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.

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.

The effect of varying process parameters during bio-catalyzed leaching of metals from end-of-life printed circuit boards (PCBs) was investigated. Fragmented PCBs (under 2 mm) were subjected to an indirect bioleaching in a stirred tank reactor while pulp density, pH and initial ferric iron content were varied. An iron oxidizing Acidithiobacillus ferrooxidans 61 microbial strain was used to generate the lixiviant through oxidizing Fe(II) to Fe(III). Chemically generated Fe(III) was tested as lixiviant under the same conditions as the biological one for comparative purposes. Cell enumeration during leaching and microscopic observations of the input and leached PCBs were conducted in parallel to shed light on the observed phenomena. The degree of bringing metals in solution was found to depend mainly on ferric iron concentration and pH. For the entire duration being always kept as 24 h, substantial portion of Cu (~87%) was extracted respectively at 1% pulp density (PD), 15.5 g/L Fe³⁺ and pH 1. For Zn and Ni, nearly 100% recovery was observed at 5% PD, 18 g/L Fe³⁺ and pH 1.1. The achieved results offer possibilities for further studies at higher pulp density, to ultimately render the bioleaching approach as enabling economical and environmentally friendly technology for urban mining of non-ferrous metals.