Mining of pyrite minerals is a major environmental issue involving both biological and geochemical processes. Here we present a study of an artificial lake of a former uranium open pit mine with the aim to connect the chemistry and bacterial community composition (454-pyrosequencing of 16S rRNA genes) in the stratified water column. A shift in the water chemistry from oxic conditions in the epilimnion to anoxic, alkaline, and metal and sulfide-rich conditions in the hypolimnion was corresponded by a strong shift in the bacterial community, with few shared operational taxonomic units (OTU) between the water layers. The epilimnetic bacterial community of the lake (similar to 20 years old) showed similarities to other temperate freshwater lakes, while the hypolimnetic bacterial community showed similarity to extreme chemical environments. The epilimnetic bacterial community had dominance of Actinobacteria and Betaproteobacteria. The hypolimnion displayed a higher bacterial diversity and was dominated by the phototrophic green sulphur bacterium of the genus Chlorobium (ca. 40 % of the total community). Deltaproteobacteria were only represented in the hypolimnion and the most abundant OTUs were affiliated with ferric iron and sulfate reducers of the genus Geobacter and Desulfobulbus, respectively. The chemistry is clearly controlling, especially the hypolimnetic, bacterial community but the community composition also indicates that the bacteria are involved in metal cycling in the lake.

In mining areas around the world, chemical and biotic processes have lead to the release of toxic substances, e.g. metals. Microbial processes influence metal speciation, while the chemical environment shapes and controls microbial populations. This thesis has two parts and the papers cover various aspects of metal-to-microbe links in a former uranium mine at Ranstad, Sweden.

The first part of the thesis examines bacteria-induced metal mobilization by the cultivation of Pseudomonas fluorescens with ore and minerals from the Ranstad mine. The presence of a mineral source stimulated bacterial growth. Pyoverdine-type siderophores typical for P. fluorescens were produced, but also other types of siderophores. The bacterial mobilization of metals is implied by the agreement in concentrations of Fe-pyoverdine complex and soluble Fe, and by the concentrations of Ni and Co in solution. U was mobilized from the ore as a result of bacterial growth raising pH.

The second part of the thesis focus on the partitioning of metals and their links to the microbial communities in the water of the former open pit mine, Lake Tranebärssjön. The lake was pH-neutral to alkaline, highly stratified, and had an anoxic hypolimnion with high concentrations of metals and SO₄^2-. Size fractionation showed the dissolved fraction dominating for most metals, while the particulate and colloidal fractions dominated for Fe. Chemical equilibrium models agreed reasonably well with these results. In the pyrosequencing study of the bacterial community, depth-related changes in water chemistry corresponded to a distinct shift in the microbial community, indicating a chemical control. The presence of metal- and SO₄^2--reducing bacteria suggests a possible microbe – chemistry connection.

The findings of this thesis could be used for the restoration and remediation of mining sites, and provides information on factors governing the establishment and control of bacterial populations in freshwaters.

Microorganisms produce chelating agents, such as siderophores and other ligands, which allow them to mobilize and scavenge essential elements from the environment when bioavailability is low. To better understand the effects of biologically mediated leaching of metals from mine waste, Pseudomonas fluorescens was cultivated in the presence of processed ore from the former uranium mine in Ranstad, southern Sweden. Light conditions, the concentration of the mineral source and oxygen availability were varied. The presence of ore in the culture flasks enhanced bacterial growth and raised the pH of the culture medium. Increasing the amount of ore or enhancing aeration of the medium further encouraged cell growth and pH rise. Bacteria mobilized Fe, Ni and Co from the ore. Fe-siderophore complexes were detected and estimated to be present at approximately 9 μm. In the presence of bacteria and light, dissolved Fe and U concentrations were higher compared to dark conditions. Increasing the amount of ore resulted in higher dissolved Ni concentrations but lower dissolved Fe, most likely due to precipitate formation. Data from this study support siderophore production by bacteria that allowed mobilization of essential nutrients from the processed ore. However, the availability of potentially toxic metals like Ni and U may also be enhanced. Microbial-promoted mobilization could contribute to leaching of toxic metals in current and historic mining areas. This process should be considered during design and implementation of remediation projects where trace metals are of environmental concern.

Lime treatment has been extensively used as a remedial measure against acidification of surface waters. In view of the decreasing acid deposition, the possible consequences of a termination of liming are discussed intensively in Sweden. This paper presents the results of the first study of long-term effects of termination of liming. The temporal trends in water chemistry were studied in two lakes after termination of liming, in a still-limed lake and in an unlimed reference lake. The lime treatment was intentionally stopped in order to evaluate the effects on water chemistry and biota. After the last liming, pH decreased steadily in both reacidifying takes until annual mean values stabilised around 5.5-6.0 and 6.2-6.5 respectively. ANC and concentrations of non-marine Ca + Mg decreased after the termination of liming. The decreasing pH resulted in increasing trends of inorganic Al (Al-i), which during recent years exceeded the lowest known effect level for fish on several occasions. This indicates that the lime treatment may have been terminated to early from an ecological perspective and with respect to the critical load of acidifying substances during the study period. However, during the same time, non-marine sulphate decreased in all lakes in the study and pH and ANC increased in the unlimed reference lake.

The alum shale in the Billingen area in southern Sweden was mined in Ranstad for 5 years during the 1960s. The crushed tailings (processed and unprocessed) were left behind when the Ranstad mine was closed that has caused leaching of metals to the surroundings. The siderophore producing bacterium Pseudomonas fluorescens (sp, SE Sweden) was grown in batch cultures for 5 to 8 days with naturally weathered (unprocessed) uranium ore (0.0029% U by weight), kolm (0.52% U by weight) and acid-leached ore (0.0099% U by weight) in chemically defined media (unbuffered and buffered). Pseudomonas fluorescens grown with ore and unbuffered medium changed the pH from 4.7 to 9.3 and leached out 0.016 to 0.9% (normalized to surface area) of the total amount of U from the different ores. Incubation of the acid-leached ore with bacteria in buffered medium leached out 0.04% of the total U. Uranium was leached out selectively at all conditions, but this could be a pH effect, as pH increased at the same time as the U concentrations did. The observed release of Fe was most likely attributed to the production of microbial siderophores (Fe³⁺ specific chelators) since Fe³⁺ has a low solubility at pH > 4. As siderophores contain a number of chelating groups they may still function as complexators even in partly degraded form also for other metals than Fe. Thus, the production of microbial chelators could contribute to the elevated metal concentrations in the drainage water from the closed Ranstad mine, as abiotic processes cannot fully explain these high metal concentrations. In the extension: ligand promoted leaching of toxic elements could also be the key to bioremediation as there is a need for nontoxic cleanup methods for metal contaminated sites.