Review of Desulfotomaculum species and proposal of the genera Desulfallas gen. nov., Desulfofundulus gen. nov., Desulfofarcimen gen. nov. and Desulfohalotomaculum gen. nov.

The genus Desulfotomaculumis a heterogeneous group of spore-forming sulfate-reducing bacteria. The type species of the genus is Desulfotomaculum nigrificans (Approved Lists 1980) emend. Visser et al. 2014. The results of phylogenetic analysis demonstrated that the genus Desulfotomaculum already has lost the clustering monophyly and was segregated into some distinct groups with low sequence similarity. Major features of the type strains in these groups were compared, and four novel genera, Desulfallas gen. nov., Desulfofundulus gen. nov., Desulfofarcimen gen. nov. and Desulfohalotomaculum gen. nov. were proposed to accommodate species transferred from the genus Desulfotomaculum.

Desulfocucumis palustris gen. nov., sp. nov., a mesophilic sulfate reducer belonging to Desulfotomaculum subcluster Ig.

A mesophilic, endospore-forming, sulfate-reducing bacterium, designated strain NAW-5T, was isolated from marsh soil. Cells of strain NAW-5T were Gram-stain-negative, curved rods that were motile. Strain NAW-5T grew at 18-48 °C (optimum 32-37 °C) and pH 5.8-8.4 (optimum pH 6.2-7.3). Electron donors utilized were various organic acids and H2 which support autotrophic growth. Fermentative growth occurred on carboxylic acids, but not on sugar. Sulfate, thiosulfate and elemental sulfur were used as electron acceptors. The respiratory isoprenoid quinone was MK-7. The genomic DNA G+C content of this strain was 46.6 mol%. Sequence analysis of the 16S rRNA gene showed that strain NAW-5T was affiliated to the family 'Desulfotomaculaceae' but the strain shared very low sequence similarity with any representatives of this family (≥89 %). Strain NAW-5T belongs to Desulfotomaculum subcluster Ig which does not include any species with validly published names. On the basis of significant differences in the phylogenetic and phenotypic properties between strain NAW-5T and related species, strain NAW-5T represents a novel species of a new genus for which the name Desulfocucumis palustris gen. nov., sp. nov. is proposed. The type strain of the type species is NAW-5T (=DSM 102911T=NBRC 112242T).

Desulfotomaculum aquiferis sp. nov. and Desulfotomaculum profundi sp. nov., isolated from a deep natural gas storage aquifer.

Two novel strictly anaerobic bacteria, strains Bs105T and Bs107T, were isolated from a deep aquifer-derived hydrocarbonoclastic community. The cells were rod-shaped, not motile and had terminal spores. Phylogenetic affiliation and physiological properties revealed that these isolates belong to two novel species of the genus Desulfotomaculum. Optimal growth temperatures for strains Bs105T and Bs107T were 42 and 45 °C, respectively. The estimated G+C content of the genomic DNA was 42.9 and 48.7 mol%. For both strains, the major cellular fatty acid was palmitate (C16 : 0). Specific carbon fatty acid signatures of Gram-positive bacteria (iso-C17 : 0) and sulfate-reducing bacteria (C17 : 0cyc) were also detected. An insertion was revealed in one of the two 16S rRNA gene copies harboured by strain Bs107T. Similar insertions have previously been highlighted among moderately thermophilic species of the genus Desulfotomaculum. Both strains shared the ability to oxidize aromatic acids (Bs105T: hydroquinone, acetophenone, para-toluic acid, 2-phenylethanol, trans-cinnamic acid, 4-hydroxybenzaldehyde, benzyl alcohol, benzoic acid 4-hydroxybutyl ester; Bs107T: ortho-toluic acid, benzoic acid 4-hydroxybutyl ester). The names Desulfotomaculum aquiferis sp. nov. and Desulfotomaculum profundi sp. nov. are proposed for the type strains Bs105T (=DSM 24088T=JCM 31386T) and Bs107T (=DSM 24093T=JCM 31387T).

Desulfotomaculum ferrireducens sp. nov., a moderately thermophilic sulfate-reducing and dissimilatory Fe(III)-reducing bacterium isolated from compost.

A novel dissimilatory Fe(III)-reducing bacterium, designated strain GSS09T, was isolated from a compost sample by using a solid medium containing acetate and ferrihydrite as electron donor and electron acceptor, respectively. Cells of strain GSS09T were anaerobic, Gram-stain-positive, motile, endospore-forming and rod-shaped. Growth occurred at 30-55 °C (optimum 50 °C), at pH 6.5-9.0 (optimum pH 7.5) and in the presence of 0-3 % (w/v) NaCl (optimum 1 %). Both sulfur compounds such as sulfate, sulfite and thiosulfate and Fe(III) oxides such as ferrihydrite could be utilized as electron acceptors. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain GSS09T was related closely to Desulfotomaculum hydrothermale Lam5T (94.5 % sequence similarity). The major fatty acids were C16 : 0 and C16 : 1ω7c/C16 : 1ω6c. The G+C content of the genomic DNA was 49.1 mol%. On the basis of phylogenetic analysis, phenotypic characterization and physiological tests, strain GSS09T is considered to represent a novel species of the genus Desulfotomaculum, for which the name Desulfotomaculum ferrireducens sp. nov. is proposed. The type strain is GSS09T (=KCTC 15523T=MCCC 1K01254T).

[Sulfate-Reducing Bacterial Communities in the Water Column of the Gdansk Deep (Baltic Sea)].

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.

Survival of Desulfotomaculum spores from estuarine sediments after serial autoclaving and high-temperature exposure.

Bacterial spores are widespread in marine sediments, including those of thermophilic, sulphate-reducing bacteria, which have a high minimum growth temperature making it unlikely that they grow in situ. These Desulfotomaculum spp. are thought to be from hot environments and are distributed by ocean currents. Their cells and spores upper temperature limit for survival is unknown, as is whether they can survive repeated high-temperature exposure that might occur in hydrothermal systems. This was investigated by incubating estuarine sediments significantly above (40-80 °C) maximum in situ temperatures (∼ 23 °C), and with and without prior triple autoclaving. Sulphate reduction occurred at 40-60 °C and at 60 °C was unaffected by autoclaving. Desulfotomaculum sp. C1A60 was isolated and was most closely related to the thermophilic D. kuznetsovii(T) (∼ 96% 16S rRNA gene sequence identity). Cultures of Desulfotomaculum sp. C1A60, D. kuznetsovii(T)and D. geothermicum B2T survived triple autoclaving while other related Desulfotomaculum spp. did not, although they did survive pasteurisation. Desulfotomaculum sp. C1A60 and D. kuznetsovii cultures also survived more extreme autoclaving (C1A60, 130 °C for 15 min; D. kuznetsovii, 135 °C for 15 min, maximum of 154 °C reached) and high-temperature conditions in an oil bath (C1A60, 130° for 30 min, D. kuznetsovii 140 °C for 15 min). Desulfotomaculum sp. C1A60 with either spores or predominantly vegetative cells demonstrated that surviving triple autoclaving was due to spores. Spores also had very high culturability compared with vegetative cells (∼ 30 × higher). Combined extreme temperature survival and high culturability of some thermophilic Desulfotomaculum spp. make them very effective colonisers of hot environments, which is consistent with their presence in subsurface geothermal waters and petroleum reservoirs.

Desulfotomaculum tongense sp. nov., a moderately thermophilic sulfate-reducing bacterium isolated from a hydrothermal vent sediment collected from the Tofua Arc in the Tonga Trench.

A novel, strictly anaerobic, moderately thermophilic, endospore-forming, sulfate-reducing bacterium, designated TGB60-1T, was isolated from a hydrothermal sediment vent collected from the Tofua Arc in the Tonga Trench. The strain was characterized phenotypically and phylogenetically. The isolated strain was observed to be Gram-positive, with slightly curved rod-shaped cells and a polar flagellum. Strain TGB60-1T was found to grow anaerobically at 37­60 °C (optimum, 50 °C), at pH 6.0­8.5 (optimum, pH 7.0) and with 1.0­4.0 % (w/v) NaCl (optimum, 3.0 %). The electron acceptors utilised were determined to be sulfate, sulfite, and thiosulfate. Strain TGB60-1T was found to utilise pyruvate and H2 as electron donors. Strain TGB60-1T was determined to be related to representatives of the genus Desulfotomaculum and the closest relatives within this genus were identified as Desulfotomaculum halophilum SEBR 3139T, Desulfotomaculum alkaliphilum S1T and Desulfotomaculum peckii LINDBHT1T (92.7, 92.1, and 91.8 % 16S rRNA gene sequence similarity, respectively). The major fatty acids (>20 %) were identified as C16:0 and C18:1 ω7c. The G+C content of the genomic DNA of this novel bacterium was determined to be 53.9 mol%. Based on this polyphasic taxonomic study, strain TGB60-1T is considered to represent a novel species in the genus Desulfotomaculum, for which the name Desulfotomaculum tongense sp. nov. is proposed. The type strain of D. tongense is strain TGB60-1T (= KTCT 4534T = JCM 18733T).

Desulfotomaculum intricatum sp. nov., a sulfate reducer isolated from freshwater lake sediment.

A novel spore-forming, sulfate-reducing bacterium, strain SR45(T), was isolated from sediment of a freshwater lake, Lake Mizugaki, in Japan. Cells of strain SR45 were rod-shaped (1.0-1.5×2.0-5.0 µm) and weakly motile; Gram staining and the KOH lysis test were negative. For growth, the optimum pH was 6.4-6.8 and the optimum temperature was 42-45 °C. Strain SR45(T) used sulfate, thiosulfate, sulfite and elemental sulfur as electron acceptors but not Fe(III). The G+C content of the genomic DNA was 41.1 mol%. Phylogenetic analyses based on genes for the 16S rRNA and DNA gyrase (gyrB) revealed that the isolated strain belonged to the family Peptococcaceae in the class Clostridia. The closest relative is Desulfotomaculum acetoxidans 5575(T), with 16S rRNA gene sequence similarity of 92-94 %. It is suggested that the strain is the second isolated member of Desulfotomaculum subcluster Ie. The isolate had multiple 16S rRNA gene copies, with 13 different sequences. On the basis of phylogenetic and phenotypic characterization, the name Desulfotomaculum intricatum sp. nov. is proposed, with the type strain SR45(T) ( = NBRC 109411(T) = DSM 26801(T)).

Desulfotomaculum peckii sp. nov., a moderately thermophilic member of the genus Desulfotomaculum, isolated from an upflow anaerobic filter treating abattoir wastewaters.

A novel anaerobic thermophilic sulfate-reducing bacterium designated strain LINDBHT1(T) was isolated from an anaerobic digester treating abattoir wastewaters in Tunisia. Strain LINDBHT1(T) grew at temperatures between 50 and 65 °C (optimum 55-60 °C), and at pH between 5.9 and 9.2 (optimum pH 6.0-6.8). Strain LINDBHT1(T) required salt for growth (1-40 g NaCl l(-1)), with an optimum of 20-30 g l(-1). In the presence of sulfate as terminal electron acceptor, strain LINDBHT1(T) used H2/CO2, propanol, butanol and ethanol as carbon and energy sources but fumarate, formate, lactate and pyruvate were not utilized. Butanol was converted to butyrate, while propanol and ethanol were oxidized to propionate and acetate, respectively. Sulfate, sulfite and thiosulfate were utilized as terminal electron acceptors but elemental sulfur, iron (III), fumarate, nitrate and nitrite were not used. The G+C content of the genomic DNA was 44.4 mol%. Phylogenetic analysis of the small-subunit rRNA gene sequence indicated that strain LINDBHT1(T) was affiliated to the genus Desulfotomaculum with the type strains of Desulfotomaculum halophilum and Desulfotomaculum alkaliphilum as its closest phylogenetic relatives (about 89% similarity). This strain represents a novel species of the genus Desulfotomaculum, Desulfotomaculum peckii sp. nov.; the type strain is LINDBHT1(T) (=DSM 23769(T)=JCM 17209(T)).

Desulfotomaculum defluvii sp. nov., a sulfate-reducing bacterium isolated from the subsurface environment of a landfill.

A novel sulfate-reducing, strictly anaerobic and endospore-forming bacterium, designated strain A5LFS102(T), was isolated from a subsurface landfill sample. The strain was characterized using a polyphasic approach. Optimal growth was observed at 37 °C and pH 7.5 with sulfate as an electron acceptor. Sulfite and thiosulfate were utilized as electron acceptors. The respiratory isoprenoid quinone was menaquinone MK-7. 16S rRNA gene sequence analysis assigned strain A5LFS102(T) to the genus Desulfotomaculum. Both 16S rRNA and dissimilatory sulfate reductase (dsr) genes were compared with those of representative members of the genus Desulfotomaculum. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain A5LFS102(T) was closely related to Desulfotomaculum aeronauticum DSM 10349(T) (94.6% sequence similarity). The G+C content of the DNA was 45.4 mol%. The total cellular fatty acid profile was dominated by C16 fatty acids. These phenotypic and genotypic data showed that strain A5LFS102(T) should be recognized as representative of a novel species of the genus Desulfotomaculum, for which the name Desulfotomaculum defluvii sp. nov. is proposed. The type strain is A5LFS102(T) (=DSM 23699(T)=JCM 14036(T)=MTCC 7767(T)).

Active sulfur cycling by diverse mesophilic and thermophilic microorganisms in terrestrial mud volcanoes of Azerbaijan.

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.

The genome of the Gram-positive metal- and sulfate-reducing bacterium Desulfotomaculum reducens strain MI-1.

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.

Metal reduction by spores of Desulfotomaculum reducens.

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.

Desulfotomaculum hydrothermale sp. nov., a thermophilic sulfate-reducing bacterium isolated from a terrestrial Tunisian hot spring.

A novel strictly anaerobic, moderately thermophilic, sulfate-reducing bacterium, designated strain Lam5(T), was isolated from a hot spring in north-east Tunisia and was characterized phenotypically and phylogenetically. The isolate stained Gram-negative but had a Gram-positive-type cell wall. The strain comprised endospore-forming, slightly curved rod-shaped cells with peritrichous flagella. It did not possess desulfoviridin. Strain Lam5(T) grew anaerobically at 40-60 degrees C (optimally at 55 degrees C) and at pH 5.8-8.2 (optimally at pH 7.1); it did not require NaCl but tolerated concentrations up to 1.5 % (w/v). It utilized lactate, pyruvate, formate, ethanol, butanol, glycerol, propanol and H(2) (plus acetate) as electron donors. Lactate was oxidized and pyruvate was fermented to acetate. Sulfate, sulfite, thiosulfate, As(V) and Fe(III) (but not elemental sulfur, fumarate, nitrate or nitrite) were used as electron acceptors. The G+C content of the genomic DNA was 46.8 mol%. Phylogenetic analysis based on 16S rRNA gene sequencing showed that strain Lam5(T) was a member of the genus Desulfotomaculum, with Desulfotomaculum putei as its closest relative (96 % similarity to the type strain). On the basis of genotypic, phenotypic and phylogenetic data, strain Lam5(T) represents a novel species of the genus Desulfotomaculum, for which the name Desulfotomaculum hydrothermale sp. nov. is proposed. The type strain is Lam5(T) (=DSM 18033(T) =JCM 13992(T)).

Desulfotomaculum alcoholivorax sp. nov., a moderately thermophilic, spore-forming, sulfate-reducer isolated from a fluidized-bed reactor treating acidic metal- and sulfate-containing wastewater.

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).

Diversity of sulfate-reducing bacteria inhabiting the rhizosphere of Phragmites australis in Lake Velencei (Hungary) revealed by a combined cultivation-based and molecular approach.

The community structure of sulfate-reducing bacteria (SRB) associated with reed (Phragmites australis) rhizosphere in Lake Velencei (Hungary) was investigated by using cultivation-based and molecular methods. The cultivation methods were restricted to recover lactate-utilizing species with the exclusion of Desulfobacter and some Desulfobacterium species presumably not being dominant members of the examined community. The most-probable-number (MPN) estimations of lactate-utilizing SRB showed that the cell counts in reed rhizosphere were at least one order of magnitude higher than that in the bulk sediment. The number of endospores was low compared to the total SRB counts. From the highest positive dilution of MPN series, 47 strains were isolated and grouped by restriction fragment length polymorphism (RFLP) analysis of the amplified 16S ribosomal RNA (rRNA) and dsrAB (dissimilatory sulfite reductase) genes. Contrary to the physiological diversity of the isolates, the combined results of RFLP analysis revealed higher diversity at species as well as at subspecies level. Based on the partial 16S rRNA sequences, the representative strains were closely affiliated with the genera Desulfovibrio and Desulfotomaculum. The partial dsrAB sequences of the clones, recovered after isolation and PCR amplification of the community DNA, were related to hitherto uncultured species of the genera Desulfovibrio and Desulfobulbus. Nevertheless, the representative of the second largest clone group was shown to be closely affiliated with the sequenced dsrAB gene of a strain isolated from the same environment and identified as Desulfovibrio alcoholivorans. Another clone sequence was closely related to a possible novel species also isolated within the scope of this work.

Methanol utilizing Desulfotomaculum species utilizes hydrogen in a methanol-fed sulfate-reducing bioreactor.

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.

Desulfotomaculum thermosubterraneum sp. nov., a thermophilic sulfate-reducer isolated from an underground mine located in a geothermally active area.

A thermophilic, Gram-positive, endospore-forming, sulfate-reducing bacterium was isolated from an underground mine in a geothermally active area in Japan. Cells of this strain, designated RL50JIIIT, were rod-shaped and motile. The temperature range for growth was 50-72 degrees C (optimum growth at 61-66 degrees C) and the pH range was 6.4-7.8 (optimum at pH 7.2-7.4). Strain RL50JIIIT tolerated up to 1.5% NaCl, but optimum growth occurred in the presence of 0-1% NaCl. Electron acceptors utilized were sulfate, sulfite, thiosulfate and elemental sulfur. Electron donors utilized were H2 in the presence of CO2, alanine, various carboxylic acids and alcohols. Fermentative growth occurred on lactate and pyruvate. The cell wall contained mesodiaminopimelic acid and the major respiratory isoprenoid quinone was menaquinone 7 (MK-7). Major whole-cell fatty acids were iso-C15:0, iso-C17:0 DMA (dimethyl acetal), iso-C15:0 DMA and iso-C17:0. Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed 98.7% similarity with Desulfotomaculum solfataricum DSM 14956T. However, DNA-DNA hybridization experiments with Desulfotomaculum kuznetsovii, Desulfotomaculum luciae and D. solfataricum and the G+C content of the DNA (54.4 mol%) allowed the differentiation of strain RL50JIIIT from the recognized species of the genus Desulfotomaculum. Strain RL50JIIIT therefore represents a novel species, for which the name Desulfotomaculum thermosubterraneum sp. nov. is proposed. The type strain is RL50JIIIT (=DSM 16057T=JCM 13837T).

Novel thermophilic sulfate-reducing bacteria from a geothermally active underground mine in Japan.

Thermophilic sulfate-reducing bacteria were enriched from samples obtained from a geothermal underground mine in Japan. The enrichment cultures contained bacteria affiliated with the genera Desulfotomaculum, Thermanaeromonas, Thermincola, Thermovenabulum, Moorella, "Natronoanaerobium," and Clostridium. Two novel thermophilic sulfate-reducing strains, RL50JIII and RL80JIV, affiliated with the genera Desulfotomaculum and Thermanaeromonas, respectively, were isolated.

Desulfotomaculum arcticum sp. nov., a novel spore-forming, moderately thermophilic, sulfate-reducing bacterium isolated from a permanently cold fjord sediment of Svalbard.

Strain 15T is a novel spore-forming, sulfate-reducing bacterium isolated from a permanently cold fjord sediment of Svalbard. Sulfate could be replaced by sulfite or thiosulfate. Hydrogen, formate, lactate, propionate, butyrate, hexanoate, methanol, ethanol, propanol, butanol, pyruvate, malate, succinate, fumarate, proline, alanine and glycine were used as electron donors in the presence of sulfate. Growth occurred with pyruvate as sole substrate. Optimal growth was observed at pH 7.1-7.5 and concentrations of 1-1.5 % NaCl and 0.4 % MgCl2. Strain 15T grew between 26 and 46.5 degrees C and optimal growth occurred at 44 degrees C. Therefore, strain 15T apparently cannot grow at in situ temperatures of Arctic sediments from where it was isolated, and it was proposed that it was present in the sediment in the form of spores. The DNA G+C content was 48.9 mol%. Strain 15T was most closely related to Desulfotomaculum thermosapovorans MLF(T) (93.5 % 16S rRNA gene sequence similarity). Strain 15T represents a novel species, for which the name Desulfotomaculum arcticum sp. nov. is proposed. The type strain is strain 15T (=DSM 17038T = JCM 12923T).