Revealing Sulfate-Reducing Microorganisms in Oilfield Waters (Flysch Carpathians,South-eastern Poland)

Sulphidogenous microorganism communities were isolated from selected oilfield waters in the Flysch Carpathians of south-eastern Poland. Organisms were incubated using the microcosms method with application of two media: minimal medium and modified Postgate C medium with yeast extract or trisodium citrate or monocyclic hydrocarbons from the BTEX group (benzene, toluene, ethylbenzene, and xylene) as the sole carbon source. Activity of sulphidogenic, autochthonous microorganism communities was noted only on the Postgate medium. Beside active sulphate reduction – max. 70%, ca. 74% biodegradation of organic compounds was also observed in the cultures. The highest content of sulphate-reducing bacteria (SRB) in the COD (ca. 83%) was noted in cultures, in which trisodium citrate and yeast extract were applied as the sole carbon source. Molecular analysis indicated not only the presence of SRB such as Desulfobacterium autothrophicum, Desulfovibrio desulfuricans, but also other microorganisms, e.g., Geobacter metallireducens. All these taxa are obligatory or facultative anaerobes, with metabolism linked mostly with elemental sulphur and/or its oxidized forms, as well as iron. Analysis of the mineral composition of the residues confirmed the presence of elemental sulphur, testifying for the active reduction of sulphates by incompletely oxidizing sulphate reducers assigned to the SRB group. Based on the obtained results, it is concluded that the physical and chemical properties of the oilfield waters are favorable for the growth and development of sulphidogenic microorganism assemblages and mineral-forming processes conducted by them.     

Microbial Communities Involved in Fe Reduction and Mobility During Soil Organic Matter (SOM) Mineralization in Two Contrasted Paddy Soils

Lowland rice fields of West Africa (Ivory Coast) and South Asia (Thailand) are affected by ferrous toxicity or salinity, respectively, and their soil waters contain large amounts of ferrous iron, depending on reducing irrigation condition and suggesting occurrence of bacterial reducing processes. To determine the involvement, dynamic and activities of bacterial communities in Fe(III) reduction and mobilization during anaerobic degradation and mineralization of soil organic matter (SOM), different experiments and analyses have been performed. Results demonstrated that the utilization of SOM as sole carbon, nutrient and energy sources favored the presence of large bacterial communities: facultative anaerobic and anaerobic bacteria, Fe(III)-reducing bacteria (FeRB) (fermentative and Fe respiring), sulfate reducing bacteria (SRB) which are involved in carbon, nitrogen, iron and sulfur cycling. The larger functional diversity is observed in the Ivory Coast paddy soils containing larger amounts of organic matter and sulfur compounds. These communities contained complementary populations (chemoorganotrophic, chemolitotrophic, aerobic, facultative anaerobic and anaerobic) that can be active at different steps of iron solubilization with simultaneous organic matter mineralization. Our results indicate that the pH controlled by bacterial activity, the nature much more than the content of organic matter, and consequently the structure and activity of bacterial communities influence significantly the availability and dynamic of iron in paddy fields which affect the soil quality.     

Anaerobic metabolic processes in the deep terrestrial subsurface

Anaerobic microorganisms were enumerated and metabolic activities measured in deep Coastal Plain sediments sampled from three water‐bearing formations at depths down to 300 m. Aseptically obtained sediment cores harbored the potential for anaerobic biodegradation of various substrates in almost all samples. Although the sediments were not predominantly anaerobic, viable methanogens and sulfate‐reducing bacteria (SRB) were present almost throughout the depth profile. Coliform organisms were also found at various locations, but were not recoverable from drilling muds or water used to slurry the muds. The anaerobic metabolism of lactate and formate was easily detected in most samples. However, acetate and benzoate were degraded only in portions of the subsurface that harbored methanogens. The water‐saturated transmissive zones harbored the highest numbers of SRB and the potential for the widest variety of anaerobic metabolic activities. Small or negligible anaerobic microbial activity was associated with thick clay layers. The accumulation of acetate and the production of methane in samples not amended with exogenous organic matter demonstrated that some strata contained reserves of fermentable carbon and suggested that environmental factors or nutrients other than carbon were potentially limiting in situ microbial activity.     

Application of aerobic microorganisms in bioremediation in situ of soil contaminated by petroleum products

Aerobic microorganisms able to biodegrade benzene, toluene, ethylbenzene, xylene (BTEX) have been isolated from an area contaminated by petroleum products. The activity of the isolated communities was tested under both laboratory and field conditions. Benzene, toluene, ethylbenzene and xylene were added to the cultures as the sole carbon source, at a concentration of 500 mg/L. In batch cultures under laboratory conditions, an 84% reduction of benzene, 86% of toluene and 82% of xylene were achieved. In cultures with ethylbenzene as the sole carbon source, the reduction was around 80%. Slightly lower values were observed under field conditions: 95% reduction of benzene and toluene, 81% of ethylbenzene and 80% of xylene. A high biodegradation activity of benzene (914 μM/L/24 h), toluene (771 μM/L/24 h), xylene (673 μM/L/24 h) and ethylbenzene (644 μM/L/24 h) was observed in the isolated communities.     

Biological fractionation of stable carbon isotopes at the aerobic/anaerobic water interface of meromictic water bodies

Mass-spectrometric investigation of carbon isotope composition (δ¹³C) was carried out for suspended organic matter and dissolved mineral compounds for the water column of some meromictic water bodies differing in salinity and trophic state. As a rule, a more pronounced carbon isotope fractionation (resulting from the metabolism of phytoplankton and anoxygenic phototrophic bacteria) was revealed in the zones of enhanced oxygenic and anoxygenic photosynthesis. Carbon isotope fractionation at the border between oxidized and reduced waters depends both on the activity of microbial communities and on the dominant species of phototrophic microorganisms. Analysis of the distribution profiles of the isotopic composition of suspended organic matter and dissolved mineral carbon revealed active mineralization of the organic matter newly formed via anoxygenic photosynthesis in the monimolimnion by microbial communities, resulting in the release of isotopically light carbon dioxide. Mineral carbon in the anaerobic zones of highly productive meromictic water bodies is therefore enriched with the light ¹²C isotope.     

Hydrocarbon biodegradation in hypersaline environments

When mineral oil, hexadecane, and glutamate were added to natural samples of varying salinity (3.3 to 28.4%) from salt evaporation ponds and Great Salt Lake, Utah, rates of metabolism of these compounds decreased as salinity increased. Rate limitations did not appear to relate to low oxygen levels or to the availability of organic nutrients. Some oxidation of l-[U-C]glutamic acid occurred even at extreme salinities, whereas oxidation of [1-C]hexadecane was too low to be detected. Gas chromatographic examination of hexane-soluble components of tar samples from natural seeps at Rozel Point in Great Salt Lake demonstrated no evidence of biological oxidation of isoprenoid alkanes subject to degradation in normal environments. Some hexane-soluble components of the same tar were altered by incubation in a low-salinity enrichment culture inoculated with garden soil. Attempts to enrich for microorganisms in saline waters able to use mineral oil as a sole source of carbon and energy were successful below, but not above, about 20% salinity. This study strongly suggests a general reduction of metabolic rate at extreme salinities and raises doubt about the biodegradation of hydrocarbons in hypersaline environments.     

Isolation and characterization of a furfural degrading sulfate-reducing bacterium from an anaerobic digester

A sulfate-reducing bacterium (SRB) was isolated from a continuous anaerobic digester, which converted the furfural-containing wastewater to methane and CO₂. This SRB isolate could use furfural, furfuryl alcohol, and 2-furoic acid as sole source of carbon and energy in a defined mineral sulfate medium. Acetic acid was the major end product of furfural degradation. This organism also used wide varieties of other carbon sources, including ethanol, pyruvate, lactate, succinate, propanol, formate, and malate. The SRB isolate contained the electron carrier desulfoviridin. It used SO₄, NO₃, and thiosulfate as electron acceptors. This isolate used ammonium chloride, nitrate and glutamate as nitrogen source. The characteristics of the SRB isolate were closely similar toDesulfovibrio sp.     

Biotransformation of phosphogypsum in wastewaters from the dairy industry

The biotransformation of phosphogypsum by stationary cultures of sulphate reducing bacteria (SRB) in dairy wastewaters (sterile and non-sterile) enriched in phosphogypsum, was studied SRB were isolated from soil contaminated with petroleum-derived products and from wastewaters from the petroleum-refining industry, taking into account that these environments are characteristic for the bacteria of the studied group. The biotransformation products formed were investigated using biological and mineralogical methods. Sulphides in the cultures were determined using the iodometric method, sulphates with the hot barium method, COD by the dichromate method. Determinations involving post-culture sediments and fluids, Ca, S, Sr and P were made using the following analytical procedures: IPC emission spectrometry with induced excitation in the medium and X-ray. Analysis of post-culture sediments was made using a DRON-2 X-ray diffractometer.

Eleven communities of microorganisms with varying degrees of effectiveness were obtained, biodegrading 58% to 98% of the organic contaminants in non-sterile wastewater, with simultaneous biotransformation of about 2.5 g phosphogypsum/L.     

Stimulation of anaerobic biodegradation of DDT and its metabolites in a muck soil: laboratory microcosm and mesocosm studies

The aim of this study was to evaluate the impact of selected electron donors and electron acceptors on the anaerobic biodegradation of DDT and its major metabolites in a muck soil with a long history of exposure to the pesticide. Loss of DDT was measured in anaerobic microcosms supplemented with H₂, lactate, and acetate. The greatest loss of DDT (approximately 87 %) was observed in microcosms amended with lactate and no additional electron acceptor compared to the no additional electron donor or acceptor sets. An increase in measureable concentrations of DDx was observed in un-amended microcosms. In larger scale mesocosms, significant increases in dissolved organic carbon (DOC) corresponded with low redox potentials. Increases in DOC corresponded with sharp increases in measured concentrations of DDx, followed by a decrease in measured DDT concentrations in lactate-amended mesocosms. Our studies indicate that sorbed DDx is released upon anaerobic incubation, and that indigenous microorganisms capable of DDx degradation respond to lactate additions. Both the potential for release of sorbed DDx and the potential for biodegradation of DDx should be considered during remediation of DDx-contaminated organic soils at low redox potentials.     

Biological souring of crude oil under anaerobic conditions

Seawater injection into oil reservoirs for purposes of secondary oil recovery is frequently accompanied by souring (increased sulfide concentrations). Production of hydrogen sulfide causes various problems, such as microbiologically influenced corrosion (MIC) and deterioration of crude oil. Sulfate-reducing bacteria (SRB) are considered to be major players in souring. Volatile fatty acids (VFAs) in oil-field water are believed to be produced by microbial degradation of crude oil. The objective of this research was to investigate mechanisms of souring, focusing specifically on VFA production via crude oil biodegradation. To this end, a microbial consortium collected from an oil–water separator was suspended in seawater; crude oil or liquid n-alkane mixture was added to the culture medium as the sole carbon source, and the culture was incubated under anaerobic conditions for 190 days. Physicochemical analysis showed that preferential toluene degradation and sulfate reduction occurred concomitantly in the culture containing crude oil. Sulfide concentrations were much lower in the alkane-supplemented culture than in the crude oil-supplemented culture. These observations suggest that SRB are related to the toluene activation and VFA consumption steps of crude oil degradation. Therefore, the electron donors for SRB are not only VFA, but many components of crude oil, especially toluene. Alkanes were also degraded by microorganisms, but did not contribute to reservoir souring.     

Phosphogypsum biotransformation in cultures of sulphate reducing bacteria in whey

Assemblages of anaerobic sulphidogenic microorganisms were isolated from soil polluted by oil-derived products and grown using the microcosms method. The cultures were grown in minimal and Postgate media with phosphogypsum (PG) as the sole electron acceptor and with lactate, casein or lactose as the sole carbon source. The most effective was the assemblage in Postgate medium with lactose as the sole carbon source. A reduction of 980 mg COD l^?1 (reduction of about 40%) and 790 mg SO₄^2? l^?1 (reduction of 53% of phosphogypsum introduced to the medium) was noted in the culture. The lowest activity was observed for minimal medium with lactose as sole carbon source (reduction of 4.4% COD and 40% PG). The selected assemblage became an inoculum for a culture in Postgate, minimal and/or distilled water medium with PG (6 g l^?1) and cheese whey (2.5 and 4.5 g l^?1).A percentage reduction of COD and SO₄^2? of PG was observed in all cultures. After growth, the residues were weighed and in all cases a distinct mass reduction of PG was observed in comparison to the 6 g l^?1 introduced to the medium. Diffractometric studies of the residues confirmed the presence of calcite and apatite. The presence of these mineral phases in the residues allows their application as agricultural fertilisers.     

Review of MTBE Biodegradation and Bioremediation

Conclusive evidence of methyl tert-butyl ether (MTBE) biotransformation and complete mineralization under aerobic conditions in environmental samples and enrichment cultures is reviewed, in addition to increasing evidence of MTBE biotransformation under anaerobic conditions. The metabolic pathway of MTBE appears to have two key intermediates, tert-butyl alcohol (TBA) and 2-hydroxy isobutyric acid (HIBA). The first enzyme in MTBE biodegradation has been identified as either a cytochrome P450 or a nonhemic monooxygenase in different isolates. Mixed and pure cultures of microorganisms have utilized MTBE as a sole carbon and energy source. Cometabolism of MTBE with n-alkanes at rates of 3.9 to 52 nmol/min/mg protein has been documented. The presence of co-contaminants such as BTEX has either not affected or seemed to limit MTBE biodegradation. Some studies of MTBE natural attenuation have attributed mass loss to biodegradation, while others have attributed mass loss to dilution and dispersion. Recent advances in the assessment of MTBE biodegradation have indicated the potential for natural anaerobic transformation of MTBE. In situ bioremediation of MTBE has been enhanced by adding air or oxygen, or by adding microorganisms and air or oxygen. Bioreactors have attained significant removal of MTBE from MTBE-contaminated influent. Despite historical concerns about the biodegradability of MTBE, several biological methods can now be used for MTBE remediation.     

Biotransformation of phosphogypsum in media containing different forms of nitrogen

Studies on the biotransformation of phosphogypsum (a waste product formed in the course of the production of phosphorous fertilizers) with the use of sulfate reducing bacteria (SRB) demonstrated that it is a good source of sulfates and biogenic elements for these bacteria, though the addition of organic carbon and nitrogen is necessary. The aim of this study was to investigate the form of nitrogen and C:N ratio in the medium on the growth of SRB community in cultures containing phosphogypsum. Batch community cultures of sulfate reducing bacteria were maintained in medium with phosphogypsum (5.0 g/l), different concentrations of sodium lactate (1.6 - 9.4 g/l) and different forms (NH4CI, CO(NH2)2, KNO3) and concentrations (0 - 250 mg/l) of nitrogen. The growth of SRB was studied in the C:N ratio of from 2:1 to 300:1. It was found that: 1 - the best source of nitrogen for SRB is urea, followed by ammonium, the worst were nitrates; 2 - the bacteria were also able to grow in medium without nitrogen but their activity was then by approximately 15% lower than in optimal growth conditions; 3 - in medium with KNO3 inhibition of sulfate reduction by approx. 50% was observed; 4 - the highest reduction of nitrates (removal of nitrate) in media with phosphogypsum and nitrates was at limiting concentrations of sodium lactate. This is probably caused by the selection under these conditions (low concentration of hydrogen sulfide) of denitrifying bacteria or sulfate reducing bacteria capable of using nitrates as an electron acceptor.     

Stable carbon isotope fractionation by sulfate-reducing bacteria

Biogeochemical transformations occurring in the anoxic zones of stratified sedimentary microbial communities can profoundly influence the isotopic and organic signatures preserved in the fossil record. Accordingly, we have determined carbon isotope discrimination that is associated with both heterotrophic and lithotrophic growth of pure cultures of sulfate-reducing bacteria (SRB). For heterotrophic-growth experiments, substrate consumption was monitored to completion. Sealed vessels containing SRB cultures were harvested at different time intervals, and delta(13)C values were determined for gaseous CO(2), organic substrates, and products such as biomass. For three of the four SRB, carbon isotope effects between the substrates, acetate or lactate and CO(2), and the cell biomass were small, ranging from 0 to 2 per thousand. However, for Desulfotomaculum acetoxidans, the carbon incorporated into biomass was isotopically heavier than the available substrates by 8 to 9 per thousand. SRB grown lithoautotrophically consumed less than 3% of the available CO(2) and exhibited substantial discrimination (calculated as isotope fractionation factors [alpha]), as follows: for Desulfobacterium autotrophicum, alpha values ranged from 1.0100 to 1.0123; for Desulfobacter hydrogenophilus, the alpha value was 0.0138, and for Desulfotomaculum acetoxidans, the alpha value was 1.0310. Mixotrophic growth of Desulfovibrio desulfuricans on acetate and CO(2) resulted in biomass with a delta(13)C composition intermediate to that of the substrates. The extent of fractionation depended on which enzymatic pathways were used, the direction in which the pathways operated, and the growth rate, but fractionation was not dependent on the growth phase. To the extent that environmental conditions affect the availability of organic substrates (e.g., acetate) and reducing power (e.g., H(2)), ecological forces can also influence carbon isotope discrimination by SRB.     

Seasonal oscillation of microbial iron and sulfate reduction in saltmarsh sediments (Sapelo Island, GA, USA)

Seasonal variations in anaerobic respiration pathways were investigated at three saltmarsh sites using chemical data, sulfate reduction rate measurements, enumerations of culturable populations of anaerobic iron-reducing bacteria (FeRB), and quantification of in situ 16S rRNA hybridization signals targeted for sulfate-reducing bacteria (SRB). Bacterial sulfate reduction in the sediments followed seasonal changes in temperature and primary production of the saltmarsh, with activity levels lowest in winter and highest in summer. In contrast, a dramatic decrease in the FeRB population size was observed during summer at all sites. The collapse of FeRB populations during summer was ascribed to high rates of sulfide production by SRB, resulting in abiotic reduction of bioavailable Fe(III) (hydr)oxides. To test this hypothesis, sediment slurry incubations at 10, 20 and 30 °C were carried out. Increases in temperature and labile organic carbon availability (acetate or lactate additions) increased rates of sulfate reduction while decreasing the abundance of culturable anaerobic FeRB. These trends were not reversed by the addition of amorphous Fe(III) (hydr)oxides to the slurries. However, when sulfate reduction was inhibited by molybdate, no decline in FeRB growth was observed with increasing temperature. Addition of dissolved sulfide adversely impacted propagation of FeRB whether molybdate was added or not. Both field and laboratory data therefore support a sulfide-mediated limitation of microbial iron respiration by SRB. When total sediment respiration rates reach their highest levels during summer, SRB force a decline in the FeRB populations. As sulfate reduction activity slows down after the summer, the FeRB are able to recover.     

Review of MTBE Biodegradation and Bioremediation

Conclusive evidence of methyl tert-butyl ether (MTBE) biotransformation and complete mineralization under aerobic conditions in environmental samples and enrichment cultures is reviewed, in addition to increasing evidence of MTBE biotransformation under anaerobic conditions. The metabolic pathway of MTBE appears to have two key intermediates, tert-butyl alcohol (TBA) and 2-hydroxy isobutyric acid (HIBA). The first enzyme in MTBE biodegradation has been identified as either a cytochrome P450 or a nonhemic monooxygenase in different isolates. Mixed and pure cultures of microorganisms have utilized MTBE as a sole carbon and energy source. Cometabolism of MTBE with n-alkanes at rates of 3.9 to 52 nmol/min/mg protein has been documented. The presence of co-contaminants such as BTEX has either not affected or seemed to limit MTBE biodegradation. Some studies of MTBE natural attenuation have attributed mass loss to biodegradation, while others have attributed mass loss to dilution and dispersion. Recent advances in the assessment of MTBE biodegradation have indicated the potential for natural anaerobic transformation of MTBE. In situ bioremediation of MTBE has been enhanced by adding air or oxygen, or by adding microorganisms and air or oxygen. Bioreactors have attained significant removal of MTBE from MTBE-contaminated influent. Despite historical concerns about the biodegradability of MTBE, several biological methods can now be used for MTBE remediation.     

Physiological characterization of strain DCB-1, a unique dehalogenating sulfidogenic bacterium

Strain DCB-1 is an obligately anaerobic bacterium which carries out the reductive dehalogenation of halobenzoates and was previously known to grow only on pyruvate plus 20% ruminal fluid. When various electron acceptors were supplied, thiosulfate and sulfite were found to stimulate growth. Sulfide was produced from thiosulfate. Cytochrome c and desulfoviridin were detected. The mol% G+C was 49 (at the thermal denaturation temperature). Of 55 carbon sources tested, only pyruvate supported growth as the sole carbon source in mineral medium. Lactate, acetate, L- and D-malate, glycerol, and L- and D-arabinose stimulated growth when supplemented with 10% ruminal fluid and 20 mM thiosulfate. In mineral medium, pyruvate was converted to acetate and lactate, with small amounts of succinate and fumarate accumulating transiently. During growth with thiosulfate, all of these products accumulated transiently. Addition of excess hydrogen to pyruvate-grown cultures resulted in diversion of carbon to formate, lactate, and butyrate, which caused a decrease in cell yield. We conclude that strain DCB-1 is a new type of sulfidogenic bacterium.     

Physiological characterization of strain DCB-1, a unique dehalogenating sulfidogenic bacterium.

Strain DCB-1 is an obligately anaerobic bacterium which carries out the reductive dehalogenation of halobenzoates and was previously known to grow only on pyruvate plus 20% ruminal fluid. When various electron acceptors were supplied, thiosulfate and sulfite were found to stimulate growth. Sulfide was produced from thiosulfate. Cytochrome c and desulfoviridin were detected. The mol% G+C was 49 (at the thermal denaturation temperature). Of 55 carbon sources tested, only pyruvate supported growth as the sole carbon source in mineral medium. Lactate, acetate, L- and D-malate, glycerol, and L- and D-arabinose stimulated growth when supplemented with 10% ruminal fluid and 20 mM thiosulfate. In mineral medium, pyruvate was converted to acetate and lactate, with small amounts of succinate and fumarate accumulating transiently. During growth with thiosulfate, all of these products accumulated transiently. Addition of excess hydrogen to pyruvate-grown cultures resulted in diversion of carbon to formate, lactate, and butyrate, which caused a decrease in cell yield. We conclude that strain DCB-1 is a new type of sulfidogenic bacterium.     

Inside the alkalinity engine: the role of electron donors in the organomineralization potential of sulfate‐reducing bacteria

Mineral precipitation in microbial mats may have been the key to their preservation as fossil stromatolites, potentially documenting evidence of the earliest life on Earth. Two factors that contribute to carbonate mineral precipitation are the saturation index (SI) and the presence of nucleation sites. Both of these can be influenced by micro‐organisms, which can either alter SI through their metabolisms, or produce and consume organic substances such as extracellular polymeric substances (EPS) that can affect nucleation. It is the balance of individual metabolisms within the mat community that determines the pH and the dissolved inorganic carbon concentration, thereby potentially increasing the alkalinity and consequently the SI. Sulfate‐reducing bacteria (SRB) are an important component of this ‘alkalinity engine.’ The activity of SRB often peaks in layers where CaCO₃ precipitates, and mineral precipitation has been demonstrated in SRB cultures; however, the effect of their metabolism on the alkalinity engine and actual contribution to mineral precipitation is the subject of controversy. Here, we show through culture experiments, theoretical calculations, and geochemical modeling studies that the pH, alkalinity, and organomineralization potential will vary depending on the type of electron donor. Specifically, hydrogen and formate can increase the pH, but electron donors like lactate and ethanol, and to a lesser extent glycolate, decrease the pH. The implication of this for the lithification of mats is that the combination of processes supplying electron donors and the utilization of these compounds by SRB may be critical to promoting mineral precipitation.     

Sulfate Reduction at a Lignite Seam: Microbial Abundance and Activity

In a combined isotope geochemical and microbiological investigation, a setting of multiple aquifers was characterized. Biologically mediated redox processes were observed in the aquifers situated in marine sands of Tertiary age and overlying Quaternary gravel deposits. Intercalated lignite seams define the aquitards, which separate the aquifers. Bacterial oxidation of organic matter is evident from dissolved inorganic carbon characterized by average carbon isotope values between ?18.4 per thousand and ?15.7 per thousand (PDB). Strongly positive sulfur isotope values of up to +50 per thousand (CTD) for residual sulfate indicate sulfate reduction under closed system conditions with respect to sulfate availability. Both, hydrochemical and isotope data are thus consistent with the recent activity of sulfate-reducing bacteria (SRB). Microbiological investigations revealed the presence of an anaerobic food chain in the aquifers. Most-probable-number (MPN) determinations for SRB and fermenting microorganisms reached highest values at the interface between aquifer and lignite seam (1.5 x 103 cells/g sediment dry mass). Five strains of SRB were isolated from highest MPN dilutions. Spore-forming bacteria appeared to dominate the SRB population. Sulfate reduction rates were determined by the 35S-radiotracer method. A detailed assessment indicates an increase in the reduction rate in proximity to the lignite seam, with a maximum turnover of 8.4 mM sulfate/a, suggesting that lignite-drived compounds represent the substrate for sulfate reduction.