Biocorrosion of 316LV steel used in oral cavity due to Desulfotomaculum nigrificans bacteria.

Corrosion processes of metallic biomaterials in the oral cavity pose a significant limitation to the life and reliable functioning of dental materials. In this article, the influence of environment bacteria Desulfotomaculum nigrificans sulfate reducing bacteria on the corrosion processes of 316LV steel was assessed. After 14 and 28 days of contact of the material with the bacterial environment, the surfaces of the tested biomaterial were observed by means of confocal scanning laser microscopy, and their chemical composition was studied using X-Ray Photoelectron Spectrometry and a scanning transmission electron microscopy. Corrosive changes, the presence of sulfur (with atomic concentration of 0.5%) on the surface of the biomaterial and the presence of a thin oxide layer (thickness of ∼20 nm) under the surface of the steel were observed. This corrosion layer with significant size reduction of grains was characterized by an increased amount of oxygen (18% mas., p < 0.001) in comparison to untreated 316LV steel (where oxygen concentration - 10% mas.). Image analysis conducted using APHELION software indicated that corrosion pits took up ∼2.8% of the total tested surface. The greatest number of corrosion pits had a surface area within the range of 100-200 µm2 . © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 222-229, 2017.

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).

Dispersal of thermophilic Desulfotomaculum endospores into Baltic Sea sediments over thousands of years.

Patterns of microbial biogeography result from a combination of dispersal, speciation and extinction, yet individual contributions exerted by each of these mechanisms are difficult to isolate and distinguish. The influx of endospores of thermophilic microorganisms to cold marine sediments offers a natural model for investigating passive dispersal in the ocean. We investigated the activity, diversity and abundance of thermophilic endospore-forming sulfate-reducing bacteria (SRB) in Aarhus Bay by incubating pasteurized sediment between 28 and 85 °C, and by subsequent molecular diversity analyses of 16S rRNA and of the dissimilatory (bi)sulfite reductase (dsrAB) genes within the endospore-forming SRB genus Desulfotomaculum. The thermophilic Desulfotomaculum community in Aarhus Bay sediments consisted of at least 23 species-level 16S rRNA sequence phylotypes. In two cases, pairs of identical 16S rRNA and dsrAB sequences in Arctic surface sediment 3000 km away showed that the same phylotypes are present in both locations. Radiotracer-enhanced most probable number analysis revealed that the abundance of endospores of thermophilic SRB in Aarhus Bay sediment was ca. 10(4) per cm(3) at the surface and decreased exponentially to 10(0) per cm(3) at 6.5 m depth, corresponding to 4500 years of sediment age. Thus, a half-life of ca. 300 years was estimated for the thermophilic SRB endospores deposited in Aarhus Bay sediments. These endospores were similarly detected in the overlying water column, indicative of passive dispersal in water masses preceding sedimentation. The sources of these thermophiles remain enigmatic, but at least one source may be common to both Aarhus Bay and Arctic sediments.

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 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).

[Description of Desulfotomaculum nigrificans subsp. salinus as a new species, Desulfotomaculum salinum sp. nov].

This study focused on the physiological, chemotaxonomic, and genotypic characteristics of two thermophilic spore-forming sulfate-reducing bacterial strains, 435T and 781, of which the former has previously been assigned to the subspecies Desulfotomaculum nigrificans subsp. salinus. Both strains reduced sulfate with the resulting production of H2S on media supplemented with H2 + CO2, formate, lactate, pyruvate, malate, fumarate, succinate, methanol, ethanol, propanol, butanol, butyrate, valerate, or palmitate. Lactate oxidation resulted in acetate accumulation; butyrate was oxidized completely, with acetate as an intermediate product. Growth on acetate was slow and weak. Sulfate, sulfite, thiosulfate, and elemental sulfur, but not nitrate, served as electron acceptors for growth with lactate. The bacteria performed dismutation of thiosulfate to sulfate and hydrogen sulfide. In the absence of sulfate, pyruvate but not lactate was fermented. Cytochromes of b and c types were present. The temperature and pH optima for both strains were 60-65 degrees C and pH 7.0. Bacteria grew at 0 to 4.5-6.0% NaCl in the medium, with the optimum being at 0.5-1.0%. Phylogenetic analysis based on a comparison of incomplete 16S rRNA sequences revealed that both strains belonged to the C cluster of the genus Desulfotomaculum, exhibiting 95.5-98.3% homology with the previously described species. The level of DNA-DNA hybridization of strains 435T and 781 with each other was 97%, while that with closely related species D. kuznetsovii 17T was 51-52%. Based on the phenotypic and genotypic properties of strains 435T and 781, it is suggested that they be assigned to a new species: Desulfotomaculum salinum sp. nov., comb. nov. (type strain 435T = VKM B 1492T).

FISH shows that Desulfotomaculum spp. are the dominating sulfate-reducing bacteria in a pristine aquifer.

The hydrochemistry and the microbial diversity of a pristine aquifer system near Garzweiler, Germany, were characterized. Hydrogeochemical and isotopic data indicate a recent activity of sulfate-reducing bacteria in the Tertiary marine sands. The community structure in the aquifer was studied by fluorescence in situ hybridization (FISH). Up to 7.3 x 10(5) cells/mL were detected by DAPI-staining. Bacteria (identified by the probe EUB338) were dominant, representing 51.9% of the total cell number (DAPI). Another 25.7% of total cell were affiliated with the domain Archaea as identified by the probe ARCH915. Within the domain Bacteria, the beta-Proteobacteria were most abundant (21.0% of total cell counts). Using genus-specific probes for sulfate-reducing bacteria (SRB), 2.5% of the total cells were identified as members of the genus Desulfotomaculum. This reflects the predominant role these microorganisms have been found to play in sulfate-reducing zones of aquifers at other sites. Previously, all SRB cultured from this site were from the spore-forming genera Desulfotomaculum and Desulfosporosinus.