The Lubin underground mine, is one of three mining divisions in the Lubin-Glogow Copper District in Lower Silesia province (Poland). It is the source of polymetallic ore that is rich in copper, silver and several heavy metals. Black shale is also significantly enriched in fossil organic matter in the form of long-chain hydrocarbons, polycyclic aromatic hydrocarbons, organic acids, esters, thiophenes and metalloporphyrins. Biological analyses have revealed that this environment is inhabited by extremophilic bacteria and fungi. Kupfershiefer black shale and samples of water, bottom and mineral sediments from the underground (below 600 m) Lubin mine were taken and 20 bacterial strains were isolated and characterized. All exhibited multi-resistant and hypertolerant phenotypes to heavy metals. We analyzed the plasmidome of these strains in order to evaluate the diversity and role of mobile DNA in adaptation to the harsh conditions of the mine environment. Experimental and bioinformatic analyses of 11 extrachromosomal replicons were performed. Three plasmids, including a broad-host-range replicon containing a Tn3 family transposon, carried genes conferring resistance to arsenic, cadmium, cobalt, mercury and zinc. Functional analysis revealed that the resistance modules exhibit host specificity, i.e., they may increase or decrease tolerance to toxic ions depending on the host strain. The other identified replicons showed diverse features. Among them we identified a catabolic plasmid encoding enzymes involved in the utilization of histidine and vanillate, a putative plasmid-like prophage carrying genes responsible for NAD biosynthesis, and two repABC-type plasmids containing virulence-associated genes. These findings provide an unique molecular insight into the pool of extrachromosomal replicons and highlight their role in the biology and adaptation of extremophilic bacteria inhabiting terrestrial deep subsurface.
Aims: The aim of this study was the isolation and characterization of micro‐organisms from Lubin copper mine potentially useful in biotechnology of metal recovery from copper bearing black shale.
Methods and Results: Eight bacterial strains were isolated from black shale ore. Phylogenetic analysis based on 16S rRNA gene homology showed that five strains belonged to the γ‐Proteobacteria, one to the Firmicutes and two to the Actinobacteria. The ability of the isolates to transform bituminous shale and use them as carbon and energy sources, as well as high resistance to metals and metalloids, esterase and lipase activities, assimilation of organic acids, degradation of phenanthrene and siderophores production were shown.
Conclusions: The indigenous bacteria exhibited a broad range of physiological properties related to geochemical parameters of the examined environment and potentially useful in biometallurgical procedures.
Significance and Impact of the Study: The results have yielded new insights into the microbiology of black shale. It can be suggested that isolated micro‐organisms might play a role in the geochemical cycle of carbon and metals occurring in the organic fraction of black shale ore and might be of potential use in biotechnological procedures for the copper recovery and other valuable metals from tailings containing black shale as well as organic rich ore.
The Upper Permian polymetallic, organic-rich Kupferschiefer black shale in the Fore-Sudetic Monocline is acknowledged to be one of the largest Cu-Ag deposits in the world. Here we report the results of the first study of bioweathering of this sedimentary rock by indigenous heterotrophic bacteria. Experiments were performed under laboratory conditions, employing both petrological and microbiological methods, which permitted the monitoring and visualization of geomicrobiological processes. The results demonstrate that bacteria play a prominent role in the weathering of black shale and in the biogeochemical cycles of elements occurring in this rock. It was shown that bacteria directly interact with black shale organic matter to produce a widespread biofilm on the Kupferschiefer shale surface. As a result of bacterial activity, the formation of pits, bioweathering of ore and rock-forming minerals, the mobilization of elements and secondary mineral precipitation were observed. The chemistry of the secondary minerals unequivocally demonstrates the mobilization of elements from minerals comprising Kupferschiefer. The redistribution of P, Al, Si, Ca, Mg, K, Fe, S, Cu and Pb was confirmed. The presence of bacterial outer membrane vesicles on the surface of black shale was observed for the first time. Biomineralization reactions occurred in both the membrane vesicles and the bacterial cells.
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