Bioleaching of metal sulfides is an interfacial process where adhesion and subsequent biofilm formation are considered to be crucial for this process. In this study, adhesion and biofilm formation by several acidophiles (Acidithiobacillus, Leptospirillum and Sulfobacillus) isolated from different biotopes with sulfide ores in Armenia were studied. Results showed that: (1) these bacteria adhere to pyrite surfaces to various extents. A correlation between pyrite biooxidation and adhesion of S. thermosulfidooxidans 6, L. ferriphilum CC, L. ferrooxidans ZC on pyrite surfaces is shown. It is supposed that bioleaching of pyrite by S. thermosulfidooxidans 6, L. ferriphilum CC, L. ferrooxidans ZC occurs by means of indirect leaching: by ferric iron of bacterial origin; (2) cells of At. ferrooxidans 61, L. ferrooxidans ZC and St. thermosulfidooxidans 6 form a monolayer biofilm on pyrite surfaces. The coverage of pyrite surfaces varies among these species. The order of the biofilm coverage is: L. ferrooxidans ZC ≥ At. ferrooxidans 61 > St. thermosulfidooxidans 6; (3) the extracellular polymeric substances (EPS) analysis indicates that the tested strains produce EPS, if grown either on soluble ferrous iron or solid pyrite. EPS are mainly composed of proteins and carbohydrates. Cells excrete higher amounts of capsular EPS than of colloidal EPS. In addition, cells grown on pyrite produce more EPS than ones grown on ferrous iron.

The aim of this study is to investigate the potential of newly isolated strain Leptospirillum (L.) ferriphilum CC for bioleaching of pyrite and chalcopyrite in pure or mixed culture with other iron- and/or sulfur-oxidizing bacteria. In this paper, kinetics of ferrous iron (Fe2+) oxidation by newly isolated strain Leptospirillum (L.) ferriphilum CC was studied. The effect of initial Fe2+ in the concentration range of 50–400 mM on bacterial growth and iron oxidation was studied. It was shown that microbial Fe2+ oxidation was competitively inhibited by Fe3+. The influence of copper, zinc, nickel and cobalt ions on the oxidation of Fe2+ by L. ferriphilum CC was also studied. Minimal inhibitory concentrations (MIC) for each metal ion were determined. The toxicity of the ions was found to be as follows: Co > Zn > Ni > Cu. The comparison of iron oxidation kinetic parameters of L. ferriphilum CC with other strains of L. ferriphilum indicates the high potential of strain L. ferriphilum CC for biogenic regeneration of concentrated ferric iron (Fe3+) in bioleaching processes of ores and ore concentrates. Bioleaching tests indicated that the newly isolated L. ferriphilum CC can be a prospective strain for the bioleaching of sulfide minerals in pure culture or in association with other iron- and/or sulfur-oxidizing bacteria.

Chemical oxidation of pyrite and chalcopyrite by ferric sulfate (Fe2(SO4)3) solution and biogenic ferric iron obtained by mixed culture of isolated thermotolerant Acidithiobacillus sp. 13Zn and Leptospirilum ferriphilum CC immobilized on natural carrierszeolite and shungite was studied. Oxidation rate of sulfide minerals was estimated by the decrease of Fe 3+ (oxidant) and increase of Fe 2+ ions in the solution. It was revealed that chemical oxidation of chalcopyrite by biogenic ferric iron occurred 2-3 times more intensively than that by Fe2(SO4)3 solution. Pyrite oxidation rate by biogenic ferric iron was twice higher than that by chemical ferric iron solution. It was shown that the treatment of pyrite and chalcopyrite by biogenic ferric iron allows to increase on average 1.5 -2 times the bioleaching of iron from pyrite and iron and copper from chalcopyrite by the associations of iron and sulfur oxidizing bacteria.

A strain of Leptospirillum sp. CC previously isolated from Akhtala polymetallic ore (Armenia) was studied. The main morphological and physiological characteristics of CC were revealed. The optimal growth temperature was 40 °C and optimal pH 1.5. A phylogenetic analysis based on 16S rRNA gene sequences (GenBank ID OM272948) showed that isolate CC was clustered with L. ferriphilum and possessed 99.8% sequence similarity with the strain L. ferriphilum OL12-2 (KF356024). The molar fraction of DNA (G+C) of the isolate was 58.5%. Bioleaching experiment indicates that L. ferriphilum CC can oxidize Fe(II) efficiently, and after 17 days, 44.1% of copper and 91.4% of iron are extracted from chalcopyrite and pyrite, respectively. The efficiency of L. ferriphilum CC in pyrite oxidation increases 1.7 times when co-cultivated with At. ferrooxidans ZnC. However, the highest activity in pyrite oxidation shows the association of L.ferriphilum CC with heterotrophic Acidocella sp. RBA bacteria. It was shown that bioleaching of copper and iron from chalcopyrite by association of L. ferriphilum CC, At. ferrooxidans ZnC, and At. albertensis SO-2 in comparison with pure culture L. ferriphilum CC for 21 days increased about 1.2 and 1.4–1.6 times, respectively.

No abstract