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.
A new strain of Leptospirillum sp. Ksh-L was isolated from a dump-bioleaching system of the Kashen copper-molybdenum mine (South Caucasus). Ksh-L is an obligate chemolithoautotroph, capable of oxidizing ferrous iron (Fe2+). Cells are Gram-negative and vibrio- or spirillum-shaped of a 0.5–3 µm size. The optimal conditions for the growth are 35 °C and pH 1.6–1.8. Cu2+ and Zn2+ have different effects on the oxidizing ability of the Leptospirillum sp. Ksh-L culture depending on the phase of growth and concentration of Fe2+. Under the conditions of gradually increasing the concentration of copper in the medium, during 4–5 successive subculturing experiments, it was possible to obtain an adapted culture of Leptospirillum sp. Ksh-L, capable of growing in the medium in the presence of up to 400 mM Cu2+. A bioleaching experiment indicates that Ksh-L can efficiently oxidize chalcopyrite. However, the bioleaching of copper from chalcopyrite by Leptospirillum ferrodiazotropum Ksh-L increased about 1.8 times in association with At. thiooxidans ATCC 19377. Phylogenetic analysis based on 16S rRNA gene sequences (GenBank ID ON226845) shows that strain Ksh-L forms a single cluster into Group III. The strain possesses 99.59%, 99.52%, and 96.60% sequence similarity with the strains YTW-96-06, YTW-66-06, and Leptospirillum ferrodiazotrophum 5C in Group III, respectively.
Bioleaching of Cu from the copper concentrate of Armanis gold-bearing polymetallic ore (Armenia) was investigated. The main objective was revealing high active bacteria and their association, as well as optimizing the bioleaching process with their application to ensure the most efficient recovery of copper from the tested concentrate. To obtain optimal bacterial associations, bottom-up and top-down approaches were used. Bioleaching of copper concentrate was carried out using pure cultures of iron- and sulfur-oxidizing bacteria and their mixed culture, as well as indigenous bacterial consortium. Comparative studies of copper bioleaching by mixed cultures of Acidithiobacillus caldus, Leptospirillum ferriphilum CC, Sulfobacillus thermosulfidooxidans 6, and indigenous consortium Arm of iron-oxidizing bacteria were performed. At the beginning of bioleaching, the amounts of extracted copper by mixed culture and Arm consortium were equal; afterward, between 20–27 days, the Arm indigenous consortium showed significantly higher activity in terms of copper extraction. In parallel, mineralogical and liberation analyses of feed material and bioleaching residues were performed.
The microbial ecology of acid mine drainage (AMD) systems in Armenia, with a long mining history, remains unexplored. This study aimed to characterize the microbial diversity and functional potential of AMD in the Syunik region and to isolate novel microorganisms with biotechnological value. A comprehensive analysis of the microbial communities’ structure of Kavart abandoned, Kapan exploring mines effluent, and Artsvanik tailing was conducted. Metagenomics revealed bacterial-dominated communities, comprising Pseudomonadota (previously “Proteobacteria”) (68–72%), with site-specific variations in genus abundance. A high abundance and diversity of metal resistance genes (MRGs), particularly for copper and arsenic, were identified. Carbohydrate-active enzyme (CAZy) analysis showed a dominance of GT2 and GT4 genes, suggesting a high potential for extracellular polymeric substances (EPS) production and biofilm formation. A novel strain of iron-oxidizing bacteria Arm-12 was isolated that shares only ~90% similarity with known Leptospirillum type species, indicating it may represent a new genus without culturable representatives. The strain exhibits enhanced copper extraction from concentrate. This study provides the first metagenomic insights into Armenian AMD systems and tailing, revealing a unique community rich in metal resistance and biofilm-forming genes. The isolation of a novel highly effective iron-oxidizer Arm-12 highlights the potential of AMD environments as a source of novel taxa with significant applications in biomining and bioremediation processes.