."
Assessing her campaign against Spitzer, Pauly said, "I win whether I lose or win, because I've done the right thing by challenging someone who's part of the entrenched good old boys network that's been selling us down the river for the past 15 to 20 years."
Spitzer said he planned to focus on public pension reform as supervisor. He also confirmed that he plans to run for Orange County district attorney if the office is open in 2014.
–Register staff writer Kiali Wong contributed to this report.
Contact the writer: 714-796-7956 or firstname.lastname@example.com
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Fredericksburg, TX
This cottage is an ode to the owner's German Heritage tracing back from Hof, Germany. Experience old world Germany through the furniture, decor and memorabilia, with some antiques being over 100 years old. This suite features a small sitting area that opens up to a wet bar. For your convenience, there is a small refrigerator, sink, microwave and a gourmet coffee maker. The large bathroom is stocked with Gilchrist and Soames bath products and decadent soft white towels. The bedroom has a king sized bed with luxurious sheets, feather pillows and uniquely crafted throw blanket. Step outside onto the back patio deck and experience some quiet solitude while you drink your coffee or enjoy a glass of wine.
Luxurious Triple-Sheeted Beds with Down Comforters and Luxury Linens
King Size Bed
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Gilchrist & Soames Bath Products including: Shower Gel, Face Soap, Shampoo, Conditioner, Body Soap, Body Lotion, Mouthwash, and a Shower Net
Fresh Flower in Room
Hair Dryer
Includes small refrigerator, microwave, and gourmet coffee maker
Alarm Clock
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On-Site Parking
Hof Haus has one King size bed accommodating 2 Adults


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On 21 December 2012, the Council of the European Union adopted Council Regulation No 1263/2012, which amends Regulation (EU) No 267/2012 (Regulation 1263/2012). Regulation 1263/2012 gives effect to the measures provided for in Council Decision 2012/635/CFSP of 15 October 2012 concerning additional restrictive measures against Iran (Decision 2012/635).
Click on the link above to download this Cleary Gottlieb briefing.


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Design by Aaron Thong
$30 $24 Womens Dolman
Super soft cotton, polyester, and rayon blend. Slouchy in the most savvy fashion, the dolman tee drapes like a vogue vixen. A round neckline and curved hemline complete the look for a casual classic. Women may want to order a size down.
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Pave diamonds sparkle at the center of a polished 18K yellow gold ring from Roberto Coin. 18K yellow gold/diamonds. Diamond:. 11 ct. T. W. Interior of jewelry piece includes signature synthetic ruby detail. All gemstone carat weights (ct. T. W. Are approximate; variance may be. 05 carat. Photo may have been enlarged and/or enhanced. Gemstone Treatment and Care. Country Of Origin: Made in Italy. Color: metallic


text: Our new kitten Bella! She loves everyone and especially those that will play with her to keep her rambunctious self entertained! She likes to give kisses and has the best purr box. She is a wonderful addition to our family and will provide many smiles for years to come :)


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Valentine Word Search iOS
- Publisher: AFKSoft
- Release Date: May 27, 2009
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Все отели Овьедо 4 звезды. Для бронирования доступно около 24 отелeй.
Гостиница в Овьедо
The 4-star Husa Gran Hotel Espana offers guests the chance to stay in one of Oviedo's oldest buildings, reflected by its fine regal architecture. The hotel offers spacious accommodation boasting cheerful decor, modern amenities, and a pictorial history of the hotel. The modern onsite facilities are also excellent quality.
This modern 4-star hotel boasts excellent central location. The hotel boasts a modern gym and an outstanding restaurant. Rooms feature modern design and free Wi-Fi internet access.
This modern 4-star hotel is located in the main shopping and leisure area of Oviedo. Main tourist attractions are within walking distance. The hotel boast a well-equipped gym and rooms with modern decor.
Отели Овьедо по звездам


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