Oxidative Dissolution of Arsenopyrite by Mesophilic and Moderately Thermophilic Acidophiles

The purpose of this work was to determine solution- and solid-phase changes associated with the oxidative leaching of arsenopyrite (FeAsS) by Thiobacillus ferrooxidans and a moderately thermoacidophilic mixed culture. Jarosite [KFe₃(SO₄)₂(OH)₆], elemental sulfur (S⁰), and amorphous ferric arsenate were detected by X-ray diffraction as solid-phase products. The oxidation was not a strongly acid-producing reaction and was accompanied by a relatively low redox level. The X-ray diffraction lines of jarosite increased considerably when ferrous sulfate was used as an additional substrate for T. ferroxidans. A moderately thermoacidophilic mixed culture oxidized arsenopyrite faster at 45°C than did T. ferroxidans at 22°C, and the oxidation was accompanied by a nearly stoichiometric release of Fe and As. The redox potential was initially low but subsequently increased during arsenopyrite oxidation by the thermoacidophiles. Jarosite, S⁰, and amorphous ferric arsenate were also formed under these conditions.     

A new look at microbial leaching patterns on sulfide minerals

Leaching patterns on sulfide minerals were investigated by high-resolution scanning electron microscopy (SEM). Our goal was to evaluate the relative contributions of inorganic surface reactions and reactions localized by attached cells to surface morphology evolution. Experiments utilized pyrite (FeS(2)), marcasite (FeS(2)) and arsenopyrite (FeAsS), and two iron-oxidizing prokaryotes in order to determine the importance of cell type, crystal structure, and mineral dissolution rate in microbially induced pit formation. Pyrite surfaces were reacted with the iron-oxidizing bacterium Acidithiobacillus ferrooxidans (at 25 degrees C), the iron-oxidizing archaeon 'Ferroplasma acidarmanus' (at 37 degrees C), and abiotically in the presence of Fe(3+) ions. In all three experiments, discrete bacillus-sized (1-2 μm) and -shaped (elliptical) pits developed on pyrite surfaces within 1 week of reaction. Results show that attaching cells are not necessary for pit formation on pyrite. Marcasite and arsenopyrite surfaces were reacted with A. ferrooxidans (at 25 degrees C) and 'F. acidarmanus' (at 37 degrees C). Cell-sized and cell-shaped dissolution pits were not observed on marcasite or arsenopyrite at any point during reaction with A. ferrooxidans, or on marcasite surfaces reacted with 'F. acidarmanus'. However, individual 'F. acidarmanus' cells were found within individual shallow (<0.5 μm deep) pits. The size and shape (round rather than elliptical) of the pits conformed closely to the shape of F. acidarmanus (cells) pits on arsenopyrite. We infer these pits to be cell-induced. We attribute the formation of pits readily detectable (by SEM) to the higher reactivity of arsenopyrite compared to pyrite and marcasite under the conditions the experiment was conducted. These pits contributed little to the overall surface topographical evolution, and most likely did not significantly increase surface area during reaction. Our results suggest that overall sulfide mineral dissolution may be dominated by surface reactions with Fe(3+) rather than by reactions at the cell-mineral interface.     

Rates of sulfide mineral oxidation by acidophilic chemolithotrophic microbial communities from various sources

A correlation was observed between the rate of oxidation of pure sulfide minerals (pyrite, pyrrhotite, and arsenopyrite) by communities of acidophilic chemolithotrophic microorganisms (ACM) and the mineral substrate where these communities were formed. The ACM community formed during continuous oxidation of the pyrite-arsenopyrite ore concentrate (Kyuchus deposit) exhibited the highest rate of pyrite oxidation. The highest rate of pyrrhotite oxidation was observed for the ACM community developed during semicontinuous oxidation of the pyrrhotite-containing pyrite-arsenopyrite ore concentrate (Olympiadinskoe deposit), by the communities isolated from the pyrrhotite concentrate, and ore of the Shanuch deposit. In the case of arsenopyrite oxidation, the ACM community isolated during oxidation of the Olympiadinskoe ore concentrate grew without a lag phase. Other communities commenced arsenopyrite oxidation at various rates only after a two-day lag phase. The similarity of the mineralogical characteristics of pure sulfide minerals with those of the minerals in the substrates where the ACM communities developed may affect the rates of oxidation.     

Chemical and biological pathways in the bacterial oxidation of arsenopyrite

Abstract: A moderately thermophilic mixed culture of bacteria catalysed the oxidative solubilization of arsenopyrite to give Fe(III), S(VI) and As(V). Toxic effects were observed in a few experiments due to teh build-up of As(III). The bacterial oxidation of arsenopyrite involved direct attack of the bacteria on the mineral to give AS(III). Subsequent oxidation of AS(III) to AS(V) occurred reaction with FE(III), but only in the presence of pyrite, which provide a catalytic surface. Arsenopyrite was unable to act as a catalyst. The pyrite- catalysed oxidation of As(III) to AS(V) by FE(III) usually only went to completion in the presence of bacteria, possibly due to their role in the provision of clean catalytic surfaces. Thus, toxic concentrations of As(III) may accumulate in reactors during the bacterial oxidation of arsenopyrite due to the absence of pyrite or a clean pyrite surface or to low concentrations of the effective oxidizing agent, Fe(III).     

Arsenopyrite Weathering and Leaching of Arsenic in an Austrian Soil

The extent of arsenopyrite weathering in relation to co-existing minerals in an Austrian soil and the leaching of arsenic from the soil has been investigated. Soil and underlying bedrock samples were collected and characterized by chemical and mineralogical analyses. The solubility of the soil arsenic under anaerobic conditions was studied by incubating the soil sample in distilled water for different periods of time using a customized lycimeter. The solubility of arsenic from pure arsenopyrite mineral and mixtures of arsenopyrite with chalcopyrite or pyrite was studied by incubating the pulverized minerals. Speciation of arsenic in the incubated and non-incubated soil samples was carried out by sequential leaching, solvent-extraction, and ion exchange chromatographic techniques.

Results of SEM analysis indicated that arsenopyite (FeAsS), the most common mineral in the area, occurs in paragenesis with pyrite (FeS₂) and chalcopyrite (CuFeS₂). The existence of these minerals with arsenopyrite was found to enhance its solubilization. From the speciation study it was found that nearly all (92%) of the arsenic in the soil exists in the inorganic form. Out of the total inorganic arsenic, the trivalent inorganic species accounted for only 3% and the remaining 89% was found to be the pentavalent form. The low solubility of As in the Graz soil is attributed to the prevalence of this pentavalent inorganic species.     

Biooxidation of a gold-bearing pyrite-arsenopyrite concentrate

Abstract: Two years of BIOX pilot plant data have been examined for steady state conditions and then correlated using logistic kinetics. It was found that the logistic equation not only predicted the performance of individual stages but also the degree of biooxidation across the entire cascade of bioreactors. It was found that the rate constant was 1.3 day^-1 in the first three stages and 0.3 day^-1 in the fourth stage. The maximum removal constant was 0.90 in stage 1 and 0.99 in the remaining stages. Plant retention time ranged from 4 to 12 days with corresponding sulphide oxidation varying from 82 to 98% respectively, and primary stage removal rates varying from 8.9 to 4.4 kg m^-3 day^-l, respectively. In addition, batch biooxidation data were obtained. The biooxidation rate was found to be about half that for the continuous bioreactors. This is in agreement with the findings of several other workers. The specific rates of bioxidation of pyrite and arsenopyrite were very similar for the bulk concentrate at about 0.15 day^-1. However, it was significant that the biooxidation of arsenopyrite in the mixed mineral preceded that of pyrite, suggesting a sequential mechanism. Gold liberation was found to be linearly related to arsenopyrite biooxidation but oxidation of pyrite appears to be preferential in the gold-rich regions.     

A mathematical model for the bacterial oxidation of a sulfide ore concentrate

The effect of dilution rate and feed solids concentration on the bacterial leaching of a pyrite/arsenopyrite ore concentrate was studied. A mathematical model was developed for the process based on the steady-state data collected over the range of dilution rates (20 to 110 h) and feed solids concentrations (6 to 18% w/v) studied. A modified Monod model with inhibition by arsenic was used to model bacterial ferrous ion oxidation rates. The model assumes that (i) pyrite and arsenopyrite leaching occurs solely by the action of ferric iron produced from the bacterial oxidation of ferrous iron and (ii) bacterial growth rates are proportional to ferrous ion oxidation rate. The equilibrium among the various ionic species present in the leach solution that are likely to have a significant effect on the bioleach process were included in the model. (c) 1994 John Wiley & Sons, Inc.     

Evaluation of process variables in bench-scale bio-oxidation of the Olympias concentrate

Abstract: A statistically designed set of eight bio-oxidation tests on the Olympias concentrate was conducted in bench-scale equipment to evaluate the effects of important variables on pyrite/arsenopyrite oxidation and gold extraction. The variables studied were total retention time, feed solid concentration and particle size. High degrees of arsenopyrite oxidation were observed in all tests, as the arsenopyrite oxidation was very fast and therefore not dependent on the variables within the studied range. Statistical analysis of the experimental data reveals that the pyrite oxidation and gold extraction are dependent mainly on the retention time and to a lesser extent on the particle size. The feed solid concentration had a small influence only on the gold extraction. Regressed equations of the experimental data can be used to predict proper operating conditions.     

Isolation of Arsenic Resistant and Arsenopyrite Oxidizing Acidithiobacillus Species from pH Neutral Colombian Mine Effluents

Abstract

Inactive mines provide a great source of bacterial diversity for studying acidophilic communities and their biotechnological applications, but prospecting of these anthropogenic environments in Colombia has been limited. Conventional microbiological methods were used to isolate acidophilic bacterial strains from effluents emanating from the Colombian gold mine ‘El Zancudo’ (Titiribí, Antioquia). Despite the drainage waters having circumneutral pH, all of the isolated strains were phylogenetically related to the extreme acidophile Acidithiobacillus genus. However, based upon 16S rRNA gene sequences the mesophilic sulfur-oxidizing indigenous strains could not be assigned to a species. Pure cultures were selected by screening in medium with soluble inorganic arsenic (III) and their mineral-oxidative activity was evaluated at 30?°C in Erlenmeyer flasks with arsenopyrite ore under rotary shaking conditions. The indigenous strains were able to catalyze arsenopyrite oxidation in a mixed culture with a pulp density of 10%, maintaining their growth in the presence of >80?mM leached arsenic. This research provides information regarding the isolation of arsenic resistant bacterial communities from neutral effluents from El Zancudo mine and the possibility of the isolated strains to be useful in the biooxidation pretreatment of refractory gold-bearing arsenopyrite ores and concentrates.     

Bacterial Oxidation of Refractory Sulfide Ores for Gold Recovery

Abstract

The microbiological leaching of refractory sulfide ores (pyrite, arsenopyrite) for recovery of gold is reviewed in this article. The underlying physiological, biochemical, and genetic fundamentals of the bacteria involved (Thiobacillus and Sulfolobus spp.) are complex and have yet to be elucidated in depth. The chemistry of acid and biological leaching of pyrite and arsenopyrite minerals is also complex, and many of the individual reactions are not known in detail. Bacterial leaching is discussed in relation to chemical speciation at acid pH values. Attempts to develop models for a better understanding of bioleaching processes are summarized. The importance of pH, redox potential, temperature, sulfur balance, and toxic metals is evaluated for optimizing conditions for bacterial activity. Gold is finely disseminated in refractory sulfide ores, thereby decreasing Au recoveries upon conventional cyanidation for gold dissolution. In the bioleaching process, bacteria remove the sulfide minerals by oxidative dissolution and thus expose Au to extraction with cyanide solution. Stirred tank reactors appear most suited for this biological leaching process. The overall oxidation of the sulfides is an important variable for gold recovery. Pilot- and commercial-scale bioleaching processes for gold-containing pyrite and arsenopyrite ores are reviewed. This application of mineral biotechnology competes favorably with pressure leaching and roasting processes, both of which are problematic and energy-intensive alternatives for pretreatment of auriferous pyrite/arsenopyrite ores.     

Migration of arsenic(III) during bacterial oxidation of arsenopyrite in chalcopyrite concentrate by Thiobacillus ferrooxidans

During bacterial oxidation of the arsenopyrite that contaminated a chalcopyrite concentrate, the bioextraction of arsenic from the concentrate was examined. A long-term constant As(III) concentration, representing a large portion of the total arsenic, occurred in the leaching medium. As(III) was not further oxidized, either under bioextraction conditions or by Fe(III) in the presence of the mesophilic bacterium Thiobacillus ferooxidans. These results are discussed in relation to the influence of leaching microorganisms on the form of arsenic in the solution. Dissolved As(III) could be reversed into a solid phase by adsorption of As(III) by forming an iron precipitate. Correspondence to: M. Mandl     

Occurrences at mineral-bacteria interface during oxidation of arsenopyrite by Thiobacillus ferrooxidans

The combination of an improved bacterial desorption method, scanning electron microscopy (SEM), diffuse reflectance and transmission infrared Fourier transform spectroscopy, and a desorption-leaching device like high-pressure liquid chromatography (HPLC) was used to analyze bacterial populations (adhering and free bacteria) and surface-oxidized phases (ferric arsenates and elemental sulfur) during the arsenopyrite biooxidation by Thiobacillus ferrooxidans. The bacterial distribution, the physicochemical composition of the leachate, the evolution of corrosion patterns, and the nature and amount of the surface-oxidized chemical species characterized different behavior for each step of arsenopyrite bioleaching. The first step is characterized by a slow but strong adhesion of bacteria to mineral surfaces, the appearance of a surface phase of elemental sulfur, the weak solubilization of Fe(II), As(III), and As(V), and the presence of the first corrosion patterns, which follow the fragility zones and the crystallographic orientation of mineral grains. After this short step, growth of the unattached bacteria begins, while ferrous ions in solution are oxidized by them. Ferric ions produced by the bacteria can oxidize the sulfide directly and are regenerated by Fe(II) bacterial oxidation. At this time, a bioleaching cycle takes place and a coarse surface phase of ferric arsenate (FeAsO(4) . xH(2)O where x approximately 2) and deep ovoid pores appear. At the end of the bioleaching cycle, the high concentration of Fe(III) and As(V) in solution promotes the precipitation of a second phase of amorphous ferric arsenate (FeAsO(4) . xH(2)O where x approximately 4) in the leachate. Then the biooxidation process ceases: The bacteria adhering to the mineral sufaces are coated by the ferric arsenates and the concentration of Fe(III) on the leachate is found to have decreased greatly. Both oxidation mechanisms (direct and indirect oxidation) have been stopped. (c) 1995 John Wiley & Sons, Inc.     

Resistance determinants of a highly arsenic-resistant strain of Leptospirillum ferriphilum isolated from a commercial biooxidation tank

Two sets of arsenic resistance genes were isolated from the highly arsenic-resistant Leptospirillum ferriphilum Fairview strain. One set is located on a transposon, TnLfArs, and is related to the previously identified TnAtcArs from Acidithiobacillus caldus isolated from the same arsenopyrite biooxidation tank as L. ferriphilum. TnLfArs conferred resistance to arsenite and arsenate and was transpositionally active in Escherichia coli. TnLfArs and TnAtcArs were sufficiently different for them not to have been transferred from one type of bacterium to the other in the biooxidation tank. The second set of arsenic resistance genes conferred very low levels of resistance in E. coli and appeared to be poorly expressed in both L. ferriphilum and E. coli.     

Mineralogical controls on the bacterial oxidation of refractory Barberton gold ores

Abstract: The effect of mineralogical characteristics of gold ore minerals on the nature of sulphide oxidation during a bacterial leaching process was investigated. Three different ore types from the South African goldmines were used, i.e. an arsenopyritic-pyritic ore (Sheba goldmine), a pyritic ore (Agnes goldmine) and a loellingitic-arsenopyritic ore (New Consort goldmine). Detailed mineralogical characterization of each ore was performed. Thereafter, polished sections of the sulphides were suspended in a bacterial leach pulp in an air-stirred vessel for various periods of time. The effect of bacterial oxidation on the sulphides was monitored. Different types of gold-bearing arsenopyrite exist, each type having its own characteristic behaviour during the bacterial oxidation process. The rate of oxidation is controlled by the amount of defects in the crystal structure, and the amount of defects is again controlled by the composition of the arsenopyrite crystal. The distribution of refractory gold in the sulphide minerals can be correlated with the presence of compositional zones and structural deviations. These same mineralogical features also control the sites and rates of bacterial oxidation. Thus. refractory gold occurs at sites which are preferentially leached by the bacteria. The rate of gold liberation from sulphides is therefore being enhanced during the early stages of bacterial oxidation. Defects in a crystal structure influence the rate of bio-oxidation, and can be related directly to the crystal structure of the sulphide mineral, the crystallographic orientation of the exposed surfaces, and differences in chemical compositional and mechanical deviations in the crytals. A combination of all of these mineralogical factors influences the bacterial oxidation process. To optimize and to understand the leaching of an individual ore it is important to establish its controlling factors.     

Construction of arsB and tetH Mutants of the Sulfur-Oxidizing Bacterium Acidithiobacillus caldus by Marker Exchange

Acidithiobacillus caldus is a moderately thermophilic, acidophilic bacterium that has been reported to be the dominant sulfur oxidizer in stirred-tank processes used to treat gold-bearing arsenopyrite ores. It is also widely distributed in heap reactors used for the extraction of metals from ores. Not only are these bacteria commercially important, they have an interesting physiology, the study of which has been restricted by the nonavailability of defined mutants. A recently reported conjugation system based on the broad-host-range IncW plasmids pSa and R388 was used to transfer mobilizable narrow-host-range suicide plasmid vectors containing inactivated and partially deleted chromosomal genes from Escherichia coli to A. caldus. Through the dual use of a selectable kanamycin resistance gene and a hybridization probe made from a deleted portion of the target chromosomal gene, single- and double-recombinant mutants of A. caldus were isolated. The functionality of the gene inactivation system was shown by the construction of A. caldus arsB and tetH mutants, and the effects of these mutations on cell growth in the presence of arsenic and by means of tetrathionate oxidation were demonstrated.     

Potential Role of Thiobacillus caldus in Arsenopyrite Bioleaching

We investigated the potential role of the three strains of Thiobacillus caldus (KU, BC13, and C-SH12) in arsenopyrite leaching in combination with a moderately thermophilic iron oxidizer, Sulfobacillus thermosulfidooxidans. Pure cultures of T. caldus and S. thermosulfidooxidans were used as well as defined mixed cultures. By measuring released iron, tetrathionate, and sulfur concentrations, we found that the presence of T. caldus KU and BC13 in the defined mixed culture lowered the concentration of sulfur, and levels of tetrathionate were comparable to or lower than those in the presence of S. thermosulfidooxidans. This suggests that T. caldus grows on the sulfur compounds that build up during leaching, increasing the arsenopyrite-leaching efficiency. This result was similar to leaching arsenopyrite with a pure culture of S. thermosulfidooxidans in the presence of yeast extract. Therefore, three possible roles of T. caldus in the leaching environment can be hypothesized: to remove the buildup of solid sulfur that can cause an inhibitory layer on the surface of the mineral, to aid heterotrophic and mixotrophic growth by the release of organic chemicals, and to solubilize solid sulfur by the production of surface-active agents. The results showed that T. caldus KU was the most efficient at leaching arsenopyrite under the conditions tested, followed by BC13, and finally C-SH12.     

Changes in biooxidation mechanism and transient biofilm characteristics by As(V) during arsenopyrite colonization with <Emphasis Type="Italic">Acidithiobacillus thiooxidans</Emphasis>

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 _n ^2- , 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).     

Mineral and iron oxidation at low temperatures by pure and mixed cultures of acidophilic microorganisms

An enrichment culture from a boreal sulfide mine environment containing a low-grade polymetallic ore was tested in column bioreactors for simulation of low temperature heap leaching. PCR-denaturing gradient gel electrophoresis and 16S rRNA gene sequencing revealed the enrichment culture contained an Acidithiobacillus ferrooxidans strain with high 16S rRNA gene similarity to the psychrotolerant strain SS3 and a mesophilic Leptospirillum ferrooxidans strain. As the mixed culture contained a strain that was within a clade with SS3, we used the SS3 pure culture to compare leaching rates with the At. ferrooxidans type strain in stirred tank reactors for mineral sulfide dissolution at various temperatures. The psychrotolerant strain SS3 catalyzed pyrite, pyrite/arsenopyrite, and chalcopyrite concentrate leaching. The rates were lower at 5 degrees C than at 30 degrees C, despite that all the available iron was in the oxidized form in the presence of At. ferrooxidans SS3. This suggests that although efficient At. ferrooxidans SS3 mediated biological oxidation of ferrous iron occurred, chemical oxidation of the sulfide minerals by ferric iron was rate limiting. In the column reactors, the leaching rates were much less affected by low temperatures than in the stirred tank reactors. A factor for the relatively high rates of mineral oxidation at 7 degrees C is that ferric iron remained in the soluble phase whereas, at 21 degrees C the ferric iron precipitated. Temperature gradient analysis of ferrous iron oxidation by this enrichment culture demonstrated two temperature optima for ferrous iron oxidation and that the mixed culture was capable of ferrous iron oxidation at 5 degrees C.     

Industrial practice and the biology of leaching of metals from ores The 1997 Pan Labs Lecture

Biomining processes have been used successfully on a commercial scale for the recovery of metals, the most important of which are copper, uranium and gold. These processes are based on the activity of chemoautolithotrophic bacteria which are able to use either iron or sulfur as their energy source and which grow in highly acid conditions. In general, low-rate dump and heap leaching processes are used for copper recovery while the biooxidation of difficult-to-treat gold-bearing arsenopyrite ores is carried out commercially in highly aerated stirred tank reactors. Because of the high levels of bacterial activity required, limitations in the growth rate of the microorganisms which were not apparent in low-rate processes have become an important factor. A key to the commercialization of the gold-bearing arsenopyrite biooxidation process was the development of a rapidly-growing, arsenic-resistant bacterial consortium. The empirical technique of mutation and selection in a continuous-flow system was used to improve the ability of the bacteria to decompose the ore. This approach resulted in a dramatic initial enhancement in growth rate but a plateau in improvement of performance has been reached. Further advances will require a more direct approach based on an understanding of the underlying physiological mechanisms and an application of the tools of molecular biology. Considerable advances have been made in our understanding of the molecular biology of Thiobacillus ferrooxidans. However much less is known about the other biomining bacteria. Recent studies using 16S rRNA analysis techniques have indicated that T. ferrooxidans may play a smaller role in continuous flow stirred tank biomining processes than was previously thought. Received 20 November 1997/ Accepted in revised form 2 March 1998     

Biotechnology in hydrometallurgical processes

The presence of microorganisms in metal sulfide leaching operations has been found to be beneficial in catalysing the dissolution process. Currently this technology is applied on a commercial scale in the recovery of copper and uranium in heap, dump and in-situ leach techniques. However, the biotechnological principles are not limited to the extraction of these two elements. Cobalt, nickel and zinc could be solubilized from their respective sulfides, and coal, silver and gold resources purified from pyrite and arsenopyrite inclusions.