Understanding a Core Pilin of the Type IVa Pili of Acidithiobacillus thiooxidans, PilV.

Pilins are protein subunits of pili. The pilins of type IV pili (T4P) in pathogenic bacteria are well characterized, but anything is known about the T4P proteins in acidophilic chemolithoautotrophic microorganisms such as the genus Acidithiobacillus. The interest in T4P of A. thiooxidans is because of their possible role in cell recruitment and bacterial aggregation on the surface of minerals during biooxidation of sulfide minerals. In this study we present a successful ad hoc methodology for the heterologous expression and purification of extracellular proteins such as the minor pilin PilV of the T4P of A. thiooxidans, a pilin exposed to extreme conditions of acidity and high oxidation-reduction potentials, and that interact with metal sulfides in an environment rich in dissolved minerals. Once obtained, the model structure of A. thiooxidans PilV revealed the core basic architecture of T4P pilins. Because of the acidophilic condition, we carried out in silico characterization of the protonation status of acidic and basic residues of PilV in order to calculate the ionization state at specific pH values and evaluated their pH stability. Further biophysical characterization was done using UV-visible and fluorescence spectroscopy and the results showed that PilV remains soluble and stable even after exposure to significant changes of pH. PilV has a unique amino acid composition that exhibits acid stability, with significant biotechnology implications such as biooxidation of sulfide minerals. The biophysics profiles of PilV open new paradigms about resilient proteins and stimulate the study of other pilins from extremophiles.

Disorder and amino acid composition in proteins: their potential role in the adaptation of extracellular pilins to the acidic media, where Acidithiobacillus thiooxidans grows.

There are few biophysical studies or structural characterizations of the type IV pilin system of extremophile bacteria, such as the acidophilic Acidithiobacillus thiooxidans. We set out to analyze their pili-comprising proteins, pilins, because these extracellular proteins are in constant interaction with protons of the acidic medium in which At. thiooxidans grows. We used the web server Operon Mapper to analyze and identify the cluster codified by the minor pilin of At. thiooxidans. In addition, we carried an in-silico characterization of such pilins using the VL-XT algorithm of PONDR® server. Our results showed that structural disorder prevails more in pilins of At. thiooxidans than in non-acidophilic bacteria. Further computational characterization showed that the pilins of At. thiooxidans are significantly enriched in hydroxy (serine and threonine) and amide (glutamine and asparagine) residues, and significantly reduced in charged residues (aspartic acid, glutamic acid, arginine and lysine). Similar results were obtained when comparing pilins from other Acidithiobacillus and other acidophilic bacteria from another genus versus neutrophilic bacteria, suggesting that these properties are intrinsic to pilins from acidic environments, most likely by maintaining solubility and stability in harsh conditions. These results give guidelines for the application of extracellular proteins of acidophiles in protein engineering.

Improvement of gold bioleaching extraction from waste telecommunication printed circuit boards using biogenic thiosulfate by Acidithiobacillus thiooxidans.

Cyanide usage in gold processing techniques has become increasingly challenging due to its toxicity and environmental impact. It is possible to develop environmentally friendly technology using thiosulfate because of its nontoxic characteristics. Thiosulfate production requires high temperatures, resulting in high greenhouse gas emissions and energy consumption. The biogenesized thiosulfate is an unstable intermediate product of Acidithiobacillus thiooxidans sulfur oxidation pathway to sulfate. A novel eco-friendly method was presented in this study to treat spent printed circuit boards (STPCBs) using biogenesized thiosulfate (Bio-Thio) obtained from Acidithiobacillus thiooxidans cultured medium. To obtain a preferable concentration of thiosulfate among other metabolites by limiting thiosulfate oxidation, optimal concentrations of inhibitor (NaN3: 3.25 mg/L) and pH adjustments (pH= 6-7) were found to be effective. Selection of the optimal conditions has led to the highest bio-production of thiosulfate (500 mg/L). The impact of STPCBs content, ammonia, ethylenediaminetetraacetic acid (EDTA), and leaching time on Cu bio-dissolution and gold bio-extraction were investigated using enriched-thiosulfate spent medium. The suitable conditions were a pulp density of 5 g/L, an ammonia concentration of 1 M, and a leaching time of 36 h, which led to the highest selective extraction of gold (65 ± 0.78%).

Quorum Sensing Signaling Molecules Positively Regulate c-di-GMP Effector PelD Encoding Gene and PEL Exopolysaccharide Biosynthesis in Extremophile Bacterium Acidithiobacillus thiooxidans.

Acidithiobacillus species are fundamental players in biofilm formation by acidophile bioleaching communities. It has been previously reported that Acidithiobacillus ferrooxidans possesses a functional quorum sensing mediated by acyl-homoserine lactones (AHL), involved in biofilm formation, and AHLs naturally produced by Acidithiobacillus species also induce biofilm formation in Acidithiobacillus thiooxidans. A c-di-GMP pathway has been characterized in Acidithiobacillus species but it has been pointed out that the c-di-GMP effector PelD and pel-like operon are only present in the sulfur oxidizers such as A. thiooxidans. PEL exopolysaccharide has been recently involved in biofilm formation in this Acidithiobacillus species. Here, by comparing wild type and ΔpelD strains through mechanical analysis of biofilm-cells detachment, fluorescence microscopy and qPCR experiments, the structural role of PEL exopolysaccharide and the molecular network involved for its biosynthesis by A. thiooxidans were tackled. Besides, the effect of AHLs on PEL exopolysaccharide production was assessed. Mechanical resistance experiments indicated that the loss of PEL exopolysaccharide produces fragile A. thiooxidans biofilms. qRT-PCR analysis established that AHLs induce the transcription of pelA and pelD genes while epifluorescence microscopy studies revealed that PEL exopolysaccharide was required for the development of AHL-induced biofilms. Altogether these results reveal for the first time that AHLs positively regulate pel genes and participate in the molecular network for PEL exopolysaccharide biosynthesis by A. thiooxidans.

Comparison of three different bioleaching systems for Li recovery from lepidolite.

Three different biological systems, the consortium of autotrophic bacteria Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans, heterotrophic fungus Aspergillus niger and heterotrophic yeast Rhodotorula mucilaginosa, were investigated for lithium extraction from lepidolite. The bacterial consortium was the most effective, 11 mg l-1 of Li was dissolved in the absence of nutrients within 336 days. Fungal and yeast bioleaching was faster (40 days), however, with lower extraction efficiency. Bioaccumulation represented a main process of Li extraction by R. mucilaginosa and A. niger, with 92 and 77% of total extracted Li accumulated in the biomass, respectively. The X-ray diffraction analysis for bioleaching residue indicated changes caused by microorganisms, however, with differences between bacterial leaching and bioleaching by fungi or yeasts. The final bioleaching yields for bacterial consortium, A. niger and R. mucilaginosa were 8.8%, 0.2% and 1.1%, respectively. Two-step bioleaching using heterotrophic organisms followed by autotrophic bioleaching could lead to the increase of the process kinetics and efficiency. Bioaccumulation of Li offers strong advantage in Li extraction from solution.

Effect of microbial nutrients supply on coal bio-desulfurization.

Research on coal desulfurization is very important for economic, social, and environmentally sustainable development. In this study, three batches of shake flask experiments were conducted for coal bio-desulfurization using Acidithiobacillus ferrooxidans to explore the relationship between microbial nutrients (iron-free M9 K medium) supply and coal bio-desulfurization efficiency. The results showed that the removal rates of pyritic sulfur and total sulfur from coal effectively increased following reintroduction of coal into the filtrate from previous batch. The removal rates of pyritic sulfur and total sulfur were 55.6% and 10.0%, 77.1% and 16.1%, and 86.5% and 28.2%, respectively, in the three batch experiments without iron-free M9 K medium addition. In contrast, the removal rates of pyritic sulfur and total sulfur reached 87.5% and 28.2%, 89.1% and 31.6%, and 92.0% and 29.1%, respectively, in the three batch experiments with 6.7% iron-free M9 K medium addition. However, addition of excessive iron-free M9 K medium was detrimental to coal bio-desulfurization because of the synthesis of jarosite (MFe3(SO4)2(OH)6, M = K+, NH4+) and gypsum (CaSO4·2H2O), which further declined the pyritic sulfur bio-oxidation efficiency and total sulfur removal efficiency.

Acidithiobacillus thiooxidans and its potential application.

Acidithiobacillus thiooxidans (A. thiooxidans) is a widespread, mesophilic, obligately aerobic, extremely acidophilic, rod-shaped, and chemolithoautotrophic gram-negative gammaproteobacterium. It can obtain energy and electrons from the oxidation of reducible sulfur, and it can fix carbon dioxide and assimilate nitrate, nitrite, and ammonium to satisfy carbon and nitrogen requirement. This bacterium exists as different genomovars and its genome size range from 3.02 to 3.97 Mb. Here, we highlight the recent advances in the understanding of the general biological features of A. thiooxidans, as well as the genetic diversity and the sulfur oxidation pathway system. Additionally, the potential applications of A. thiooxidans were summarized including the recycling of metals from metal-bearing ores, electric wastes, and sludge, the improvement of alkali-salinity soils, and the removal of sulfur from sulfur-containing solids and gases.

Adaptive mechanism of Acidithiobacillus thiooxidans CCTCC M 2012104 under stress during bioleaching of low-grade chalcopyrite based on physiological and comparative transcriptomic analysis.

Acidithiobacillus thiooxidans (A. thiooxidans) is often used for sulfur-bearing ores bioleaching, but its adaptive mechanism to harsh environments remains unclear. Here, we explored the adaptive mechanism of A. thiooxidans in the process of low-grade chalcopyrite bioleaching based on the physiology and comparative transcriptome analysis. It was indicated that A. thiooxidans maintains intracellular pH homeostasis by regulating unsaturated fatty acids, especially cyclopropane fatty acids, intracellular ATP, amino acid metabolism, and antioxidant factors. Comparative transcriptome analysis indicated that the key genes involved in sulfur oxidation, sor and soxABXYZ, were significantly up-regulated, generating more energy to resist extreme environmental stress by more active sulfur metabolism. Confocal laser scanning microscope analysis found that down-regulation of flagellar-related genes was likely to promote the biofilm formation. System-level understanding of leaching microorganisms under extreme stress can contribute to the evolution of these extremophiles via genetic engineering modification work, which further improves bioleaching in future.

Mechanism underlying the bioleaching process of LiCoO2 by sulfur-oxidizing and iron-oxidizing bacteria.

Benefiting from lower operational costs and energy requirements than do hydrometallurgical and pyrometallurgical processes in metal recovery, the bioleaching of LiCoO2 through the use of sulfur-oxidizing and iron-oxidizing bacteria has drawn increasing attention. However, the bioleaching mechanism of LiCoO2 has not been clearly elaborated. In the present study, the effects of the energy source of bacteria, such as Fe2+, pyrite and S0, and the products of bacterial oxidation, such as Fe3+ and sulfuric acid, on the chemical leaching of LiCoO2 were studied. The results indicated that lithium was dissolved by acid, and cobalt was released by the reduction of Fe2+ and acid dissolution. The recovery of Li+ and Co2+ could be significantly improved by pH adjustment. Finally, optimal recoveries of Li+ and Co2+ were observed in the pyrite group, reaching 91.4% and 94.2%, respectively. By using pyrite as the energy source, the role of bacteria in bioleaching of LiCoO2 was investigated. The results showed that bacteria could produce sulfuric acid by oxidizing pyrite to promote the mobilization of Li+ and Co2+. The recovery of lithium and cobalt could be increased to 100.0% and 99.3% by bacteria. Moreover, extracellular polymeric substances secreted by bacteria were found to be a factor for the improvement of Li+ and Co2+ recovery.

Electrochemical monitoring of Acidithiobacillus thiooxidans biofilm formation on graphite surface with elemental sulfur.

Inorganic wastewaters and sediments from the mining industry and mineral bioleaching processes have not been fully explored in bioelectrochemical systems (BES). Knowledge of interfacial changes due to biofilm evolution under acidic conditions may improve applications in electrochemical processes, specifically those related to sulfur compounds. Biofilm evolution of Acidithiobacillus thiooxidans on a graphite plate was monitored by electrochemical techniques, using the graphite plate as biofilm support and elemental sulfur as the only energy source. Even though the elemental sulfur was in suspension, S0 particles adhered to the graphite surface favoring biofilm development. The biofilms grown at different incubation times (without electric perturbation) were characterized in a classical three electrode electrochemical cell (sulfur and bacteria free culture medium) by non-invasive electrochemical impedance spectroscopy (EIS) and cyclic voltammetry. The biofilm structure was confirmed by Environmental Scanning Electrode Microscopy, while the relative fractions of exopolysaccharides and extracellular hydrophobic compounds at different incubation times were evaluated by Confocal Laser Scanning Microscopy. The experimental conditions chosen in this work allowed the EIS monitoring of the biofilm growth as well as the modification of Extracellular Polymeric Substances (EPS) composition (hydrophobic/ exopolysaccharides EPS ratio). This strategy could be useful to control biofilms for BES operation under acidic conditions.

Leaching of metals from end-of-life solar cells.

The issue of recycling waste solar cells is critical with regard to the expanded use of these cells, which increases waste production. Technology establishment for this recycling process is essential with respect to the valuable and hazardous metals present therein. In the present study, the leaching potentials of Acidithiobacillus thiooxidans, Acidithiobacillus ferrooxidans, Penicillium chrysogenum, and Penicillium simplicissimum were assessed for the recovery of metals from spent solar cells, with a focus on retrieval of the valuable metal Te. Batch experiments were performed to explore and compare the metal removal efficiencies of the aforementioned microorganisms using spent media. P. chrysogenum spent medium was found to be most effective, recovering 100% of B, Mg, Si, V, Ni, Zn, and Sr along with 93% of Te at 30 °C, 150 rpm and 1% (w/v) pulp density. Further optimization of the process parameters increased the leaching efficiency, and 100% of Te was recovered at the optimum conditions of 20 °C, 200 rpm shaking speed and 1% (w/v) pulp density. In addition, the recovery of aluminum increased from 31 to 89% upon process optimization. Thus, the process has considerable potential for metal recovery and is environmentally beneficial.

Changes in biooxidation mechanism and transient biofilm characteristics by As(V) during arsenopyrite colonization with Acidithiobacillus thiooxidans.

Chemical and surface analyses are carried out using Raman spectroscopy, X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM-EDS), atomic force microscopy (AFM), confocal laser scanning microscopy (CLSM), glow discharge spectroscopy (GDS) and extracellular surface protein quantification to thoroughly investigate the effect of supplementary As(V) during biooxidation of arsenopyrite by Acidithiobacillus thiooxidans. It is revealed that arsenic can enhance bacterial reactions during bioleaching, which can strongly influence its mobility. Biofilms occur as compact-flattened microcolonies, being progressively covered by a significant amount of secondary compounds (S n2- , S0, pyrite-like). Biooxidation mechanism is modified in the presence of supplementary As(V), as indicated by spectroscopic and microscopic studies. GDS confirms significant variations between abiotic control and biooxidized arsenopyrite in terms of surface reactivity and amount of secondary compounds with and without As(V) (i.e. 6 µm depth). CLSM and protein analyses indicate a rapid modification in biofilm from hydrophilic to hydrophobic character (i.e. 1-12 h), in spite of the decrease in extracellular surface proteins in the presence of supplementary As(V) (i.e. stressed biofilms).

Optimization of kinetics and operating parameters for the bioleaching of heavy metals from sewage sludge, using co-inoculation of two Acidithiobacillus species.

This study explores the potential for synchronous extraction of Cu, Cr, Ni and Zn during sewage sludge bioleaching processes, using three types of bacterial cultures: a pure culture of Acidithiobacillus ferrooxidans (A. ferrooxidans); a pure culture of Acidithiobacillus thiooxidans (A. thiooxidans); and a mixed culture of A. ferrooxidans and A. thiooxidans. Variable operating parameters included initial pH, solids concentration, sulfur concentration and ferrous iron concentration, with optimization via Box-Behnken design of response surface methodology. Results indicate that the mixed culture of A. ferrooxidans and A. thiooxidans, was the most effective at bioleaching heavy metals from sewage sludge. The optimal operating conditions were as follows: an initial pH of 2.0, with concentrations of 3% solids, 6.14 g L-1 sulfur and 4.55 g L-1 ferrous iron. Maximum extraction efficiencies obtained after 14 days of bioleaching under optimal conditions, were 98.54% Cu, 57.99% Cr, 60.06% Ni and 95.60% Zn. Bioleaching kinetics were effectively simulated using a shrinking core model to explain the leaching reaction, with modelling results suggesting that the rate was determined by the diffusion step.

Transport-Induced Spatial Patterns of Sulfur Isotopes (δ34S) as Biosignatures.

Cave minerals deposited in the presence of microbes may host geochemical biosignatures that can be utilized to detect subsurface life on Earth, Mars, or other habitable worlds. The sulfur isotopic composition of gypsum (CaSO4·2H2O) formed in the presence of sulfur-oxidizing microbes in the Frasassi cave system, Italy, was evaluated as a biosignature. Sulfur isotopic compositions (δ34SV-CDT) of gypsum sampled from cave rooms with sulfidic air varied from -11 to -24‰, with minor deposits of elemental sulfur having δ34S values between -17 and -19‰. Over centimeter-length scales, the δ34S values of gypsum varied by up to 8.5‰. Complementary laboratory experiments showed negligible fractionation during the oxidation of elemental sulfur to sulfate by Acidithiobacillus thiooxidans isolated from the caves. Additionally, gypsum precipitated in the presence and absence of microbes at acidic pH characteristic of the sulfidic cave walls has δ34S values that are on average 1‰ higher than sulfate. We therefore interpret the 8.5‰ variation in cave gypsum δ34S (toward more negative values) to reflect the isotopic effect of microbial sulfide oxidation directly to sulfate or via elemental sulfur intermediate. This range is similar to that expected by abiotic sulfide oxidation with oxygen, thus complicating the use of sulfur isotopes as a biosignature at centimeter-length scales. However, at the cave room (meter-length) scale, reactive transport modeling suggests that the overall ∼13‰ variability in gypsum δ34S reflects isotopic distillation of circulating H2S gas due to microbial sulfide oxidation occurring along the cave wall-atmosphere interface. Systematic variations of gypsum δ34S along gas flow paths can thus be interpreted as biogenic given that slow, abiotic oxidation cannot produce the same spatial patterns over similar length scales. The expression and preservation potential of this biosignature is dependent on gas flow parameters and diagenetic processes that modify gypsum δ34S values over geological timescales. Key Words: Gypsum-Sulfur isotopes-Biosignature-Sulfide oxidation-Cave. Astrobiology 18, 59-72.

Bioelectrochemical system for the biooxidation of a chalcopyrite concentrate by acidophilic bacteria coupled to energy current generation and cathodic copper recovery.

OBJECTIVES: To develop a bioelectrochemical system (BES) to couple the biooxidation of chalcopyrite (CuFeS2), bioelectrogenesis, and the cathodic Cu2+ reduction, bioanodes of acidophilic (pH < 2) and aerobic chemolithoautotrophic bacteria Acidithiobacillus thiooxidans (sulfur oxidizing) and Leptospirillum sp. (Fe2+ oxidizing) were used. RESULTS: CuFeS2 biooxidation increases the charge transfer from the media due to the bioleaching of Cu and Fe. The biofilm on a graphite bar endows a more electropositive (anodic) character to the bioelectrode. By adding the bioleachate generated by both bacteria into the anodic chamber, the acidic bioleachate provides the faradaic intensity. The maximum current density was 0.86 ± 19 mA cm-2 due to the low potential of the BES of 0.18 ± 0.02 V. Such low potential was sufficient for the cathodic deposit of Cu2+. CONCLUSIONS: This work demonstrates a proof of concept for energy savings for mining industries: bioanodes of A. thiooxidans and Leptospirillum sp. are electroactive during the biooxidation of CuFeS2.

Solubilization of phosphorus from phosphate rocks with Acidithiobacillus thiooxidans following a growing-then-recovery process.

Phosphorus is an essential nutrient for the synthesis of biomolecules and is particularly important in agriculture, as soils must be constantly supplemented with its inorganic form to ensure high yields and productivity. In this paper, we propose a process to solubilize phosphorus from phosphate rocks, where Acidithiobacillus thiooxidans cultures are pre-cultivated to foster the acidic conditions for bioleaching-two-step "growing-then-recovery"-. Our method solubilizes 100% of phosphorus, whereas the traditional process without pre-cultivation-single-step "growing-and-recovery"-results in a maximum of 56% solubilization. As a proof of principle, we demonstrate that even at low concentrations of the phosphate rock, 1% w/v, the bacterial culture is unviable and biological activity is not observed during the single-step process. On the other hand, in our method, the bacteria are grown without the rock, ensuring high acid production. Once pH levels are below 0.7, the mineral is added to the culture, resulting in high yields of biological solubilization. According to the Fourier Transform Infrared Spectroscopy spectrums, gypsum is the dominant phosphate phase after both the single- and two-step methods. However, calcite and fluorapatite, dominant in the un-treated rock, are still present after the single-step, highlighting the differences between the chemical and the biological methods. Our process opens new avenues for biotechnologies to recover phosphorus in tropical soils and in low-grade phosphate rock reservoirs.

Initial stage of the biofilm formation on the NiTi and Ti6Al4V surface by the sulphur-oxidizing bacteria and sulphate-reducing bacteria.

The susceptibility to the fouling of the NiTi and Ti6Al4V alloys due to the adhesion of microorganisms and the biofilm formation is very significant, especially in the context of an inflammatory state induced by implants contaminated by bacteria, and the implants corrosion stimulated by bacteria. The aim of this work was to examine the differences between the sulphur-oxidizing bacteria (SOB) and sulphate-reducing bacteria (SRB) strains in their affinity for NiTi and Ti6Al4V alloys. The biofilms formed on alloy surfaces by the cells of five bacterial strains (aerobic SOB Acidithiobacillus thiooxidans and Acidithiobacillus ferrooxidans, and anaerobic SRB Desulfovibrio desulfuricans-3 strains) were studied using scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). The protein concentrations in liquid media have also been analyzed. The results indicate that both alloys tested may be colonized by SOB and SRB strains. In the initial stage of the biofilm formation, the higher affinity of SRB to both the alloys has been documented. However, the SOB strains have indicated the higher (although differentiated) adaptability to changing environment as compared with SRB. Stimulation of the SRB growth on the alloys surface was observed during incubation in the liquid culture media supplemented with artificial saliva, especially of lower pH (imitated conditions under the inflammatory state, for example in the periodontitis course). The results point to the possible threat to the human health resulting from the contamination of the titanium implant alloys surface by the SOB (A. thiooxidans and A. ferrooxidans) and SRB (D. desulfuricans).

Properties of realgar bioleaching using an extremely acidophilic bacterium and its antitumor mechanism as an anticancer agent.

Realgar is a naturally occurring arsenic sulfide (or Xionghuang, in Chinese). It contains over 90% tetra-arsenic tetra-sulfide (As4S4). Currently, realgar has been confirmed the antitumor activities, both in vitro and in vivo, of realgar extracted using Acidithiobacillus ferrooxidans (A. ferrooxidans). Bioleaching, a new technology to greatly improve the use rate of arsenic extraction from realgar using bacteria, is a novel methodology that addressed a limitation of the traditional method for realgar preparation. The present systematic review reports on the research progress in realgar bioleaching and its antitumor mechanism as an anticancer agent. A total of 93 research articles that report on the biological activity of extracts from realgar using bacteria and its preparation were presented in this review. The realgar bioleaching solution (RBS) works by inducing apoptosis when it is used to treat tumor cells in vitro and in vivo. When it is used to treat animal model organisms in vivo, such as mice and Caenorhabditis elegans, tumor tissues grew more slowly, with mass necrosis. Meanwhile, the agent also showed obvious inhibition of tumor cell growth. Bioleaching technology greatly improves the utilization of realgar and is a novel methodology to improve the traditional method.

Sulfur and Zinc Availability from Co-granulated Zn-Enriched Elemental Sulfur Fertilizers.

Acidification by oxidation of elemental sulfur (ES) can solubilize ZnO, providing slow release of both sulfur (S) and zinc (Zn) in soil. For this study, a new granular fertilizer with ES and ZnO was produced and evaluated. The effect of incorporating microorganisms or a carbon source in the granule was also evaluated. Four granulated ES-Zn fertilizers with and without S-oxidizing microorganisms, a commercial ES pastille, ZnSO4, and ZnO were applied to the center of Petri dishes containing two contrasting pH soils. Soil pH, CaCl2-extractable S and Zn, and remaining ES were evaluated at 30 and 60 days in two soil sections (0-5 and 5-9 mm from the fertilizer application site). A visualization test was performed to evaluate Zn diffusion over time. A significant pH decrease was observed in the acidic soil for all ES-Zn fertilizer treatments and in the alkaline soil for the Acidithiobacillus thiooxidans-inoculated treatment only. In agreement with Zn visualization tests, extractable-Zn concentrations were higher from the point of application in the acidic (62.9 mg dm-3) compared to the alkaline soil (5.5 mg dm-3). Elemental S oxidation was greater in the acidic soil (20.9%) than slightly alkaline soil (12%). The ES-Zn granular fertilizers increased S and Zn concentrations in soil and can provide a strategically slow release of nutrients to the soil.

Bioleaching combined brine leaching of heavy metals from lead-zinc mine tailings: Transformations during the leaching process.

During the process of bioleaching, lead (Pb) recovery is low. This low recovery is caused by a problem with the bioleaching technique. This research investigated the bioleaching combination of bioleaching with brine leaching to remove heavy metals from lead-zinc mine tailings. The impact of different parameters were studied, including the effects of initial pH (1.5-3.0) and solid concentration (5-20%) for bioleaching, and the effects of sodium chloride (NaCl) concentration (10-200 g/L) and temperature (25 and 50 °C) for brine leaching. Complementary characterization experiments (Sequential extraction, X-ray diffractometer (XRD), scanning electronic microscope (SEM)) were also conducted to explore the transformation of tailings during the leaching process. The results showed that bioleaching efficiency was significantly influenced by initial pH and solid concentration. Approximately 85.45% of iron (Fe), 4.12% of Pb, and 97.85% of zinc (Zn) were recovered through bioleaching in optimum conditions. Increasing the brine concentration and temperature promoted lead recovery. Lead was recovered from the bioleaching residues at a rate of 94.70% at 25 °C and at a rate of 99.46% at 50 °C when the NaCl concentration was 150 g/L. The study showed that bioleaching significantly changed the speciation of heavy metals and the formation and surface morphology of tailings. The metals were mainly bound in stable fractions after bioleaching.