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Nickel-iron composites are efficient in catalyzing oxygen evolution. Here, we develop a microorganism corrosion approach to construct nickel-iron hydroxides. The anaerobic sulfate-reducing bacteria, using sulfate as the electron acceptor, play a significant role in the formation of iron sulfide decorated nickel-iron hydroxides, which exhibit excellent electrocatalytic performance for oxygen evolution. Experimental and theoretical investigations suggest that the synergistic effect between oxyhydroxides and sulfide species accounts for the high activity. This microorganism corrosion strategy not only provides efficient candidate electrocatalysts but also bridges traditional corrosion engineering and emerging electrochemical energy technologies.

Around the world, several dozen deep sedimentary aquifers are being used for storage of natural gas. Ad hoc studies of the microbial ecology of some of them have suggested that sulfate reducing and methanogenic microorganisms play a key role in how these aquifers' communities function. Here, we investigate the influence of gas storage on these two metabolic groups by using high-throughput sequencing and show the importance of sulfate-reducing Desulfotomaculum and a new monophyletic methanogenic group. Aquifer microbial diversity was significantly related to the geological level. The distance to the stored natural gas affects the ratio of sulfate-reducing Firmicutes to deltaproteobacteria. In only one aquifer, the methanogenic archaea dominate the sulfate-reducers. This aquifer was used to store town gas (containing at least 50% H 2 ) around 50 years ago. The observed decrease of sulfates in this aquifer could be related to stimulation of subsurface sulfate-reducers. These results suggest that the composition of the microbial communities is impacted by decades old transient gas storage activity. The tremendous stability of these gas-impacted deep subsurface microbial ecosystems suggests that in situ biotic methanation projects in geological reservoirs may be sustainable over time., (© 2019 Society for Applied Microbiology and John Wiley & Sons Ltd.)

Recent studies have shown that interspecies electron transfer between chemoheterotrophic bacteria and methanogenic archaea can be mediated by electric currents flowing through conductive iron oxides, a process termed electric syntrophy. In this study, we conducted enrichment experiments with methanogenic microbial communities from rice paddy soil in the presence of ferrihydrite and/or sulfate to determine whether electric syntrophy could be enabled by biogenic iron sulfides. Although supplementation with either ferrihydrite or sulfate alone suppressed methanogenesis, supplementation with both ferrihydrite and sulfate enhanced methanogenesis. In the presence of sulfate, ferrihydrite was transformed into black precipitates consisting mainly of poorly crystalline iron sulfides. Microbial community analysis revealed that a methanogenic archaeon and iron- and sulfate-reducing bacteria (Methanosarcina, Geobacter, and Desulfotomaculum, respectively) predominated in the enrichment culture supplemented with both ferrihydrite and sulfate. Addition of an inhibitor specific for methanogenic archaea decreased the abundance of Geobacter, but not Desulfotomaculum, indicating that Geobacter acquired energy via syntrophic interaction with methanogenic archaea. Although electron acceptor compounds such as sulfate and iron oxides have been thought to suppress methanogenesis, this study revealed that coexistence of sulfate and iron oxide can promote methanogenesis by biomineralization of (semi)conductive iron sulfides that enable methanogenesis via electric syntrophy., (© 2018 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.)

Anaerobic sludge from a sewage treatment plant was used to acclimatize microbial colonies capable of anaerobic oxidation of methane (AOM) coupled to sulfate reduction. Clone libraries and fluorescence in situ hybridization were used to investigate the microbial population. Sulfate-reducing bacteria (SRB) (e.g., Desulfotomaculum arcticum and Desulfobulbus propionicus) and anaerobic methanotrophic archaea (ANME) (e.g., Methanosaeta sp. and Methanolinea sp.) coexisted in the enrichment. The archaeal and bacterial cells were randomly or evenly distributed throughout the consortia. Accompanied by sulfate reduction, methane was oxidized anaerobically by the consortia of methane-oxidizing archaea and SRB. Moreover, CH 4 and SO 4 2- were consumed by methanotrophs and sulfate reducers with CO 2 and H 2 S as products. The H 3 CSH produced by methanotrophy was an intermediate product during the process. The methanotrophic enrichment was inoculated in a down-flow biofilter for the treatment of methane and H 2 S from a landfill site. On average, 93.33% of H 2 S and 10.71% of methane was successfully reduced in the biofilter. This study tries to provide effective method for the synergistic treatment of waste gas containing sulfur compounds and CH 4 ., (Copyright © 2018. Published by Elsevier B.V.)

We established Fe(III)-reducing co-cultures of two species of metal-reducing bacteria, the Gram-positive Desulfotomaculum reducens MI-1 and the Gram-negative Geobacter sulfurreducens PCA. Co-cultures were given pyruvate, a substrate that D. reducens can ferment and use as electron donor for Fe(III) reduction. G. sulfurreducens relied upon products of pyruvate oxidation by D. reducens (acetate, hydrogen) for use as electron donor in the co-culture. Co-cultures reduced Fe(III) to Fe(II) robustly, and Fe(II) was consistently detected earlier in co-cultures than pure cultures. Notably, faster cell growth, and correspondingly faster pyruvate oxidation, was observed by D. reducens in co-cultures. Global comparative proteomic analysis was performed to observe differential protein abundance during co-culture vs. pure culture growth. Proteins previously associated with Fe(III) reduction in G. sulfurreducens, namely c-type cytochromes and type IV pili proteins, were significantly increased in abundance in co-cultures relative to pure cultures. D. reducens ribosomal proteins were significantly increased in co-cultures, likely a reflection of faster growth rates observed for D. reducens cells while in co-culture. Furthermore, we developed multiple reaction monitoring (MRM) assays to quantitate specific biomarker peptides. The assays were validated in pure and co-cultures, and protein abundance ratios from targeted MRM and global proteomic analysis correlate significantly., (© 2018 John Wiley & Sons Ltd.)

Mesotoga prima strain PhosAc3 is a mesophilic representative of the phylum Thermotogae comprising only fermentative bacteria so far. We show that while unable to ferment glucose, this bacterium is able to couple its oxidation to reduction of elemental sulfur. We demonstrate furthermore that M. prima strain PhosAc3 as well as M. prima strain MesG1 and Mesotoga infera are able to grow in syntrophic association with sulfate-reducing bacteria (SRB) acting as hydrogen scavengers through interspecies hydrogen transfer. Hydrogen production was higher in M. prima strain PhosAc3 cells co-cultured with SRB than in cells cultured alone in the presence of elemental sulfur. We propose that the efficient sugar-oxidizing metabolism by M. prima strain PhosAc3 in syntrophic association with a hydrogenotrophic sulfate-reducing bacterium can be extrapolated to all members of the Mesotoga genus. Genome comparison of Thermotogae members suggests that the metabolic difference between Mesotoga and Thermotoga species (sugar oxidation versus fermentation) is mainly due to the absence of the bifurcating [FeFe]-hydrogenase in the former. Such an obligate oxidative process for using sugars, unusual within prokaryotes, is the first reported within the Thermotogae. It is hypothesized to be of primary ecological importance for growth of Mesotoga spp. in the environments that they inhabit., (© 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.)

In many organisms, the UGA stop codon is recoded to insert selenocysteine (Sec) into proteins. Sec incorporation in bacteria is directed by an mRNA element, known as the Sec-insertion sequence (SECIS), located downstream of the Sec codon. Unlike other aminoacyl-tRNAs, Sec-tRNA Sec is delivered to the ribosome by a dedicated elongation factor, SelB. We recently identified a series of tRNA Sec -like tRNA genes distributed across Bacteria that also encode a canonical tRNA Sec . These tRNAs contain sequence elements generally recognized by cysteinyl-tRNA synthetase (CysRS). While some of these tRNAs contain a UCA Sec anticodon, most have a GCA Cys anticodon. tRNA Sec with GCA anticodons are known to recode UGA codons. Here we investigate the clostridial Desulfotomaculum nigrificans tRNA Sec -like tRNA Cys , and show that this tRNA is acylated by CysRS, recognized by SelB, and capable of UGA recoding with Cys in Escherichia coli. We named this non-canonical group of tRNA Cys as 'tRNA ReC ' (Recoding with Cys). We performed a comprehensive survey of tRNA ReC genes to establish their phylogenetic distribution, and found that, in a particular lineage of clostridial Pelotomaculum, the Cys identity elements of tRNA ReC had mutated. This novel tRNA, which contains a UCA anticodon, is capable of Sec incorporation in E. coli, albeit with lower efficiency relative to Pelotomaculum tRNA Sec . We renamed this unusual tRNA Sec derived from tRNA ReC as 'tRNA ReU ' (Recoding with Sec). Together, our results suggest that tRNA ReC and tRNA ReU may serve as safeguards in the production of selenoproteins and - to our knowledge - they provide the first example of programmed codon-anticodon mispairing in bacteria.

Although iron- and sulfate-reducing bacteria in subsurface environments have crucial roles in biogeochemical cycling of C, Fe, and S, how specific electron donors impact the compositional structure and activity of native iron- and/or sulfate-reducing communities is largely unknown. To understand this better, we created bicarbonate-buffered batch systems in duplicate with three different electron donors (acetate, lactate, or glucose) paired with ferrihydrite and sulfate as the electron acceptors and inoculated them with subsurface sediment as the microbial inoculum. Sulfate and ferrihydrite reduction occurred simultaneously and were faster with lactate than with acetate. 16S rRNA-based sequence analysis of the communities over time revealed that Desulfotomaculum was the major driver for sulfate reduction coupled with propionate oxidation in lactate-amended incubations. The reduction of sulfate resulted in sulfide production and subsequent abiotic reduction of ferrihydrite. In contrast, glucose promoted faster reduction of ferrihydrite, but without reduction of sulfate. Interestingly, the glucose-amended incubations led to two different biogeochemical trajectories among replicate bottles that resulted in distinct coloration (white and brown). The two outcomes in geochemical evolution might be due to the stochastic evolution of the microbial communities or subtle differences in the initial composition of the fermenting microbial community and its development via the use of different glucose fermentation pathways available within the community. Synchrotron-based x-ray analysis indicated that siderite and amorphous Fe(II) were formed in the replicate bottles with glucose, while ferrous sulfide and vivianite were formed with lactate or acetate. These data sets reveal that use of different C utilization pathways projects significant changes in microbial community composition over time that uniquely impact both the geochemistry and mineralogy of subsurface environments.

Purpose: Degradation processes of metallic biomaterials in the oral cavity limit the stability and reliability of dental materials. The influence of environment bacteria Desulfotomaculum nigrificans sulfate reducing bacteria on the corrosion processes of Co-Cr-Mo and Ti-6Al-4V alloys was assessed., Methods: After 28 and 56 days of contact of the materials with the bacterial environment, the surfaces of the biomaterials tested were observed by means of confocal scanning laser microscopy (CSLM), and their chemical composition was studied using X-Ray Photoelectron Spectrometry (XPS)., Results: Corrosive changes and the presence of sulfur (with medium atomic concentration of 0.5% for Co-Cr-Mo and 0.3% for Ti-6AL-4V) were observed on the surface of the biomaterials. Image analysis conducted using Aphelion software indicated that corrosion pits took up approx. 2.3% and 1.8% (after 28 days) and 4.2% and 3.1% (after 56 days) of the total test surfaces of cobalt and titanium alloys respectively. The greatest number of corrosion pits had a surface area within the range of 1-50 m2. They constituted from 37% up to 83% of all changes, depending on the type of material., Conclusions: An evident influence of the SRB on the surfaces of cobalt and titanium alloys was observed. Significant corrosive losses caused by the activity of microorganisms were observed on the metallic surfaces under study. The results of this study have much cognitive and utilitarian significance.

Petrochemical wastewater often contains high concentrations of phenol and sulfate that must be properly treated to meet discharge standards. This study acclimated anaerobic-activated sludge to treat saline phenolic wastewater with sulfate reduction and clarified the diversity and degradation mechanism of the microbial community. The active sludge in an upflow anaerobic sludge blanket (UASB) reactor could remove 90 % of phenol and maintain the effluent concentration of SO4 (2-) below 400 mg/L. Cloning and sequencing showed that Clostridium spp. and Desulfotomaculum spp. were major phenol-degrading bacteria. Phenol was probably degraded through the carboxylation pathway and sulfate reduction catalyzed by adenosine-5'-phosphosulfate (APS) reductase and dissimilatory sulfite reductase (DSR). A real-time polymerase chain reaction (RT-PCR) showed that as phenol concentration increased, the quantities of 16S rRNA gene, dsrB, and mcrA in the sludge all decreased. The relative abundance of dsrB dropped to 12.46 %, while that of mcrA increased to 56.18 %. The change in the electron flow ratio suggested that the chemical oxygen demand (COD) was removed mainly by sulfate-reducing bacteria under a phenol concentration of 420 mg/L, whereas it was removed mainly by methanogens above 630 mg/L.

Sulphate-reducing bacteria Desulfomicrobium sp. CrR3 and Desulfotomaculum. sp. are able to use fumarate as electron donor and acceptor. When they use fumarate as an electron acceptor succinate accumulates in the medium. If fumarate serves as electron donor, minor amounts of citrate, isocitrate and acetate are detected except succinate. In the case of simultaneous introduction of fumarate, SO4(2-) and Cr2O7(2-), the last inhibits usage of fumarate and SO4(2-).

Understanding of microbial metal reduction is based almost solely on studies of Gram-negative organisms. In this study, we focus on Desulfotomaculum reducens MI-1, a Gram-positive metal reducer whose genome lacks genes with similarity to any characterized metal reductase. Using non-denaturing separations and mass spectrometry identification, in combination with a colorimetric screen for chelated Fe(III)-NTA reduction with NADH as electron donor, we have identified proteins from the D. reducens proteome not previously characterized as iron reductases. Their function was confirmed by heterologous expression in Escherichia coli. Furthermore, we show that these proteins have the capability to reduce soluble Cr(VI) and U(VI) with NADH as electron donor. The proteins identified are NADH : flavin oxidoreductase (Dred_2421) and a protein complex composed of oxidoreductase flavin adenine dinucleotide/NAD(P)-binding subunit (Dred_1685) and dihydroorotate dehydrogenase 1B (Dred_1686). Dred_2421 was identified in the soluble proteome and is predicted to be a cytoplasmic protein. Dred_1685 and Dred_1686 were identified in both the soluble as well as the insoluble protein fraction, suggesting a type of membrane association, although PSORTb predicts both proteins are cytoplasmic. This study is the first functional proteomic analysis of D. reducens and one of the first analyses of metal and radionuclide reduction in an environmentally relevant Gram-positive bacterium., (© 2014 Society for Applied Microbiology and John Wiley & Sons Ltd.)

Biodiversity of sulfate-reducing bacterial communities in the water column of the Gdansk Deep, Baltic Sea, where H2S had been detected in near-bottom layers, was analyzed by PCR with primers for the 16S rRNA genes of six major phylogenetic subgroups of sulfate-reducing bacteria (SRB). Using denaturing gradient gel electrophoresis followed by sequencing, the nucleotide sequences of reamplified dsrB gene fragments from investigated water samples were determined. For the first time the presence of nucleotide sequences of the dsrB gene was detected by PCR in the water samples from all hydrochemical layers, including subsurface oxic waters. The presence of the 16S rRNA genes of representatives of Desulfotomaculum, Desulfococcus-Desulfonema-Desulfosarcina, and Desulfovibrio-Desulfomicrobium SRB subgroups was also revealed throughout the water column of the Gdansk Deep. Analysis of translated amino acid sequences encoded by the dsrB gene demonstrated the highest homology with the relevant sequences of uncultured SRB from various marine habitats.

Microbial community structure, and niche and neutral processes can all influence response to disturbance. Here, we provide experimental evidence for niche versus neutral and founding community effects during a bioremediation-related organic carbon disturbance. Subsurface sediment, partitioned into 22 flow-through columns, was stimulated in situ by the addition of acetate as a carbon and electron donor source. This drove the system into a new transient biogeochemical state characterized by iron reduction and enriched Desulfuromonadales, Comamonadaceae and Bacteroidetes lineages. After approximately 1 month conditions favoured sulfate reduction, and were accompanied by a substantial increase in the relative abundance of Desulfobulbus, Desulfosporosinus, Desulfitobacterium and Desulfotomaculum. Two subsets of four to five columns each were switched from acetate to lactate amendment during either iron (earlier) or sulfate (later) reduction. Hence, subsets had significantly different founding communities. All lactate treatments exhibited lower relative abundances of Desulfotomaculum and Bacteroidetes, enrichments of Clostridiales and Psychrosinus species, and a temporal succession from highly abundant Clostridium sensu stricto to Psychrosinus. Regardless of starting point, lactate-switch communities followed comparable structural trajectories, whereby convergence was evident 9 to 16 days after each switch, and significant after 29 to 34 days of lactate addition. Results imply that neither the founding community nor neutral processes influenced succession following perturbation., (© 2014 Society for Applied Microbiology and John Wiley & Sons Ltd.)

Desulfotomaculum reducens MI-1 is a Gram-positive, sulfate-reducing bacterium also capable of reducing several metals, among which is Fe(III). Very limited knowledge is available on the potential mechanism(s) of metal reduction among Gram-positive bacteria, despite their preponderance in the microbial communities that inhabit some inhospitable environments (e.g., thermal or hyperthermal ecosystems, extreme pH or salinity environments, heavy metal or radionuclide contaminated sediments). Here, we show that in the presence of pyruvate, this micro-organism is capable of reducing both soluble Fe(III)-citrate and solid-phase hydrous ferric oxide, although growth is sustained by pyruvate fermentation rather than Fe(III) respiration. Despite the fact that Fe(III) reduction does not support direct energy conservation, D. reducens uses it as a complementary means of discarding excess reducing equivalent after H2 accumulation in the culture headspace renders proton reduction unfavorable. Thus, Fe(III) reduction permits the oxidation of greater amounts of pyruvate than fermentation alone. Fe(III) reduction by D. reducens is mediated by a soluble electron carrier, most likely riboflavin. Additionally, an intracellular electron storage molecule acts as a capacitor and accumulates electrons during pyruvate oxidation for slow release to Fe(III). The reductase responsible for the transfer of electrons from the capacitor to the soluble carrier has not been identified, but data presented here argue against the involvement of c-type cytochromes., (© 2013 John Wiley & Sons Ltd.)

Terrestrial mud volcanoes (TMVs) represent geochemically diverse habitats with varying sulfur sources and yet sulfur cycling in these environments remains largely unexplored. Here we characterized the sulfur-metabolizing microorganisms and activity in four TMVs in Azerbaijan. A combination of geochemical analyses, biological rate measurements and molecular diversity surveys (targeting metabolic genes aprA and dsrA and SSU ribosomal RNA) supported the presence of active sulfur-oxidizing and sulfate-reducing guilds in all four TMVs across a range of physiochemical conditions, with diversity of these guilds being unique to each TMV. The TMVs varied in potential sulfate reduction rates (SRR) by up to four orders of magnitude with highest SRR observed in sediments where in situ sulfate concentrations were highest. Maximum temperatures at which SRR were measured was 60°C in two TMVs. Corresponding with these trends in SRR, members of the potentially thermophilic, spore-forming, Desulfotomaculum were detected in these TMVs by targeted 16S rRNA analysis. Additional sulfate-reducing bacterial lineages included members of the Desulfobacteraceae and Desulfobulbaceae detected by aprA and dsrA analyses and likely contributing to the mesophilic SRR measured. Phylotypes affiliated with sulfide-oxidizing Gamma- and Betaproteobacteria were abundant in aprA libraries from low sulfate TMVs, while the highest sulfate TMV harboured 16S rRNA phylotypes associated with sulfur-oxidizing Epsilonproteobacteria. Altogether, the biogeochemical and microbiological data indicate these unique terrestrial habitats support diverse active sulfur-cycling microorganisms reflecting the in situ geochemical environment., (© 2012 Society for Applied Microbiology and Blackwell Publishing Ltd.)

The effects of 50 Hz magnetic fields on sulphate- reducing bacteria viability were studied electrochemically. Two types of graphite electrodes (pyrolytic and glassy carbon) covered with whole bacterial cells behind a dialysis membrane were used for electrochemical measurements. We found about 15% decrease of reduction peak current density (which indicates desulphurization activity of the bacterial cells - their metabolic activity) on cyclic voltammograms after magnetic field exposure compared to the control samples. We suppose that the magnetic field does not influence the metabolic activity (desulphurization) of sulphate-reducing bacteria but most probably causes bacterial death.

A promising remediation approach to mitigate subsurface uranium contamination is the stimulation of indigenous bacteria to reduce mobile U(VI) to sparingly soluble U(IV). The product of microbial uranium reduction is often reported as the mineral uraninite. Here, we show that the end products of uranium reduction by several environmentally relevant bacteria (Gram-positive and Gram-negative) and their spores include a variety of U(IV) species other than uraninite. U(IV) products were prepared in chemically variable media and characterized using transmission electron microscopy (TEM) and X-ray absorption spectroscopy (XAS) to elucidate the factors favoring/inhibiting uraninite formation and to constrain molecular structure/composition of the non-uraninite reduction products. Molecular complexes of U(IV) were found to be bound to biomass, most likely through P-containing ligands. Minor U(IV)-orthophosphates such as ningyoite [CaU(PO(4))(2)], U(2)O(PO(4))(2), and U(2)(PO(4))(P(3)O(10)) were observed in addition to uraninite. Although factors controlling the predominance of these species are complex, the presence of various solutes was found to generally inhibit uraninite formation. These results suggest a new paradigm for U(IV) in the subsurface, i.e., that non-uraninite U(IV) products may be found more commonly than anticipated. These findings are relevant for bioremediation strategies and underscore the need for characterizing the stability of non-uraninite U(IV) species in natural settings.

Spore-forming, Gram-positive sulfate-reducing bacteria (SRB) represent a group of SRB that dominates the deep subsurface as well as niches in which resistance to oxygen and dessication is an advantage. Desulfotomaculum reducens strain MI-1 is one of the few cultured representatives of that group with a complete genome sequence available. The metabolic versatility of this organism is reflected in the presence of genes encoding for the oxidation of various electron donors, including three- and four-carbon fatty acids and alcohols. Synteny in genes involved in sulfate reduction across all four sequenced Gram-positive SRB suggests a distinct sulfate-reduction mechanism for this group of bacteria. Based on the genomic information obtained for sulfate reduction in D. reducens, the transfer of electrons to the sulfite and APS reductases is proposed to take place via the quinone pool and heterodisulfide reductases respectively. In addition, both H(2) -evolving and H(2) -consuming cytoplasmic hydrogenases were identified in the genome, pointing to potential cytoplasmic H(2) cycling in the bacterium. The mechanism of metal reduction remains unknown., (© 2010 Society for Applied Microbiology and Blackwell Publishing Ltd.)

Marine sediments harbour diverse populations of dormant thermophilic bacterial spores that become active in sediment incubation experiments at much higher than in situ temperature. This response was investigated in the presence of natural complex organic matter in sediments of two Arctic fjords, as well as with the addition of freeze-dried Spirulina or individual high-molecular-weight polysaccharides. During 50 degrees C incubation experiments, Arctic thermophiles catalysed extensive mineralization of the organic matter via extracellular enzymatic hydrolysis, fermentation and sulfate reduction. This high temperature-induced food chain mirrors sediment microbial processes occurring at cold in situ temperatures (near 0 degrees C), yet it is catalysed by a completely different set of microorganisms. Using sulfate reduction rates (SRR) as a proxy for organic matter mineralization showed that differences in organic matter reactivity determined the extent of the thermophilic response. Fjord sediments with higher in situ SRR also supported higher SRR at 50 degrees C. Amendment with Spirulina significantly increased volatile fatty acids production and SRR relative to unamended sediment in 50 degrees C incubations. Spirulina amendment also revealed temporally distinct sulfate reduction phases, consistent with 16S rRNA clone library detection of multiple thermophilic Desulfotomaculum spp. enriched at 50 degrees C. Incubations with four different fluorescently labelled polysaccharides at 4 degrees C and 50 degrees C showed that the thermophilic population in Arctic sediments produce a different suite of polymer-hydrolysing enzymes than those used in situ by the cold-adapted microbial community. Over time, dormant marine microorganisms like these are buried in marine sediments and might eventually encounter warmer conditions that favour their activation. Distinct enzymatic capacities for organic polymer degradation could allow specific heterotrophic populations like these to play a role in sustaining microbial metabolism in the deep, warm, marine biosphere.

A fast, sensitive and reliable quartz crystal microbalance (QCM) biosensor is described for the selective detection of the marine pathogenic sulphate-reducing bacterium (SRB), D. desulfotomaculum. Based on the amplification of the response of vancomycin-functionalized magnetic nanoparticles (Van-mNPs), under an external magnetic field, the bacteria-mNPs conjugates attach to the surface of an Au electrode. The QCM biosensor gave a distinct response to the vancomycin-sensitive, D. desulfotomaculum, but had no obvious response to the vancomycin-resistant bacterium, Vibrio anguillarum. The effects of the optimization conditions such as the incubation time and pH on the detection were also investigated, respectively. Optimised assays showed that the biosensor could obtain the best response with a 30 min incubation of the bacteria with the Van-mNPs. A linear relationship between the QCM response and the logarithm of the bacterial concentration was observed in the range of 1.8 x 10(4) to 1.8 x 10(7)cfu/ml. The sensor system has a potential for further applications and provides a facile and sample method for detection of pathogenic bacteria., ((c) 2009 Elsevier B.V. All rights reserved.)

Earlier research demonstrated the secretion of benzoate, which must be oxygenated to its 4-hydroxy derivative in order to be included in further sulfate uptake processes. The present study on Desulfotomaculum acetoxidans DSM 771 was designed to determine the activity and catalytic specificity of the enzyme (most probably peroxidase) catalyzing the hydroxylation of secreted benzoate. Peroxidase activity measured with ABTS (2,2'-azino-bis (3-ethylbenzathiazoline-6-sulfonic acid) during cultivation indicated the greatest activity on the third and thirteen days (3.4 and 2.3 nkat per ml sample respectively). The highest (0.7979) correlation coefficient was calculated between peroxidase activity and hydrogen peroxide levels. The cell walls from 3- and 13-day cultures were subjected to an isolation procedure, PIPES (piperazine-N,N'-bis (2-ethane-sulfonic acid) extract followed by preparative electrophoresis. The extracts of a approximately 30 kDa band on the gel were analyzed by Western blotting and the membrane was stained with TMB (3,3',5,5'-tetramethylbenzidine-specific for the presence of peroxidase). This same protein was incubated for 6 h with benzoate, H2O2, Na2SO4. The product formed a complex with Fe3+, whose maximum absorption spectra (501.7 nm) corresponded with a ferric complex of synthetic 4-hydroxy-3-sulfo-benzoate. The H2S level during the cultivation was higher in culture grown with 15.5 mM 4-hydroxy-3-sulfo-benzoate than in culture with lactate supplemented with 15.5 mM sulfate. The role of peroxidase in oxygen utilization and sulfate uptake is discussed.

The bioremediation of uranium-contaminated sites is designed to stimulate the activity of microorganisms able to catalyze the reduction of soluble U(VI) to the less soluble mineral UO(2). U(VI) reduction does not necessarily support growth in previously studied bacteria, but it typically involves viable vegetative cells and the presence of an appropriate electron donor. We characterized U(VI) reduction by the sulfate-reducing bacterium Desulfotomaculum reducens strain MI-1 grown fermentatively on pyruvate and observed that spores were capable of U(VI) reduction. Hydrogen gas - a product of pyruvate fermentation - rather than pyruvate, served as the electron donor. The presence of spent growth medium was required for the process, suggesting that an unknown factor produced by the cells was necessary for reduction. Ultrafiltration of the spent medium followed by U(VI) reduction assays revealed that the factor's molecular size was below 3 kDa. Pre-reduced spent medium displayed short-term U(VI) reduction activity, suggesting that the missing factor may be an electron shuttle, but neither anthraquinone-2,6-disulfonic acid nor riboflavin rescued spore activity in fresh medium. Spores of D. reducens also reduced Fe(III)-citrate under experimental conditions similar to those for U(VI) reduction. This is the first report of a bacterium able to reduce metals while in a sporulated state and underscores the novel nature of the mechanism of metal reduction by strain MI-1.

Values of Delta(34)S (=delta(34)S(HS)-delta(34)S(SO(4)), where delta(34)S(HS) and delta(34)S(SO(4)) indicate the differences in the isotopic compositions of the HS(-) and SO(4)(2-) in the eluent, respectively) for many modern marine sediments are in the range of -55 to -75 per thousand, much greater than the -2 to -46 per thousand epsilon(34)S (kinetic isotope enrichment) values commonly observed for microbial sulfate reduction in laboratory batch culture and chemostat experiments. It has been proposed that at extremely low sulfate reduction rates under hypersulfidic conditions with a nonlimited supply of sulfate, isotopic enrichment in laboratory culture experiments should increase to the levels recorded in nature. We examined the effect of extremely low sulfate reduction rates and electron donor limitation on S isotope fractionation by culturing a thermophilic, sulfate-reducing bacterium, Desulfotomaculum putei, in a biomass-recycling culture vessel, or "retentostat." The cell-specific rate of sulfate reduction and the specific growth rate decreased progressively from the exponential phase to the maintenance phase, yielding average maintenance coefficients of 10(-16) to 10(-18) mol of SO(4) cell(-1) h(-1) toward the end of the experiments. Overall S mass and isotopic balance were conserved during the experiment. The differences in the delta(34)S values of the sulfate and sulfide eluting from the retentostat were significantly larger, attaining a maximum Delta(34)S of -20.9 per thousand, than the -9.7 per thousand observed during the batch culture experiment, but differences did not attain the values observed in marine sediments.

The efficacy of the bark of Eucalyptus tereticornis (Smith) as an adsorbent for the removal of metal ions and sulphate from acid mine water was assessed. About 96% of Fe, 75% of Zn, 92% of Cu and 41% of sulphate removal was achieved from the acid mine water of pH 2.3 with a concomitant increase in pH value by about two units after interaction with the tree bark, under appropriate conditions. The adsorption isotherms adhered to Freundlich and Langmuir relationships and were exothermic in nature. The free energy of the adsorption process was found to be negative attesting to the feasibility of the reaction. The adsorption kinetics followed the first-order Lagergren rate equation. The filtrate obtained after treatment with E. tereticornis (Sm) bark was found to contain essential elements like potassium, magnesium, calcium, sodium and phosphate apart from carbon which served as a successful growth medium for the sulphate reducing bacteria (SRB) namely Desulfotomaculum nigrificans. Bacterial growth studies showed that about 57% and 72% of sulphate reduction could be achieved at initial pH values of 4.1 and 5.5 respectively of the acid mine water. Pretreatment of the acid mine water with tree bark followed by bioremoval using Dsm. nigrificans resulted in about 75% and 84% respectively of sulphate reduction at pH 4.1 and 5.5, cumulatively by biosorption and bioreduction. The mechanisms of metal ion removal using tree bark and sulphate reduction using Dsm. nigrificans are discussed.

A moderately thermophilic, Gram-positive, endospore-forming, sulfate-reducing bacterium was isolated from a fluidized-bed reactor treating acidic water containing metal and sulfate. The strain, designated RE35E1T, was rod-shaped and motile. The temperature range for growth was 33-51 degrees C (optimum 44-46 degrees C) and the pH range was 6.0-7.5 (optimum pH 6.4-7.3). The strain grew optimally without additional NaCl. The electron acceptors were 10 mM sulfate, thiosulfate and elemental sulfur and 1 mM (but not 10 mM) sulfite. Various alcohols and carboxylic acids were utilized as electron donors. Fermentative growth occurred on pyruvate. The cell wall contained meso-diaminopimelic acid, and the major respiratory isoprenoid quinone was menaquinone MK-7. The major whole-cell fatty acids were iso-C15 : 0, iso-C17 : 1 omega 10c and iso-C17 : 0. Strain RE35E1T was related to representatives of the genera Desulfotomaculum and Sporotomaculum, the closest relatives being Desulfotomaculum arcticum DSM 17038T (96.3 % 16S rRNA gene sequence similarity) and Sporotomaculum hydroxybenzoicum DSM 5475T (92.0 % similarity). Strain RE35E1T represents a novel species, for which the name Desulfotomaculum alcoholivorax sp. nov. is proposed. The type strain is RE35E1T (=DSM 16058T=JCM 14019T).

Desulfotomaculum carboxydivorans, recently isolated from a full-scale anaerobic wastewater treatment facility, is a sulfate reducer capable of hydrogenogenic growth on carbon monoxide (CO). In the presence of sulfate, the hydrogen formed is used for sulfate reduction. The organism grows rapidly at 200 kPa CO, pH 7.0, and 55 degrees C, with a generation time of 100 min, producing nearly equimolar amounts of H(2) and CO(2) from CO and H(2)O. The high specific CO conversion rates, exceeding 0.8 mol CO (g protein)(-1) h(-1), makes this bacterium an interesting candidate for a biological alternative of the currently employed chemical catalytic water-gas shift reaction to purify synthesis gas (contains mainly H(2), CO, and CO(2)). Furthermore, as D. carboxydivorans is capable of hydrogenotrophic sulfate reduction at partial CO pressures exceeding 100 kPa, it is also a good candidate for biodesulfurization processes using synthesis gas as electron donor at elevated temperatures, e.g., in biological flue gas desulfurization. Although high maximal specific sulfate reduction rates (32 mmol (g protein)(-1) h(-1)) can be obtained, its sulfide tolerance is rather low and pH dependent, i.e., maximally 9 and 5 mM sulfide at pH 7.2 and pH 6.5, respectively.

The performance of a fluidized-bed reactor (FBR) based sulfate reducing bioprocess was predicted using artificial neural network (ANN). The FBR was operated at high (65 degrees C) temperature and it was fed with iron (40-90 mg/L) and sulfate (1,000-1,500 mg/L) containing acidic (pH = 3.5-6) synthetic wastewater. Ethanol was supplemented as carbon and electron source for sulfate reducing bacteria (SRB). The wastewater pH of 4.3-4.4 was neutralized by the alkalinity produced in acetate oxidation and the average effluent pH was 7.8 +/- 0.8. The oxidation of acetate is the rate-limiting step in the sulfidogenic ethanol oxidation by thermophilic SRB, which resulted in acetate accumulation. Sulfate reduction and acetate oxidation rates showed variation depending on the operational conditions with the maximum rates of 1 g/L/d (0.2 g/g volatile solids (VS)/d) and 0.3 g/L/d (0.06 g/g VS/d), respectively. This study presents an ANN model predicting the performance of the reactor and determining the optimal architecture of this model; such as best back-propagation (BP) algorithm and neuron numbers. The Levenberg-Marquardt algorithm was selected as the best of 12 BP algorithms and optimal neuron number was determined as 20. The developed ANN model predicted acetate (R=0.91), sulfate (R=0.95), sulfide (R=0.97), and alkalinity (R=0.94) in the FBR effluent. Hence, the ANN based model can be used to predict the FBR performance, to control the operational conditions for improved process performance., ((c) 2006 Wiley Periodicals, Inc.)

Ulcerative colitis(UC) is colon localized disease. Broad epithelial cell damage, crypt abscesses and accumulation of neutrophils are recognized for UC. Although the cause of UC is indistinct at this time, there is a growing consensus that abnormal intestinal microflora would be related with UC. There have been several evidences that excessive production of hydrogen sulfide by bacteria in colon would be associated with UC. Sulfate reducing bacteria are able to utilize sulfate as an electron receptor for dissimilation of organic substrate and hydrogen gas, resulting in generating toxic hydrogen sulfide. This review is dealt with the association between sulfate reducing bacteria and UC in aetiology and bacterial pathogenesis.

A sulfate-reducing bacterium, strain WW1, was isolated from a thermophilic bioreactor operated at 65 degrees C with methanol as sole energy source in the presence of sulfate. Growth of strain WW1 on methanol or acetate was inhibited at a sulfide concentration of 200 mg l(-1), while on H2/CO2, no apparent inhibition occurred up to a concentration of 500 mg l(-1). When strain WW1 was co-cultured under the same conditions with the methanol-utilizing, non-sulfate-reducing bacteria, Thermotoga lettingae and Moorella mulderi, both originating from the same bioreactor, growth and sulfide formation were observed up to 430 mg l(-1). These results indicated that in the co-cultures, a major part of the electron flow was directed from methanol via H2/CO2 to the reduction of sulfate to sulfide. Besides methanol, acetate, and hydrogen, strain WW1 was also able to use formate, malate, fumarate, propionate, succinate, butyrate, ethanol, propanol, butanol, isobutanol, with concomitant reduction of sulfate to sulfide. In the absence of sulfate, strain WW1 grew only on pyruvate and lactate. On the basis of 16S rRNA analysis, strain WW1 was most closely related to Desulfotomaculum thermocisternum and Desulfotomaculum australicum. However, physiological properties of strain WW1 differed in some aspects from those of the two related bacteria.

Three independent 28 or 32-day stationary cultures of Desulfotomaculum acetoxidans DSM 771 strain were carried out under anoxic conditions in acetate or lactate-containing media. The acids were the sole carbon and energy sources in these media. During cultivation the turbidity (for calculation of cell division index) and hydrogen sulfide contents were determined in culture broth and reduced glutathione and protein concentrations were assayed in culture broth supernatant. In these three successive cultures, the bacterium initially grew much faster on lactate than on acetate. However, after two weeks of culture this difference disappeared and in fact the growth rate was higher on acetate than on lactate. The level of H2S formed (product of the dissimilatory pathway of sulfate reduction) demonstrated that this pathway was more effective when lactate was a carbon source and the average H2S concentration was from over 3-fold to about 9-fold greater in lactate than in acetate cultures. Also GSH (glutathione, product of the assimilatory sulfate reduction pathway) average level was about 2-fold higher in lactate-grown cultures. The high negative values of the correlation coefficients between GSH and O2 levels, especially during the first 4 days of cultivation, indicate that GSH is a very important antioxidizing extracellular agent of D. acetoxidans. The rapid increase in GSH level, preceding the release of H2S, indicates the metabolic priority of the assimilation pathway of sulfate reduction. For both carbon sources the highest coefficient of correlation was found between protein and H2S levels. These results suggest that hydrogen sulfide is bound by proteins (which contain cysteinyl residues) secreted by D. acetoxidans cells. Indicated way of H2S bounding could result in its accumulation. This coefficient of correlation increased gradually in the successive cultures. The ratio of H2S concentration to protein concentration increased gradually in the successive cultures, too.

Desulfotomaculum nigrificans, a typical sulfate reducing bacterium (SRB) was successfully grown in the presence of 12-210 mg/L of zinc. Complete bioremoval of zinc was achieved in 2 days for 12 mg/L while the bioremoval efficiency was about 70% in 40 days in the presence of 210 mg/L initial concentration of zinc, attesting to the inhibition of bacterial cell growth at higher zinc concentrations. The bioremoval mechanism was predominantly governed by bioprecipitation with biosorption contributing to a minor extent. The amount of protein present in the extracellular secretions obtained by growth of SRB in modified Baars' medium devoid of iron was the highest followed by those obtained in the presence of zinc or iron, in that order. Bioremediation studies carried out using a specially designed set-up, facilitating the transfer of biogenically produced hydrogen sulfide gas to a separate precipitation assembly, confirmed that zinc could be successfully precipitated from its corresponding sulfate solution, varying in concentration from 10 to 20,000 mg/L. Detailed characterization of the various zinc sulfide precipitates by EDAX and X-ray diffraction analysis conformed to wurtzite structure. The isoelectric points of high purity zinc sulfide and that of chemically synthesized, biogenically produced and zinc sulfide precipitated using bacterially produced hydrogen sulfide gas (BPH-ZnS) were located at pH 3, 7.8, 2.8 and 8, respectively.

The utility of rice husk as an adsorbent for metal ions such as iron, zinc and copper from acid mine water was assessed. The adsorption isotherms exhibited Langmuirian behavior and were endothermic in nature. The free energy values for adsorption of the chosen metal ions onto rice husk were found to be highly negative attesting to favorable interaction. Over 99% Fe(3+), 98% of Fe(2+) and Zn(2+) and 95% Cu(2+) uptake was achieved from acid mine water, with a concomitant increase in the pH value by two units using rice husk. The remediation studies carried out on acid mine water and simulated acid mine water pretreated with rice husk indicated successful growth of Desulfotomaculum nigrificans (D. nigrificans). The amount of sulphate bioreduction in acid mine water at an initial pH of 5.3 was enhanced by D. nigrificans from 21% to 40% in the presence of rice husk filtrate supplemented with carbon and nitrogen. In simulated acid mine water with fortified husk filtrate, the sulphate reduction was even more extensive, with an enhancement to 73%. Concurrently, almost 90% Fe(2+), 89% Zn(2+) and 75% Cu(2+) bioremoval was attained from simulated acid mine water. Metal adsorption by rice husk was confirmed in desorption experiments in which almost complete removal of metal ions from the rice husk was achieved after two elutions using 1M HCl. The possible mechanisms of metal ion adsorption onto rice husk and sulphate reduction using D. nigrificans are discussed.

The filtrate obtained by interacting a known amount of rice husk with deionised, Milli-Q water was assessed as a carbon source and nutrient medium for the growth of Desulfotomaculum nigrificans, a typical sulfate-reducing bacterium. The filtrate contained essential growth constituents such as magnesium, potassium, phosphorous apart from calcium, sodium, chloride and sulfate ions. Based on the 1H and 13C NMR characterization studies, the organic composition of the components dissolved from the rice husk, was found to be: (i) 66% lignocellulosic material, (ii) 24% xylose+arabinose and (iii) 10% galactose. The growth studies indicated a 15-fold increase in the bacterial cell number in about 20 days. Nearly 81% and 66% reduction in sulfate concentration could be achieved in about 28 days, from the solutions containing initial sulfate concentrations of 550 mg/l and 1200 mg/l respectively. In both the cases studied, the iron concentration could be reduced by over 85%.

Sulfate-reducing bacteria (e.g. Desulfotomaculum acetoxidans) exist in animal intestine. These bacteria are able to bind heavy metals (e.g. cadmium or lead). Comparative investigations on the composition of cellular walls of Desulfotomaculum acetoxidans--depending on the initial Fe2+ supplement in the medium (7.5, 57.5 and 507.5 M) were performed. Iron(II) was cumulated as FeS or as pyrite (FeS2). However, if the initial amount of iron was higher, its majority (46% 85%) was transported onto the membrane. It was determined that the siderophore found in Desulfotomaculum acetoxidans was deferroxamine as in animals.

A thermophilic terephthalate-degrading methanogenic consortium was successfully enriched for 272 days in an anaerobic hybrid reactor, and the microbial structure was characterized using terminal RFLPs, clone libraries and fluorescence in-situ hybridization with rRNA-targeted oligonucleotide probes. All the results suggested that Methanothrix thermophila-related methanogens, Desulfotomaculum-related bacterial populations in the Gram-positive low-G + C group, and OP5-related populations were the key members responsible for terephthalate degradation under thermophilic methanogenic conditions except during periods when the reactor experienced heat shock and pump failure. These perturbations caused a significant shift in bacterial population structure in sludge samples taken from the sludge bed but not from the surface of the packing materials. After system recovery, many other bacterial populations emerged, which belonged mainly to the Gram-positive low-G + C group and Cytophaga-Flexibacter-Bacteroides, as well as beta-Proteobacteria, Planctomycetes and Nitrospira. These newly emerged populations were probably also capable of degrading terephthalate in the hybrid system, but were out-competed by those bacterial populations before perturbations.

Deep subsurface sandstones in the area of Berlin (Germany) located 600 to 1060 m below the surface were examined for the presence of viable microorganisms. The in situ temperatures at the sampling sites ranged from 37 to 45 degrees C. Investigations focussed on sulfate-reducing bacteria able to grow on methanol and triethylene glycol, which are added as chemicals to facilitate the long-term underground storage of natural gas. Seven strains were isolated from porewater brines in the porous sandstone. Three of them were obtained with methanol (strains H1M, H3M, and B1M), three strains with triethylene glycol (strains H1T, B1T, and B2T) and one strain with a mixture of lactate, acetate and butyrate (strain H1-13). Due to phenotypic properties six isolates could be identified as members of the genus Desulfovibrio, and strain B2T as a Desulfotomaculum. The salt tolerance and temperature range for growth indicated that the isolates originated from the indigenous deep subsurface sandstones. They grew in mineral media reflecting the in situ ionic composition of the different brines, which contained 1.5 to 190 g NaCl x l(-1) and high calcium and magnesium concentrations. The Desulfovibrio strains grew at temperatures between 20 and 50 degrees C, while the Desulfotomaculum strain was thermophilic and grew between 30 and 65 degrees C. The strains utilized a broad spectrum of electron donors and acceptors. They grew with carbon compounds like lactate, pyruvate, formate, n-alcohols (C1-C5), glycerol, ethylene glycol, malate, succinate, and fumarate. Some strains even utilized glucose as electron donor and carbon source. All strains were able to use sulfate, sulfite and nitrate as electron acceptors. Additionally, three Desulfovibrio strains reduced manganese oxide, the Desulfotomaculum strain reduced manganese oxide, iron oxide, and elemental sulfur. The 16S rRNA analysis revealed that the isolates belong to three different species. The strains H1T, H3M and B1M could be identified as Desulfovibrio indonesiensis, and strain B2T as Desulfotomaculum geothermicum. The other Desulfovibrio strains (H1M, H1-13, and B1T) showed identical 16S rDNA sequences and similarities as low as 93% to their closest relative, Desulfovibrio aminophilusT. Therefore, these isolates were assigned to a new species, Desulfovibrio cavernae sp. nov., with strain H1M as the type strain.

A strictly anaerobic bacterium, strain OX39, was isolated with o-xylene as organic substrate and sulfate as electron acceptor from an aquifer at a former gasworks plant contaminated with aromatic hydrocarbons. Apart from o-xylene, strain OX39 grew on m-xylene and toluene and all three substrates were oxidized completely to CO(2). Induction experiments indicated that o-xylene, m-xylene, and toluene degradation were initiated by different specific enzymes. Methylbenzylsuccinate was identified in supernatants of cultures grown on o-xylene and m-xylene, and benzylsuccinate was detected in supernatants of toluene-grown cells, thus indicating that degradation was initiated in all three cases by fumarate addition to the methyl group. Strain OX39 was sensitive towards sulfide and depended on Fe(II) in the medium as a scavenger of the produced sulfide. Analysis of the PCR-amplified 16S rRNA gene revealed that strain OX39 affiliates with the gram-positive endospore-forming sulfate reducers of the genus Desulfotomaculum and is the first hydrocarbon-oxidizing bacterium in this genus.

(5Z)-4-bromo-5-(bromomethylene)-3-butyl-2(5H)-furanone (furanone) from the red marine alga Delisea pulchra was found previously to inhibit the growth, swarming and biofilm formation of Gram-positive bacteria (Ren et al., 2002, Lett Appl Microbiol 34: 293-299). In the present study, the Gram-positive sulphate-reducing bacterium (SRB), Desulfotomaculum orientis, was used to study the inhibition of mild steel corrosion due to the addition of furanone. The weight loss from batch coupon experiments incubated with 40 microg x ml(-1) furanone was reduced fivefold compared with samples that lacked furanone. Analysis of the metal surface with environmental scanning electron microscopy further confirmed the protection afforded by the addition of furanone. In agreement with the corrosion inhibition, most probable number (MPN) analysis showed that 20 and 40 microg x ml(-1) furanone inhibited 58% and 96% of the D. orientis growth respectively. Hence, furanone has the potential to inhibit microbial-induced corrosion related to Gram-positive bacteria.

The microbial populations responsible for anaerobic degradation of phthalate isomers were investigated by enrichment and isolation of those microbes from anaerobic sludge treating wastewater from the manufacturing of terephthalic acid. Primary enrichments were made with each of three phthalate isomers (ortho-, iso-, and terephthalate) as the sole energy source at 37 degrees C with two sources of anaerobic sludge (both had been used to treat wastewater containing high concentrations of phthalate isomers) as the inoculum. Six methanogenic enrichment cultures were obtained which not only degraded the isomer used for the enrichment but also had the potential to degrade part of other phthalate isomers as well as benzoate with concomitant production of methane, presumably involving strictly syntrophic substrate degradation. Our 16S rRNA gene-cloning analysis combined with fluorescence in situ hybridization revealed that the predominant bacteria in the enrichment cultures were affiliated with a recently recognized non-sulfate-reducing subcluster (subcluster Ih) in the group 'Desulfotomaculum lineage I' or a clone cluster (group TA) in the class delta-PROTEOBACTERIA: Several attempts were made to isolate these microbes, resulting in the isolation of a terephthalate-degrading bacterium, designated strain JT, in pure culture. A coculture of the strain with the hydrogenotrophic methanogen Methanospirillum hungatei converted terephthalate to acetate and methane within 3 months of incubation, whereas strain JT could not degrade terephthalate in pure culture. During the degradation of terephthalate, a small amount of benzoate was transiently accumulated as an intermediate, indicative of decarboxylation of terephthalate to benzoate as the initial step of the degradation. 16S rRNA gene sequence analysis revealed that the strain was a member of subcluster Ih of the group 'Desulfotomaculum lineage I', but it was only distantly related to other known species.

Four strains of thermophilic, endospore-forming, sulfate-reducing bacteria were enriched and isolated from hot solfataric fields in the Krafla area of north-east Iceland, using methanol and sulfite as substrates. Morphologically, these strains resembled thermophilic Desulfotomaculum species. The strains grew with alcohols, including methanol, with glucose and fructose as electron donors, and with sulfate, sulfite or thiosulfate as electron acceptors. For all four strains, the optimum temperature and pH for growth were 60 degrees C and pH 7.3, respectively; no added NaCl was required. Phylogenetic analysis based on partial 16S rRNA gene sequence comparisons showed high levels of similarity of the novel strains (>92 %) with Desulfotomaculum kuznetsovii and Desulfotomaculum luciae. However, DNA-DNA hybridization studies with D. kuznetsovii revealed that the four strains belonged to one novel species. A representative of this group of isolates, strain V21(T), is proposed as the type strain of a novel species of the spore-forming, sulfate-reducing genus Desulfotomaculum, namely Desulfotomaculum solfataricum (type strain V21(T)=DSM 14956(T)=CIP 107984(T)).