Rare earth element sequestration by Aspergillus oryzae biomass.

The fungus Aspergillus oryzae could be shown to be a viable alternative for biosorption of valuable metals from solution. Fungal biomass can be obtained easily in high quantities as a waste of biofermentation processes, and used in a complex, multi-phase solution mimicking naturally occurring, mining-affected water samples. With test solution formulated after natural conditions, formation of secondary Al and Fe phases co-precipitating Ce was recorded in addition to specific biosorption of rare earth elements. Remarkably, the latter were removed from the solution despite the presence of high concentrations of interfering Fe and Al. The biomass was viable even after prolonged incubation in the metal solution, and minimal inhibitory concentrations for single metals were higher than those in the test solution. While precipitation/biosorption of Ce (maximal biosorption efficiency was 58.0 ± 22.3% after 6 h of incubation) coincided with the gross removal of Fe from the metal solution, Y (81.5 ± 11.3% efficiency, 24 h incubation) and Nd (87.4 ± 9.1% efficiency, 24 h incubation) were sequestered later, similarly to Ni and Zn. The biphasic binding pattern specific to single metals could be connected to dynamically changing pH and NH4+ concentrations, which were attributed to the physiological changes taking place in starving A. oryzae biomass. The metals were found extracellularly in minerals associated with the cell wall, and intracellularly precipitated in the vacuoles. The latter process was explained with intracellular metal detoxification resulting in metal resistance.

Implications of spatial heterogeneity of tailing material and time scale of vegetation growth processes for the design of phytostabilisation.

Phytostabilisation projects for tailing dams depend on processes occurring at spatial scales of 106 m2 and at decadal time scales. Most experiments supporting the design and monitoring of such projects have much smaller spatial and time scales. Usually, they are only designed for one single scale. Here, we report the results of three coupled experiments performed at pot, lysimeter and field plot scales using six sampling periodstimes from 3 to 20 months. The work explicitly accounts for the sampling times when evaluating the effects of amendments on the performance of plants grown in tailing substrates. Two treatments with potentially complementary roles were applied: zeolites to decrease availability of Cd, Cu, Pb and Zn and green fertilizer to increase the availability of nutrients. Zeolites have a positive influence on plant development, especially in the early stages. Analyses of the pooled datasets for all sampling times revealed the possibility of predicting plant physiological variables, such as protein concentrations, pigments and oxidative stress enzyme activities, as a function of the factors extracted by principal component analysis from the metal concentrations in plants, phosphorus concentrations in plants, and sampling times. Two potentially general methodological rules were extracted: account for the spatial geochemical variability of tailings, and cover the broadest possible range of time scales by experiments. The proposed experimental methodology can be of general use for the design of tailing dam remediation technologies with improvements involving the set of measured variables and sampling frequency and by carefully relating the costs to the institutional aspects of tailing dam management.

Microbiomes in an acidic rock-water cave system.

Belowground ecosystems are accessible by mining, where a specific microbial community can be discovered. The biodiversity of a former alum mine rich in carbon, but with a low pH of 2.6-3.7, was evaluated by DNA- and cultivation-dependent methods using samples of the black slate rock material, secondary mineralization phases and seepage water. Pyrite oxidation within the low-grade metamorphic Silurian black slate established high concentrations of Fe and $\rm{SO}_4^{2-}$ forming the extreme conditions visible with acidophilic and Fe-oxidizing microorganisms. In addition, an unexpected predominance of fungi in this C-rich and acidic cave ecosystem, including high numbers of Mucoromycota and Mortierellomycota, was detected. Therefore, fungal cultures were obtained, mainly from the secondary mineral phases that are iron phosphates. Hence, the fungi might well have been involved in phosphate mobilization there. The rock material itself is rich in organic carbon that can be used by oxidase activity. The cultivation setup mimicked the cave conditions (low temperature, low pH, oxic conditions), with one oligotrophic and one medium rich in nutrients that allowed for isolation of different fungal (and eutrophic bacterial) groups. The acidic conditions prevented the occurrence of many basidiomycetes, while the isolated fungi could survive these adverse conditions.

Element distribution in fruiting bodies of Lactarius pubescens with focus on rare earth elements.

During growth and senescence, fungal fruiting bodies accumulate essential and non-essential elements to different extent in their compartments. This study bases on a dataset of 32 basidiocarps of the ectomycorrhizal Lactarius pubescens sampled in a former U mining area. Statistical analyses were combined with rare earth element (REE, La-Lu) patterns to study the element distribution within sporocarp compartments and between three different age classes. For this purpose, fruiting bodies were separated into stipe, pileus trama, pileipelles and lamellae, dried and digested with HNO3. While macronutrient (e.g. K, Mg, P, S) contents resemble those of a non-mining affected site, several elements (e.g. Co, Mn) were site-specifically taken up relative to elevated soil contents. With statistics, two main element distribution groups for L. pubescens were revealed: mainly essential (Cu, Mg, Mn, P, S, Zn, Cd, Co, Ni) and mainly non-essential elements (Al, Ca, Fe, Sr, U, REE). The highest REE contents were found in pileipelles and lamellae, corresponding to relatively small cell sizes. Stipes and pileus trama had low REE contents due to their function as transport systems. During growth, light REE (La-Nd) were strongly enriched in lamellae and pileipelles. Middle REE (Sm-Dy) enrichment was found both in soil and fungal biomass. Contents of nutrients decrease with age, while non-essential elements are enriched especially in pileipelles and lamellae. A weak positive Ce anomaly appeared in the bioavailable soil fraction and in the pileipelles of younger individuals. Substrate dependent uptake thus gets reduced with sporocarp senescence, possibly due to redistribution.

Origin of middle rare earth element enrichment in acid mine drainage-impacted areas.

The commonly observed enrichment of middle rare earth elements (MREE) in water sampled in acid mine drainage (AMD)-impacted areas was found to be the result of preferential release from the widespread mineral pyrite (FeS2). Three different mining-impacted sites in Europe were sampled for water, and various pyrite samples were used in batch experiments with diluted sulphuric acid simulating AMD-impacted water with high sulphate concentration and high acidity. All water samples independent on their origin from groundwater, creek water or lake water as well as on the surrounding rock types showed MREE enrichment. Also the pyrite samples showed MREE enrichment in the respective acidic leachate but not always in their total contents indicating a process-controlled release. It is discussed that most probably complexation to sulphite (SO3 (2-)) or another intermediate S-species during pyrite oxidation is the reason for the MREE enrichment in the normalized REE patterns.

Microbially assisted phytoremediation approaches for two multi-element contaminated sites.

Phytoremediation is an environmental friendly, cost-effective technology for a soft restoration of abandoned mine sites. The grasses Agrostis capillaris, Deschampsia flexuosa and Festuca rubra, and the annual herb Helianthus annuus were combined with microbial consortia in pot experiments on multi-metal polluted substrates collected at a former uranium mine near Ronneburg, Germany, and a historic copper mine in Kopparberg, Sweden, to test for phytoextraction versus phytostabilization abilities. Metal uptake into plant biomass was evaluated to identify optimal plant-microbe combinations for each substrate. Metal bioavailability was found to be plant species and element specific, and influenced by the applied bacterial consortia of 10 strains, each isolated from the same soil to which it was applied. H. annuus showed high extraction capacity for several metals on the German soil independent of inoculation. Our study could also show a significant enhancement of extraction for F. rubra and A. capillaris when combined with the bacterial consortium, although usually grasses are considered metal excluder species. On the Swedish mixed substrate, due to its toxicity, with 30 % bark compost, A. capillaris inoculated with the respective consortium was able to extract multi-metal contaminants.

Neutralisation of an acidic pit lake by alkaline waste products.

A former open pit where black shale (alum shale) was excavated during 1942-1965 has been water filled since 1966. The water chemistry was dominated by calcium and sulphate and had a pH of 3.2-3.4 until 1997-1998, when pH was gradually increasing. This was due to the intrusion of leachates from alkaline cement waste deposited close to the lake. A stable pH of around 7.5 was obtained after 6-7 years. The chemistry of the pit lake has changed due to the neutralisation. Concentrations of some dissolved metals, notably zinc and nickel, have gone down, as a result of adsorption/co-precipitation on solid phases (most likely iron and aluminium hydroxides), while other metals, notably uranium and molybdenum, are present at elevated levels. Uranium concentration is reaching a minimum of around pH 6.5 and is increasing at higher pH, which may indicate a formation of neutral and anionic uranyl carbonate species at high pH (and total carbonate levels around 1 mM). Weathering of the water-exposed shale is still in progress.