Enrichment Of Sulfate-reducing Bacteria Research Articles

Comparison of Nitrate and Perchlorate in Controlling Sulfidogenesis in Heavy Oil-Containing Bioreactors.

Control of microbial reduction of sulfate to sulfide in oil reservoirs (a process referred to as souring) with nitrate has been researched extensively. Nitrate is reduced to nitrite, which is a strong inhibitor of sulfate-reducing bacteria (SRB). Perchlorate has been proposed as an alternative souring control agent. It is reduced to chlorate (ClO3-) and chlorite (ClO2-), which is dismutated to chloride and O2. These can react with sulfide to form sulfur. Chlorite is also highly biocidal. Here we compared the effectiveness of perchlorate and nitrate in inhibiting SRB activity in medium containing heavy oil from the Medicine Hat Glauconitic C (MHGC) field, which has a low reservoir temperature and is injected with nitrate to control souring. Using acetate, propionate and butyrate as electron donors, perchlorate-reducing bacteria (PRB) were obtained in enrichment culture and perchlorate-reducing Magnetospirillum spp. were isolated from MHGC produced waters. In batch experiments with MHGC oil as the electron donor, nitrate was reduced to nitrite and inhibited sulfate reduction. However, perchlorate was not reduced and did not inhibit sulfate reduction in these incubations. Bioreactor experiments were conducted with sand-packed glass columns, containing MHGC oil and inoculated with an oil-grown mesophilic SRB enrichment. Once active souring (reduction of 2 mM sulfate to sulfide) was observed, these were treated with nitrate and/or perchlorate. As in the batch experiments, 4 mM nitrate completely inhibited sulfide production, while partial inhibition occurred with 1 and 2 mM nitrate, but injection of 4 mM perchlorate did not inhibit sulfate reduction and perchlorate was not reduced. The enriched and isolated PRB were unable to use heavy oil components, like alkylbenzenes, which were readily used by nitrate-reducing bacteria. Hence perchlorate, injected into a low temperature heavy oil reservoir like the MHGC, may not be reduced to toxic intermediates making nitrate a preferable souring control agent.

Long-Term Feeding of Elemental Sulfur Alters Microbial Community Structure and Eliminates Mercury Methylation Potential in Sulfate-Reducing Bacteria Abundant Activated Sludge.

This study reported a novel observation that the long-term cultivation of sulfur-reducing bacteria (S0RB) from a sulfate-reducing bacteria (SRB)-abundant seeding sludge with elemental sulfur feeding significantly shaped the microbial community structure and eliminated the mercury methylation potential in the S0RB-enriched sludge. In this study, the enrichments of SRB and S0RB from activated sludge were obtained through long-term cultivations. Subsequently, the batch tests showed that approximately 5000 μg/L Hg (II) was completely removed from the solution by both the SRB-enriched and S0RB-enriched sludge. Extremely low or no MeHg production was observed in the S0RB-enriched sludge (less than the limit of detection, 0.01 μg/L), while 1.49 μg/L MeHg accumulated in the SRB-enriched sludge. Other batch tests using the sludge samples from a replication of the cultivation showed that the methylation capability of the S0RB-enriching sludge gradually diminished to a negligible level over a 6 month cultivation time. However, some mercury-methylation-related bacteria were present in the enrichment of S0RB such as Geobacter. The absence of MeHg in the S0RB-enriched sludge may be attributed to the dissolved organic matter (DOM) instead of the sulfur- and sulfate-reduction pathway or MeHg demethylation when exposed to Hg (II). The cultivated S0RB could be used for mercury-contaminated wastewater treatment without MeHg concern.

Effects of plant downtime on the microbial community composition in the highly saline brine of a geothermal plant in the North German Basin.

The microbial biocenosis in highly saline fluids produced from the cold well of a deep geothermal heat store located in the North German Basin was characterized during regular plant operation and immediately after plant downtime phases. Genetic fingerprinting revealed the dominance of sulfate-reducing bacteria (SRB) and fermentative Halanaerobiaceae during regular plant operation, whereas after shutdown phases, sequences of sulfur-oxidizing bacteria (SOB) were also detected. The detection of SOB indicated oxygen ingress into the well during the downtime phase. High 16S ribosomal RNA (rRNA) and dsrA gene copy numbers at the beginning of the restart process showed an enrichment of bacteria, SRB, and SOB during stagnant conditions consistent with higher concentrations of dissolved organic carbon (DOC), sulfate, and hydrogen sulfide in the produced fluids. The interaction of SRB and SOB during plant downtimes might have enhanced the corrosion processes occurring in the well. It was shown that scale content of fluids was significantly increased after stagnant phases. Moreover, the sulfur isotopic signature of the mineral scales indicated microbial influence on scale formation.

Enrichment and isolation of acidophilic sulfate‐reducing bacteria from Tinto River sediments

Although some acidophilic and alkaliphilic species have been described recently, most of the known sulfate-reducing bacteria (SRB) grow optimally at neutral pH. In this study, sulfate reduction was studied with sediment samples from the extremely acidic Tinto River basin. Stable enrichments of SRB were obtained at pH 4 with glycerol, methanol and hydrogen; at pH 4.5 with lactate and at pH 5.5 with succinate as substrates. Inhibition of sulfate reduction by organic acids below their pKa was observed. Cloning and sequencing of 16S rRNA gene showed that fermentative bacteria (Paludibacter spp., Oscillibacter spp.) and SRB (Thermodesulfobium spp., Desulfosporosinus spp., Desulfitobacterium spp., Desulfotomaculum spp.) were co-enriched. By repeated serial dilutions and streaking on agar plates, four strains of SRB belonging to the Firmicutes phylum were obtained. Two of them show 96% 16S rRNA gene sequence similarity with Desulfosporosinus acidophilus, and a third one with Desulfosporosinus orientis. Another isolate has just 93% rRNA gene sequence similarity with the Desulfosporosinus/Desulfitobacterium cluster and might represent a novel species within a novel genus. One of the Desulfosporosinus strains was further investigated showing maximum growth at pH 5.5, and a pH-dependent inhibitory effect of organic acids and sulfide.

Enrichment of sulfate-reducing bacteria and resulting mineral formation in media mimicking pore water metal ion concentrations and pH conditions of acidic pit lakes

Acid mine drainage sites are extreme environments with high acidity and metal ion concentrations. Under anoxic conditions, microbial sulfate reduction may trigger the formation of secondary minerals as a result of H2S production and pH increase. This process was studied in batch experiments with enrichment cultures from acidic sediments of a pit lake using growth media set at different pH values and containing elevated concentrations of Fe²⁺ and Al³⁺. At initial pH values of 5 and 6, sulfate reduction occurred shortly after inoculation. Sulfate- reducing bacteria affiliated to the genus Desulfosporosinus predominated the microbial communities as shown by 16S rRNA gene analysis performed at the end of the incubation. At initial pH values of 3 and 4, sulfate reduction and cell growth occurred only after an extended lag phase, however, at a higher rate than in the less acidic assays. At the end of the growth phase, enrichments were dominated by Thermodesulfobium spp. suggesting that these sulfate reducers were better adapted to acidic conditions. Iron sulfides in the bulk phase were common in all assays, but specific aluminum precipitates formed in close association with cell surfaces and may function as a detoxification mechanism of dissolved Al species at low pH.

English

A nitrate-reducing bacteria (NRB) strain with vigorous growth, strong nitrate reduction ability, strain B9 2-1, was isolated from Suizhong36-1 oilfield, its routine identification and analysis of 16S rRNA and also the competitive inhibition experiments with the enrichment of sulfate-reducing bacteria (SRB) were carried out. The results showed that only the dosing of nitrate, nitrite as electron acceptors, the activation of nitrate-reducing bacteria, as well as the inhibition of sulfide production resulted from a limited capacity, while addition of NRB isolated from the produced fluid, growth and sulfide production activity of sulfate reducing bacteria produced a significant inhibition and antibacterial effects of nitrite, which was better than nitrate. At the same time, the small amount of molybdate dosing showed better results, which will be of significance when applied to shipping and state-defending industries.   Key words: Nitrate-reducing bacteria, sulfate-reducing bacteria, restriction fragment length polymorphism, nitrates, nitrite, oil field, competitive inhibition.

Isolation of a nitrate-reducing bacteria strain from oil field brine and the inhibition of sulfate-reducing bacteria

A nitrate- reducing bacteria (NRB) strain with vigorous growth, strong nitrate reduction ability, strain B9 2-1, was isolated from Suizhong36-1 oilfield, its routine identification and analysis of 16S rRNA and also the competitive inhibition experiments with the enrichment of sulfate-reducing bacteria (SRB) were carried out. The results showed that only the dosing of nitrate, nitrite as electron acceptors, the activation of nitrate- reducing bacteria, as well as the inhibition of sulfide production resulted from a limited capacity, while addition of NRB isolated from the produced fluid, growth and sulfide production activity of sulfate reducing bacteria produced a significant inhibition and antibacterial effects of nitrite, which was better than nitrate. At the same time, the small amount of molybdate dosing showed better results, which will be of significance when applied to shipping and state-defending industries.

Microbial community of reduced pockmark sediments (Gdansk Deep, Baltic Sea)

The microbial community of reduced pockmark sediments in the Russian sector of the Gdansk Deep, Baltic Sea, was investigated by molecular biological techniques. Fluorescent in situ hybridization was used to determine the numbers of eubacteria, archaea, and sulfate-reducing bacteria. Eubacteria were found to predominate in the upper 10 cm of the sediment (up to 5.3 × 109 cells/g wet sediment), while the number of archaea increased in the 10- to 30-cm layers (up to 2.8 × 109 cells/g wet sediment, which is higher than the number of eubacteria in the same horizons). Analysis of 16S rRNA gene fragments revealed members of the following phyla: Proteobacteria, Chloroflexi, Firmicutes, Planctomycetales, and high-G + C gram-positive bacteria. Sulfate-reducing bacteria (SRBs) of the families Syntrophaceae, Desulfuromonadaceae, and Actinobacteria of the genera Kocuria and Rothia were the predominant groups. Molecular probes were used to determine predominance of Desulfovibrionales in the SRB enrichment cultures obtained from different horizons of pockmark sediments. Three archaeal phylotypes were revealed, belonging to Euryarchaeota. One of these fell into the group of uncultured methanotrophic archaea (ANME-1a), while the other two were most closely related to uncultured methanogens.

Microbial links between sulfate reduction and metal retention in uranium- and heavy metal-contaminated soil.

Sulfate-reducing bacteria (SRB) can affect metal mobility either directly by reductive transformation of metal ions, e.g., uranium, into their insoluble forms or indirectly by formation of metal sulfides. This study evaluated in situ and biostimulated activity of SRB in groundwater-influenced soils from a creek bank contaminated with heavy metals and radionuclides within the former uranium mining district of Ronneburg, Germany. In situ activity of SRB, measured by the (35)SO(4)(2-) radiotracer method, was restricted to reduced soil horizons with rates of < or =142 +/- 20 nmol cm(-3) day(-1). Concentrations of heavy metals were enriched in the solid phase of the reduced horizons, whereas pore water concentrations were low. X-ray absorption near-edge structure (XANES) measurements demonstrated that approximately 80% of uranium was present as reduced uranium but appeared to occur as a sorbed complex. Soil-based dsrAB clone libraries were dominated by sequences affiliated with members of the Desulfobacterales but also the Desulfovibrionales, Syntrophobacteraceae, and Clostridiales. [(13)C]acetate- and [(13)C]lactate-biostimulated soil microcosms were dominated by sulfate and Fe(III) reduction. These processes were associated with enrichment of SRB and Geobacteraceae; enriched SRB were closely related to organisms detected in soils by using the dsrAB marker. Concentrations of soluble nickel, cobalt, and occasionally zinc declined < or =100% during anoxic soil incubations. In contrast to results in other studies, soluble uranium increased in carbon-amended treatments, reaching < or =1,407 nM in solution. Our results suggest that (i) ongoing sulfate reduction in contaminated soil resulted in in situ metal attenuation and (ii) the fate of uranium mobility is not predictable and may lead to downstream contamination of adjacent ecosystems.

Evaluation of enrichments of sulfate reducing bacteria from pristine hydrothermal vents sediments as potential inoculum for reducing trichloroethylene

The evaluation of enrichments from pristine hydrothermal vents sediments on its capability of reducing trichloroethylene (TCE) under sulfate reducing conditions with lactate and volatile fatty acids (VFAs) as substrates was performed. Effect of the possible TCE biodegradation intermediates cis and trans 1,2 dichloroethenes on sulfate reduction (SR) was also evaluated. The influence of cyanocobalamin (CNB12) and riboflavin (RF) on the SR and biodegradation of TCE was also determined. Sediments from the vents were incubated at 37°C and supplemented with 4 g l−1 SO42−, lactate or VFAs and amended in the corresponding treatments with either CNB12 or RF in separated experiments. A percentage of TCE removal of 86 (150 μmol l−1 initial concentration) was attained coupled to 48% sulfate depletion with lactate as substrate. Up to 93% removal of TCE (300 μmol l−1 initial concentration) and 40% of sulfate was reached for VFAs as electron donor. A combination of lactate and CNB12 yielded the best SR. The overall results suggest a syntrophic association in this microbial community in which sulfate reducers, dehalogenating, and probably halorespiring bacteria may be interacting and taking advantage of the fermentation of substrates differently, but without interruption of SR in spite of the fact that TCE was always present. It was also clear that sulfate reduction must be established in the cultures before any degradation can occur. The microbial community present in these hydrothermal vents sediments could be a new source of inoculum for bioreactors designed for dechlorination purposes.

Effect of oxidized leachate on degradation of lignin by sulfate-reducing bacteria

Municipal solid waste materials (MSWs) in landfills need a long period of stabilization because lignin compounds in MSWs and leachate are not readily biodegraded, but inhibit methanogenic metabolism. Recirculation of leachate into the landfill offers the potential advantage of increasing the rate of decomposition of organic matter. However, the degradation of lignin by leachate recirculation alone is quite difficult. Several recent studies have demonstrated that sulfate-reducing bacteria (SRB) were able to degrade lignin compounds. In this study, batch tests were conducted to investigate the impacts of SRB enrichment on lignin decomposition rates as well as the decomposition of other biodegradable organics. Further, the effects of nitrite and nitrate on lignin degradation rates were also studied. A 16S rRNA assay showed that the SRB used herein, which were obtained by enriching solid waste collected from a closed MSW landfill, were Thaurea sp. and Desulfovibrio sp. Lignin was found to be biodegraded by the SRB and the rate of lignin removal per unit of waste volatile suspended solid was 2.9 mg lignin g(-1) VSS day(- 1). It was found that the initial degradation rate increased under higher initial lignin concentrations. However, the degradation rate during days 6-19 became slower than that during the initial 9 days because lignin consisted of complexly bonded aromatic compounds that were not readily biodegradable. Adding other organics such as lactate seemed to improve the rate and amount of lignin degradation, probably due to the increase in SRB associated with consumption of the additional organics. The lignin removal percentage decreased with increases in oxidized nitrogen (nitrite or nitrate) concentrations, indicating that oxidized nitrogen could inhibit SRB activity. Conclusively, the study verified the existence of SRB in the landfill and showed that the SRB could be activated for the degradation of lignin by the recirculation of the leachate, which is consistent with other studies showing that leachate recirculation could shorten the stabilization period of the landfill.

Characterization of sulfate-reducing bacteria dominated surface communities during start-up of a down-flow fluidized bed reactor

An anaerobic down-flow fluidized bed reactor was inoculated with granular sludge and started-up with sulfate containing synthetic wastewater to promote the formation of a biofilm enriched in sulfate-reducing bacteria (SRB), to produce biogenic sulfide. The start-up was done in two stages operating the reactor in batch for 45 days followed by 85 days of continuous operation. Low-density polyethylene was used as support. The biofilm formation was followed up by biochemical and electron microscopy analyses and the composition of the community was examined by 16S rDNA sequence analysis. Maximum immobilized volatile solids (1.2 g IVS/L(support)) were obtained after 14 days in batch regime. During the 85 days of continuous operation, the reactor removed up to 80% of chemical oxygen demand (COD), up to 28% of the supplied sulfate and acetate was present in the effluent. Sulfate-reducing activity determined in the biofilm with ethanol or lactate as substrate was 11.7 and 15.3 g COD/g IVS per day, respectively. These results suggested the immobilization of sulfate reducers that incompletely oxidize the substrate to acetate; the phylogenetic analysis of the cloned 16S rDNA gene sequences showed high identity to the genus Desulfovibrio that oxidizes the substrates incompletely. In contrast, in the granular sludge used as inoculum a considerable number of clones showed homology to Methanobacterium and just few clones were close to SRB. The starting-up approach allowed the enrichment of SRB within the diverse community developed over the polyethylene support.

Polysulfide reduction using sulfate-reducing bacteria in a photocatalytic hydrogen generation system

A hydrogen generation process using photocatalytic reactions has been proposed. In this process, hydrogen sulfide is a source of hydrogen and is turned into polysulfide. In order to establish the cyclic operation of a photocatalytic hydrogen generation system, it is necessary to convert polysulfide back into hydrogen sulfide with a small energy input. This paper proposes the use of sulfate-reducing bacteria (SRB) for the regeneration of hydrogen sulfide. Batch cultivation of natural source SRB samples were carried out using a culture medium containing polysulfide as the only sulfur compound source. SRB produced hydrogen sulfide from several kinds of polysulfide sources, including a photocatalytic hydrogen generation-produces solution. Production lag phase and production rate of hydrogen sulfide were affected by initial polysulfide concentration. SRB activity was inhibited at high initial polysulfide concentrations. SRB enrichment culture T2, exhibited the highest hydrogen sulfide production rate, and was able to utilize several kinds of organic matter as the electron donor. The results suggest the possibility of using large biomass sources, such as sewage sludge and the raw garbage in a hydrogen generation system. We developed speculative estimates that an SRB based hydrogen generation system is feasible.

Diversity of sulfate-reducing bacteria from an extreme hypersaline sediment, Great Salt Lake (Utah)

The diversity of sulfate-reducing bacteria (SRB) inhabiting the extreme hypersaline sediment (270 g L(-1) NaCl) of the northern arm of Great Salt Lake was studied by integrating cultivation and genotypic identification approaches involving PCR-based retrieval of 16S rRNA and dsrAB genes, the latter encoding major subunits of dissimilatory (bi) sulfite reductase. The majority (85%) of dsrAB sequences retrieved directly from the sediment formed a lineage of high (micro) diversity affiliated with the genus Desulfohalobium, while others represented novel lineages within the families Desulfohalobiaceae and Desulfobacteraceae or among Gram-positive SRB. Using the same sediment, SRB enrichment cultures were established in parallel at 100 and at 190 g L(-1) NaCl using different electron donors. After 5-6 transfers, dsrAB and 16S rRNA gene-based profiling of these enrichment cultures recovered a SRB community composition congruent with the cultivation-independent profiling of the sediment. Pure culture representatives of the predominant Desulfohalobium-related lineage and of one of the Desulfobacteraceae-affilated lineages were successfully obtained. The growth performance of these isolates and of the enrichment cultures suggests that the sediment SRB community of the northern arm of Great Salt Lake consists of moderate halophiles, which are salt-stressed at the in situ salinity of 27%.

Biological and chemical interactions with U(VI) during anaerobic enrichment in the presence of iron oxide coated quartz

Microcosm experiments were performed to understand chemical and biological interactions with hexavalent uranium (U(VI)) in the presence of iron oxide bearing minerals and trichloroethylene (TCE) as a co-contaminant. Interactions of U(VI) and hydrous iron oxide moieties on the mineral oxide surfaces were studied during enrichments for dissimilatory iron reducing (DIRB) and sulfate reducing bacteria (SRB). Microbes enriched from groundwater taken from the Test Area North (TAN) site at the Idaho National Laboratory (INL) were able to reduce the U(VI) in the adsorption medium as well as the iron on quartz surfaces. Early in the experiment disappearance of U(VI) from solution was a function of chemical interactions since no microbial activity was evident. Abiotic removal of U(VI) was enhanced in the presence of carbonate. As the experiment proceeded, further removal of U(VI) from solution was associated with the fermentation of lactate to propionate and acetate. During later phases of the experiment when lactate was depleted from the growth medium in the microcosm containing the DIRB enrichments, U(VI) concentrations in the solution phase increased until additional lactate was added. When additional lactate was added and fermentation proceeded, U(VI) concentrations in the liquid phase again returned to near zero. Similar results were shown for the SRB enrichment but lower uranium concentrations were seen in the liquid phase, while in the enrichment with carbonate a similar increase in uranium concentration was not seen. Chemical and biological interactions appear to be important on the mobilization/immobilization of U(VI) in an iron oxide system when TCE is present as a co-contaminant. Interestingly, TCE present in the microcosm experiments was not dechlorinated which was probably an effect of redox conditions that were unsuitable for reductive dechlorination by the microbial culture tested.

Microbiological and Geochemical Heterogeneity in an In Situ Uranium Bioremediation Field Site

The geochemistry and microbiology of a uranium-contaminated subsurface environment that had undergone two seasons of acetate addition to stimulate microbial U(VI) reduction was examined. There were distinct horizontal and vertical geochemical gradients that could be attributed in large part to the manner in which acetate was distributed in the aquifer, with more reduction of Fe(III) and sulfate occurring at greater depths and closer to the point of acetate injection. Clone libraries of 16S rRNA genes derived from sediments and groundwater indicated an enrichment of sulfate-reducing bacteria in the order Desulfobacterales in sediment and groundwater samples. These samples were collected nearest the injection gallery where microbially reducible Fe(III) oxides were highly depleted, groundwater sulfate concentrations were low, and increases in acid volatile sulfide were observed in the sediment. Further down-gradient, metal-reducing conditions were present as indicated by intermediate Fe(II)/Fe(total) ratios, lower acid volatile sulfide values, and increased abundance of 16S rRNA gene sequences belonging to the dissimilatory Fe(III)- and U(VI)-reducing family Geobacteraceae. Maximal Fe(III) and U(VI) reduction correlated with maximal recovery of Geobacteraceae 16S rRNA gene sequences in both groundwater and sediment; however, the sites at which these maxima occurred were spatially separated within the aquifer. The substantial microbial and geochemical heterogeneity at this site demonstrates that attempts should be made to deliver acetate in a more uniform manner and that closely spaced sampling intervals, horizontally and vertically, in both sediment and groundwater are necessary in order to obtain a more in-depth understanding of microbial processes and the relative contribution of attached and planktonic populations to in situ uranium bioremediation.

Sulfate-reducing bacterial community structure and their contribution to carbon mineralization in a wastewater biofilm growing under microaerophilic conditions.

The community structure of sulfate-reducing bacteria (SRB) and the contribution of SRB to carbon mineralization in a wastewater biofilm growing under microaerophilic conditions were investigated by combining molecular techniques, molybdate inhibition batch experiments, and microelectrode measurements. A 16S rDNA clone library of bacteria populations was constructed from the biofilm sample. The 102 clones analyzed were grouped into 53 operational taxonomic units (OTUs), where the clone distribution was as follows: Cytophaga- Flexibacter- Bacteroides (41%), Proteobacteria (41%), low-G+C Gram-positive bacteria (18%), and other phyla (3%). Three additional bacterial clone libraries were also constructed from SRB enrichment cultures with propionate, acetate, and H(2) as electron donors to further investigate the differences in SRB community structure due to amendments of different carbon sources. These libraries revealed that SRB clones were phylogenetically diverse and affiliated with six major SRB genera in the delta-subclass of the Proteobacteria. Fluorescent in situ hybridization (FISH) analysis revealed that Desulfobulbus and Desulfonema were the most abundant SRB species in this biofilm, and this higher abundance (ca. 2-4x10(9) cells cm(-3) and 5x10(7) filaments cm(-3), respectively) was detected in the surface of the biofilm. Microelectrode measurements showed that a high sulfate-reducing activity was localized in a narrow zone located just below the oxic/anoxic interface when the biofilm was cultured in a synthetic medium with acetate as the sole carbon source. In contrast, a broad sulfate-reducing zone was found in the entire anoxic strata when the biofilm was cultured in the supernatant of the primary settling tank effluent. This is probably because organic carbon sources diffused into the biofilm from the bulk water and an unknown amount of volatile fatty acids was produced in the biofilm. A combined approach of molecular techniques and batch experiments with a specific inhibitor (molybdate) clearly demonstrated that Desulfobulbus is a numerically important member of SRB populations and the main contributor to the oxidation of propionate to acetate in this biofilm. However, acetate was preferentially utilized by nitrate-reducing bacteria but not by acetate-utilizing SRB.

Identification and Phylogenetic Analysis of Thermophilic Sulfate-Reducing Bacteria in Oil Field Samples by 16S rDNA Gene Cloning and Sequencing

Thermophilic sulfate-reducing bacteria (SRB) have been recognized as an important source of hydrogen sulfide (H2S) in hydrocarbon reservoirs and in production systems. Four thermophilic SRB enrichment cultures from three different oil field samples (sandstone core, drilling mud, and production water) were investigated using 16S rDNA sequence comparative analysis. In total, 15 different clones were identified. We found spore-forming, low G+C content, thermophilic, sulfate-reducingDesulfotomaculum-related sequences present in all oil field samples, and additionally a clone originating from sandstone core which was assigned to the mesophilicDesulfomicrobiumgroup. Furthermore, three clones related to Gram-positive, non-sulfate-reducingThermoanaerobacterspecies and four clones close toClostridium thermocopriaewere found in enrichment cultures from sandstone core and from production water, respectively. In addition, the deeply rooted lineage of two of the clones suggested previously undescribed, Gram-positive, low G+C content, thermophilic, obligately anaerobic bacteria present in production water. Such thermophilic, non-sulfate-reducing microorganisms may play an important ecological role alongside SRB in oil field environments.

Anaerobic degradation of indolic compounds by sulfate-reducing enrichment cultures, and description of Desulfobacterium indolicum gen. nov., sp. nov.

Indole (1.5 mmol/l) added to suflate-rich marine mud or sulfate-free sewage digestor sludge was anaerobically degraded within one week. Enrichments from sludge samples in defined indole-containing media with or without sulfate were selective for sulfate-reducing bacteria or mixed methanogenic associations, respectively. Other enrichments of sulfate-reducing bacteria were obtained with skatole, indoleacetate, indolepropionate, quinoline, and pyridine. From a marine enrichment with indole as sole electron donor and carbon source, an oval to rod-shaped, Gram-negative, nonsporing sulfate-reducing bacterium (strain In04) was isolated. Growth occurred in defined bicarbonate-buffered, sulfide-reduced media supplemented with vitamin B12. Furthen aromatic compounds utilized as electron donors and carbon sources were anthranilic acid and quinoline. Nonaromatic compounds used as substrates were formate, acetate, propionate, ethanol, propanol, butanol, pyruvate, malate, fumarate, and succinate. However, growth with substrates other than indole was rather slow. Thiosulfate served as an alternative electron acceptor. Complete oxidation of indole to CO2 was shown by stoichiometric measurements in batch culture with sulfate as electron acceptor. An average growth yield of 31.3 g cell dry weight was obtained per mol of indole oxidized. Pigment analysis revealed that cytochromes and menaquinone MK-7 (H2) were present. Desulfoviridin could not be detected. Strain In04 is described as new species of the new genus Desulfobacterium indolicum.