Sulfate-Reducing Bacteria Release Barium and Radium from Naturally Occurring Radioactive Material in Oil-Field Barite

Scale and sludge deposits formed during oil production can contain elevated levels of Ra, often coprecipitated with barium sulfate (barite). The potential for sulfate-reducing bacteria to release ²²⁶Ra and Ba (a Ra analog) from oil-field barite was evaluated. The concentration of dissolved Ba increased when samples containing pipe scale, tank sludge, or oil-field brine pond sediment were incubated with sulfate-reducing bacteria Desulfovibrio sp., Str LZK1, isolated from an oil-field brine pond. However, Ba release was not stoichiometric with sulfide production in oil-field samples, and < 0.1% of the Ba was released. Potential for the release of ²²⁶Ra was demonstrated, and the ²²⁶Ra release associated with sulfate-reducing activity was predictable from the amount of Ba released. As with Ba, only a fraction of the ²²⁶Ra expected from the amount of sulfide produced was released, and most of the Ra remained associated with the solid material.     

复合垂直流构建湿地基质酶活性与污水净化效果

构建湿地是20世纪70年代兴起的处理污水的生态环境,由于其具有建造,运行和日常管理费用低廉,处理效果稳定,且适应面广,越来越受到世界各国的重视,这种污水处理技术在发展中国家有着十分广泛的应用前景,本文研究与揭示了复合垂直流构建湿地基质中的磷酸酶和脲酶活性及与污水净化效果的关系;（1）不同类型湿地基质酶活性不同。甚至不同月份的酶活性也不相同。（2）不同深度基质中的酶活性是不相同的。（3）基质磷酸酶的活性与复合垂直流构建湿地对污水中总磷（TP）,无机磷（IP）以及化学需氧量（CODCr)的去除率有很显著的相关关系。（4）脲酶的活性与凯氏氮（KN）的去除率相关性极显著,这些为利用酶活性强度作为评价净化效果和挑选合适湿地植物的指标提供了理论依据。     

Involvement of Sulfur- and Iron-Transforming Bacteria in Heaving of House Foundations

About 1,000 houses built on excavated nonweathered mudstone sediments, originally deposited in the Neogene, have been damaged by microbially induced heaving of foundations. The maximal height of the heaving was 48 cm. The presence of sulfate-reducing, sulfur-oxidizing, and acidophilic iron-oxidizing bacteria in the mudstone indicated that the joint activity of these three types of bacteria could account for the heaving. A hypothesis is presented in which, first, the temperature of the newly exposed mudstone sediments increased above 25 °C, which stimulated the sulfate-reducing bacteria in the mudstone to actively reduce sulfate to hydrogen sulfide. The mudstone sediments under the houses gradually dried, and became permeable to air. Consequently, sulfur-oxidizing bacteria oxidized the hydrogen sulfide to sulfuric acid and the environmental pH decreased to approximately 3. Next, the acidophilic iron-oxidizing bacteria actively oxidized the sulfur in pyrite to produce much more acid. The resulting sulfuric acid reacted with calcium carbonate and with ferric and potassium ions to produce gypsum and jarosite, respectively. A combination of the increased volume of gypsum and jarosite crystals and the production of CO 2 as a by-product of their formation made the mudstone sediments bulky. The end result was widespread heaving.     

湿地系统中植物和土壤在治理重金属污染中的作用

重金属污染环境的治理是目前环境工程的核心课题。湿地作为水陆相互作用形成的独特生态系统,在重金属污染治理中的作用倍受关注。对湿地植物、土壤在治理重金属污染中所起的关键作用及其机理做一综述,并对治理重金属污染的湿地构建提出几点建议。     

人工湿地中降解有机污染物细菌的分离筛选

采用平板划线法从人工湿地污水处理系统底泥和污水中分离出 2 3株细菌 ,在实验室条件下检测了这些细菌对灭菌污水和自然污水CODcr的去除效果 ,结果表明其中 8株细菌 (DN -4、DN -5、DN- 6、DN -10、DN -11、DN -12、DN- 13、WS- 5 )对灭菌污水和自然污水的CODcr均有较高的去除率 ,胞外酶检测表明该 8株细菌均能产生淀粉酶和接触酶 ,这些细菌具有潜在的应用价值。     

Occurrence of Pathogenic Free‐Living Amoebae and Bacterial Indicators in a Constructed Wetland Treating Domestic Wastewater from a Single Household

Constructed wetlands are effective wastewater treatment systems because of their ability to remove large amounts of organic matter and pathogens. The goals of this study were to characterize the presence of pathogenic free‐living amoebae and bacterial indicators (total and fecal coliforms), and to ascertain the removal efficiencies of physical and chemical pollutants, in a constructed wetland treating domestic wastewater from a single household. Influent and effluent samples were collected monthly over a ten‐month period for biological, physical and chemical analyses. Thirty‐two species of free‐living amoebae were isolated from the system. The genus Acanthamoeba was the most frequently encountered (59 %) and was removed from the wastewater with the greatest efficiency (80 %). Removal of bacteria was low, the highest removal rates were found in August (4 logarithmic units) and January (3 logarithmic units). The average removal efficiencies of suspended solids, BOD₅ and ammoniacal nitrogen were 71.5 %, 50.6 % and 13.1 %, respectively. The relatively low removal efficiencies of the various bacteriological, physical and chemical parameters suggest that the hydraulic retention time was probably insufficient for optimal treatment to occur. The effluent quality was unacceptable for unrestricted irrigation of crops that are eaten uncooked.     

Coral-Associated Bacteria and Their Role in the Biogeochemical Cycling of Sulfur

Marine bacteria play a central role in the degradation of dimethylsulfoniopropionate (DMSP) to dimethyl sulfide (DMS) and acrylic acid, DMS being critical to cloud formation and thereby cooling effects on the climate. High concentrations of DMSP and DMS have been reported in scleractinian coral tissues although, to date, there have been no investigations into the influence of these organic sulfur compounds on coral-associated bacteria. Two coral species, Montipora aequituberculata and Acropora millepora, were sampled and their bacterial communities were characterized by both culture-dependent and molecular techniques. Four genera, Roseobacter, Spongiobacter, Vibrio, and Alteromonas, which were isolated on media with either DMSP or DMS as the sole carbon source, comprised the majority of clones retrieved from coral mucus and tissue 16S rRNA gene clone libraries. Clones affiliated with Roseobacter sp. constituted 28% of the M. aequituberculata tissue libraries, while 59% of the clones from the A. millepora libraries were affiliated with sequences related to the Spongiobacter genus. Vibrio spp. were commonly isolated from DMS and acrylic acid enrichments and were also present in 16S rRNA gene libraries from coral mucus, suggesting that under “normal” environmental conditions, they are a natural component of coral-associated communities. Genes homologous to dddD, and dddL, previously implicated in DMSP degradation, were also characterized from isolated strains, confirming that bacteria associated with corals have the potential to metabolize this sulfur compound when present in coral tissues. Our results demonstrate that DMSP, DMS, and acrylic acid potentially act as nutrient sources for coral-associated bacteria and that these sulfur compounds are likely to play a role in structuring bacterial communities in corals, with important consequences for the health of both corals and coral reef ecosystems.Dimethylsulfoniopropionate (DMSP) is an organic sulfur compound implicated in the formation of clouds via its cleavage product dimethyl sulfide (DMS) and therefore has the potential to exert major cooling effects on climate (9, 38). The production of DMSP is mainly restricted to a few classes of marine macro- and microalgae (27, 68), with the main producers being phytoplankton species belonging to prymnesiophyte and dinoflagellate taxa (28, 62, 67). Recently, significant concentrations of DMSP and DMS have been recorded in association with animals that harbor symbiotic algae such as scleractinian corals and giant clams (7, 8, 68), raising questions about the role of coral reefs in sulfur cycling. The densities of symbiotic dinoflagellates (genus Symbiodinium, commonly known as zooxanthellae) in coral tissues are similar to those recorded for dinoflagellates in phytoplankton blooms (11, 68). Since dinoflagellates are among the most significant producers of DMSP and high intracellular concentrations of DMSP have been found in both cultured zooxanthellae (26) and scleractinian corals (6-8, 25), these observations suggest that endosymbiotic zooxanthellae have an integral role in sulfur cycling in oligotrophic reef waters.Most of the DMSP produced by planktonic dinoflagellates is exuded into the surrounding water, where it is degraded by bacteria via two possible pathways: the first one converts a large fraction (ca. 75%) of dissolved DMSP to methylmercaptopropionate, which is subsequently incorporated into the biomass of microbial cells (22, 27, 66). The second pathway transforms the remaining part of the dissolved DMSP to equimolar concentrations of DMS and acrylic acid (43, 66, 72). This metabolic pathway for DMSP degradation has been identified in the alphaproteobacterial species Sulfitobacter sp. and the enzyme involved (DMSP-dependent DMS lyase [DddL]) characterized (10). Another pathway for DMS formation (without production of acrylate) has been described for Marinomonas sp. and the gene responsible, dddD, identified. In addition, the protein DddR has been directly implicated in the regulation of the gene encoding DddD (66). The DMS produced by these enzymes are then released into the surrounding water (27). Prior to the 1980s, diffusion of supersaturated DMS from the oceans to the atmosphere was thought to be the major removal pathway of this compound from the oceans (35, 72). More recently, however, it has been estimated that between 50 and 80% of the DMS produced by DMSP-degrading bacteria is degraded directly by other types of bacteria (58, 59), although the populations and metabolic pathways involved in the degradation of DMS are still poorly understood.Coral-associated bacterial communities are known to be diverse and highly abundant (12, 30, 48, 49, 52). These dynamic communities exploit a number of habitats associated with corals, including mucus on coral surfaces (48), intracellular niches within coral tissues (3, 16, 45, 47, 52), spaces within coral skeletons (15, 51), and seawater surrounding corals (16, 61). Each of these habitats is believed to harbor different bacterial populations (4, 52). Despite high bacterial diversity, corals have been reported to harbor species-specific microbial communities for beneficial effects; however, their role in coral health is poorly understood (47-50). In coral reef environments, bacteria are dependent upon organic compounds produced by photoautotrophic organisms such as endosymbiotic zooxanthellae (48); therefore, photosynthates translocated to coral tissues and mucus may determine microbial communities closely associated with corals (48, 52). The high levels of DMSP and DMS produced by corals, coupled with the dependence of DMSP and DMS conversion on processes typically involving bacteria, suggest that corals are likely to harbor bacterial species involved in the cycling of these compounds. To investigate the potential of the organosulfur compound DMSP and its breakdown products, DMS and acrylic acid, to drive coral-associated microbial communities, we used these compounds as sole carbon sources to isolate bacteria from two coral species (Montipora aequituberculata and Acropora millepora) and then directly compared these microbial communities with coral-associated microbiota identified using culture-independent analyses. Genes implicated in the metabolism of DMSP were also characterized from isolated strains, confirming that bacteria associated with corals have the potential to metabolize organic sulfur compounds present in coral tissues.     

Determination of Manganese Stability in a Constructed Wetland Sediment Using Redox Gel Probes

Redox gel probes containing immobilized particulate manganese compounds (MnO 2 , MnCO 3 , and MnS) were placed on a surveyed grid in the sediment of a wetland receiving coal mine drainage in western Pennsylvania (USA). The stability of these compounds in the wetland was shown to be highly variable both temporally and spatially, indicating that apparent manganese removal based on water quality data did not result in long-term manganese retention in sediments. Contour maps of the gel probe data revealed the importance of local environmental conditions, such as surface water velocity, on geochemical conditions influencing manganese compound stability in sediments, as well as seasonal changes in the ability of the wetland to retain MnO 2 in sediments. Estimates of in situ MnO 2 reduction rates using gel probe data agree with earlier published estimates based on laboratory studies. Although the factors influencing particulate metal stability in sediments are extremely complex and difficult to study, the redox gel probe method is demonstrated to be a cost-effective means of obtaining an areal and depth-related picture of that stability during a particular period of time.     

Activity and Diversity of Sulfate-Reducing Bacteria in a Petroleum Hydrocarbon-Contaminated Aquifer

Microbial sulfate reduction is an important metabolic activity in petroleum hydrocarbon (PHC)-contaminated aquifers. We quantified carbon source-enhanced microbial SO₄²⁻ reduction in a PHC-contaminated aquifer by using single-well push-pull tests and related the consumption of sulfate and added carbon sources to the presence of certain genera of sulfate-reducing bacteria (SRB). We also used molecular methods to assess suspended SRB diversity. In four consecutive tests, we injected anoxic test solutions (1,000 liters) containing bromide as a conservative tracer, sulfate, and either propionate, butyrate, lactate, or acetate as reactants into an existing monitoring well. After an initial incubation period, 1,000 liters of test solution-groundwater mixture was extracted from the same well. Average total test duration was 71 h. We measured concentrations of bromide, sulfate, and carbon sources in native groundwater as well as in injection and extraction phase samples and characterized the SRB population by using fluorescence in situ hybridization (FISH) and denaturing gradient gel electrophoresis (DGGE). Enhanced sulfate reduction concomitant with carbon source degradation was observed in all tests. Computed first-order rate coefficients ranged from 0.19 to 0.32 day⁻¹ for sulfate reduction and from 0.13 to 0.60 day⁻¹ for carbon source degradation. Sulfur isotope fractionation in unconsumed sulfate indicated that sulfate reduction was microbially mediated. Enhancement of sulfate reduction due to carbon source additions in all tests and variability of rate coefficients suggested the presence of specific SRB genera and a high diversity of SRB. We confirmed this by using FISH and DGGE. A large fraction of suspended bacteria hybridized with SRB-targeting probes SRB385 plus SRB385-Db (11 to 24% of total cells). FISH results showed that the activity of these bacteria was enhanced by addition of sulfate and carbon sources during push-pull tests. However, DGGE profiles indicated that the bacterial community structure of the dominant species did not change during the tests. Thus, the combination of push-pull tests with molecular methods provided valuable insights into microbial processes, activities, and diversity in the sulfate-reducing zone of a PHC-contaminated aquifer.     

Influence of Plants and Organic Matter on the Nitrogen Removal in Laboratory‐Scale Model Subsurface Flow Constructed Wetlands Inoculated with Anaerobic Ammonium Oxidizing Bacteria

Anaerobic oxidation of ammonium has become an alternative for the treatment of wastewater with high ammonium loads, and it was also suggested to be involved in the nitrogen removal process in constructed wetlands. Nonetheless, its role has not been well evaluated as yet. In this paper, results of a lab‐scale study are presented focusing on the evaluation of the role of Anammox bacteria, plants, applied ammonia, nitrite nitrogen loads, and the presence of organic matter in nitrogen transformation processes in subsurface‐flow constructed wetlands. The inoculation of the experimental model wetlands with active Anammox biomass increased the total nitrogen and ammonium removal rates to values up to 5.7 g N/m² d, which is almost 10 times higher than those values reported for subsurface flow constructed wetlands. Although the presence of plants caused a higher removal rate, the role of the plants became less important with high nitrite influent concentration. Because the unplanted experimental system without the addition of any organic carbon source showed also high nitrogen removal rates, it can be concluded that beside the potential for “conventional” denitrification in the planted systems the main mechanism for explaining the high nitrogen removal rates obtained during the experiments was the anaerobic ammonia oxidation. The assay of the formation of hydrazine from hydroxylamine and the findings of the molecular biology tests fitted with the positive results for potential Anammox activity obtained in the bottle test. The addition of organic carbon, specifically acetate, apparently had no great influence on Anammox activity, which is in agreement with the findings reported by other authors. Nevertheless, the addition influenced the redox potential. Some questions are still left open, which are mainly associated with the scaling up of these results and the inoculation of Anammox biomass in full‐scale systems.     

Phosphorus release from sediments in a treatment wetland: Contrast between DET and EPC0 methodologies

Wetlands are capable of reducing nutrient loadings to receiving water bodies, and hence many artificial wetlands have been constructed for wastewater nutrient removal. In this study, diffusive equilibrium in thin films (DETs) and equilibrium phosphorus concentration (EPC₀) analysis were used to examine the role of sediment as a nutrient source or sink in a constructed treatment wetland in summer. The effect of dredging on sediment-water nutrient exchange was also studied. Soluble reactive phosphorus (SRP), ammonium (NH₄⁺) and sulphate (SO₄²⁻) concentration profiles were measured by DET across the sediment-water interface (SWI) in both a settling pond and iris reed bed within the wetland. The SRP concentrations in the sediment pore-waters of the settling pond were extremely high (up to 29,500 μg l⁻¹) near the SWI. This is over an order of magnitude higher than the levels found in the water column, which in turn are over an order of magnitude higher than environmental levels proposed to limit eutrophication in rivers. The profiles demonstrated an average net release of SRP and NH₄⁺ from the settling pond sediment to the overlying water of 58 mg m⁻² d⁻¹ (±32 mg m⁻² d⁻¹ (1 sd)) and 16 mg m⁻² d⁻¹ (±25 mg m⁻² d⁻¹ (1 sd)), respectively. The DET SO₄²⁻ concentration profiles revealed that the sediment was anoxic within 2 cm of the SWI. Dredging of the reed bed made no significant difference to the P release characteristics across the SWI. The EPC₀s were much lower than the SRP concentration of the overlying water, indicating that the sediment had the potential to act as a phosphate sink. The apparent contradiction of the DET and EPC₀ results is attributed to the fact that DET measurements are made in situ, where as EPC₀ measurements are ex situ. These results show that substantial releases of P can occur from wetland sediments, and also highlight the need for caution when interpreting ex situ EPC₀ analytical results.     

Influence of Season and Plant Species on the Abundance and Diversity of Sulfate Reducing Bacteria and Ammonia Oxidizing Bacteria in Constructed Wetland Microcosms

Constructed wetlands offer an effective means for treatment of wastewater from a variety of sources. An understanding of the microbial ecology controlling nitrogen, carbon and sulfur cycles in constructed wetlands has been identified as the greatest gap for optimizing performance of these promising treatment systems. It is suspected that operational factors such as plant types and hydraulic operation influence the subsurface wetland environment, especially redox, and that the observed variation in effluent quality is due to shifts in the microbial populations and/or their activity. This study investigated the biofilm associated sulfate reducing bacteria and ammonia oxidizing bacteria (using the dsrB and amoA genes, respectively) by examining a variety of surfaces within a model wetland (gravel, thick roots, fine roots, effluent), and the changes in activity (gene abundance) of these functional groups as influenced by plant species and season. Molecular techniques were used including quantitative PCR and denaturing gradient gel electrophoresis (DGGE), both with and without propidium monoazide (PMA) treatment. PMA treatment is a method for excluding from further analysis those cells with compromised membranes. Rigorous statistical analysis showed an interaction between the abundance of these two functional groups with the type of plant and season (p?<?0.05). The richness of the sulfate reducing bacterial community, as indicated by DGGE profiles, increased in planted vs. unplanted microcosms. For ammonia oxidizing bacteria, season had the greatest impact on gene abundance and diversity (higher in summer than in winter). Overall, the primary influence of plant presence is believed to be related to root oxygen loss and its effect on rhizosphere redox.     

扎龙湿地昆虫群落结构及动态

湿地是介于水体与陆地之间的特殊的生态系统,其中昆虫扮演着重要的角色。通过选取扎龙村、烟筒屯、土木台和育苇场为样地以诱集夜间活动的昆虫为主,对扎龙湿地昆虫群落结构进行了探讨。结果表明:扎龙湿地夜间活动的昆虫分属14目54科139种,以鳞翅目、鞘翅目和双翅目为优势类群。各区域昆虫群落种-多度关系均表现为对数正态分布。物种丰富度为扎龙村＞烟筒屯＞育苇场＞土木台,而群落多样性和均匀度均为烟筒屯＞扎龙村＞土木台＞育苇场,Shannon-Wiener 多样性指数(H') 与均匀度(J')和物种丰富度(S)时间动态关系表现为:烟筒屯Shannon-Wiener 多样性指数(H')与均匀度(J')和物种丰富度(S)均一致;扎龙村和育苇场Shannon-Wiener 多样性指数(H')与均匀度(J')一致,而与物种丰富度(S)弱相关;土木台 Shannon-Wiener 多样性指数(H')与均匀度(J')和物种丰富度(S)均表现为弱相关。研究得出扎龙湿地总体环境质量较好,但局部地区(如土木台)有退化的趋势,并分析造成上述结果的原因。     

Growth of Anaerobic Methane-Oxidizing Archaea and Sulfate-Reducing Bacteria in a High-Pressure Membrane Capsule Bioreactor

Communities of anaerobic methane-oxidizing archaea (ANME) and sulfate-reducing bacteria (SRB) grow slowly, which limits the ability to perform physiological studies. High methane partial pressure was previously successfully applied to stimulate growth, but it is not clear how different ANME subtypes and associated SRB are affected by it. Here, we report on the growth of ANME-SRB in a membrane capsule bioreactor inoculated with Eckernförde Bay sediment that combines high-pressure incubation (10.1 MPa methane) and thorough mixing (100 rpm) with complete cell retention by a 0.2-μm-pore-size membrane. The results were compared to previously obtained data from an ambient-pressure (0.101 MPa methane) bioreactor inoculated with the same sediment. The rates of oxidation of labeled methane were not higher at 10.1 MPa, likely because measurements were done at ambient pressure. The subtype ANME-2a/b was abundant in both reactors, but subtype ANME-2c was enriched only at 10.1 MPa. SRB at 10.1 MPa mainly belonged to the SEEP-SRB2 and Eel-1 groups and the Desulfuromonadales and not to the typically found SEEP-SRB1 group. The increase of ANME-2a/b occurred in parallel with the increase of SEEP-SRB2, which was previously found to be associated only with ANME-2c. Our results imply that the syntrophic association is flexible and that methane pressure and sulfide concentration influence the growth of different ANME-SRB consortia. We also studied the effect of elevated methane pressure on methane production and oxidation by a mixture of methanogenic and sulfate-reducing sludge. Here, methane oxidation rates decreased and were not coupled to sulfide production, indicating trace methane oxidation during net methanogenesis and not anaerobic methane oxidation, even at a high methane partial pressure.     

N-Substituted indole-3-thiolate [4Fe–4S] clusters with a unique and tunable combination of spectral and redox properties

A series of N-substituted indole-3-thiols, synthesized by sequential alkylation, thiouronium salt formation, and hydrolysis, are used to generate a novel family of [4Fe–4S] clusters. The redox transitions of the clusters deviate from those of other [4Fe–4S] cluster families, with half-wave potentials lying in a range midway between those of [4Fe–4S] clusters bound by aliphatic thiolate ligands and those bound by thiophenolate-based ligands. In UV–vis spectroscopy, the new cluster family shows absorption maxima that are among the most red-shifted reported thus far in [4Fe–4S] cluster chemistry. The indole-3-thiolate ligand thus leads to a highly specific and uncommon combination of [4Fe–4S] cluster properties, which can be fine-tuned by facile derivatization at the indole nitrogen atom.     

Identification, isolation, and analysis of a gene cluster involved in iron acquisition by Pseudomonas mendocina ymp

Microbial acquisition of iron from natural sources in aerobic environments is a little-studied process that may lead to mineral instability and trace metal mobilization. Pseudomonas mendocina ymp was isolated from the Yucca Mountain Site for long-term nuclear waste storage. Its ability to solubilize a variety of Fe-containing minerals under aerobic conditions has been previously investigated but its molecular and genetic potential remained uncharacterized. Here, we have shown that the organism produces a hydroxamate and not a catecholate-based siderophore that is synthesized via non-ribosomal peptide synthetases. Gene clustering patterns observed in other Pseudomonads suggested that hybridizing multiple probes to the same library could allow for the identification of one or more clusters of syntenic siderophore-associated genes. Using this approach, two independent clusters were identified. An unfinished draft genome sequence of P. mendocina ymp indicated that these mapped to two independent contigs. The sequenced clusters were investigated informatically and shown to contain respectively a potentially complete set of genes responsible for siderophore biosynthesis, uptake, and regulation, and an incomplete set of genes with low individual homology to siderophore-associated genes. A mutation in the cluster’s pvdA homolog (pmhA) resulted in a siderophore-null phenotype, which could be reversed by complementation. The organism likely produces one siderophore with possibly different isoforms and a peptide backbone structure containing seven residues (predicted sequence: Acyl-Asp-Dab-Ser-fOHOrn-Ser-fOHorn). A similar approach could be applied for discovery of Fe− and siderophore-associated genes in unsequenced or poorly annotated organisms.     

Diversity and Characterization of Sulfate-Reducing Bacteria in Groundwater at a Uranium Mill Tailings Site

Microbially mediated reduction and immobilization of U(VI) to U(IV) plays a role in both natural attenuation and accelerated bioremediation of uranium-contaminated sites. To realize bioremediation potential and accurately predict natural attenuation, it is important to first understand the microbial diversity of such sites. In this paper, the distribution of sulfate-reducing bacteria (SRB) in contaminated groundwater associated with a uranium mill tailings disposal site at Shiprock, N.Mex., was investigated. Two culture-independent analyses were employed: sequencing of clone libraries of PCR-amplified dissimilatory sulfite reductase (DSR) gene fragments and phospholipid fatty acid (PLFA) biomarker analysis. A remarkable diversity among the DSR sequences was revealed, including sequences from δ-Proteobacteria, gram-positive organisms, and the Nitrospira division. PLFA analysis detected at least 52 different mid-chain-branched saturate PLFA and included a high proportion of 10me16:0. Desulfotomaculum and Desulfotomaculum-like sequences were the most dominant DSR genes detected. Those belonging to SRB within δ-Proteobacteria were mainly recovered from low-uranium (≤302 ppb) samples. One Desulfotomaculum-like sequence cluster overwhelmingly dominated high-U (>1,500 ppb) sites. Logistic regression showed a significant influence of uranium concentration over the dominance of this cluster of sequences (P = 0.0001). This strong association indicates that Desulfotomaculum has remarkable tolerance and adaptation to high levels of uranium and suggests the organism's possible involvement in natural attenuation of uranium. The in situ activity level of Desulfotomaculum in uranium-contaminated environments and its comparison to the activities of other SRB and other functional groups should be an important area for future research.     

Growth and Population Dynamics of Anaerobic Methane-Oxidizing Archaea and Sulfate-Reducing Bacteria in a Continuous-Flow Bioreactor

The consumption of methane in anoxic marine sediments is a biogeochemical phenomenon mediated by two archaeal groups (ANME-1 and ANME-2) that exist syntrophically with sulfate-reducing bacteria. These anaerobic methanotrophs have yet to be recovered in pure culture, and key aspects of their ecology and physiology remain poorly understood. To characterize the growth and physiology of these anaerobic methanotrophs and the syntrophic sulfate-reducing bacteria, we incubated marine sediments using an anoxic, continuous-flow bioreactor during two experiments at different advective porewater flow rates. We examined the growth kinetics of anaerobic methanotrophs and Desulfosarcina-like sulfate-reducing bacteria using quantitative PCR as a proxy for cell counts, and measured methane oxidation rates using membrane-inlet mass spectrometry. Our data show that the specific growth rates of ANME-1 and ANME-2 archaea differed in response to porewater flow rates. ANME-2 methanotrophs had the highest rates in lower-flow regimes (μ_ANME-2 = 0.167 · week⁻¹), whereas ANME-1 methanotrophs had the highest rates in higher-flow regimes (μ_ANME-1 = 0.218 · week⁻¹). In both incubations, Desulfosarcina-like sulfate-reducing bacterial growth rates were approximately 0.3 · week⁻¹, and their growth dynamics suggested that sulfate-reducing bacterial growth might be facilitated by, but not dependent upon, an established anaerobic methanotrophic population. ANME-1 growth rates corroborate field observations that ANME-1 archaea flourish in higher-flow regimes. Our growth and methane oxidation rates jointly demonstrate that anaerobic methanotrophs are capable of attaining substantial growth over a range of environmental conditions used in these experiments, including relatively low methane partial pressures.