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In this work, the plasmonic and photothermal effects of CuS nanoparticles biosynthesized from acid mine drainage (AMD) were studied. CuS were formed by delivering the H2S generated by a sulfidogenic bioreactor to an off-line system containing the AMD. The precipitates collected after contact for an hour were washed and physico-chemically characterized, showing a nanoparticle with a mean diameter of 33 nm, crystalline nature and semiconductor behavior with a direct band gap of 2.2 eV. Moreover, the CuS nanoparticles exhibited localized surface plasmonic resonance in the near infrared range, with a high absorption band centered at 973 nm of wavelength, which allowed an increase in the temperature of the surrounding media under irradiation. Finally, the cytotoxicity of the CuS nanoparticles as well as their potential use as part of drug delivery platforms were investigated.

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In the present work, CuS nanoparticles were biorecovered from a real acid mine drainage (AMD) and its photocatalytic and antibacterial activities were studied. CuS were formed by delivering biogenic H2S produced by a continuous sulfidogenic bioreactor to an off-line vessel containing the AMD. The main physico-chemical properties of CuS nanoparticles were analyzed by UV-vis spectroscopy, TEM, FE-SEM, XRD and XPS. Moreover, its photocatalytic activity on the photodegradation of organic dyes in water and its antibacterial activity against several bacterial strains were studied and compared with CuS nanoparticles synthetized from a CuSO4 aqueous solution based on the same synthesis method. CuS nanoparticles from the real AMD showed similar physico-chemical properties and photocatalytic and antibacterial activities in comparison to CuS nanoparticles formed with the copper solutions. These results open the way to recover valorous CuS nanoparticles from AMD with potential industrial applications using a metal bioremediation process based on sulfidogenic bioreactors.

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Recently, rare-earth elements (REEs) have attracted great interest due to their importance in several fields, such as the high-technology and medicine industries. Due to the recent intensification of the use of REEs in the world and the resulting potential impact on the environment, new analytical approaches for their determination, fractionation and speciation are needed. Diffusive gradients in thin films are a passive technique already used for sampling labile REEs, providing in situ analyte concentration, fractionation and, consequently, remarkable information on REE geochemistry. However, data based on DGT measurements until now have been based exclusively on the use of a single binding phase (Chelex-100, immobilized in APA gel). The present work proposes a new method for the determination of rare earth elements using an inductively coupled plasma‒mass spectrometry technique and a diffusive gradients in thin films (DGT) technique for application in aquatic environments. New binding gels were tested for DGT using carminic acid as the binding agent. It was concluded that acid dispersion directly in agarose gel presented the best performance, offering a simpler, faster, and greener method for measuring labile REEs compared to the existing DGT binding phase. Deployment curves obtained by immersion tests in the laboratory show that 13 REEs had linearity in their retention by the developed binding agent (retention x time), confirming the main premise of the DGT technique obeying the first Fick's diffusion law. For the first time, the diffusion coefficients were obtained in agarose gels (diffusion medium) and carminic acid immobilized in agarose as the binding phase for La, Ce, Pr, Nd, Sm, Eu, Gd, Dy, Ho, Er, Tm, Yb and Lu, which were 3.94 × 10-6, 3.87 × 10-6, 3.90 × 10-6, 3.79 × 10-6, 3.71 × 10-6, 4.13 × 10-6, 3.75 × 10-6, 3.94 × 10-6, 3.45 × 10-6, 3.97 × 10-6, 3.25 × 10-6, 4.06 × 10-6, and 3.50 × 10-6 cm2 s-1, respectively. Furthermore, the proposed DGT devices were tested in solutions with different pH values (3.5, 5.0, 6.5 and 8) and ionic strengths (I = 0.005 mol L-1, 0.01 mol L-1, 0.05 mol L-1 and 0.1 mol L-1 - NaNO3). The results of these studies showed an average variation in the analyte retention for all elements at a maximum of approximately 20% in the pH tests. This variation is considerably lower than those previously reported when using Chelex resin as a binding agent, particularly for lower pH values. For the ionic strength, the maximum average variation was approximately 20% for all elements (except for I = 0.005 mol L-1). These results indicate the possibility of a wide range of the proposed approach to be used for in situ deployment without the use of correction based on apparent diffusion coefficients (as required for using the conventional approach). In laboratory deployments using acid mine drainage water samples (treated and untreated), it was shown that the proposed approach presents excellent accuracy compared with data obtained from Chelex resin as a binding agent.

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Sulfate-rich effluents have been successfully treated in anaerobic reactors using sulfate-reducing bacteria (SRB). Many authors have demonstrated that these systems require nitrogen and phosphorous supplementation to achieve high sulfate removal rates. However, the resource ratio theory assumes that some species can be dominant according to the nutritional relations used or even without external nutrient supplementation. Thus, this study evaluated the SRB communities in batch reactors without external nitrogen and phosphorus sources based on most probable number (MPN) quantification, denaturing gradient gel electrophoresis (DGGE) analyses and sequencing. The sulfate and chemical oxygen demand (COD) removal and kinetic parameters were also determined. After 100 days of operation, the sulfate and COD removal achieved 71.8 ± 10% and 86.5 ± 10%, respectively. The SRB population increased from 8.106 to 4 × 1012 MPN 100 mL-1, and the richness of SRB bands was much higher at the end of the experiment compared to the inoculum. In addition, the sequenced bands from SRB-DGGE showed similarities to Desulfacinum infernum, Desulfobulbus sp, Syntrophobacter and Desulfomicrobium aestuarii-related sequences. Therefore, biological treatment of acid mine drainage wastewater was effective in the absence of nutrients, lowering costs and providing high sulfate removal efficiency.

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Uranium mining causes several radiological impacts on the surrounding environment, notably in the water bodies, mainly due to the release of long half-life radionuclides from the 238U and 232Th series. The Ore Treatment Unit, an old uranium mine undergoing decommissioning, has three points of liquid effluent release (#014, #025, and #076). For current study, 78 samples of water were collected at #014, 33 samples at #025, and 63 samples at #076. The radionuclides were analyzed by gross alpha count, gross beta count, and by arsenazo spectrophotometry. Analyses were carried out using the radiological water quality criterion established by World Health Organization and other organizations, together with the Brazilian legislation, to assess if the released effluents may be used unrestrictedly by the individuals of the public. At #014, the mean values of activity concentration (AC), in Bq·L-1, were as follows: Unat = 0.107, 226Ra = 0.035, 210Pb = 0.031, 232Th = 0.007, and 228Ra = 0.049. At #025 the mean values of AC, in Bq·L-1, were as follows: Unat = 0.086, 226Ra = 0.015, 210Pb = 0.028, 232Th = 0.006, and 228Ra = 0.032. Finally, at point #076, the mean AC values, in Bq·L-1, were as follows: Unat = 3.624, 226Ra = 0.074, 210Pb = 0.054, 232Th = 0.013, and 228Ra = 0.069. The current study showed that natural radionuclides were not in secular equilibrium. Despite uranium presented its values outside the limits of guidance levels, it can be state that the unrestricted use of effluents released in the three water bodies is authorized from the radiological point of view. In terms of dose rate, the releases at three points were within the radiological limits of potability. On the other hand, in an additional analysis, #76 presented chemical toxicity above the authorized value, pointing the need of restricted use of water from the point of view of chemical toxicity.

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Xichú River is a Mexican river located in an environmental preservation area called Sierra Gorda Biosphere Reserve. Around it, there are tons of abandoned mine residues that represent a serious environmental issue. Sediment samples of Xichú River, visibly contaminated by flows of an acid mine drainage, were collected to study their prokaryotic diversity. The study was based on both cultural and non-cultural approaches. The analysis of total 16S rRNA gene by MiSEQ sequencing allowed to identify 182 Operational Taxonomic Units. The community was dominated by Pseudomonadota, Bacteroidota, "Desulfobacterota" and Acidobacteriota (27, 21, 19 and 16%, respectively). Different culture conditions were used focusing on the isolation of anaerobic bacteria, including sulfate-reducing bacteria (SRB) and arsenate-reducing bacteria (ARB). Finally, 16 strains were isolated. Among them, 12 were phylogenetically identified, with two strains being SRB, belonging to the genus Solidesulfovibrio ("Desulfobacterota"), while ten are ARB belonging to the genera Azospira (Pseudomonadota), Peribacillus (Bacillota), Raineyella and Propionicimonas (Actinomycetota). The isolate representative of Raineyella genus probably corresponds to a new species, which, besides arsenate, also reduces nitrate, nitrite, and fumarate.

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Sulfate-reducing bioreactors are a biotechnological alternative for the treatment of acid mine drainage (AMD). In this study, two separate bioreactors with pH and temperature-controlled (Bio I and II) were operated with two different acidophilic microbial consortia to determine their efficiencies in sulfate removal from a synthetic acidic mine water. The bioreactors were operated for 302 days in continuous flow mode under the same parameters: fed with a sulfate solution of ∼30 mM with a pH of 2.5, the temperature at 30°C, stirred gently at 40 rpm and using a continuous stream of nitrogen to help remove the H2S produced in the bioreactor. The glycerol consumption, acetate production, and sulfate removal were monitored throughout the course of the experiment. The community composition and potential metabolic functional groups were analyzed via 16S rRNA partial gene sequencing. Bio I consortium reduced the sulfate, achieving a range of sulfate concentration from 4.7 to 19 mM in the effluent liquor. The removal of sulfate in Bio II was between 5.6 and 18 mM. Both bioreactors' communities showed the presence of the genus De sulfosporosinus as the main sulfate-reducing bacteria (SRB). Despite differences in microbial composition, both bioreactors have similar potential metabolism, with a higher percentage of microorganisms that can use sulfate in respiration. Overall, both bioreactors showed similar performance in treating acidic mine water containing mostly sulfate using two different acidophilic sulfidogenic consortia obtained from different global locations.

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Acid mine drainage (AMD) causes major environmental problems and consequently, several treatments are proposed, favoring the passive systems because of their many advantages. The main goal of these procedures is the neutralization and removal of potentially toxic elements (PTE), yet little is known about the changes in the microbial assemblages in response to the hydrochemical variations during the treatments. Therefore, the main objective of this research was to determine the changes in the diversity and structure of the prokaryotic assemblages in a hybrid abiotic and biological (wetland) passive treatment system. The 16S rRNA gene survey showed that the AMD coming from the mine (pH 2.6) was mainly composed of acidophilic genera such as Acidithiobacillus, Leptospirillum, Ferritrophicum, and Cuniculiplasma (up to 76 % relative abundance). In the abiotic treatment, Acidiphilium was dominant in the sections with limestone filters (pH 2.2-4.8), followed by Limnobacter in the subsequent dolomite/limestone and phosphoric rock filters (pH 5.2-5.8). In these abiotic passive treatment sections, the microbial assemblage showed a limited diversity and richness. However, when the treated AMD reached the two final wetlands (pH ~6.8), the microbial diversity and richness increased, suggesting that further bioattenuation mechanisms might be occurring. Limnobacter and Novosphingobium were the main bacterial genera in the water samples of the wetland sections (Arundo donax). These changes in the composition of the microbial assemblages were highly correlated with the pH and Eh values during the treatment (p-value <0.001); however, the concentration of metal(loid)s such as Al, Cd, Fe, Mn, Ni, and Zn were also significantly related (p-value <0.05). In conclusion, the studied passive AMD treatment system enhanced the chemical quality of the treated AMD, showing high removal efficiencies for Al and Fe (> 99 %), and increasing the microbial diversity and richness in the effluent.

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RESUMEN En esta investigación bibliográfica se encontraron reportes sobre una gran variedad de especies responsables de precipitar a cuatro metales de interés (Cu, Pb, Zn y Fe). En la mayoría de las investigaciones no solamente se considera la precipitación de estos metales, sino también la de otros elementos que están presentes en cada efluente estudiado. Los artículos aquí mencionados tienen una relación directa con el efluente proveniente de la operación unitaria de flotación. Aportan conocimiento acerca del proceso de sulfato-reducción, comprendiendo el mecanismo mediante microorganismos con características específicas, especialmente su versatilidad pues se desarrollan en diferentes ecosistemas. Se muestra que varias especies, como Desulfobacter o Desulfovibrio son comunes pues tienen condiciones relativamente sencillas para desarrollarse. Los microorganismos sulfato reductores (MSR) son eficientes para reducir la acidez del agua (de la operación unitaria de flotación de una mina, de cocinas, de corrientes marinas, etc.). También lo son para precipitar diferentes elementos pues no requieren de algún agente externo salvo en contadas ocasiones donde debe actuar un catalizador. Hay investigaciones sobre los nutrientes que deben adicionarse para incrementar su actividad. Los reportes de investigación revisados identificaron las variables a controlar para obtener buenos resultados en la remoción de metales y menores impactos en el ambiente. Es de gran importancia el desarrollo de proyectos que tomen en cuenta un sistema natural, como la degradación anaerobia, para alcanzar un punto en el cual la tecnología y el ambiente puedan complementarse logrando bienes de consumo necesarios para la población sin causar daños irreparables a la naturaleza.

ABSTRACT In this bibliographical research, reports were found on a great variety of species responsible for precipitating four metals of interest (Cu, Pb, Zn and Fe). In most of the investigations, not only the precipitation of these metals is considered, but also that of other elements that are present in each effluent studied. The items mentioned here have a direct relationship with the effluent from the flotation unit operation. They provide knowledge about the sulfate-reduction process, understanding the mechanism through microorganisms with specific characteristics, especially their versatility as they develop in different ecosystems. It is shown that several species, such as Desulfobacter or Desulfovibrio, are common because they have relatively simple conditions to develop. Sulfate-reducing microorganisms (SRM) are efficient in reducing the acidity of water (from the flotation unit operation of a mine, kitchens, ocean currents, etc.). They are also used to precipitate different elements since they do not require any external agent except on rare occasions when a catalyst must act. There is research on the nutrients that should be added to increase its activity. The research reports reviewed identified the variables to control to obtain good results in the removal of metals and less impact on the environment. The development of projects that take into account a natural system, such as anaerobic degradation, is of great importance in order to reach a point where technology and the environment can complement each other, achieving necessary consumer goods for the population without causing irreparable damage to nature.

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The Ore Treatment Unit was a uranium mining company that is currently being decommissioned. The local rainfall index makes it necessary to release effluents into the environment. After releasing, the wastewater is available for unrestricted use. Current study aims to use national and international recommendations to assess the radiological potability of released effluents at one of the three points of company's interface with the environment. Twenty-four samples of water were collected and activity concentrations (AC) were obtained by gross alpha count, gross beta count, and for arsenazo spectrophotometry. Statistical analysis techniques were applied to the data with the purpose of understanding the results for the soluble, particulate, and total fractions. The mean AC for effluents were 3.580, 0.082, 0.103, 0.063, and 0.090 Bq L-1 for Unat, 226Ra, 210Pb, 232Th, and 228Ra, respectively, for the total fraction. The analysis of variance pointed to Unat as a critical radionuclide, since it presented more than 90% of the total AC released into the environment. Pearson's R2 pointed to soluble fraction as a major contributor to the total AC released. The guidance level proposed by WHO was used to assess the radiological potability of the effluents. The results obtained indicated the need for trigger other analyses. Committed effective dose was estimated due to the unrestricted use of effluents and the value obtained, 0.23 mSv year-1, was below the maximum allowed limit. Finally, the radiotoxicity of the released effluent was evaluated and the value obtained was ~ 50% of the maximum allowed limit. In conclusion, the present study showed that the level of radioactivity released into the environment by the Ore Treatment Unit does not present a radiological risk to the surrounding population.

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Reactive transport models have proven abilities to simulate the quantity and quality of drainage from mine waste rock. Tracer experiments indicate the presence of fast and slow flow regimes in many heterogeneous waste-rock piles. Although multidomain models have been developed specifically for systems with such distinctive hydrodynamics, there have been limited applications of multidomain reactive transport models to simulate composite drainage chemistries from waste-rock piles to date. This work evaluated the ability of dual-domain multicomponent reactive transport models (DDMRTMs) to reproduce breakthrough curves of conservative (chloride) and reactive (molybdenum) solutes observed at a well-characterized experimental waste-rock pile at the Antamina Mine, Peru. We found that the DDMRTM simulations quantitatively matched eight-year-long records of conservative transport through the waste-rock pile when parameterized mainly with field-measured properties obtained from the site and limited calibration. The DDMRTM model also provided a reasonable match to field observations of the reactive solute. The limited calibrated parameters are physically realistic, corroborating the ability of these multidomain models to reproduce the complex reactive-transport processes governing polluted rock drainage from large-scale waste-rock piles.

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Resumo A drenagem ácida de mina da mineração de carvão é um dos mais graves problemas ambientais que existem atualmente e é caracterizada, principalmente, por apresentar elevada acidez, baixo pH e expressiva concentração de metais tóxicos, como ferro, Mn e muitos outros, afetando diretamente mananciais e rios. Em busca de uma alternativa que pudesse melhorar, de forma eficiente e econômica, os níveis de acidez e ferro da água impactada pela drenagem ácida de mina, foi desenvolvido um adsorvente geopolimérico à base de materiais residuais da indústria cerâmica e do beneficiamento de arroz (cinzas da casca de arroz). O objetivo desta pesquisa foi avaliar a eficiência do geopolímero na remoção de íons ferro em água contaminada com drenagem ácida de mina. Foram avaliados aspectos de dosagem do adsorvente, efeito da temperatura, concentrações iniciais de ferro, cinética e parâmetros termodinâmicos do processo de adsorção. O percentual de ferro removido foi de 92,76%, à temperatura de 25 °C, em um período de 20 min, com uma concentração de adsorvente de 4 g L-1. A capacidade máxima de adsorção de ferro pelo geopolímero foi de 7,18 mg.g-1. O principal mecanismo de adsorção ocorreu em razão da quimissorção, que segue o modelo cinético de pseudossegunda ordem. O geopolímero se mostrou como uma alternativa eficiente ao tratamento de água contaminada com drenagem ácida de mina.

Abstract Acid mine drainage is a worldwide problem and is characterized by high acidity, low pH and expressive concentration of heavy metals, such as iron, Mn and many others, directly affecting water sources and rivers. In search of an alternative that could efficiently and economically improve the levels of acidity and water iron impacted by acid mine drainage, a geopolymeric adsorbent based on residual materials was developed: from the ceramic industry and rice processing (rice husk ash). In this work, it was evaluated the efficiency of the geopolymer in removing iron ions in water contaminated with acid mine drainage. Aspects of adsorbent dosage, temperature effect, initial iron concentrations, kinetics and thermodynamic parameters of the adsorption process were evaluated. The percentage of iron removed was 92.76%, at a temperature of 25 °C, for 20 min, with an adsorbent concentration of 4 g L-1, with the maximum capacity for adsorption of iron by the geopolymer being 7.18 mg.g-1. The main mechanism of adsorption occurred due to chemisorption, which follows the kinetic model of pseudo-second order. Geopolymer appears potentially useful an efficient alternative in the treatment of water contaminated with acid mine drainage.

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In southern South America, Brazil, in the state of Santa Catarina, the neglect and lack of environmental supervision during coal mining caused the contamination of surface and groundwater by acid mine drainage (AMD). By the force of the local law, environmental reclamation actions in these abandoned areas have been carried. A scientific work of monitoring and assessment of the water resources within these areas has never been developed, as the efficacy of the reclamation strategies has never been discussed. This work aims to fill this gap by presenting and analyzing the environmental reclamation strategy of a former degraded coal mining area and its impacts on local water resources. The water monitoring plan in Area IV's was carried out in groundwater, and in lentic (ponds) and lotic (rivers) environments of surface waters, fourteen monitoring campaigns were conducted. The results showed that upstream and downstream river points have different water qualities, with the downstream points having poorer water quality, still affected by past mining activities. From the surface water perspective, the reclaiming method adopted was effective in three of the four ponds, presenting problems only in the downstream one. Two hypotheses were proposed; the first hypothesis is that contamination happens due to leaching of the material that still remains on the ponds' banks into the water. Another hypothesis is that the contamination comes from the upstream groundwater inflow into the pond, which runs through the entire area before reaching the pond. Those results serve to further access the actual monitoring perspectives as well as to better develop future reclaiming strategies.

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Pollution due to acidic and metal-enriched waters affects the quality of surface and groundwater resources, limiting their uses for various purposes. Particularly, manganese pollution has attracted attention due to its impact on human health and its negative effects on ecosystems. Applications of nanomaterials such as graphene oxide (GO) have emerged as potential candidates for removing complex contaminants. In this study, we present the preliminary results of the removal of Mn(II) ions from acidic waters by using GO functionalized with zinc oxide nanoparticles (ZnO). Batch adsorption experiments were performed under two different acidity conditions (pH1 = 5.0 and pH2 = 4.0), in order to evaluate the impact of acid pH on the adsorption capacity. We observed that the adsorption of Mn(II) was independent of the pHPZC value of the nanoadsorbents. The qmax with GO/ZnO nanocomposites was 5.6 mg/g (34.1% removal) at pH = 5.0, while with more acidic conditions (pH = 4.0) it reached 12.6 mg/g (61.2% removal). In turn, the results show that GO/ZnO nanocomposites were more efficient to remove Mn(II) compared with non-functionalized GO under the pH2 condition (pH2 = 4.0). Both Langmuir and Freundlich models fit well with the adsorption process, suggesting that both mechanisms are involved in the removal of Mn(II) with GO and GO/ZnO nanocomposites. Furthermore, adsorption isotherms were efficiently modeled with the pseudo-second-order kinetic model. These results indicate that the removal of Mn(II) by GO/ZnO is strongly influenced by the pH of the solution, and the decoration with ZnO significantly increases the adsorption capacity of Mn(II) ions. These findings can provide valuable information for optimizing the design and configuration of wastewater treatment technologies based on GO nanomaterials for the removal of Mn(II) from natural and industrial waters.

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Acid mine drainage-affected environments are interesting microbial niches for the isolation of metal-resistant microorganisms. In this sense, the aim of the present work is to isolate and characterize metal-resistant microorganisms from sediments of an abandoned gold mine located in San Luis (Argentina). For these purposes, the metal removal capacity and the microelemental composition of the biomass exposed to metals were evaluated. Likewise, proteomic techniques were applied to understand the removal and resistance mechanisms. Fusarium tricinctum M6 was isolated and identified as tolerant to Cu(II), Fe(II) and Cr(VI). When faced with 40 µg mL-1 Cu(II), the growth was affected by 60% and the removal capacity was 30-35%. Copper was found uniformly distributed in the biomass (5.23% w/w) and variations in the proportion of other biomass constituent elements were detected. When exposed to Cu(II), F. tricinctum M6 showed differential expression of intra and extracellular proteins involved in different metabolic processes. A large number of proteins with metal ion binding sites were detected both at intra and extracellular levels. The results obtained in the present work indicated bioadsorption of the metal on the cell surface and an important readjustment of the protein expression to counteract the stress produced by Cu(II).

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The risk of generation of acid drainages in the tailings of the Pan de Azúcar mine that closed its activities more than three decades ago, was evaluated through biooxidation studies using iron- and sulfur-oxidizing extremophilic leaching consortia. Most of tailings showed a high potential for generating acid drainage, in agreement with the results from net acid generation (NAG) assays. In addition, molecular analysis of the microbial consortia obtained by enrichment of the samples, demonstrated that native leaching microorganisms are ubiquitous in the area and they seemed to be more efficient in the biooxidation of the tailings than the collection microorganisms. The acid drainages detected at the site and those formed by oxidation of the tailings, produced a significant ecotoxicological effect demonstrated by a bioassay. These drainages, even at high dilutions, could seriously affect a nearby Ramsar site (Laguna de Pozuelos) that is connected to the Pan de Azúcar mine through a hydrological route (Cincel River).

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Acid mine drainage occurs due to the chemical and microbiological oxidation of sulfide minerals and can be a source of potentially toxic elements contamination of groundwater and surface water. The objective of this study was to identify microorganisms involved in sulfide oxidation in the tailings of a Bahia Gold Belt mine (Brazil). Samples of solids and water were collected at the mine tailings dam and characterized. The microorganisms were isolated after enrichment and subsequent purification. The major constituents of the tailings are Si, Fe, Al, S, and K. The sulfur content of the tailings is 0.98%. The major phases are quartz, muscovite, and clinochlore. Gravity concentrates of the tailings show several particles of pyrite, that is, the major sulfide phase. Molecular analysis identified the microorganisms isolated in the acid mine drainage process in this region. Five bacterium species were found: Acidithiobacillus spp., Acidithiobacillus ferrooxidans, Acidiphilium spp., Leptospirillum type II, and Sulfobacillus spp. No organisms of the archaea or eukaryote domains were found. The isolate was used in the bioleaching of copper sulfide ore, and the copper extraction was about 60% in 60 days for ground ore.

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The environmental contamination due to mining activities in the Andean region of Bolivia is a serious concern, as it leads to highly acidic (pH 2.4) acid mine drainage (AMD), severely polluted by sulfate (>12,000 mg L-1). Passive bioreactors entailing biological sulfate reduction and removal of metals through sulfide precipitation have been recognized as a promising biotechnology. The reactivity of mixtures containing locally available substrates: sheep manure, compost and straw, was assessed through batch experiments conducted with a synthetic solution simulating the composition of AMD from San José mine (Oruro). The removal of sulfate and metals was successful in all reactors, at the end of the experiment (56 days) sulfate concentrations dropped to 1378-2081 mg L-1, corresponding to a removal efficiency between 84% and 89%, while average removal for Fe, Zn, Pb, and Cd were 99.8%, 98.5%, 94.7%, 98.6%, respectively. The sulfate and metal removal showed three phases. In the first phase, the removal was independent of the organic composition and attributable to pH-controlled mechanisms i.e. adsorption, precipitation of oxy(hydroxides) and co-precipitation. During the second phase, sulfate and metals concentrations remained rather constant; while in the third phase, the removal was affected by the organic matter composition. Sulfate removal rate attained the highest values (227-243 mg L-1 d-1) in the third phase, and it was attributable to biological reduction with not sulfate limitation. The depletion of nutrients rather than the sulfate availability may have limited the sulfate removal at the end of the experiment.

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Extreme acidophiles thrive in environments rich in protons (pH values <3) and often high levels of dissolved heavy metals. They are distributed across the three domains of the Tree of Life including members of the Proteobacteria. The Acidithiobacillia class is formed by the neutrophilic genus Thermithiobacillus along with the extremely acidophilic genera Fervidacidithiobacillus, Igneacidithiobacillus, Ambacidithiobacillus, and Acidithiobacillus. Phylogenomic reconstruction revealed a division in the Acidithiobacillia class correlating with the different pH optima that suggested that the acidophilic genera evolved from an ancestral neutrophile within the Acidithiobacillia. Genes and mechanisms denominated as "first line of defense" were key to explaining the Acidithiobacillia acidophilic lifestyle including preventing proton influx that allows the cell to maintain a near-neutral cytoplasmic pH and differ from the neutrophilic Acidithiobacillia ancestors that lacked these systems. Additional differences between the neutrophilic and acidophilic Acidithiobacillia included the higher number of gene copies in the acidophilic genera coding for "second line of defense" systems that neutralize and/or expel protons from cell. Gain of genes such as hopanoid biosynthesis involved in membrane stabilization at low pH and the functional redundancy for generating an internal positive membrane potential revealed the transition from neutrophilic properties to a new acidophilic lifestyle by shaping the Acidithiobacillaceae genomic structure. The presence of a pool of accessory genes with functional redundancy provides the opportunity to "hedge bet" in rapidly changing acidic environments. Although a core of mechanisms for acid resistance was inherited vertically from an inferred neutrophilic ancestor, the majority of mechanisms, especially those potentially involved in resistance to extremely low pH, were obtained from other extreme acidophiles by horizontal gene transfer (HGT) events.

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Adsorption technologies are a focus of interest for the removal of pollutants in water treatment systems. These removal methods offer several design, operation and efficiency advantages over other wastewater remediation technologies. Particularly, graphene oxide (GO) has attracted great attention due to its high surface area and its effectiveness in removing heavy metals. In this work, we study the functionalization of GO with zinc oxide nanoparticles (ZnO) to improve the removal capacity of aluminum (Al) and copper (Cu) in acidic waters. Experiments were performed at different pH conditions (with and without pH adjustment). In both cases, decorated GO (GO/ZnO) nanocomposites showed an improvement in the removal capacity compared with non-functionalized GO, even when the pH of zero charge (pHPZC) was higher for GO/ZnO (5.57) than for GO (3.98). In adsorption experiments without pH adjustment, the maximum removal capacities for Al and Cu were 29.1 mg/g and 45.5 mg/g, respectively. The maximum removal percentages of the studied cations (Al and Cu) were higher than 88%. Further, under more acidic conditions (pH 4), the maximum sorption capacities using GO/ZnO as adsorbent were 19.9 mg/g and 33.5 mg/g for Al and Cu, respectively. Moreover, the removal percentages reach 95.6% for Al and 92.9% for Cu. This shows that decoration with ZnO nanoparticles is a good option for improving the sorption capacity of GO for Cu removal and to a lesser extent for Al, even when the pH was not favorable in terms of electrostatic affinity for cations. These findings contribute to a better understanding of the potential and effectiveness of GO functionalization with ZnO nanoparticles to treat acidic waters contaminated with heavy metals and its applicability for wastewater remediation.

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