Spatialized Audio in a Custom-Built OpenGL-Based Ear Training Virtual Environment.

Interval recognition is an important part of ear training-the key aspect of music education. Once trained, the musician can identify pitches, melodies, chords, and rhythms by listening to music segments. In a conventional setting, the tutor would teach a trainee the intervals using a musical instrument, typically a piano. However, this is expensive, time consuming, and nonengaging for either party. With the emergence of new technologies, including virtual reality (VR) and areas such as edutainment, this and similar trainings can be transformed into more engaging, more accessible, customizable (virtual) environments, with the addition of new cues and bespoke progression settings. In this work, we designed and implemented a VR ear training system for interval recognition. The usability, user experience, and the effect of multimodal integration through the addition of a perceptual cue, spatial audio, was investigated in two experiments with 46 participants. The results show that the system is highly acceptable and provides a very good experience for users. Furthermore, we show that the added spatial auditory cues provided in the VR application give users significantly more information for judging the musical intervals, something that is not possible in a non-VR environment.

Potential for hydrogen-oxidizing chemolithoautotrophic and diazotrophic populations to initiate biofilm formation in oligotrophic, deep terrestrial subsurface waters.

BACKGROUND: Deep terrestrial biosphere waters are separated from the light-driven surface by the time required to percolate to the subsurface. Despite biofilms being the dominant form of microbial life in many natural environments, they have received little attention in the oligotrophic and anaerobic waters found in deep bedrock fractures. This study is the first to use community DNA sequencing to describe biofilm formation under in situ conditions in the deep terrestrial biosphere. RESULTS: In this study, flow cells were attached to boreholes containing either "modern marine" or "old saline" waters of different origin and degree of isolation from the light-driven surface of the earth. Using 16S rRNA gene sequencing, we showed that planktonic and attached populations were dissimilar while gene frequencies in the metagenomes suggested that hydrogen-fed, carbon dioxide- and nitrogen-fixing populations were responsible for biofilm formation across the two aquifers. Metagenome analyses further suggested that only a subset of the populations were able to attach and produce an extracellular polysaccharide matrix. Initial biofilm formation is thus likely to be mediated by a few bacterial populations which were similar to Epsilonproteobacteria, Deltaproteobacteria, Betaproteobacteria, Verrucomicrobia, and unclassified bacteria. CONCLUSIONS: Populations potentially capable of attaching to a surface and to produce extracellular polysaccharide matrix for attachment were identified in the terrestrial deep biosphere. Our results suggest that the biofilm populations were taxonomically distinct from the planktonic community and were enriched in populations with a chemolithoautotrophic and diazotrophic metabolism coupling hydrogen oxidation to energy conservation under oligotrophic conditions.

Complete Genome Sequence of the Subsurface, Mesophilic Sulfate-Reducing Bacterium Desulfovibrio aespoeensis Aspo-2.

Desulfovibrio aespoeensis Aspo-2, DSM 10631(T), is a mesophilic, hydrogenotrophic sulfate-reducing bacterium sampled from a 600-m-deep subsurface aquifer in hard rock under the island of Äspö in southeastern Sweden. We report the genome sequence of this bacterium, which is a 3,629,109-bp chromosome; plasmids were not found.

Metabolic activity of subterranean microbial communities in deep granitic groundwater supplemented with methane and H(2).

It was previously concluded that opposing gradients of sulphate and methane, observations of 16S ribosomal DNA sequences displaying great similarity to those of anaerobic methane-oxidizing Archaea and a peak in sulphide concentration in groundwater from a depth of 250-350 m in Olkiluoto, Finland, indicated proper conditions for methane oxidation with sulphate. In the present research, pressure-resistant, gas-tight circulating systems were constructed to enable the investigation of attached and unattached anaerobic microbial populations from a depth of 327 m in Olkiluoto under in situ pressure (2.4 MPa), diversity, dissolved gas and chemistry conditions. Three parallel flow cell cabinets were configured to allow observation of the influence on microbial metabolic activity of 11 mM methane, 11 mM methane plus 10 mM H2 or 2.1 mM O2 plus 7.9 mM N2 (that is, air). The concentrations of these gases and of organic acids and carbon, sulphur chemistry, pH and Eh, ATP, numbers of cultivable micro-organisms, and total numbers of cells and bacteriophages were subsequently recorded under batch conditions for 105 days. The system containing H2 and methane displayed microbial reduction of 0.7 mM sulphate to sulphide, whereas the system containing only methane resulted in 0.2 mM reduced sulphate. The system containing added air became inhibited and displayed no signs of microbial activity. Added H2 and methane induced increasing numbers of lysogenic bacteriophages per cell. It appears likely that a microbial anaerobic methane-oxidizing process coupled to acetate formation and sulphate reduction may be ongoing in aquifers at a depth of 250-350 m in Olkiluoto.

Influence of H(2) and O(2) on sulphate-reducing activity of a subterranean community and the coupled response in redox potential.

Deep Fennoscandian groundwater is anaerobic, reducing in character and populated by a large diversity of obligate and facultative anaerobic microorganisms. Concentrations of H(2) and carbon monoxide are often 0.01-1 µM and of dissolved organic carbon (DOC) and methane 0.01-1 mM. Microbial activity involving these electron and energy donors may help keep deep groundwater anaerobic and reduced. H(2) was added in concentrations of 0.1-10 mM to a sulphate-reducing community attached to crushed rock in groundwater under a pressure of 2.0 MPa and in situ geochemical conditions. Experiments reported a threshold concentration of approximately 1 µM H(2) at which sulphate reduction ceased, despite the presence of DOC and acetate, suggesting that H(2) was needed for sulphate-reducing activity. δ(13)C values of acetate and DOC data suggested that organic material was degraded to acetate by means of a heterotrophic process. New pressure-resistant micro-sensors for measuring E(h) indicated an H(2)-concentration-dependent decrease in E(h). The investigated community rapidly mitigated the increase in E(h) caused by repeated additions of 0.1-0.2 mM pulses of O(2) as long as H(2) was available. The results imply that sulphate reduction to sulphide with H(2) may dominate sulphate-rich groundwater, which may have implications for metallic underground constructions.

Subterranean microbial populations metabolize hydrogen and acetate under in situ conditions in granitic groundwater at 450 m depth in the Äspö Hard Rock Laboratory, Sweden.

Pressure-resistant circulating systems were constructed to enable the investigation of attached and unattached microbial populations under in situ pressure (2.5 MPa), diversity, dissolved gas and chemistry conditions. Three parallel flow cell cabinets were configured to allow observation of the effect on microbial metabolic activity of adding 3 mM hydrogen or 2.4 mM acetate, compared with an untreated control. Hydrogen addition reduced the generation time fourfold to 2 weeks, doubled the sulphide production rate and increased acetate production by approximately 50%. The acetate addition induced acetate consumption. The studied subterranean microbial processes appeared to proceed very slowly in terms of volume and time, although the results suggest that individual cells could be very active. Lytic bacteriophages are hypothesized to have caused this contradictive observation. Phages may consequently significantly reduce the rates of subterranean microbial processes. Furthermore, the results suggest that hydrogen from corroding underground constructions could induce significant local microbial activity and that the low concentrations of hydrogen often observed in pristine subterranean environments may support slow but sustainable microbial activity in deep groundwater.

Culture-dependent comparison of microbial diversity in deep granitic groundwater from two sites considered for a Swedish final repository of spent nuclear fuel.

Site selection for a spent nuclear fuel (SNF) repository required analysis of microbial abundance and diversity at two Swedish sites, Forsmark and Laxemar-Simpevarp. Information about sulphate-reducing bacteria (SRB) was required, as sulphide could corrode copper SNF canisters. Total number of cells (TNC) and ATP were analysed, and plate counts and most probable number (MPN) analyses were conducted using eight media based on different electron donors and acceptors for specific microorganism physiological groups. Groundwater chemical composition and E(h) were analysed; sampling depths were 112-978 m below sea level. TNC was 5.5 × 10(3) to 4.7 × 10(5) cells mL(-1), correlating with ATP concentrations. Culturability in TNC percentage was 0.01-35.9, averaging 5.12. Culturable numbers varied greatly between sample positions and uncorrelated with depth. SRB were found in 29 samples and were below detection in three; the MPN of SRB correlated negatively with E(h), as did the MPN of acetogens. Data indicated that microbial sulphate reduction was ongoing in many sampled aquifers; published stable isotope data and modelling results supported this observation. The sites did not differ significantly, but the large data range suggested that analysis of more samples would enable detailed evaluation of microbial processes and their relationship with geochemical information.

Abundance and diversity of biofilms in natural and artificial aquifers of the Äspö Hard Rock Laboratory, Sweden.

Six cores were drilled and retrieved from 186-m depth in the Äspö Hard Rock Laboratory (HRL) tunnel to investigate whether indigenous biofilms develop on fracture surfaces in groundwater-conducting aquifers in granitic rock. A clone library was constructed from fracture surface material (FSM), for community composition analysis. Quantitative polymerase chain reaction (qPCR) was applied to quantify gene copies using the 16S rRNA gene for domain Bacteria and the adenosine-phosphosulfate reductase gene (apsA) for sulfate-reducing bacteria (SRB). Results were compared with three groundwater systems with biofilms in laminar flow reactors (LFRs) at 450-m depth in the Äspö HRL. The total number of cells, counted microscopically, was approximately 2 × 10(5) cells cm(-2) in the LFR systems, consistent with the obtained qPCR 16S rRNA gene copies. qPCR analysis reported ∼1 × 10(2) up to ∼1 × 10(4) gene copies cm(-2) on the FSM from the drill cores. In the FSM biofilms, 33% of the sequenced clones were related to the iron-reducing bacterium Stenotrophomonas maltophilia, while in the LFR biofilms, 41% of the sequenced clones were affiliated with the genera Desulfovibrio, Desulforhopalus, Desulfomicrobium, and Desulfobulbus. The community composition of the FSM biofilms differed from the drill water community, excluding drill water contamination. This work reports significant numbers of microorganisms on natural hard rock aquifer fracture surfaces with site-specific community compositions. The probability that biofilms are generally present in groundwater-conducting aquifers in deep granitic rock is consequently great.

The bacteriological analysis and health risks in the urban estuary of St. George's Bay, Grenada, West Indies.

The dilution rates of indicators Enterococcus faecalis and E. coli were studied from the St. John's River estuary in Grenada, West Indies. Health risk zones were established based on the levels of bacteriological pollution. In accordance with the World Health Organization (WHO) health risk guidelines, risks were in the range of <1% gastrointestinal (GI); <0.3% acute febrile respiratory illness (AFRI) to a 1%-5% GI; and 0.9%-1.9% AFRI within 100 m from the St. John's River outflow site in St. George's Bay. These values were the result of river water dilution, where the most probable number (MPN) levels for both indicator organisms from the river were equivalent to that of raw sewage with an AFRI health risk of >3.9% and a GI risk of >10%. The distance intervals farther than 100 m showed fluctuating values and corresponding health risks. E. faecalis and E. coli strains isolated were resistant to 35.7% and 42.9% of the antibiotics tested, respectively.

Bacteriophage lytic to Desulfovibrio aespoeensis isolated from deep groundwater.

Viruses were earlier found to be 10-fold more abundant than prokaryotes in deep granitic groundwater at the Aspö Hard Rock Laboratory (HRL). Using a most probable number (MPN) method, 8-30 000 cells of sulphate-reducing bacteria per ml were found in groundwater from seven boreholes at the Aspö HRL. The content of lytic phages infecting the indigenous bacterium Desulfovibrio aespoeensis in Aspö groundwater was analysed using the MPN technique for phages. In four of 10 boreholes, 0.2-80 phages per ml were found at depths of 342-450 m. Isolates of lytic phages were made from five cultures. Using transmission electron microscopy, these were characterized and found to be in the Podoviridae morphology group. The isolated phages were further analysed regarding host range and were found not to infect five other species of Desulfovibrio or 10 Desulfovibrio isolates with up to 99.9% 16S rRNA gene sequence identity to D. aespoeensis. To further analyse phage-host interactions, using a direct count method, growth of the phages and their host was followed in batch cultures, and the viral burst size was calculated to be approximately 170 phages per lytic event, after a latent period of approximately 70 h. When surviving cells from infected D. aespoeensis batch cultures were inoculated into new cultures and reinfected, immunity to the phages was found. The parasite-prey system found implies that viruses are important for microbial ecosystem diversity and activity, and for microbial numbers in deep subsurface groundwater.

Numbers, biomass and cultivable diversity of microbial populations relate to depth and borehole-specific conditions in groundwater from depths of 4-450 m in Olkiluoto, Finland.

Microbiology, chemistry and dissolved gas in groundwater from Olkiluoto, Finland, were analysed over 3 years; samples came from 16 shallow observation tubes and boreholes from depths of 3.9-16.2 m and 14 deep boreholes from depths of 35-742 m. The average total number of cells (TNC) was 3.9 x 10(5) cells per ml in the shallow groundwater and 5.7 x 10(4) cells per ml in the deep groundwater. There was a significant correlation between the amount of biomass, analysed as ATP concentration, and TNC. ATP concentration also correlated with the stacked output of anaerobic most probable number cultivations of nitrate-, iron-, manganese- and sulphate-reducing bacteria, and acetogenic bacteria and methanogens. The numbers and biomass varied at most by approximately three orders of magnitude between boreholes, and TNC and ATP were positively related to the concentration of dissolved organic carbon. Two depth zones were found where the numbers, biomass and diversity of the microbial populations peaked. Shallow groundwater down to a depth of 16.2 m on average contained more biomass and cultivable microorganisms than did deep groundwater, except in a zone at a depth of approximately 300 m where the average biomass and number of cultivable microorganisms approached those of shallow groundwater. Starting at a depth of approximately 300 m, there were steep gradients of decreasing sulphate and increasing methane concentrations with depth; together with the peaks in biomass and sulphide concentration at this depth, these suggest that anaerobic methane oxidation may be a significant process at depth in Olkiluoto.

Viruses in granitic groundwater from 69 to 450 m depth of the Aspö hard rock laboratory, Sweden.

The objectives of this study were to determine if viruses exist in deep granitic groundwater and to analyse their abundance and morphological diversity. Fluorescent microscopy counts on 10 groundwater samples ranging from 69 to 450 m depth were in the range of 10(4)-10(6) TNC ml(-1) (TNC, total number of prokaryotic cells) and 10(5)-10(7) VLP ml(-1) (VLP, virus-like particles). A good positive correlation of VLP with TNC (r=0.91, P=0.0003) was found with an average VLP/TNC ratio of 12. Transmission electron microscopy revealed four distinct bacteriophage groups (polyhedral, tailed, filamentous and pleomorphic) with at least seven phage families of which some are known to be lytic. Our results suggest the presence of viruses in deep granitic groundwater up to 450 m depth. If they are active and lytic, they will constitute an important group of predators that might control the numbers of microorganisms in the analysed groundwater.

Curium(III) complexation with pyoverdins secreted by a groundwater strain of Pseudomonas fluorescens.

Pyoverdins, bacterial siderophores produced by ubiquitous fluorescent Pseudomonas species, have great potential to bind and thus transport actinides in the environment. Therefore, the influence of pyoverdins secreted by microbes on the migration processes of actinides must be taken into account in strategies for the risk assessment of potential nuclear waste disposal sites. The unknown interaction between curium(III) and the pyoverdins released by Pseudomonas fluorescens (CCUG 32456) isolated from the granitic rock aquifers at the Aspö Hard Rock Laboratory (Aspö HRL), Sweden, is the subject of this paper. The interaction between soluble species of curium(III) and pyoverdins was studied at trace curium(III) concentrations (3 x 10(-7)M) using time-resolved laser-induced fluorescence spectroscopy (TRLFS). Three Cm(3+)-P. fluorescens (CCUG 32456) pyoverdin species, M(p)H(q)L(r), could be identified from the fluorescence emission spectra, CmH(2)L(+), CmHL, and CmL(-), having peak maxima at 601, 607, and 611 nm, respectively. The large formation constants, log beta(121 )= 32.50 +/- 0.06, log beta(111) = 27.40 +/- 0.11, and log beta(101) = 19.30 +/- 0.17, compared to those of other chelating agents illustrate the unique complexation properties of pyoverdin-type siderophores. An indirect excitation mechanism for the curium(III) fluorescence was observed in the presence of the pyoverdin molecules.

Siderophore production by Pseudomonas stutzeri under aerobic and anaerobic conditions.

The siderophore production of the facultative anaerobe Pseudomonas stutzeri, strain CCUG 36651, grown under both aerobic and anaerobic conditions, was investigated by liquid chromatography and mass spectrometry. The bacterial strain has been isolated at a 626-m depth at the Aspö Hard Rock Laboratory, where experiments concerning the geological disposal of nuclear waste are performed. In bacterial culture extracts, the iron in the siderophore complexes was replaced by gallium to facilitate siderophore identification by mass spectrometry. P. stutzeri was shown to produce ferrioxamine E (nocardamine) as the main siderophore together with ferrioxamine G and two cyclic ferrioxamines having molecular masses 14 and 28 atomic mass units lower than that of ferrioxamine E, suggested to be ferrioxamine D(2) and ferrioxamine X(1), respectively. In contrast, no siderophores were observed from anaerobically grown P. stutzeri. None of the siderophores produced by aerobically grown P. stutzeri were found in anaerobic natural water samples from the Aspö Hard Rock Laboratory.

Influence of in situ biofilm coverage on the radionuclide adsorption capacity of subsurface granite.

Any migration of radionuclides from nuclear waste repositories is expected to be mitigated by adsorption to the host rocks surrounding hydraulically conductive fractures. Fluid rock interfaces are considered to be important barriers for nuclear waste disposal schemes but their adsorptive capacity can be affected by the growth of microbial biofilms. This study indicates that biofilms growing on fracture surfaces decrease the rocks adsorption capacity for migrating radionuclides except for trivalent species. Potential suppression of adsorption by biofilms should, therefore, be accounted for in performance safety assessment models. In this study, the adsorptive capacity of in situ anaerobic biofilms grown 450 m underground on either glass or granite slides was compared to the capacity of the same surfaces without biofilms. Surfaces were exposed to the radiotracers 60Co(II), 147Pm(III), 241Am(III), 234Th(IV), and 237Np(V) for a period of 660 h in a pH neutral anaerobic synthetic groundwater. Adsorption was investigated at multiple time points over the 660 h using liquid scintillation and ICP-MS. Results indicate that these surfaces adsorb between 0 and 85% of the added tracers under the conditions of the specific experiments. After 660 h, the distribution coefficients, R (ratio between what is sorbed and what is left in the aqueous phase), approached 3 x 10(4) m for 60Co, 3 x 10(5) m for 147Pm and 241Am, 1 x 10(6)m for 234Th, and 1 x 10(3) m for 237Np. The highest rate of adsorption was during the first 200 h of the adsorption experiments and started to approach equilibrium after 500 h. Adsorption to colloids and precipitates contributed to decreases of up to 20% in the available 60Co, 147Pm, 241Am, and 237Np in the adsorption systems. In the 234Th system 95% of the aqueous 234Th was removed by adsorbing to colloids. Although the range of Rvalues for each surface tested generally overlapped, the biofilms consistently demonstrated lower R values except for the trivalant 147Pm and 241Am adsorption systems.

Cadmium complexation by bacteriogenic iron oxides from a subterranean environment.

This study quantifies the metal sorption characteristics of subterranean bacteriogenic iron oxides (BIOS) and their organic phases (intermixed intact and fragmented bacteria). A Cd2+ ion-selective electrode was used to generate high-resolution metal sorption data as a function of increasing pH. A multisite Langmuir model, along with a linear programming regression method (LPM), was applied to fit experimental data. This approach found two discrete Cd2+ binding sites for the BIOS with average -log10 equilibrium constants (pK(S,j)) of 1.06 +/- 0.19 and 2.24 +/- 0.28. Three discrete sites were obtained for the bacterial fraction, with pK(S,j) values of -0.05 +/- 0.12, 1.18 +/- 0.02, and 3.81 +/- 0.16. This indicated that the BIOS surface had a lower affinity for Cd2+ than that of the bacteria. pK(S,j) values for the BIOS were similar to those reported for pure iron oxide phases, while the organic fraction pK(S,j) spectrum was consistent with previous spectra for intact bacteria. Individual binding site densities of 0.04 +/- 0.01 and 0.05 +/- 0.02 and 0.29 +/- 0.05, 0.11 +/- 0.01, and 0.09 +/- 0.02 micromol/mg of BIOS corresponded to the iron oxide mixture and bacteria fraction, respectively. These values indicated high concentrations of strong affinity Cd2+ complexing groups on the bacterial surface. Comparison of total site densities of 0.08 +/- 0.02 and 0.48 +/- 0.06 micromol/mg of BIOS for the mixture and the bacterial phase, respectively, suggested a nonadditive character for the BIOS surface reactivity. This was emphasized by a higher affinity for Cd2+, as well as an increase in total site concentration observed for the bacterial phase. LPM was able to distinguish between the BIOS mixture and its organic fraction Cd2+ complexation characteristics. This approach is therefore a useful tool for the study of natural sorbent materials controlling metal partitioning in contaminated and pristine environments.

Distribution, diversity and activity of microorganisms in the hyper-alkaline spring waters of Maqarin in Jordan.

The hyper-alkaline, high-Ca(2+) springs of Maqarin, Jordan, were investigated as an analogue for various microbial processes at the extremely high pH generated by cement and concrete in some underground radioactive waste repositories. Leaching of metamorphic, cementitious phases in Maqarin has produced current, hyper-alkaline groundwater with a maximum pH of 12.9. Six consecutive expeditions were undertaken to the area during 1994-2000. The total number of microorganisms in the alkaline waters was 10(3)-10(5) cells/ml. Analysis of the 16S-ribosomal ribonucleic acid (rRNA) diversity revealed microorganisms mainly belonging to the Proteobacteria. Obvious similarities between the obtained sequences and sequences from other alkaline sites could not be found. Numerous combinations of culture media compositions were inoculated with spring, seepage and groundwaters and incubated under aerobic and anaerobic conditions with various carbon sources. Assimilation studies were performed using identical radio-labeled carbon sources. Glucose seemed to be the preferred carbon source for assimilation, followed by acetate, lactate, and leucine. The results demonstrate that microorganisms from the hyper-alkaline springs of Maqarin could grow and be metabolically active under aerobic and anaerobic hyper-alkaline conditions. However, the growth and activity found were not vigorous; instead, slow growth, low numbers, and a generally low metabolic activity were found. This suggests that microbial activity will be low during the hyper-alkaline phase of cementitious repositories.

Surface chemical heterogeneity of bacteriogenic iron oxides from a subterranean environment.

This study quantifies the surface chemical heterogeneity of bacteriogenic iron oxides (BIOS) and its end-members (2-line ferrihydrite and intermixed intact and fragmented bacteria). On a dry weight basis, BIOS consisted of 64.5 +/- 1.8% ferrihydrite and 34.5 +/- 1.8% organic matter. Enrichment of Al, Cu, Cr, Mn, Sr, and Zn was shown in the solid versus the aqueous phase (1.9 < log Kd < 4.2). Within the solid-phase Al (69.5%), Cu (78.7%), and Zn (77.9%) were associated with the bacteria, whereas Cr (59.8%), Mn (99.8%), and Sr (79.4%) preferred ferrihydrite. Acid-base titration data from the BIOS and bacteria were fitted using FOCUS pKa spectroscopy. The bacteria spectrum with pKa's of 4.18 +/- 0.37, 4.80 +/- 0.54, 6.98 +/- 0.45, and 9.75 +/- 0.68 was similar to discrete and continuous spectra for intact and fragmented bacteria. The BIOS spectrum recorded pKa's of 4.27 +/- 0.51, 6.61 +/- 0.51, 7.89 +/- 1.10, and 9.65 +/- 0.66 and was deconvoluted to remove overlapping binding site contributions from the bacteria. The resulting residual iron oxide spectrum coincided with discrete MUSIC spectra for goethite and lepidocrocite with pKa values of 4.10 +/- 0.43, 6.53 +/- 0.45, 7.81 +/- 0.76, and 9.51 +/- 0.68. Surface site density analysis showed that acidic sites (pKa < 6) were contributed by the bacteria (37%), whereas neutral sites (6 < pKa < 8) were characteristic of the iron oxide fraction (35%). Basic sites (8 < pKa) were higher in the bacteria (57%), than in the BIOS (44%) or iron oxide fractions (47%). This analysis suggested a high degree of bacterial group masking and a similarity between the BIOS and goethite surface reactivity. An understanding of the BIOS surface chemical heterogeneity and inherent proton and metal binding capacity was obtained through the use of FOCUS apparent pKa spectroscopy.

Microbially mediated redox processes in natural analogues for radioactive waste.

Natural analogues allow scientists to investigate biogeochemical processes relevant to radioactive waste disposal that occur on time scales longer than those that may be studied by time-limited laboratory experiments. The Palmottu U-Th deposit in Finland and the Bangombé natural nuclear reactor in Gabon involve the study of natural uranium, and are both considered natural analogues for subsurface radioactive waste disposal. The microbial population naturally present in groundwater may affect the redox conditions, and hence, the radionuclide solubility and migration. Therefore, groundwater samples from the two sites were investigated for microbial populations. The total numbers of cells ranged from 10(4) to 10(6) cells ml(-1). Iron-reducing bacteria (IRB) were the largest culturable microbial population in the Palmottu groundwater and were present at up to 1.3 x 10(5) cells ml(-1). Sulfate-reducing bacteria (SRB) and acetogens could also be cultured from the Palmottu groundwater. The numbers of IRB and SRB were largest in groundwater with the lowest uranium concentrations. Removal of dissolved U(VI) from solution was concomitant with the growth of IRB enrichment cultures and the reduction of iron. The redox buffer in the Palmottu groundwater consists of iron and uranium species, both of which are affected by IRB. IRB and aerobic heterotrophs were cultured from the Bangombé groundwater, where redox potentials are buffered by iron and organic carbon species. Microbial populations similar to those found at Palmottu and Bangombé are found throughout the Fennoscandian Shield, a potential host rock for subsurface radioactive waste disposal. These results confirm that microorganisms can be expected to play a role in stabilizing radioactive waste disposed of in the subsurface by lowering redox potential and immobilizing radionuclides.

Distribution of culturable microorganisms in Fennoscandian Shield groundwater.

Microbial populations in 16 groundwater samples from six Fennoscandian Shield sites in Finland and Sweden were investigated. The average total cell number was 3.7x10(5) cells ml(-1), and there was no change in the mean of the total cell numbers to a depth of 1390 m. Culture media were designed based on the chemical composition of each groundwater sample and used successfully to culture anaerobic microorganisms from all samples between 65 and 1350 m depth. Between 0.0084 and 14.8% of total cells were cultured from groundwater samples. Sulfate-reducing bacteria, iron-reducing bacteria and heterotrophic acetogenic bacteria were cultured from groundwater sampled at 65-686 m depth in geographically distant sites. Different microbial populations were cultured from deeper, older and more saline groundwater from 863 to 1350 m depth. Principal component analysis of groundwater chemistry data showed that sulfate- and iron-reducing bacteria were not detected in the most saline groundwater. Iron-reducing bacteria and acetogens were cultured from deep groundwater that contained 0.35-3.5 mM sulfate, while methanogens and acetogens were cultured from deep sulfate-depleted groundwater. In one borehole from which autotrophic methanogens were cultured, dissolved inorganic carbon was enriched in (13)C compared to other Fennoscandian Shield groundwater samples, suggesting that autotrophs were active. It can be concluded that a diverse microbial community is present from the surface to over 1300 m depth in the Fennoscandian Shield.