Organic matter degradation in the deep, sulfidic waters of the Black Sea: insights into the ecophysiology of novel anaerobic bacteria.

BACKGROUND: Recent studies have reported the identity and functions of key anaerobes involved in the degradation of organic matter (OM) in deep (> 1000 m) sulfidic marine habitats. However, due to the lack of available isolates, detailed investigation of their physiology has been precluded. In this study, we cultivated and characterized the ecophysiology of a wide range of novel anaerobes potentially involved in OM degradation in deep (2000 m depth) sulfidic waters of the Black Sea. RESULTS: We have successfully cultivated a diverse group of novel anaerobes belonging to various phyla, including Fusobacteriota (strain S5), Bacillota (strains A1T and A2), Spirochaetota (strains M1T, M2, and S2), Bacteroidota (strains B1T, B2, S6, L6, SYP, and M2P), Cloacimonadota (Cloa-SY6), Planctomycetota (Plnct-SY6), Mycoplasmatota (Izemo-BS), Chloroflexota (Chflx-SY6), and Desulfobacterota (strains S3T and S3-i). These microorganisms were able to grow at an elevated hydrostatic pressure of up to 50 MPa. Moreover, this study revealed that different anaerobes were specialized in degrading specific types of OM. Strains affiliated with the phyla Fusobacteriota, Bacillota, Planctomycetota, and Mycoplasmatota were found to be specialized in the degradation of cellulose, cellobiose, chitin, and DNA, respectively, while strains affiliated with Spirochaetota, Bacteroidota, Cloacimonadota, and Chloroflexota preferred to ferment less complex forms of OM. We also identified members of the phylum Desulfobacterota as terminal oxidizers, potentially involved in the consumption of hydrogen produced during fermentation. These results were supported by the identification of genes in the (meta)genomes of the cultivated microbial taxa which encode proteins of specific metabolic pathways. Additionally, we analyzed the composition of membrane lipids of selected taxa, which could be critical for their survival in the harsh environment of the deep sulfidic waters and could potentially be used as biosignatures for these strains in the sulfidic waters of the Black Sea. CONCLUSIONS: This is the first report that demonstrates the cultivation and ecophysiology of such a diverse group of microorganisms from any sulfidic marine habitat. Collectively, this study provides a step forward in our understanding of the microbes thriving in the extreme conditions of the deep sulfidic waters of the Black Sea. Video Abstract.

Characteristics of submerged and partially submerged caves (habitat type 8330) in Romania.

Underwater sea caves form a relatively under-examined habitat type within the marine regions of Europe, although they provide unique physical conditions such as reduced light and wave energy, in addition to reduced temperature amplitude. This study aimed at revealing the characteristics of submerged cavities on the southern Romanian continental shelf where six protected areas exist. We used high-resolution bathymetry data and side-scan sonar imaging to identify limestone outcrops where cavities would be most probable to form and then performed visual observation during SCUBA diving activities. We found that submerged cavities appear in all marine-protected areas and their neighboring unprotected areas from the shore to ~20-m depth mostly in the form of caverns, niches, and overhangs that meet the physical requirements of the habitat type 8330 as defined by the European Commission. We propose that habitat type 8330 should be listed in the Natura 2000 Standard Data Forms of the protected areas where it is missing, and we also propose that some protected areas should be expanded to include important areas with the occurrence of this habitat type. Finally, we note here for the first time the occurrence of Movile-type karst and an associated sulfidic water spring on the southern shore of Lake Techirghiol, similar to the region near Mangalia where it was first described in the literature. Although our finding prompts for a more detailed study, it indicates the existence of an underground ecosystem similar to that from the well-studied Movile Cave, but isolated by a distance of over 20 km.

Compost-assisted revegetation of highly phytotoxic sulfidic tailings with Medicago sativa L. plants grown from the seed to seedpod stage under greenhouse experimental mesocosms conditions.

The revegetation of highly phytotoxic sulfidic tailings is a challenging task which may often be successfully accomplished only following the addition of soil amendments. This study evaluated the use of green compost at increasing rates (10, 25 and 50% v/v) for the revegetation of extremely acidic sulfidic tailings of the North Mathiatis mine, Cyprus, with the use of alfalfa (Medicago sativa L.) plants, under greenhouse conditions. Alfalfa seeds were successfully germinated in tailings amended either with 25% or 50% (v/v) compost (52 and 85%, respectively). Plants managed to complete their life cycle and produce seeds only in the tailings amended with 50% (v/v) compost, since plants grown in tailings amended with lower rates of compost (i.e., 10 or 25% v/v) showed severe symptoms of phytotoxicity and eventually died. The amendment of tailings with 50% (v/v) green compost resulted in increased pH values, water holding capacity and organic content levels, soil respiration rates, as well as changes in soil elemental composition compared with tailings alone treatment, which in turn facilitated the growth and development of alfalfa plants during the whole experimental period (140 days). Plants managed to reach the late seedpod growth stage, indicating their potential regeneration and continual existence to the amended tailings, simultaneously uncovering the development of favorable conditions in the rhizosphere for the successful revegetation of studied tailings.

Sulfur-cycling chemolithoautotrophic microbial community dominates a cold, anoxic, hypersaline Arctic spring.

BACKGROUND: Gypsum Hill Spring, located in Nunavut in the Canadian High Arctic, is a rare example of a cold saline spring arising through thick permafrost. It perennially discharges cold (~ 7 °C), hypersaline (7-8% salinity), anoxic (~ 0.04 ppm O2), and highly reducing (~ - 430 mV) brines rich in sulfate (2.2 g.L-1) and sulfide (9.5 ppm), making Gypsum Hill an analog to putative sulfate-rich briny habitats on extraterrestrial bodies such as Mars. RESULTS: Genome-resolved metagenomics and metatranscriptomics were utilized to describe an active microbial community containing novel metagenome-assembled genomes and dominated by sulfur-cycling Desulfobacterota and Gammaproteobacteria. Sulfate reduction was dominated by hydrogen-oxidizing chemolithoautotrophic Desulfovibrionaceae sp. and was identified in phyla not typically associated with sulfate reduction in novel lineages of Spirochaetota and Bacteroidota. Highly abundant and active sulfur-reducing Desulfuromusa sp. highly transcribed non-coding RNAs associated with transcriptional regulation, showing potential evidence of putative metabolic flexibility in response to substrate availability. Despite low oxygen availability, sulfide oxidation was primarily attributed to aerobic chemolithoautotrophic Halothiobacillaceae. Low abundance and transcription of photoautotrophs indicated sulfur-based chemolithoautotrophy drives primary productivity even during periods of constant illumination. CONCLUSIONS: We identified a rare surficial chemolithoautotrophic, sulfur-cycling microbial community active in a unique anoxic, cold, hypersaline Arctic spring. We detected Mars-relevant metabolisms including hydrogenotrophic sulfate reduction, sulfur reduction, and sulfide oxidation, which indicate the potential for microbial life in analogous S-rich brines on past and present Mars. Video Abstract.

5-year leaching experiments to evaluate a modified bauxite residue: remediation of sulfidic mine tailings.

The ALTEO company produces approximately 300,000 tons per year of bauxite residue after alumina extraction, which is washed and dried in a press filter to produce Bauxaline®. In this study, different ways for recovering and reusing this residue were explored, namely transformation into a vegetated soil, use in acid mine drainage depollution, and application in sulfide-mine tailings remediation. The Bauxaline® was therefore transformed into modified bauxite residue (MBR), resulting in reduced alkalinity, salinity, and sodicity. To counterbalance the net acid generation potential of two sulfidic mine tailings with 1 mol H+ kg-1 (1.5% sulfide) and 3.3 mol H+ kg-1 (5.3% sulfide), respectively, various treatments were applied. These treatments included the addition of 10% MBR or 10% MBR plus limestone, or by limestone only, within 40-l lysimeters. Six lysimeters were monitored over a 5-year period to assess the long-term emissions from treated materials. Vegetation was tested under various conditions, and its impact on emission was evaluated. The emissions of mine tailings treated with MBR and limestone were very low. The mine tailings with limestone showed intermittent peaks of emission, probably due to the coating of calcite grain by ferric oxide, hindering contact with percolating water. Vegetation successfully grew in the treated tailings. This study demonstrated that the alkalinity of limestone can temporarily immobilize elements in sulfidic mine tailings, with a reduction factor of emissions of 300 and 40 for the two mine tailings, respectively. For long-term immobilization, the alkalinity provided by both limestone and MBR and the Al and Fe oxides of MBR are more effective and necessary for long-term immobilization, with a reduction factor of 300 and 900, respectively.

Competition-cooperation in the chemoautotrophic ecosystem of Movile Cave: first metagenomic approach on sediments.

BACKGROUND: Movile Cave (SE Romania) is a chemoautotrophically-based ecosystem fed by hydrogen sulfide-rich groundwater serving as a primary energy source analogous to the deep-sea hydrothermal ecosystems. Our current understanding of Movile Cave microbiology has been confined to the sulfidic water and its proximity, as most studies focused on the water-floating microbial mat and planktonic accumulations likely acting as the primary production powerhouse of this unique subterranean ecosystem. By employing comprehensive genomic-resolved metagenomics, we questioned the spatial variation, chemoautotrophic abilities, ecological interactions and trophic roles of Movile Cave's microbiome thriving beyond the sulfidic-rich water. RESULTS: A customized bioinformatics pipeline led to the recovery of 106 high-quality metagenome-assembled genomes from 7 cave sediment metagenomes. Assemblies' taxonomy spanned 19 bacterial and three archaeal phyla with Acidobacteriota, Chloroflexota, Proteobacteria, Planctomycetota, Ca. Patescibacteria, Thermoproteota, Methylomirabilota, and Ca. Zixibacteria as prevalent phyla. Functional gene analyses predicted the presence of CO2 fixation, methanotrophy, sulfur and ammonia oxidation in the explored sediments. Species Metabolic Coupling Analysis of metagenome-scale metabolic models revealed the highest competition-cooperation interactions in the sediments collected away from the water. Simulated metabolic interactions indicated autotrophs and methanotrophs as major donors of metabolites in the sediment communities. Cross-feeding dependencies were assumed only towards 'currency' molecules and inorganic compounds (O2, PO43-, H+, Fe2+, Cu2+) in the water proximity sediment, whereas hydrogen sulfide and methanol were assumedly traded exclusively among distant gallery communities. CONCLUSIONS: These findings suggest that the primary production potential of Movile Cave expands way beyond its hydrothermal waters, enhancing our understanding of the functioning and ecological interactions within chemolithoautotrophically-based subterranean ecosystems.

Suppression of reductive characters in white wine by Cu fractions: Efficiency and duration of protection during bottle aging.

Cu in wine can suppress sulfidic-odours, but the active forms and duration of protection are uncertain. Additions of 0, 0.3 or 0.6 mg/L Cu(II) were made to Chardonnay and Pinot Grigio at bottling. Throughout a 12- or 14-month storage period, Cu fractions were determined by colorimetry, and sulfhydryl compounds by gas chromatography with sulfur chemiluminescence detection. After Cu(II) addition, the dominant Cu fractions were associated with Cu(II)-organic acids (fraction I) and Cu(I)-thiol complexes (fraction II), and over 8-months their concentrations gradually fell below 0.015 mg/L. During this time, a fraction of Cu, predominantly attributed to sulfide-bound Cu, increased in concentration. Suppression of free hydrogen sulfide was assured when the combined Cu fractions I and II concentrations were above 0.015 mg/L, while free methanethiol suppression required Cu fraction I concentration above 0.035 mg/L. Decay rates for Cu fractions demonstrated that the duration that Cu can actively suppress sulfidic odours is wine-dependent.

Hydrolysis of Argyrodite Sulfide-Based Separator Sheets for Industrial All-Solid-State Battery Production.

Researchers have been working for many years to find new material and cell systems that can be used as potential post-lithium-ion batteries. Among these, the all-solid-state battery is considered a promising candidate, with sulfide-based materials having essential advantages over other solid electrolyte materials, particularly in terms of their high ionic conductivity. A great challenge, however, is their high reactivity in contact with water, where harmful hydrogen sulfide (H2S) is formed. Since H2S formation has implications for both worker safety and material quality, it is important to quantify its impact. For this reason, this paper examines the relationship between the product properties and the H2S formation as well as influences resulting from the production environment. Exemplary material states along the process chain of a wet coating process route are analyzed for the steps of storage, mixing, coating, drying, and densifying with Li6PS5Cl (LPSCl) as a solid electrolyte material. By determining the H2S formation rate for sulfide-based separator sheets, it is shown that the water content in the surrounding atmosphere has the highest impact, while other investigated parameters are negligibly small in comparison. Among the product properties, the geometric surface and pore surface have a great influence. These results demonstrate the need for a controlled atmosphere in the production facilities at dew points of -40 to -50 °C. At those moisture levels, occupational safety and product quality are ensured for the investigated solid electrolyte sheets of LPSCl. This study is the first to provide quantitative data from the point of view of the production environment on the formation of H2S gas when using solid sulfide electrolytes and can therefore serve as a guideline for equipment, material, and cell manufacturers.

Shifts in benthic bacterial communities associated with farming stages and a microbiological proxy for assessing sulfidic sediment conditions at fish farms.

To assess the aquaculture-induced sediment conditions associated with sulfur cycles, shifts in bacterial communities across farming stages were investigated. The sulfate reduction rate (SRR), and concentrations of acid volatile sulfide (AVS) and H2S were significantly higher at the mid- and post-farming stages than at the early stage, indicating that the aquaculture effects persist even after harvest. Incomplete organic carbon-oxidizing sulfate-reducing bacteria (IO-SRB) affiliated with Desulfobulbaceae, and gammaproteobacterial sulfur oxidizing bacteria (SOB) (Thiohalobacter, Thioprofundum, and Thiohalomonas) were dominant during the early stage, whereas fermenting bacteria (Bacteroidetes and Firmicutes) and complete oxidizing SRB (CO-SRB) belonging to Desulfobacteraceae, and epsilonproteobacterial SOB (Sulfurovum) dominated during the mid- and post-stages. The shift in SRB and SOB communities well reflected the anoxic and sulfidic conditions of farm sediment. Especially, the Sulfurovum-like SOB correlated highly and positively with H2S, AVS, and SRR, suggesting that they could be relevant microbiological proxies to assess sulfidic conditions in farm sediment.

Application of Silicate-Based Coating on Pyrite and Arsenopyrite to Inhibit Acid Mine Drainage.

The prevention of acid generation from sulfidic mine wastes is a problem that challenges the global scientific community for decades. A promising strategy is related to the formation of coating layer around sulfides for inhibiting surface oxidation. In the current research, the conditions favoring the formation of an efficient silicate-based coating around pyrite and arsenopyrite were studied, using batch tests. The coating solutions contained silicate-oxyanions, an oxidant (H2O2) and buffered at pH 6. The effect of Si concentration (0.1-50 mM), liquid/solid ratio (5-100 mL/g) and contact time (up to 24 h) was investigated. Pyrite tailings treated with a solution of 1 mM Si/0.1 M H2O2 at L/S:100 mL/g for 24 h resulted in the optimum formation of a coating, which reduced the amount of SO4-2-released by 72%, compared to the sample treated in the absence of Si. However, silicate treatment had a negative effect on arsenopyrite tailings inducing As mobilization.

Delving into the bacterial diversity of spoiled green Manzanilla Spanish-style table olive fermentations.

This work applies metataxonomic, standard statistics, and compositional data (CoDa) techniques to study the bacterial diversity of spoiled and normal Spanish-style table olive fermentations, analysing a total of 10-tons of industrial fermentation containers from two processing yards. Forty percent were affected by butyric, sulfidic, or putrid spoilage, while 60% followed the ordinary fermentation course. The samples were obtained at 30 days of fermentation, determining their 16S rRNA gene Amplicon Sequence Variant compositions (ASVs). The butyric containers showed a bacterial profile strongly associated with the genera Enterococcus, Leuconostoc, and Atlantibacter, but also with Lactiplantibacillus and Melissococcus, and less confident to Raoultella, Enterobacter, Serratia, and Celerinatantimonas. The sulfidic fermentation was linked to Alkalibacterium and, to a lesser extent, Marinilactibacillus and the absence of Lactiplantibacillus. Putrid spoilage was mainly related to Halolactibacillus and Alkalibacterium. Sulfidic/putrid (together) differed from butyric spoilage by the presence of Alkalibacterium/Marinilactibacillus as well as by Halomonas/Halanaerobium. Lactiplantibacillus dominated normal fermentations, but Vibrio was also frequently found (0-46%), apparently not causing any alteration. These results contribute to a better microbial characterisation of non-zapatera spoiled table olive fermentations. They also suggest using several statistical techniques to discriminate normal vs spoiled fermentations adequately.

Epigenetic inheritance of DNA methylation changes in fish living in hydrogen sulfide-rich springs.

Environmental factors can promote phenotypic variation through alterations in the epigenome and facilitate adaptation of an organism to the environment. Although hydrogen sulfide is toxic to most organisms, the fish Poecilia mexicana has adapted to survive in environments with high levels that exceed toxicity thresholds by orders of magnitude. Epigenetic changes in response to this environmental stressor were examined by assessing DNA methylation alterations in red blood cells, which are nucleated in fish. Males and females were sampled from sulfidic and nonsulfidic natural environments; individuals were also propagated for two generations in a nonsulfidic laboratory environment. We compared epimutations between the sexes as well as field and laboratory populations. For both the wild-caught (F0) and the laboratory-reared (F2) fish, comparing the sulfidic and nonsulfidic populations revealed evidence for significant differential DNA methylation regions (DMRs). More importantly, there was over 80% overlap in DMRs across generations, suggesting that the DMRs have stable generational inheritance in the absence of the sulfidic environment. This is an example of epigenetic generational stability after the removal of an environmental stressor. The DMR-associated genes were related to sulfur toxicity and metabolic processes. These findings suggest that adaptation of P. mexicana to sulfidic environments in southern Mexico may, in part, be promoted through epigenetic DNA methylation alterations that become stable and are inherited by subsequent generations independent of the environment.

Multi-Statistical Approach for the Study of Volatile Compounds of Industrial Spoiled Manzanilla Spanish-Style Table Olive Fermentations.

Table olives can suffer different types of spoilage during fermentation. In this work, a multi-statistical approach (standard and compositional data analysis) was used for the study of the volatile organic compounds (VOCs) associated with altered (butyric, sulfidic, and putrid) and non-altered (normal) Manzanilla Spanish-style table olive fermentations. Samples were collected from two industrial fermentation yards in Seville (Spain) in the 2019/2020 season. The VOC profiles of altered (n = 4) and non-altered (n = 6) samples were obtained by headspace solid-phase microextraction combined with gas chromatography-mass spectrometry (HS-SPME-GC-MS). Ninety-one VOCs were identified and grouped into alcohols (30), esters (21), carbonyl compounds (12), acids (10), terpenes (6), phenols (6), sulfur compounds (2), and others (4). The association of the VOCs with spoilage samples depended on the standard or compositional statistical methodology used. However, butyric spoilage was strongly linked by several techniques to methyl butanoate, ethyl butanoate, and butanoic acid; sulfidic spoilage with 2-propyl-1-pentanol, dimethyl sulfide, methanol, 2-methylbutanal, 2-methyl-2-butenal, ethanol, 2-methyl-3-buten-2-ol, and isopentanol, while putrid was mainly related to D-limonene and 2-pentanol. Our data contribute to a better characterisation of non-zapatera spoiled table olive fermentations and show the convenience of using diverse statistical techniques for a most robust selection of spoilage VOC markers.

Bioleaching of Gold from Sulfidic Gold Ore Concentrate and Electronic Waste by Roseovarius tolerans and Roseovarius mucosus.

Gold bioleaching mediated by iodide oxidizing bacteria (IOB) has been proposed as a sustainable alternative to conventional technologies such as cyanidation. This study evaluated the ability of two IOB sourced from a commercial culture collection, Roseovarius (R.) tolerans DSM 11457T and R. mucosus DSM 17069T, to bioleach gold from electronic waste (e-waste) (1030 ppm gold) and sulfidic gold ore concentrate (45 ppm gold) using one-step, two-step and spent medium leaching at 1% pulp density over 10 days. Two-step bioleaching of ore concentrate resulted in the highest gold leaching yields (approximately ~100% and 34% for R. tolerans and R. mucosus, respectively), followed by spent medium leaching and one-step leaching. The yields remained low for e-waste with both strains (maximum 0.93% and 1.6% for R. tolerans and R. mucosus, respectively) and decreased over time, likely due to the instability of the solubilized gold at relatively low redox potentials (<300 mV vs. Ag/AgCl). Another limiting factor may be the partial inhibition of bacterial growth in the presence of the ore concentrate and e-waste. Therefore, future studies should evaluate the pre-treatment of the ore concentrate and e-waste to remove inhibitory and oxidant consuming compounds before bioleaching with IOB to optimize leaching yields.

Role of microbial diversity for sustainable pyrite oxidation control in acid and metalliferous drainage prevention.

Acid and metalliferous drainage (AMD) remains a challenging issue for the mining sector. AMD management strategies have attempted to shift from treatment of acid leachates post-generation to more sustainable at-source prevention. Here, the efficacy of microbial-geochemical at-source control approach was investigated over a period of 84 weeks. Diverse microbial communities were stimulated using organic carbon amendment in a simulated silicate-containing sulfidic mine waste rock environment. Mineral waste in the unamended leach system generated AMD quickly and throughout the study, with known lithotrophic iron- and sulfur-oxidising microbes dominating column communities. The organic-amended mineral waste column showed suppressed metal dissolution and AMD generation. Molecular DNA-based next generation sequencing confirmed a less diverse lithotrophic community in the acid-producing control, with a more diverse microbial community under organic amendment comprising organotrophic iron/sulfur-reducers, autotrophs, hydrogenotrophs and heterotrophs. Time-series multivariate statistical analyses displayed distinct ecological patterns in microbial diversity between AMD- and non-AMD-environments. Focused ion beam-TEM micrographs and elemental mapping showed that silicate-stabilised passivation layers were successfully established across pyrite surfaces in organic-amended treatments, with these layers absent in unamended controls. Organic amendment and resulting increases in microbial abundance and diversity played an important role in sustaining these passivating layers in the long-term.

Rhizosphere modifications of iron-rich minerals and forms of heavy metals encapsulated in sulfidic tailings hardpan.

Hardpan caps formed after extensive weathering of the top layer of sulfidic tailings have been advocated to serve as physical barriers separating reactive tailings in depth and root zones above. However, in a hardpan-based root zone reconstructed with the soil cover, roots growing into contact with hardpan surfaces may induce the transformation of Fe-rich minerals and release potentially toxic elements for plant uptake. For evaluating this potential risk, two representative native species, Turpentine bush (Acacia chisholmii, AC) and Red Flinders grass (Iseilema vaginiflorum, RF), of which pre-cultured root mats were interfaced with thin discs of crushed hardpan minerals in the rhizosphere (RHIZO) test. After 35 days, the surface dissolution of hardpan minerals occurred and Fe-rich cement minerals were transformed from ferrihydrite-like minerals to goethite-like and Fe(III)-carboxylic complexes, as revealed by scanning electron microscopy coupled with energy dispersive spectroscopy (SEM-EDS) and synchrotron-based X-ray absorption fine structure spectroscopy (XAFS) analysis. This transformation may result from the functions of root exudates. The transformation of hardpan cement minerals caused the co-dissolution of Cu and Zn initially encapsulated in the cements and their uptake by plants. Nevertheless, only was the minority of the plant Cu and Zn transported into shoots.

Prevention of sulfide oxidation in waste rock by the addition of lime kiln dust.

During the operation of a mine, waste rock is often deposited in heaps and usually left under ambient conditions allowing sulfides to oxidize. To focus on waste rock management for preventing acid rock drainage (ARD) formation rather than ARD treatment could avoid its generation and reduce lime consumption, costs, and sludge treatment. Leachates from 10 L laboratory test cells containing sulfide-rich (> 60% pyrite) waste rock with and without the addition of lime kiln dust (LKD) (5 wt.%) were compared to each other to evaluate the LKD's ability to maintain near neutral pH and reduce the sulfide oxidation. Leaching of solely waste rock generated an acidic leachate (pH < 1.3) with high concentrations of As (21 mg/L), Cu (20 mg/L), Fe (18 g/L), Mn (45 mg/L), Pb (856 µg/L), Sb (967 µg/L), S (17 g/L), and Zn (23 mg/L). Conversely, the addition of 5 wt.% LKD generated and maintained a near neutral pH along with decreasing of metal and metalloid concentrations by more than 99.9%. Decreased concentrations were most pronounced for As, Cu, Pb, and Zn while S was relatively high (100 mg/L) but decreasing throughout the time of leaching. The results from sequential extraction combined with element release, geochemical calculations, and Raman analysis suggest that S concentrations decreased due to decreasing sulfide oxidation rate, which led to gypsum dissolution. The result from this study shows that a limited amount of LKD, corresponding to 4% of the net neutralizing potential of the waste rock, can prevent the acceleration of sulfide oxidation and subsequent release of sulfate, metals, and metalloids but the quantity and long-term stability of secondary minerals formed needs to be evaluated and understood before this method can be applied at a larger scale.

Zinc and lead encapsulated in amorphous ferric cements within hardpans in situ formed from sulfidic Cu-Pb-Zn tailings.

Hardpans are massively indurated layers formed at the top layer of sulfidic tailings dams, which develop cementation structures and result in heavy metal immobilization. However, the micro-structural and complex forms of the cementing materials are not fully understood, as well as the mechanisms by which Zn and Pb are stabilized in the hardpans. The present study deployed synchrotron-based X-ray fluorescence microscopy (XFM) to have characterized the cementing structures, examined the distribution of Fe, Zn and Pb, and obtained laterally-resolved speciation of Zn within the hardpans using fluorescence X-ray absorption near-edge structure (XANES) imaging. The XFM analyses revealed that the Fe-rich cement layers consisted of Fe (oxyhydr)oxides coupled with amorphous Si materials, immobilizing Zn and Pb. Through laterally-resolved XANES imaging analyses, Zn-ferrihydrite-like precipitates were predicted to account for >76% of the total Zn within the Fe-rich cement layers. In contrast, outside of the cement layers, 9-63% of the Zn was estimated as labile ZnSO4.7H2O, with the remainder in the form of Zn-sulfide. These findings demonstrated that the Fe-rich cement layers were critical in immobilizing Zn and Pb within hardpans via mineral passivation and encapsulation, as the basis for long-term geochemical stability in the hardpan layer of sulfidic mine tailings.

Phosphate treatment alleviated acute phytotoxicity of heavy metals in sulfidic Pb-Zn mine tailings.

Phytostabilization of sulfidic PbZn tailing landscapes may be one of interim options of tailings management, but which is limited by acute phytotoxicity of heavy metals in the tailings. The present study aimed to investigate the effectiveness of soluble phosphate (i.e., K2HPO4) in immobilizing soluble Pb, Cd and Zn and lowering their acute phytotoxicity. The addition of soluble phosphate improved the growth of native plants Acacia chisholmii and survival rate of A. ligulata, where the latter exhibited 100% survival rate. This was in contrast to effects of conventional organic amendment in the tailings on metal solubility (e.g., elevated metal levels in porewater) and plant survival (e.g., only 42%). Organic amendment with mulch did not lower the levels of water-soluble Cd, Pb and Zn and their concentrations in plant tissues after 56 days of plant growth in the treatment. In contrast, the tailings amended with K2HPO4 significantly decreased metal concentrations in the porewater and plant tissues by about 80-92% and 56-88%, respectively. The metal immobilization by phosphate was due to the formation of insoluble or sparingly soluble metal (Pb, Cd and Zn)-phosphate minerals in the tailings with circumneutral pH conditions, as revealed by using X-ray diffraction and scanning electron microanalyses. The reduced metal concentrations in roots and shoots of Acacia species after direct root contact with the K2HPO4 amended tailings suggested that metals (i.e., Pb, Cd and Zn) were effectively immobilized by the phosphate treatment of the tailings. These findings indicate that addition of high dosage of soluble phosphate may provide a low cost option to treat sulfidic PbZn tailings for rapid phytostabilization of the tailings surface, as an interim option to manage environmental risks of sulfidic PbZn tailings.

Adsorption and transformation of thioarsenite at hematite/water interface under anaerobic condition in the presence of sulfide.

The adsorption behavior of thioarsenite (TAsIII) on the surface of hematite (α-Fe2O3) is unknown at present. In the present study, we have investigated the transformation and reactions of TAsIII [monothioarsenite (MTAsIII) and dithioarsneite (DTAsIII)] on the surface of α-Fe2O3 in the presence of sulfide at S/As = 1 and 3 by X-ray absorption spectroscopy (XAS) and Raman spectroscopy. The adsorption envelopes reveal that the adsorption of TAsIII on α-Fe2O3 is significantly less than that of arsenite (AsIII) in the pH range from 7 to 11 with the initial As concentration of 25 mg L-1. However, at the initial As concentration of 135 mg L-1, the uptake of TAsIII by α-Fe2O3 is higher at pH 7 but lower at pH 8-11 than that of AsIII. The adsorption isotherms show that the adsorption of As on α-Fe2O3 is largely inhibited by the presence of aqueous sulfide at pH 7 with low As equilibrium concentration (<40 mg L-1). Whereas the uptake of As by α-Fe2O3 is highly elevated compared with the value predicted by Langmuir model at pH 7 with high As equilibrium concentration (>40 mg L-1), implying the formation of As-bearing (surface) precipitate. The As and S K-edge XAS as well as Raman spectroscopy confirm the formation of As sulfide precipitate on the surface of α-Fe2O3 in MTAsIII system. It is worth to note that the oxidation of (thio)AsIII occurs on the surface of α-Fe2O3 in DTAsIII system under strictly anaerobic conditions. These results shed new light on the understanding of the interfacial behavior of As and point to the potential implication in immobilization and removal of arsenic in sulfidic environment.