Strains Of Sulfate-reducing Bacteria Research Articles

Sulfate-reducing Bacteria in Rice Field Soil and on Rice Roots

Rice plants that were grown in flooded rice soil microcosms were examined for their ability to exhibit sulfate reducing activity. Washed excised rice roots showed sulfate reduction potential when incubated in anaerobic medium indicating the presence of sulfate-reducing bacteria. Rice plants, that were incubated in a double-chamber (phylloshpere and rhizosphere separated), showed potential sulfate reduction rates in the anoxic rhizosphere compartment. These rates decreased when oxygen was allowed to penetrate through the aerenchyma system of the plants into the anoxic root compartment, indicating that sulfate reducers on the roots were partially inhibited by oxygen or that sulfate was regenerated by oxidation of reduced S-compounds. The potential activity of sulfate reducers on rice roots was consistent with MPN enumerations showing that H2-utilizing sulfate-reducing bacteria were present in high numbers on the rhizoplane (4.1 x 10(7) g-1 root fresh weight) and in the adjacent rhizosperic soil (2.5 x 10(7) g-1 soil dry weight). Acetate-oxidizing sulfate reducers, on the other hand, showed highest numbers in the unplanted bulk soil (1.9 x 10(6) g-1 soil dry weight). Two sulfate reducing bacteria were isolated from the highest dilutions of the MPN series and were characterized physiologically and phylogenetically. Strain F1-7b which was isolated from the rhizoplane with H2 as electron donor was related to subgroup II of the family Desulfovibrionaceae. Strain EZ-2C2, isolated from the rhizoplane on acetate, grouped together with Desulforhabdus sp. and Syntrophobacter wolinii. Other strains of sulfate-reducing bacteria originated from bulk soil of rice soil microcosms and were isolated using different electron donors. From these isolates, strains R-AcA1, R-IbutA1, R-PimA1 and R-AcetonA170 were Gram-positive bacteria which were affiliated with the genus Desulfotomaculum. The other isolates were members of subgroup II of the Desulfovibrionaceae (R-SucA1 and R-LacA1), were related to Desulforhabdus sp. (strain BKA11), Desulfobulbus (R-PropA1), or culstered between Desulfobotulus sapovorans and Desulfosarcina variabilis (R-ButA1 and R-CaprA1).

Desulfovibrio aminophilus sp. nov., a Novel Amino Acid Degrading and Sulfate Reducing Bacterium from an Anaerobic Dairy Wastewater Lagoon

A mesophilic strain of sulfate-reducing bacterium, designated ALA-3T (T = type strain), was isolated from an anaerobic lagoon of a dairy wastewater treatment plant. The curved, Gram-negative, non-sporeforming cells (0.2 x 3.0-4.0 microns) existed singly or in chains, and were motile by single polar flagella. Optimum growth occurred at 35 degrees C and pH 7.5 on a medium containing lactate and sulfate. Thiosulfate or sulfite but not elemental sulfur, nitrate, or fumarate could also replace sulfate as an electron acceptor. Formate, alanine, aspartate, leucine, isoleucine, valine, and methionine, H2/CO2 and ethanol also served as electron donors with sulfate as an electron acceptor. Pyruvate, casamino acids, peptone, serine, glycine, cysteine and threonine were fermented. Sulfite and thiosulfate were disproportionated to sulfate and sulfide. The G + C content of the DNA was 66 mol % G + C. Phylogenetic analysis revealed that Desulfovibrio africanus was the nearest relative (similarity of 89%). Strain ALA-3T is physiologically and phylogenetically different from other Desulfovibrio species, and is designated Desulfovibrio aminophilus sp. nov. (DSM 12254).

Tetrahydrofolate serves as a methyl acceptor in the demethylation of dimethylsulfoniopropionate in cell extracts of sulfate-reducing bacteria.

Tetrahydrofolate was shown to function as a methyl acceptor in the anaerobic demethylation of dimethylsulfoniopropionate to methylthiopropionate in cell extracts of the sulfate-reducing bacterium strain WN. Dimethylsulfoniopropionate-dependent activities were 0.56 micromol methyltetrahydrofolate min-1 (mg protein)-1 and were higher than required to explain the growth rate of strain WN on dimethylsulfoniopropionate. The reaction did not require ATP or reductive activation by titanium(III)-nitrilotriacetic acid. Preincubation of the extract under air significantly decreased the activity (35% loss in 3 h). Three other dimethylsulfoniopropionate-demethylating sulfate reducers, Desulfobacterium niacini, Desulfobacterium vacuolatum, and Desulfobacterium strain PM4, had dimethylsulfoniopropionate:tetrahydrofolate methyltransferase activities of 0.16, 0.05, and 0.24 micromol min-1 (mg protein)-1, respectively. No methyltransferase activity to tetrahydrofolate was found with betaine as a substrate, not even in extracts of betaine-grown cells of these sulfate reducers. Dimethylsulfoniopropionate demethylation in cell extracts of strain WN was completely inhibited by 0.5 mM propyl iodide; in the light, the inhibition was far less strong, indicating involvement of a corrinoid-dependent methyltransferase.

Bioremediation of acid mine water using facultatively methylotrophic metal-tolerant sulfate-reducing bacteria

A microbial process is proposed for the decontamination of acid uranium mine water high in sulfates and metals. Sulfate reducers are suitable for such a process. Anaerobic reduction of sulfate results in the formation of H 2S which leads to an increase in pH and the precipitation of the metals. As cheap an readily available carbon and energy source methanol was chosen. Six strains of sulfate-reducing bacteria were isolated from a waste water pond, one from a copper mine. All strains were found to utilize methanol as electron donor and carbon source. Other electron donors included lactate, pyruvate and acetate. The strains were not uniform in their morphology, cells were of different sizes and rod-shaped or curved. All strains stained gram-negative. Desulfovibrio salexigens DSM 2538, D. africanus DSM 2603 and D. carbinolicus DSM 3852 were used as reference strains. One of the isolates named UFZ B 378 was found to grow at pH 4.0, D. salexigens at pH 4.5, all other strains did not grow below pH 6.0. Culture media with varying sulfate and metal concentrations at different pH values were used to simulate the composition of the mine waters.

Phylogenetic positions of Desulfofustis glycolicus gen. nov., sp. nov., and Syntrophobotulus glycolicus gen. nov., sp. nov., two new strict anaerobes growing with glycolic acid.

The glycolate-oxidizing, sulfate-reducing bacterium strain PerGlyS and the syntrophically glycolate-oxidizing bacterium strain FlGlyR were studied with respect to their phylogenetic relationships on the basis of in vitro amplification and direct sequencing of 16S rRNA-encoding DNA. Strain PerGlyS clustered with representatives of the delta subclass of the class Proteobacteria, close to "Desulforhopalus vacuolatus" but sufficiently distinct to preclude its assignment to this genus. These organisms, together with Desulfobulbus propionicus, represent a phylogenetic subgroup among members of the delta subclass of Proteobacteria. Strain FlGlyR was found to cluster with the gram-positive bacteria with low-G + C DNA, and Desulfitobacterium dehalogenans and Desulfotomaculum orientis are its closest relatives. Other species of the genus Desulfotomaculum are phylogenetically only moderately closely related to these organisms. These results necessitate the establishment of new genera and species for these two strains. Strain PerGlyS was designated the type strain of Desulfofustis glycolicus gen. nov., sp. nov., and strain FlGlyR was designated the type strain of Syntrophobotulus glycolicus gen. nov., sp. nov.

Characterization of sulfate-reducing bacteria isolated from oil-field waters.

The occurrence and metabolic capacities of sulfate-reducing bacteria (SRB) were studied in 23 water samples taken from producing wells at 14 different sites. Oil fields in France, the North Sea, and the Gulf of Guinea were selected and classified according to physicochemical parameters (salinity ranging from 0.3 to 120 g.L-1 and temperature between 29 and 85 degrees C). After the distribution of SRB within oil fields was studied, several strains of SRB were isolated and characterized metabolically. Twenty of the thirty-seven strains were not related to any known species. Most of the identified strains were members of the genera Desulfovibrio and Desulfotomaculum by molecular, morphological, and physiological properties.

Dimethylsulfoxide reduction by marine sulfate-reducing bacteria

Dimethylsulfoxide (DMSO) reduction occurred in five out of nine strains of sulfate-reducing bacteria from marine or saline environments, but not in three freshwater isolates. DMSO reduction supported growth in all positive strains. In Desulfovibrio desulfuricans strain PA2805, DMSO reduction occurred simultaneously with sulfate reduction and was not effectively inhibited by molybdate, a specific inhibitor of sulfate reduction. The growth yield per mol lactate was 26% higher with DMSO than with sulfate as electron acceptor. In extracts of cells of strain PA2805 grown on sulfate, a low level of DMSO-reducing activity was present (0.013 µmol (mg protein)− min−); higher levels were found in cells grown on DMSO (0.56 µmol (mg protein)− min−). In anoxic marine environments DMSO reduction by sulfate-reducing bacteria may lead to enhanced dimethylsulfide emission rates.

Dimethylsulfoxide reduction by marine sulfate-reducing bacteria

Dimethylsulfoxide (DMSO) reduction occurred in five out of nine strains of sulfate-reducing bacteria from marine or saline environments, but not in three freshwater isolates. DMSO reduction supported growth in all positive strains. In Desulfovibrio desulfuricans strain PA2805, DMSO reduction occurred simultaneously with sulfate reduction and was not effectively inhibited by molybdate, a specific inhibitor of sulfate reduction. The growth yield per mol lactate was 26% higher with DMSO than with sulfate as electron acceptor. In extracts of cells of strain PA2805 grown on sulfate, a low level of DMSO-reducing activity was present (0.013 μmol (mg protein) − min −); higher levels were found in cells grown on DMSO (0.56 μmol (mg protein) − min −). In anoxic marine environments DMSO reduction by sulfate-reducing bacteria may lead to enhanced dimethylsulfide emission rates.

Production of sulfur from gypsum as an industrial byproduct

Biological sulfate reduction was investigated at the bench and pilot scales in order to determine optimum culture conditions. Efficient strains of sulfate-reducing bacteria (SRB) were selected by classical microbiological methods and by mutagenesis. Improvement factors, including stripping, scale-up, sulfate, and organic substrate concentrations, have been studied in batch bioreactors.Two types of pilot-scale bioreactors have been adopted, the first being completely mixed with free cells and the second having two stages with immobilized cells on a fixed bed. An overall bioconversion capacity of 11 kg/m3·d of gypsum and 1.2 kg/m3·d of dissolved organic carbon has been achieved in the two-stage bioreactor.

Physiological properties of a sulfate-reducing bacterium isolated from municipal sewage sludge and its possible role as a syntrophic acidogen in the ecosystem.

Three strains of sulfate-reducing bacteria utilizing H2 as the electron donor of sulfate reduction were isolated from anaerobic digester slurry of municipal sewage sludge. The isolates were Gram-negative, non-spore- forming, motile, curved rods with a single polar flagellum. Cytochrome c3 and desulfoviridin were present. The DNA base compositions (mol% Three strains of sulfate-reducing bacteria utilizing H2 as the electron donor of sulfate reduction were isolated from anaerobic digester slurry of municipal sewage sludge. The isolates were Gram-negative, non-spore-forming, motile, curved rods with a single polar flagellum. Cytochrome c3 and desulfoviridin were present. The DNA base compositions (mol%G+C) were 59.6±0.7). The isolates used H2, formate, lactate, pyruvate, fumarate, malate, alcohols and amino acids as the electron donor for sulfate reduction. Alcohols were oxidized to corresponding monocarboxylates, and other organic compounds were to acetate. Sulfate, Three strains of sulfate-reducing bacteria utilizing H2 as the electron donor of sulfate reduction were isolated from anaerobic digester slurry ofmunicipal sewage sludge. The isolates were Gram-negative, non-spore-forming, motile, curved rods with a single polar flagellum. Cytochrome c3 and desulfoviridin were present. The DNA base compositions (mol% G+C) were 59.6±0.7. The isolates used H2, formate, lactate, pyruvate, fumarate, malate, alcohols and amino acids as the electron donor for sulfate reduction. Alcohols were oxidized to corresponding monocarboxylates, and other organic compounds were to acetate. Sulfate, sulfite and thiosulfate, but not nitrate, were used as the electron acceptor and reduced to sulfide. The isolates grew with pyruvate, fumarate, or malate as the energy source in the absence of oxidized sulfur compounds. yruvate was oxidized to acetate with the production of formate and H2. Fumarate and malate were oxidized to acetate, and the oxidation was coupled with the reduction of fumarate or malate to succinate. From the morphological and physiological properties, the isolates were considered to belong to a Desulfovibrio sp. In association with Methanobacterium formicicum as a hydrogen-consuming partner, one of the isolates could also grow in the absence of oxidized sulfur compounds by utilizing organic substrates other than pyruvate, malate and fumarate. In the coculture of the isolate and the methanogen, all the organic compounds other than malate and fumarate, which could serve as the electron donor for sulfate reduction in the pure culture of isolates, were syntrophically oxidized with the production of methane. These results indicate that the isolates of sulfate-reducing bacteria can play a role as a syntrophic degrader of a wide range of organic compounds in the methanogenic sewage sludge devoid of sulfate.

The effect of industrial biocides on sulphate-reducing bacteria under high pressure

This study was undertaken to determine the influence of temperature (20, 37, and 50°C) and pressure (1, 100 and 200 atm) on a strain of sulphate-reducing bacteria (SRB), isolated from an oil reservoir in Alaska. The effect of different concentrations (100, 200 and 500 ppm) of biocides isothiazolone (ITZ) and formaldehyde (FA) on planktonic population of SRB was tested in order to determine the efficacy of biocides under these conditions. The highest bacterial growth rate was 0.26±0.03 h−1 at 37°C under pressure of 100 atm. Statistical evaluation showed that although both temperature and pressure had exerted an effect on bacteria by significantly increasing their growth rate; temperature rather than pressure had greater influence on bacterial proliferation. The effectiveness of both FA and ITZ in controlling planktonic populations of SRB was comparable except at 37°C/200 atm, under which conditions FA proved to be more potent. The effectiveness of both biocides decreased with an increase in cell number, as observed at 37°C/100 atm.

Formation of thionates by freshwater and marine strains of sulfate-reducing bacteria

The formation of thionates (thiosulfate, trithionate and tetrahionate) during the reduction of sulfate or sulfite was studied with four marine and four freshwater strains of sulfate-reducing bacteria. Growing cultures of two strains of the freshwater species Desulfovibrio desulfuricans formed up to 400 μM thiosulfate and 100 μM trithionate under conditions of electron donor limitation. Tetrathionate was observed in lower concentrations of up to 30 μM. Uncoupler-treated washed cells of the four freshwater strains formed thiosulfate and trithionate at low electron donor concentrations with sulfite in excess. In contrast, only one of four marine strains formed thionates. The freshwater strain Desulfobulbus propionicus transformed sulfite almost completely to thiosulfate and trithionate. The amounts produced increased with time, concentration of added sulfite and cell density. Tetrathionate was detected only occasionally and in low concentrations, and was probably formed by chemical oxidation of thiosulfate. The results confirm the diversity of the sulfite reduction pathways in sulfate-reducing bacteria, and suggest that thiosulfate and trithionate are normal by-products of sulfate reduction.

Anaerobic degradation of aniline and dihydroxybenzenes by newly isolated sulfate-reducing bacteria and description of Desulfobacterium anilini.

A new, rod-shaped, Gram-negative, non-sporing sulfate reducer (strain Ani1) was enriched and isolated from marine sediment with aniline as sole electron donor and carbon source. The strain degraded aniline completely to CO2 and NH3 with stoichiometric reduction of sulfate to sulfide. Strain Ani1 also degraded aminobenzoates and further aromatic and aliphatic compounds. The strain grew in sulfide-reduced mineral medium supplemented only with vitamin B12 and thiamine. Cells contained cytochromes, carbon monoxide dehydrogenase, and sulfite reductase P582, but no desulfoviridin. Strain Ani1 is described as a new species of the genus Desulfobacterium D. anilini. Marine enrichments with the three dihydroxybenzene isomers led to three different strains of sulfate-reducing bacteria; each of them could grow only with the isomer used for enrichment. Two strains isolated with catechol (strain Cat2) or resorcinol (strain Re10) were studied in detail. Both strains oxidized their substrates completely to CO2, and contained cytochromes, carbon monoxide dehydrogenase, and sulfite reductase P 582. Desulfoviridin was not present. Whereas the rod-shaped catechol oxidizer (strain Cat2) was able to grow on 18 aromatic compounds and several aliphatic substrates, the coccoid resorcinol-degrading bacterium (strain Re10) utilized only resorcinol, 2,4-dihydroxybenzoate and 1,3-cyclohexanedion. These strains could not be affiliated with existing species of sulfate-reducing bacteria. A further coccoid sulfate-reducing bacterium (strain Hy5) was isolated with hydroquinone and identified as a subspecies of Desulfococcus multivorans. Most-probable-number enumerations with catechol, phenol, and resorcinol showed relatively large numbers (10(4)-10(6) per ml) of aryl compound-degrading sulfate reducers in marine sediment samples.

Analysis of the transcriptional unit encoding the genes for rubredoxin (rub) and a putative rubredoxin oxidoreductase (rbo) in Desulfovibrio vulgaris Hildenborough

The nucleotide sequence of a 2.0-kilobase-pair EcoRI restriction fragment upstream from the gene (rub, 162 base pairs) encoding rubredoxin from Desulfovibrio vulgaris Hildenborough indicates that it is part of a larger transcriptional unit, containing an additional 378-base-pair open reading frame which terminates 16 nucleotides from the translational start of the rub gene and could encode a polypeptide of 14 kilodaltons (kDa). Northern (RNA) blotting of RNA isolated from both D. vulgaris Hildenborough and Escherichia coli TG2 transformed with plasmid pJK29, which contains both genes on a 1.1-kilobase-pair SalI insert, confirms that the genes for this 14-kDa polypeptide and rubredoxin are present on a single transcript of 680 nucleotides. Strong evidence that the 14-kDa polypeptide is also a redox protein is provided by the fact that its NH2 terminus is homologous to desulforedoxin, which has been isolated from D. gigas as a small dimeric redox protein (36 amino acids per monomer), coordinating two iron atoms. Since rubredoxin is a potential redox partner for the 14-kDa protein, it has been tentatively named rubredoxin oxidoreductase, produced by the rbo gene. Southern blotting indicates that the rbo-rub operon is present in several species and strains of sulfate-reducing bacteria.

Transfer of broad host-range plasmids to sulphate-reducing bacteria

The broad-host-range, IncQ, plasmid R300B (Sm, Su) has been stably transferred to two strains of sulphate-reducing bacteria (Desulfovibrio sp. 8301 and Desulfovibrio desulfuricans 8312), using the IncP1 transfer system of the helper plasmid pRK2013 and cocultivation of sulphate-reducing bacteria with facultative anaerobes in media provided with sulphate and nitrate ions as electron acceptors. R300B was transferred at a frequency of 10−2 to 1 per acceptor cell. The SmR marker was expressed in both sulphate-reducing bacteria strains while the SuR was expressed only in strain 8301. R300B can also be transferred back to E. coli strains provided with IncP1 plasmids taking advantage of the retrotransfer ability of these plasmids. This occurs at a frequency up to 10−4 by recipient E. coli cell.

Metabolism of l-alanine in Desulfotomaculum ruminis and two marine Desulfovibrio strains

The degradation of l-alanine by three strains of sulfate-reducing bacteria that can grow with l-alanine as an energy source was investigated. In Desulfotomaculum ruminis and most likely also in two marine Desulfovibrio strains alanine is converted to pyruvate via an NAD-dependent alanine dehydrogenase. D. ruminis contained high activities of soluble NADH and NADPH dehydrogenases. In the marine strains the activities were much lower and the NADH dehydrogenase was partly associated with the membrane fraction.

Occurrence of polyglucose as a storage polymer in Desulfovibrio species and Desulfobulbus propionicus

The occurrence of organic storage compounds in six strains of sulfate-reducing bacteria was investigated.

Ultrastructural and biochemical characterization of Miyazaki strains of Desulfovibrio vulgaris.

Ultrastructural examinations and biochemical characterizations were carried out on "Miyazaki" strains of sulfate-reducing bacteria, the biochemistry of which has been extensively studied by Japanese scientists, the identification of which, however, has not yet been done satisfactorily. Miyazaki strains, MK, MF, and MY, were all strictly anaerobic, had a single polar flagellum, and contained desulfoviridin and cytochrome c3. Cells of strain MK were typical comma-shaped, short, curved rods (0.5× 1.5-2μm), while cells of strains MF and MY were longer (2-5μm) and were either curved, sigmoid, or spiral. Occasionally the "Y-shape" or blebbing of the cells was observed among cells of MF and MY. The shorter blebs usually had a single flagellum.Carbon sources, buoyant density of DNA, and electrophoretic mobilities of desulfoviridin, ATP sulfurylase, and adenosine phosphosulfate reductase were also determined. Although cells of strains MF and MY were clearly distinguished from other cells of Desulfovibrio spp. in morphology, all three strains were identified as Desulfovibrio vulgaris.

The oxygen tolerance of sulfate-reducing bacteria isolated from North Sea waters

Five strains of sulfate-reducing bacteria, previously isolated from North Sea waters and identified asDesulfovibrio vulgaris, were investigated for their abilities to survive in aerobic natural seawater. Viable organisms of all strains were recoverable after exposure to oxygen for more than 72 h. The level of the protective enzymes superoxide dismutase and catalase detectable in these strains and the low rates at which oxygen was reduced probably account at least in part for their considerable abilities to survive in aerobic environments. The autoxidizable nature of cytochromec 3 and KCN-inhibitable cytochromec oxidase activity present in these bacteria are postulated to act as possible oxygen-scavenging mechanisms analogous to the activity of NADH oxidase present in certain other strict anaerobes.

Structure-function relationship in hemoproteins: The role of cytochrome c 3 in the reduction of colloidal sulfur by sulfate-reducing bacteria

Cytochromes c3 of different strains of sulfate-reducing bacteria have been purified and tested for their capacity to reduce colloidal sulfur to hydrogen sulfide. The results are in good agreement with the activities reported for the whole cells. Cytochrome c3 is the sulfur reductase of some strains of sulfate-reducing bacteria such as Desulfovibrio desulfuricans Norway 4 and sulfate-reducing bacterium strain 9974 from which the sulfur reductase activity can be purified with the cytochrome c3. In contrast, Desulfovibrio vulgaris Hildenborough cytochrome c3 is inhibited by the product of the reaction namely hydrogen sulfide. Chloramphenicol has no effect on the sulfur reductase activity of D. desulfuricans Norway 4 when resting cells grown on lactate-sulfate medium are put in the presence of colloidal sulfur. This shows that the sulfur reductase activity is constitutive and corresponds to the fact that colloidal sulfur grown cells do not contain more cytochrome c3 (or another sulfur reductase) than lactate-sulfate-grown cells.