Dissimilatory Sulfate-reducing Bacteria Research Articles

Microbial communities in paddy soils: differences in abundance and functionality between rhizosphere and pore water, influence of different soil organic carbon, sulfate fertilization, and cultivation time, and contribution to arsenic mobility and speciation.

Abiotic factors and rhizosphere microbial populations influence arsenic accumulation in rice grains. Despite mineral and organic surfaces are keystones in element cycling, localization of specific microbial reactions in the root/soil/pore water system is still unclear. Here, we tested if original unplanted soil, rhizosphere soil, and pore water represented distinct ecological microniches for arsenic-, sulfur- and iron-cycling microorganisms and compared the influence of relevant factors such as soil type, sulfate fertilization, and cultivation time. In rice open-air-mesocosms with two paddy soils (2.0% and 4.7% organic carbon), Illumina 16S rRNA gene sequencing demonstrated little significant effects of cultivation time and sulfate fertilization that decreased Archaea-driven microbial networks and incremented sulfate reducing and sulfur oxidizing bacteria. Different compartments, characterized by different bacterial and archaeal compositions, had the strongest effect with higher microbial abundances, bacterial biodiversity and interconnections in the rhizosphere versus pore water. Within each compartment, a significant soil type effect was observed. Higher percentage contributions of rhizosphere dissimilatory arsenate- and iron-reducing, arsenite-oxidizing, and, surprisingly, dissimilatory sulfate-reducing bacteria as well as pore water iron-oxidizing bacteria in the lower organic carbon soil supported previous chemistry-based interpretations of a more active S-cycling, a higher percentage of thioarsenates, and lower arsenic mobility by sorption to mixed Fe(II)Fe(III)-minerals in this soil.

Diversity and biogenesis contribution of sulfate-reducing bacteria in arsenic-contaminated soils from realgar deposits.

Microbial sulfate reduction, a vital mechanism for microorganisms living in anaerobic, sulfate-rich environments, is an essential aspect of the sulfur biogeochemical cycle. However, there has been no detailed investigation of the diversity and biogenesis contribution of sulfate-reducing bacteria in arsenic-contaminated soils from realgar deposits. To elucidate this issue, soil samples from representative abandoned realgar deposits were collected. Microcosm assays illustrated that all three samples (2-1, 2-2, and 2-3) displayed efficient sulfate and As(V)-respiring activities. Furthermore, a total of 28 novel sequence variants of dissimilatory sulfite reductase genes and 2 new families of dsrAB genes were successfully identified. A novel dissimilatory sulfate-reducing bacterium, Desulfotomaculum sp. JL1, was also isolated from soils, and can efficiently respiratory reduce As(V) and sulfate in 4 and 5days, respectively. JL1 can promote the generation of yellow precipitates in the presence of multiple electron acceptors (both contain sulfate and As(V) in the cultures), which indicated the biogenesis contribution of sulfate-reducing bacteria to the realgar mine. Moreover, this area had unique microbial communities; the most abundant populations belonged to the phyla Proteobacteria, Chloroflexi, and Acidobacteriota, which were attributed to the unique geochemistry characteristics, such as total organic carbon, total As, NO3-, and SO42-. The results of this study provide new insight into the diversity and biogenesis contributions of sulfate-reducing bacteria in arsenic-contaminated soils from realgar deposits.

Biogeochemical dynamics and microbial community development under sulfate- and iron-reducing conditions based on electron shuttle amendment.

Iron reduction and sulfate reduction are two of the major biogeochemical processes that occur in anoxic sediments. Microbes that catalyze these reactions are therefore some of the most abundant organisms in the subsurface, and some of the most important. Due to the variety of mechanisms that microbes employ to derive energy from these reactions, including the use of soluble electron shuttles, the dynamics between iron- and sulfate-reducing populations under changing biogeochemical conditions still elude complete characterization. Here, we amended experimental bioreactors comprised of freshwater aquifer sediment with ferric iron, sulfate, acetate, and the model electron shuttle AQDS (9,10-anthraquinone-2,6-disulfonate) and monitored both the changing redox conditions as well as changes in the microbial community over time. The addition of the electron shuttle AQDS did increase the initial rate of FeIII reduction; however, it had little effect on the composition of the microbial community. Our results show that in both AQDS- and AQDS+ systems there was an initial dominance of organisms classified as Geobacter (a genus of dissimilatory FeIII-reducing bacteria), after which sequences classified as Desulfosporosinus (a genus of dissimilatory sulfate-reducing bacteria) came to dominate both experimental systems. Furthermore, most of the ferric iron reduction occurred under this later, ostensibly “sulfate-reducing” phase of the experiment. This calls into question the usefulness of classifying subsurface sediments by the dominant microbial process alone because of their interrelated biogeochemical consequences. To better inform models of microbially-catalyzed subsurface processes, such interactions must be more thoroughly understood under a broad range of conditions.

Isolation and cultivation of a novel sulfate-reducing magnetotactic bacterium belonging to the genus Desulfovibrio

Magnetotactic bacteria (MTB) synthesize magnetosomes composed of membrane-enveloped magnetite (Fe3O4) and/or greigite (Fe3S4) nanoparticles in the cells. It is known that the magnetotactic Deltaproteobacteria are ubiquitous and inhabit worldwide in the sediments of freshwater and marine environments. Mostly known MTB belonging to the Deltaproteobacteria are dissimilatory sulfate-reducing bacteria that biomineralize bullet-shaped magnetite nanoparticles, but only a few axenic cultures have been obtained so far. Here, we report the isolation, cultivation and characterization of a dissimilatory sulfate-reducing magnetotactic bacterium, which we designate “strain FSS-1”. We found that the strain FSS-1 is a strict anaerobe and uses casamino acids as electron donors and sulfate as an electron acceptor to reduce sulfate to hydrogen sulfide. The strain FSS-1 produced bullet-shaped magnetite nanoparticles in the cells and responded to external magnetic fields. On the basis of 16S rRNA gene sequence analysis, the strain FSS-1 is a member of the genus Desulfovibrio, showing a 96.7% sequence similarity to Desulfovibrio putealis strain B7-43T. Futhermore, the magnetosome gene cluster of strain FSS-1 was different from that of Desulfovibrio magneticus strain RS-1. Thus, the strain FSS-1 is considered to be a novel sulfate-reducing magnetotactic bacterium belonging to the genus Desulfovibrio.

Stable isotopes (H, O, S) signatures evidencing evolutionary trends of Brazilian spas groundwaters

This paper reports a stable isotope (H, O, S) study of groundwater samples (25) in 10 spas from southeast Brazil. They are cold (25 °C, 36%), hypothermal (25–33 °C, 60%) and mesothermal (33–36 °C, 4%) waters; and are predominantly composed of bicarbonate, carbonate and sodium. Dissolved sulfate is the dominant anion in one water sample and its concentration increased with increase in pH, electrical conductivity (EC) and total dissolved solids (TDS) values in the samples. 2H and 18O data reported in the literature for circa 700 rainwater samples collected in São Paulo State and Brasília airport, Brazil, were used to construct the following regional meteoric waterline (RMWL): δ2H V-SMOW (‰) = 8.06 δ18Owater V-SMOW (‰) + 12.85. The HO isotopes of the studied groundwater samples show a δ18Owater variation of −14.1 to −5.3‰ V-SMOW, whereas the δ2H range was −66.5 to −31.7‰ V-SMOW, which was plotted on or near the RMWL, suggesting a meteoric origin for them. However, one hypothermal water sample (29 °C) plotted away from this RMWL, and is from a deep fractured aquifer which mainly occurs in fenitization aureole rocks surrounding the carbonatite complex at Araxá spa, Minas Gerais State, and a magmatic hydrothermal water may contribute to this water sample. The use of both δ34Ssulfate and δ18Osulfate values in the selected sites permitted the plotting of a mixture line of the dissolved sulfate in the different aquifer systems of this study, considering two major sources as endmembers: sulfide oxidation, SOX (δ34Ssulfate = +1‰ and δ18Osulfate = +1‰) and sulfates possessing δ34Ssulfate = +9‰ and δ18Osulfate = +21‰ (OSW). This is an unusual isotopic composition opposite from that of sea water. The δ34Ssulfate and δ18Osulfate values tend to increase with increase in dissolved sulfate concentration. The OS isotopes signatures that deviated from the general mixture line could be due to several processes, including atmospheric deposition, SO2 oxidation in the atmosphere, soil-derived sulfate, sulfide oxidation, evaporites dissolution and the presence of dissimilatory sulfate-reducing bacteria. Three sulfate isotope composition of spring waters from Águas da Prata spa in the Poços de Caldas alkaline massif (PCAM) represented endmember sources that defined a triangle around the remaining PCAM samples, as well as the samples from the crystalline basement, São Francisco craton, and Alto Paranaíba igneous province (APIP). Such triangle allows for estimation of the relative isotopic contribution in the different spas groundwaters, which is a new approach that focuses on the use of δ34Ssulfate-δ18Osulfate pair in environmental studies, and shows its usefulness in addition to other conventional hydrogeochemical fingerprints.

Water management impacts the soil microbial communities and total arsenic and methylated arsenicals in rice grains

The bioavailability of the metalloid arsenic (As) in paddy soil is controlled by microbial cycling of As and other elements such as iron (Fe) and sulfur (S), which are strongly influenced by water management in paddy fields. In this study, we evaluated how water management affects As bioavailability by growing rice plants in a geogenic As-contaminated soil. We determined As speciation in soil porewater and the diversity of the associated microbial community. Continuous flooding enhanced the release of Fe and As and increased arsenite (As(III)) and methylated As species concentrations in the rice grain compared with aerobic treatment. Total inorganic and organic As in the grain was 84% and 81% lower, respectively, in the aerobic treatment compared with the continuous flooding treatment. The amounts of Fe(III)-reducing bacteria (FeRB) increased in the flooded rhizosphere soil. The abundance of FeRB in the soil correlated with the dissolution of Fe and As. Among the As-transformation genes quantified, the aioA gene for As(III) oxidation and arsM gene for As(III) methylation were most abundant. The arsM copy number correlated positively with the levels of dsrB (dissimilatory (bi) sulfite reductase β-subunit), suggesting that dissimilatory sulfate-reducing bacteria (SRB) may play an important role in dimethylarsenate (DMAs(V)) production in soil. Our results show that decreased populations of rhizosphere FeRB and SRB contributed to a lower bioavailability of As, and decreased production of methylated arsenicals under oxic conditions.

The effect of gram-positive (Desulfosporosinus orientis) and gram-negative (Desulfovibrio desulfuricans) sulfate-reducing bacteria on iron sulfide mineral precipitation.

Growth of two dissimilatory sulfate-reducing bacteria, Desulfosporosinus orientis (gram-positive) and Desulfovibrio desulfuricans (gram-negative), in a chemically defined culture medium resulted in similar growth rates (doubling times for each culture = 2.8 h) and comparable rates of H2S generation (D. orientis = 0.19 nmol/L S2- per cell per h; D. desulfuricans = 0.12 nmol/L S2- per cell per h). Transmission electron microscopy of whole mounts and thin sections revealed that the iron sulfide mineral precipitates produced by the two cultures were morphologically different. The D. orientis culture flocculated, with the minerals occurring as subhedral plate-like precipitates, which nucleated on the cell wall during exponential growth producing extensive mineral aggregates following cell autolysis and endospore release. In contrast, the D. desulfuricans culture produced fine-grained colloidal or platy iron sulfide precipitates primarily within the bulk solution. Mineral analysis by scanning electron microscopy - energy dispersive spectroscopy indicated that neither culture promoted advanced mineral development beyond a 1:1 Fe:S stoichiometry. This analysis did not detect pyrite (FeS2). The average Fe:S ratios were 1 : 1.09 ± 0.03 at 24 h and 1 : 1.08 ± 0.03 at 72 h for D. orientis and 1 : 1.05 ± 0.02 at 24 h and 1 : 1.09 ± 0.07 at 72 h for D. desulfuricans. The formation of "biogenic" iron sulfides by dissimilatory sulfate-reducing bacteria is influenced by bacterial cell surface structure, chemistry, and growth strategy, i.e., mineral aggregation occurred with cell autolysis of the gram-positive bacterium.

The shift of sulfate-reducing bacterial communities from the upland to the paddy stage in a rapeseed-rice rotation system, and the effect from the long-term straw returning

Sulfate reduction is an essential process in the biogeochemical sulfur cycle in soils. It is mainly driven by dissimilatory sulfate-reducing bacteria (DSRB). The responses of the DSRB community in a rapeseed-rice rotation system to the environmental variability caused by the shift from the upland to the paddy stages, and the effects of the long-term straw returning on this shift remains unclear. We surveyed the sulfate reducing potential (SRP) and the structure of the DSRB community by high-throughput sequencing that targeted dsrB (and the xenologous dsrB) in soils. The SRP, ranging from 1.2 to 5.8μmold−1dwg−1, was increased in the paddy stage, and it was likely to be enhanced by the long-term straw returning. Highly abundant DSRB optimal taxonomic units (OTUs) were found to be affiliated with the Nitrospirae supercluster (including the uncultured DsrAB lineage WX, lineage 10, and an unknown clade with GU372064), the Environmental supercluster 1 (including the uncultured DsrAB lineage 8 and an unknown lineage related to EF065019), and the Firmicutes group (including the uncultured DsrAB lineage 6 and an unknown lineage between the uncultured DsrAB lineages 2 and 3). The environmental transformation from the upland to the paddy stages led to a decrease in the α-diversity and the number of detectable OTUs. The abundance of a few dominant DSRB OTUs was changed by either the environmental transformation from the upland to the paddy stages or the effects caused by the long-term straw returning. In this study, the changes in the DSRB community structure correlated with the decrease in soil pH and total phosphorus content, and the increase in available sulfate and moisture contents.

Growth of magnetotactic sulfate-reducing bacteria in oxygen concentration gradient medium.

Although dissimilatory sulfate-reducing bacteria (SRB) are generally described as strictly anaerobic organisms with regard to growth, several reports have shown that some SRB, particularly Desulfovibrio species, are quite resistant to O2 . For example, SRB remain viable in many aerobic environments while some even reduce O2 to H2 O. However, reproducible aerobic growth of SRB has not been unequivocally documented. Desulfovibrio magneticus is a SRB that is also a magnetotactic bacterium (MTB). MTB biomineralize magnetosomes which are intracellular, membrane-bounded, magnetic iron mineral crystals. The ability of D. magneticus to grow aerobically in several different media under air where an O2 concentration gradient formed, or under O2 -free N2 gas was tested. Under air, cells grew as a microaerophilic band of cells at the oxic-anoxic interface in media lacking sulfate. These results show that D. magneticus is capable of aerobic growth with O2 as a terminal electron acceptor. This is the first report of consistent, reproducible aerobic growth of SRB. This finding is critical in determining important ecological roles SRB play in the environment. Interestingly, the crystal structure of the magnetite crystals of D. magneticus grown under microaerobic conditions showed significant differences compared with those produced anaerobically providing more evidence that environmental parameters influence magnetosome formation.

Sulfur-34S Stable Isotope Labeling of Amino Acids for Quantification (SULAQ34) of Proteomic Changes in Pseudomonas fluorescens during Naphthalene Degradation

The relative quantification of proteins is one of the major techniques used to elucidate physiological reactions. Because it allows one to avoid artifacts due to chemical labeling, the metabolic introduction of heavy isotopes into proteins and peptides is the preferred method for relative quantification. For eukaryotic cells, stable isotope labeling by amino acids in cell culture (SILAC) has become the gold standard and can be readily applied in a vast number of scenarios. In the microbial realm, with its highly versatile metabolic capabilities, SILAC is often not feasible, and the use of other (13)C or (15)N labeled substrates might not be practical. Here, the incorporation of heavy sulfur isotopes is shown to be a useful alternative. We introduce (34)S stable isotope labeling of amino acids for quantification and the corresponding tools required for spectra extraction and disintegration of the isotopic overlaps caused by the small mass shift. As proof of principle, we investigated the proteomic changes related to naphthalene degradation in P. fluorescens ATCC 17483 and uncovered a specific oxidative-stress-like response.

Sulfate-Reducing Bacterium Desulfovibrio desulfuricans ND132 as a Model for Understanding Bacterial Mercury Methylation

We propose the use of Desulfovibrio desulfuricans ND132 as a model species for understanding the mechanism of microbial Hg methylation. Strain ND132 is an anaerobic dissimilatory sulfate-reducing bacterium (DSRB), isolated from estuarine mid-Chesapeake Bay sediments. It was chosen for study because of its exceptionally high rates of Hg methylation in culture and its metabolic similarity to the lost strain D. desulfuricans LS, the only organism for which methylation pathways have been partially defined. Strain ND132 is an incomplete oxidizer of short-chain fatty acids. It is capable of respiratory growth using fumarate as an electron acceptor, supporting growth without sulfide production. We used enriched stable Hg isotopes to show that ND132 simultaneously produces and degrades methylmercury (MeHg) during growth but does not produce elemental Hg. MeHg produced by cells is mainly excreted, and no MeHg is produced in spent medium. Mass balances for Hg and MeHg during the growth of cultures, including the distribution between filterable and particulate phases, illustrate how medium chemistry and growth phase dramatically affect Hg solubility and availability for methylation. The available information on Hg methylation among strains in the genus Desulfovibrio is summarized, and we present methylation rates for several previously untested species. About 50% of Desulfovibrio strains tested to date have the ability to produce MeHg. Importantly, the ability to produce MeHg is constitutive and does not confer Hg resistance. A 16S rRNA-based alignment of the genus Desulfovibrio allows the very preliminary assessment that there may be some evolutionary basis for the ability to produce MeHg within this genus.

Distinct Microbial Behavior of Re Compared to Tc: Evidence Against Microbial Re Fixation in Aquatic Sediments

Rhenium is enriched in suboxic and anoxic sediments relative to oxic sediments, a characteristic that is being exploited in its use as a paleoredox indicator. Rhenium is fixed at sediment depths where iron reduction and sulfate reduction are the dominant microbial terminal electron-accepting processes. In order to explore mechanisms of its fixation, we investigated perrhenate behavior in pure, batch cultures of two dissimilatory sulfate-reducing strains (Desulfovibrio desulfuricans subsp. desulfuricans and Desulfovibrio desulfuricans ND132) and two iron-reducing strains (Geobacter metallireducens GS-15 and Shewanella oneidensis MR-1). Perrhenate concentrations tested ranged from 0.04 to 12 μM, roughly 4 to 7 orders of magnitude larger than seawater Re concentrations. Within this broad concentration range, none of the organisms tested actively removed Re from solution during one week's growth to stationary phase. Despite these results, the sulfate-reducing cultures appeared to have reached supersaturation relative to ReS2(s), and the iron-reducing cultures may have reached supersaturation relative to ReO2(s). We conclude that neither direct nor short-term indirect microbial processes involving these bacteria are likely to explain Re fixation in sediments. Our results cannot exclude the possibility that microbial metabolites, such as Fe(II) or sulfide, do drive abiotic Re fixation over longer periods of time. The lack of perrhenate reduction by dissimilatory sulfate-reducing bacteria and iron-reducing bacteria contrasts with published reports of pertechnetate behavior. Despite many qualitative similarities between Re and Tc, it is clear that these two elements are quantitatively dissimilar, with Re fixation requiring more intensely reducing conditions.

Characterization of halophiles in natural MgSO 4 salts and laboratory enrichment samples: Astrobiological implications for Mars

The presence of sulfate salts and limited subsurface water (ice) on Mars suggests that any liquid water on Mars today will occur as (magnesium) sulfate-rich brines in regions containing sources of magnesium and sulfur. The Basque Lakes of British Columbia, Canada, represent a hypersaline terrestrial analogue site, which possesses chemical and physical properties similar to those observed on Mars. The Basque Lakes also contain diverse halophilic organisms representing all three Kingdoms of life, growing in surface and near-subsurface environments. Of interest from an astrobiological perspective, crushed magnesium sulfate samples that were analyzed using a modified Lowry protein assay contained biomass in every crystal inspected, with biomass values from 0.078 to 4.21 mg biomass/g salt; average=0.74±0.7 mg biomass/g salt. Bacteria and Archaea cells were easily observed even in low-biomass samples using light microscopy, and bacteria trapped within magnesium sulfate crystals were observed using confocal microscopy. Regions within the salt also contained bacterial pigments, e.g., carotenoids, which were separate from the cells, indicating that cell lysis might have occurred during entrapment within the salt matrix. These biosignatures, cells, and any ‘soluble’ organic constituents were primarily found trapped within fluid inclusions or fluid-filled void spaces between intergrown crystals. Diffuse reflectance infrared Fourier transform spectroscopy (reflectance IR) analysis of enrichment cultures, containing cyanobacteria, Archaea, or dissimilatory sulfate-reducing bacteria, highlighted molecular biosignature features between 550–1650 and 2400–3000 cm −1. Spectra from natural salts demonstrated that we can detect biomass within salt crystals using the most sensitive biosignatures, which are the 1530–1570 cm −1, C–N, N–H, –COOH absorptions and the 1030–1050 cm −1 C–OH, C–N, PO 4 3− bond features. The lowest detection limit for a biosignature absorption feature using reflectance IR was with a natural sample that possessed 0.78 mg biomass/g salt. In a model cell, i.e., a 0.5 by 1 μm bacillus, this biomass value corresponds to approximately 7.8×10 8 cells/g salt. Based on its ability to detect biomass entrapped within natural sulfate salts, reflectance IR may make an effective remote-sensing tool for finding enrichments of organic carbon within outcrops and surficial sedimentary deposits on Mars.

Photosynthetic sulfur bacteria from Zimapan Reservoir (Mexico): Seasonal variations and toxic effects of zinc in a microcosm system

Photosynthetic sulfur bacteria oxidize the sulfide produced by dissimilatory sulfate-reducing bacteria, thus preventing the occurrence of pollution by these compounds in the oxygenic zone of aquatic systems. Zinc is one of the most abundant heavy metals. Its toxic effects have been documented in various organisms that can photosynthesize, but a lack of information prevails in this respect, about photosynthetic sulfur bacteria. The aim of the present study was to evaluate the toxic effects of zinc on green sulfur bacteria (Chlorobiaceae) and purple sulfur bacteria (Chromatiaceae) in a microcosm as well to explore the possible relationships between the bacterial population growths with the physicochemical properties of the metalimnion of Zimapan Reservoir (Mexico) during a 1-year period. Hydrosoluble organic carbon (HOC) and light availability in the metalimnion were the limiting factors for growth in these bacteria. Higher HOC levels occurred in Spring, at the end of the wet season and in Winter. Current zinc levels in the reservoir seem to elicit toxic effects. At lethal zinc concentrations, the best concentration-response relationship was shown. The biomarker for purple sulfur bacteria was the bacteriochlorophyll content, while total reducing sugar was the best biomarker for green sulfur bacteria. At sublethal concentrations, zinc alters adenosine triphosphate levels, total reducing sugar, total protein content, and bacteriochlorophyll content. The observed responses indicate that green sulfur bacteria are more sensitive than purple sulfur bacteria and are suitable target organisms for use in establishing the accepted threshold concentration of zinc (NOEC 0.056–0.097 mg L−1 of Zn) in anoxic deep water.

‘Candidatus Magnetoglobus multicellularis’, a multicellular, magnetotactic prokaryote from a hypersaline environment

Phylogenetic analysis and phenotypic characterization were used to assign a multicellular magnetotactic prokaryote the name 'Candidatus Magnetoglobus multicellularis'. 'Candidatus Magnetoglobus multicellularis' lives in a large hypersaline coastal lagoon from Brazil and has properties that are unique among prokaryotes. It consists of a compact assembly or aggregate of flagellated bacterial cells, highly organized in a sphere, that swim in either helical or straight trajectories. The life cycle of 'Candidatus Magnetoglobus multicellularis' is completely multicellular, in which one aggregate grows by enlarging the size of its cells and approximately doubling the volume of the whole organism. Cells then divide synchronously, maintaining the spherical arrangement; finally the cells separate into two identical aggregates. Phylogenetic 16S rRNA gene sequence analysis showed that 'Candidatus Magnetoglobus multicellularis' is related to the dissimilatory sulfate-reducing bacteria within the Deltaproteobacteria and to other previously described, but not yet well characterized, multicellular magnetotactic prokaryotes.

Hydrocarbon Biomarkers in the Topla-Mezica Zinc-Lead Deposits, Northern Karavanke/Drau Range, Slovenia: Paleoenvironment at the Site of Ore Formation

The Mississippi Valley-type zinc and lead deposits at Topla (250,150 metric tons (t) of ore grading 10 wt % Zn and 3.3 wt % Pb) and Mežica (19 million metric tons (Mt) of ore grading 5.3 wt % Pb and 2.7 wt % Zn) occur within the Middle to Upper Triassic platform carbonate rocks of the northern Karavanke/Drau Range geotectonic units of the Eastern Alps, Slovenia. The ore and host rocks of these deposits have been investigated by a combination of inorganic and organic geochemical methods to determine major, trace, and rare earth element (REE) concentrations, hydrocarbon distribution, and stable isotope ratios of carbonates, kerogen, extractable organic matter, and individual hydrocarbons. These data combined with sedimentological evidence provide insight into the paleoenvironmental conditions at the site of ore formation. The carbonate isotope composition, the REE patterns, and the distribution of hydrocarbon biomarkers (normal alkanes and steranes) suggest a marine depositional environment. At Topla, a relatively high concentration of redox sensitive trace elements (V, Mo, U) in the host dolostones and REE patterns parallel to that of the North American shale composite suggest that sediments were deposited in a reducing environment. Anoxic conditions enhanced the preservation of organic matter and resulted in relatively higher total organic carbon contents (up to 0.4 wt %). The isotopic composition of the kerogen (δ 13 C kerogen = −29.4 to −25.0‰, δ 15 N kerogen = −13.6 to 6.8‰) suggests that marine algae and/or bacteria were the main source of organic carbon with a very minor contribution from detrital continental plants and a varying degree of alteration. Extractable organic matter from Topla ore is generally depleted in 13 C compared to the associated kerogen, which is consistent with an indigenous source of the bitumens. The mineralization correlates with δ 15 N kerogen values around 0 per mil, 13C depleted kerogen, 13C enriched n -heptadecane, and relatively high concentrations of bacterial hydrocarbon biomarkers, indicating a high cyanobacterial biomass at the site of ore formation. Abundant dissimilatory sulfate-reducing bacteria, feeding on the cyanobacterial remains, led to accumulation of biogenic H 2 S in the pore water of the sediments. This biogenic H 2 S was mainly incorporated into sedimentary organic matter and diagenetic pyrite. Higher bacterial activity at the ore site also is indicated by specific concentration ratios of hydrocarbons, which are roughly correlated with total Pb plus Zn contents. This correlation is consistent with mixing of hydrothermal metal-rich fluids and local bacteriogenic sulfide sulfur. The new geochemical data provide supporting evidence that Topla is a low-temperature Mississippi Valley-type deposit formed in an anoxic supratidal saline to hypersaline environment. A laminated cyanobacterial mat, with abundant sulfate-reducing bacteria was the main site of sulfate reduction.

Sulfide fluxes in a microbial mat from the Ebro Delta, Spain.

The sulfur cycle of Ebro Delta microbial mats was studied in order to determine sulfide production and sulfide consumption. Vertical distribution of two major functional groups involved in the sulfur cycle, anoxygenic phototrophic bacteria and dissimilatory sulfate-reducing bacteria (SRB), was also studied. The former reached up to 2.2x10(8) cfu cm(-3) sediment in the purple layer, and the latter reached about 1.8x10(5) SRB cm(-3) sediment in the black layer. From the changes in sulfide concentrations under light-dark cycles it can be inferred that the rate of H(2)S production was 6.2 micro mol H(2)S cm(-3) day(-1) at 2.6 mm, and 7.6 micro mol H(2)S cm(-3) day(-1) at 6 mm. Furthermore, sulfide consumption was also assessed, determining rates of 0.04, 0.13 and 0.005 mmol l(-1) of sulfide oxidized at depths of 2.6, 3 and 6 mm, respectively.

Desulfovibrio magneticus sp. nov., a novel sulfate-reducing bacterium that produces intracellular single-domain-sized magnetite particles.

A novel type of dissimilatory sulfate-reducing bacterium, designated strain RS-1T, capable of producing intracellular magnetite particles (magnetosomes) was isolated from freshwater sulfide-rich sediments. Phylogenetic analysis based on 16S rDNA sequences revealed that RS-1T is a member of the genus Desulfovibrio. Its closest known relative is Desulfovibrio burkinensis (sequence similarity of 98.7%). Strain RS-1T contains desulfoviridin, c-type cytochromes and, unlike other Desulfovibrio spp., it possesses menaquinone MK-7(H2) instead of MK-6 or MK-6(H2). Strain RS-1T is also unique compared with other members of Desulfovibrio in its ability to synthesize intracellular magnetite particles. A novel species, Desulfovibrio magneticus sp. nov., is proposed for RS-1T (= ATCC 700980T = DSM 13731T), a sulfate-reducing magnetotactic bacterium.

Volatile sulfur production by pig cecal bacteria in batch culture and screening inhibitors of sulfate reducing bacteria.

We studied the effects of specific inhibitors of methanogenesis (2-bromoethane sulfonate, BES) and sulfate reduction (sodium molybdate) on volatile sulfur production in batch cultures of pig cecal bacteria. The volatile sulfur concentration in headspace gas was determined by flame-photometric detector gas chromatography. BES stimulated production of hydrogen sulfide (H2S) and methanethiol, and sodium molybdate completely inhibited the production of these volatile sulfur compounds. The results indicated that dissimilate sulfate reduction is mainly responsible for volatile sulfur production in the hindgut. Therefore the extracts of herbs, food colors, and aroma chemicals were tested for their inhibitory effects on H2S production by a dissimilatory sulfate-reducing bacteria. Desulfovibrio desulfuricans DSM642. H2S was measured by the chromatography of the headspace gas, using a flame photometric detector. Of 306 herbal extracts tested, 69 extracts from 38 herbs inhibited H2S production at 1.0 mg/mL. Sisymbrium officinale (hedge mustard) was the most potent inhibitor. Six pigments inhibited H2S release. Erythrosine and rose bengal showed inhibitory effects at 0.01 mg/mL. Peppermint oil and 96 aroma chemicals were assayed for their effects on H2S release. Thirty-two aroma chemicals suppressed H2S production at 0.1 mg/mL, and camphene, 1-decanol, and 2-nonanone were effective at 0.01 mg/mL.

Identification of a new class of 5'-adenylylsulfate (APS) reductases from sulfate-assimilating bacteria.

A gene was cloned from Burkholderia cepacia DBO1 that is homologous with Escherichia coli cysH encoding 3'-phosphoadenylylsulfate (PAPS) reductase. The B. cepacia gene is the most recent addition to a growing list of cysH homologs from a diverse group of sulfate-assimilating bacteria whose products show greater homology to plant 5'-adenylylsulfate (APS) reductase than they do to E. coli CysH. The evidence reported here shows that the cysH from one of the species, Pseudomonas aeruginosa, encodes APS reductase. It is able to complement an E. coli cysH mutant and a cysC mutant, indicating that the enzyme is able to bypass PAPS, synthesized by the cysC product. Insertional knockout mutation of P. aeruginosa cysH produced cysteine auxotrophy, indicating its role in sulfate assimilation. Purified P. aeruginosa CysH expressed as a His-tagged recombinant protein is able to reduce APS, but not PAPS. The enzyme has a specific activity of 5.8 micromol. min(-1). mg of protein(-1) at pH 8.5 and 30 degrees C with thioredoxin supplied as an electron donor. APS reductase activity was detected in several bacterial species from which the novel type of cysH has been cloned, indicating that this enzyme may be widespread. Although an APS reductase from dissimilatory sulfate-reducing bacteria is known, it shows no structural or sequence homology with the assimilatory-type APS reductase reported here. The results suggest that the dissimilatory and assimilatory APS reductases evolved convergently.