Stability of copper tolerance in Thiobacillus ferrooxidans

A strain of Thiobacillus ferrooxidans MAL-4-1 was adapted to grow at higher concentrations of copper by repeated subculturing in the presence of increasing levels of added cupric ions in 9K medium. The strains adapted to copper were found to be more efficient in bioleaching of copper from concentrates. When copper tolerant strains were back cultured repeatedly in 9K medium without cupric ions, the initially developed metal tolerance was observed to be lost. This indicates that the copper tolerance developed is stress-dependent and not a permanent trait of the adapted strain.

Differential influence of ions on the copy number of plasmids in Thiobacillus ferrooxidans

Thiobacillus ferrooxidans MAL4-1, an isolate from Malanjkhand copper mines, India, was adapted to grow in the presence of high concentration (30 gL(-1)) of Cu2+, resulting in a 15-fold increase in its tolerance to Cu2+. While wild-type T. ferrooxidans MAL4-1 contained multiple plasmids, cultures adapted to Cu2+ concentrations of 20 gL(-1) or more showed a drastic reduction in the copy number of the plasmids. The reduction for three of the plasmids was estimated to be over 50-fold. Examination of the plasmid profiles of the strains adapted to high concentration of SO42- anion (as Na2SO4 or ZnSO4) indicated that the reduction in plasmid copy number is not owing to SO42- anion, but is specific for Cu2+. The effect of mercury on the plasmids was similar to that of copper. Deadaptation of the Cu2+- Or Hg2+-adapted T. ferrooxidans resulted in restoration of the plasmids to the original level within the first passage. The fact that the plasmid copy number, in general, is drastically reduced in Cu2+-adapted T. ferrooxidans suggests that resistance to copper is chromosome mediated. This is the first report of a selective negative influence of copper ions on the copy number of plasmids in T. ferrooxidans.

Role of bacterial growth conditions and adhesion in the bioleaching of chalcopyrite by Thiobacillus ferrooxidans

The role of growth conditions and adhesion of Thiobacillus ferrooxidans on the leaching of chalcopyrite was investigated. Thiobacillus ferrooxidans grown on sulfur, thiosulfate and ferrous ion substrates was used in this comparative study. Growth on sulfur, a solid substrate, requires bacterial adhesion unlike that required in the presence of soluble thiosulfate and ferrous ion in a mineral-salts medium. Solid substrate-grown cells showed higher rates of leaching than those grown in liquid media. An initial lag period noticed during leaching by solution-grown cells was absent when solid substrate-grown cells were used. Such a behavior is attributed to the presence of an inducible proteinaceous cell-surface appendage on the sulfur-grown cells. This appendage aids in bacterial adhesion onto the mineral surfaces. Such an appendage is absent in solution-grown cells, as substantiated by electrophoretic measurements. The importance of bacterial adhesion and the direct mechanism in leaching by Thiobacillus ferrooxidans are demonstrated.

Growth and development of Thiobacillus ferrooxidans for engineering applications

A bioprocessing approach for the extraction of base, nuclear and precious metals from refractory and lean grade ores has been reviewed in this paper. Characteristic morphological features of Thiobacillus ferrooxidans, the organism which has been extensively used for biooxidation of sulphide ores have been discussed. Mechanisms of chemoautotrophy and mineral oxidation have been illustrated. The current engineering applications of this microorganism have also been brought out. Various methods for accelerating the growth of Thiobacillus ferrooxidans for faster biooxidation and genetic manipulation for development of desired strains have been outlined.

Studies on multi-metal ion tolerance of Thiobacillus ferrooxidans

The influence of different concentrations of base metal ions, such as CU2+, Zn2+ and Fe3+, when present either alone or in different possible binary and ternary combinations in a 9K medium, on the fel rous ion oxidation ability of Thiobacillus ferrooxidans was studied. Levels and degree of toxicity of these ions have been quantified in terms of toxicity index (TI). Copper and zinc tolerant strains of the bacteria were developed through serial subculturing and their activity tested in the presence of the above metal ions in comparison with the behavior of wild unadapted cells under similar conditions. Copper tolerant strains (25 g/L Cu2+) were found to be more efficient in the bioleaching of both copper and zinc concentrates than wild unadapted strains, while zinc tolerant strains (40 g/L Zn2+) exhibited better leaching efficiency only in the bioleaching of sphalerite concentrates. The significance and relevance of multi-metal ion tolerance in Thiobacillus ferrooxidans has been highlighted with respect to bioleaching of sulphide mineral concentrates. (C) 1997 Published by Elsevier Science Ltd.

Electrochemical aspects of leaching of ocean nodules in the presence and absence of microorganisms

instead of using chemical-reducing agents to facilitate the reduction and dissolution of manganese and iron oxide in the ocean nodule, electrochemical reduction based on two approaches, namely, cathodic polarization and galvanic interaction, can also be considered as attractive alternatives. Galvanic leaching of ocean nodules in the presence of pyrite and pyrolusite for complete recovery of Cu, Ni and Co has been discussed. The key for successful and efficient dissolution of copper, nickel and cobalt from ocean nodules depends on prior reduction of the manganese and ferric oxides with which the above valuable nonferrous metals are interlocked. Polarization studies using a slurry electrode system indicated that maximum dissolution of iron and manganese due to electrochemical reduction occurred at negative DC potentials of -600 mV (SCE) and -1400 mV (SCE). The present work is also relevant to galvanic bioleaching of ocean nodules using autotrophic microorganisms, such as Thiobacillus ferrooxidans and T thiooxidans, which resulted in significant dissolution of copper, nickel and cobalt at the expense of microbiologically generated acids. Various electrochemical and biochemical mechanisms are outlined and the electroleaching and galvanic processes so developed are shown to yield almost complete dissolution of all metal values. (C) 2002 Elsevier Science B.V. All rights reserved.

Surface chemical and adsorption studies using Thiobacillus ferrooxidans with reference to bacterial adhesion to sulfide minerals

Adhesion of Thiobacillus ferrooxidans to pyrite and chalcopyrite in relation to its importance in bioleaching and bioflotation has been studied. Electrokinetic studies as well as FT-IR spectra suggest that the surface chemistry of Thiobacillus ferrooxidans depends on bacterial growth conditions. Sulfur-,Pyrite- and chalcopyrite-grown Thiobacillus ferrooxidans were found to be relatively more hydrophobic. The altered surface chemistry of Thiobacillus ferrooxidans was due to secretion of newer and specific proteinaceous compounds. The adsorption density corresponds to a monolayer coverage in a horizontal orientation of the cells. The xanthate flotation of pyrite in presence of Thiobacillus ferrooxidans is strongly depressed where as the cells have insignificant effect on chalcopyrite flotation. This study demonstrate that: (a)Thiobacillus ferrooxidans cells can be used for selective flotation of chalcopyrite from pyrite and importantly at natural pH values. (b)Sulfur-grown cells exhibits higher leaching kinetics than ferrous ion-grown cells.

Oxidación biológica (Thiobacillus) de gases azufrados y uso del efluente como fertilizante agrícola

Tesis (Doctorado en Ciencias Agrícolas con Especialidad en Agua-Suelo) UANL

Aislamiento y caracterización de cepas de Thiobacillus Ferroxidans con alta resistencia a arsénico y su utilización en la recuperación de oro a partir de concentrados de pirita y arsenopirita

Tesis (Doctorado en Ciencias con Especialidad en Biotecnología) UANL

Analyse du métabolisme du soufre de la bactérie autotrophique acidophile Acidithiobacillus thiooxidans ATCC 19377

Les impacts environnementaux dues à l'extraction minière sont considérables. C'est l'action des microorganismes, en utilisant leur métabolisme du soufre sur les déchets miniers, qui engendre les plus grands défis. Jusqu'à présent, peu de recherches ont été effectués sur les microorganismes environnementaux pour la compréhension globale de l'action du métabolisme du soufre dans une optique de prévention et de rémédiation des impacts environnementaux de l'extraction minière. Dans cette étude, nous avons étudié une bactérie environnementale, Acidithiobacillus thiooxidans, dans le but de comprendre le métabolisme du soufre selon le milieu de culture et le niveau d'acidité du milieu. Nous avons utilisé la transcriptomique à haut débit, RNA-seq, en association avec des techniques de biogéochimie et de microscopie à électrons pour déterminer l'expression des gènes codants les enzymes du métabolisme du soufre. Nous avons trouvé que l'expression des gènes des enzymes du métabolisme du soufre chez ce microorganisme sont dépendantes du milieu, de la phase de croissance et du niveau d'acidité présent dans le milieu. De plus, les analyses biogéochimiques montrent la présence de composés de soufre réduits et d'acide sulfurique dans le milieu. Finalement, une analyse par microscopie électronique révèle que la bactérie emmagasine des réserves de soufre dans son cytoplasme. Ces résultats permettent une meilleure compréhension de son métabolisme et nous rapprochent de la possibilité de développer une technique de prédiction des réactions ayant le potentiel de causer des impacts environnementaux dus à l'extraction minière.

Aplicação de técnicas eletroquímicas no estudo da dissolução oxidativa da covelita (CuS) por Thiobacillus ferrooxidans

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Differentiation of Acidithiobacillus ferrooxidans and A-thiooxidans strains based on 16S-23S rDNA spacer polymorphism analysis

Restriction fragment length polymorphism (RFLP) and sequence analyses of the PCR-amplified 16S-23S rDNA intergenic spacer (ITS) were used for differentiating Acidithiobacillus thiooxidans strains from other related acidithiobacilli, including A. ferrooxidans and A. caldus. RFLP fingerprints obtained with AluI, DdeI, HaeIII, HinfI and MspI enabled the differentiation of all Acidithiobacillus reference strains into species groups. The A. thiooxidans strains investigated (metal mine isolates) yielded identical RFLP patterns to the A. thiooxidans type strain (ATCC 19377(T)), except for strain DAMS, which had a distinct pattern for all enzymes tested. Fourteen A. ferrooxidans mine strains were assigned to 3 RFLP groups, the majority of which were grouped with A. ferrooxidans ATCC 23270(T). The spacer region of one representative strain from each of the RFLP groups obtained was subjected to sequence analysis, in addition to eleven additional A. thiooxidans strains isolated from sediment and water samples, and A. caldus DSM 8584(T). The tRNA(IIe) and tRNA(Ala) genes, present in all strains analyzed, showed high sequence similarity. Phylogenetic analysis of the ITS sequences differentiated all three Acidithiobacillus species. Inter- and infraspecific genetic variations detected were mainly due to the size and sequence polymorphism of the ITS3 region. Mantel tests showed no significant correlation between ITS sequence similarity and the geographical origin of strains. The results showed that the 16S-23S rDNA spacer region is a useful target for the development of molecular-based methods aimed at the detection, rapid differentiation and identification of acidithiobacilli. (C) 2004 Elsevier SAS. All rights reserved.

Effect of anaerobic digestion and initial pH on metal bioleaching from sewage sludge

The effects of anaerobic digestion and initial pH on the bioleaching of metals from sewage sludge were investigated in shake flask experiments. A strain of Acidithiobacillus thiooxidans was employed in the assays using secondary and anaerobic sludges, which resulted in similar solubilization yields of the metals chromium, copper, lead, nickel, and zinc for both the sludges investigated. The effect of initial pH (7.0 and 4.0) on metal bioleaching was assayed by using the anaerobic sludge inoculated with indigenous sulfur-oxidizing thiobacilli. Although the time required to reach the end of the experiment (final pH close to 1.0) was shortened at initial pH of 4.0, final metal solubilization was not significantly different for both initial pH values, resulting in higher solubilization yields for copper, nickel, and zinc (higher than 80%). Chromium and lead presented solubilization yields close to 50%. The results obtained in this work showed that the metal bioleaching process can be applied to sewage sludge regardless of the type of sludge and without the requirement of pH adjustment.

Biological leaching of Mn, Al, Zn, Cu and Ti in an anaerobic sewage sludge effectuated by Thiobacillus ferrooxidans and its effect on metal partitioning

The chemical fractionation and bioleaching of Mn, At, Zn, Cu and Ti in municipal sewage sludge were investigated using Thiobacillus ferrooxidans as leaching microorganism. As a result of the bacterial activity, ORP increase and pH reduction were observed. Metal solubilization was accomplished only in experimental systems supplemented with energy source (Fe(II)). The solubilization efficiency approached similar to80% for Mn and Zn, 24% for Cu, 10% for At and 0.2% for Ti. The chemical fractionation of Mn, At, Zn, Cu and Ti was investigated using a five-step sequential extraction procedure employing KNO3. KF, Na4P2O7, EDTA and HNO3. The results show that the bioleaching process affected the partitioning of Mn and Zn, increasing its percentage of elution in the KNO3 fraction while reducing it in the KF, Na4P2O7 and EDTA fractions. No significant effect was detected on the partitioning of Cu and Al. However, quantitatively the metals Mn, Zn, Cu and At were extracted with higher efficiency after the bacterial activity. Titanium was unaffected by the bioleaching process in both qualitative and quantitative aspects. (C) 2002 Elsevier B.V. Ltd. All rights reserved.