Availability Of Electron Donors Research Articles

Fluid chemistry and impact of different operating modes on microbial community at Neubrandenburg heat storage (Northeast German Basin)

The efficient use of energy is an important issue of public interest. In the Neubrandenburg heat storage surplus heat from a gas and steam cogeneration plant is stored in an aquifer system for use of the stored energy during times of high heat demand. The reliability of such a plant can strongly be influenced by microbial communities. Therefore, biogeochemical monitoring of the heat storage of Neubrandenburg was conducted from March 2006 to January 2010 to characterize the natural variability of prokaryotic life by way of phospholipid fatty acid (PLFA) analysis and the availability of electron acceptors (e.g. sulfate) and donors [e.g. dissolved organic carbon (DOC)] under different operating modes of the plant. Analysis of the fluid chemistry showed that a sufficient amount of electron acceptors (sulfate ca. 1g/l) and donors (DOC up to 19mg/l) for potential microbial respiration and energy consumption is present. Phospholipid analysis of filter samples from the heat storage revealed a viable microbial community in the plant with adaptation to changes in the operating mode (charge/discharge) and associated variation in temperature (45–73°C). The PLFAs mainly influenced were saturated and branched FAs, most likely reflecting temperature adaptation by a variable microbial community in different parts of the heat storage. Furthermore, branched monoenoic FAs indicated the presence of sulfate-reducing bacteria within the plant.

FdhTU-Modulated Formate Dehydrogenase Expression and Electron Donor Availability Enhance Recovery ofCampylobacter jejunifollowing Host Cell Infection

Campylobacter jejuni is a food-borne bacterial pathogen that colonizes the intestinal tract and causes severe gastroenteritis. Interaction with host epithelial cells is thought to enhance severity of disease, and the ability of C. jejuni to modulate its metabolism in different in vivo and environmental niches contributes to its success as a pathogen. A C. jejuni operon comprising two genes that we designated fdhT (CJJ81176_1492) and fdhU (CJJ81176_1493) is conserved in many bacterial species. Deletion of fdhT or fdhU in C. jejuni resulted in apparent defects in adherence and/or invasion of Caco-2 epithelial cells when assessed by CFU enumeration on standard Mueller-Hinton agar. However, fluorescence microscopy indicated that each mutant invaded cells at wild-type levels, instead suggesting roles for FdhTU in either intracellular survival or postinvasion recovery. The loss of fdhU caused reduced mRNA levels of formate dehydrogenase (FDH) genes and a severe defect in FDH activity. Cell infection phenotypes of a mutant deleted for the FdhA subunit of FDH and an ΔfdhU ΔfdhA double mutant were similar to those of a ΔfdhU mutant, which likewise suggested that FdhU and FdhA function in the same pathway. Cell infection assays followed by CFU enumeration on plates supplemented with sodium sulfite abolished the ΔfdhU and ΔfdhA mutant defects and resulted in significantly enhanced recovery of all strains, including wild type, at the invasion and intracellular survival time points. Collectively, our data indicate that FdhTU and FDH are required for optimal recovery following cell infection and suggest that C. jejuni alters its metabolic potential in the intracellular environment.

Chlorate reduction capacity and characterisation of chlorate reducing bacteria communities in sediments of the rio Cruces wetland in southern Chile

This study investigated chlorate reduction kinetics in multiple samples of sediments from a longitudinal profile of a wetland located downstream of the effluent discharge of a cellulose plant, including characterisation of the bacterial communities involved. The sediments were exposed to different initial chlorate concentrations in microcosm tests, with and without the addition of acetate as an external electron donor, and in a matrix of natural water or a defined medium. At a high initial chlorate concentration of 100 mg/L, in the absence of an external electron source, the degradation curves presented first-order kinetics, influenced by electron donor availability. The first-order kinetic constant varied between 0.05 and 0.17 day−1. Subsequently, when the initial chlorate concentration was reduced to 7 mg/L, a zero-order kinetic was obtained, with the kinetic constant presenting values between 1.1 and 1.3 mg/L-day. No correlation was observed between chlorate degradation kinetics and the location of the sampling points or the previous history of exposure to chlorate. Other factors evaluated, such as the availability of organic matter or the chlorate reducing bacteria count, also proved not to have any incidence on the results. The richness of chlorate reducing bacteria species in the different samples analysed were also similar, with the greatest similarity being found between cld genes in the samples from the upstream or downstream sampling points. Additionally, cld genes most similar to those present in PCRB like Dechlorospirillum sp., Alicycliphilus denitrificans, Dechloromonas agitata, Dechloromonas sp. LT1 and Ideonella dechloratans were detected. This study showed that the anaerobic sediments of the Cruces river wetland present a high potential for chlorate natural attenuation, regardless of the previous history of exposure to chlorate. This capacity is associated with the presence of a diverse community of chlorate reducing bacteria.

Sulfur bacteria in wastewater stabilization ponds periodically affected by the ‘red-water’ phenomenon

Several wastewater stabilization ponds (WSP) in Tunisia suffer periodically from the ‘red-water’ phenomenon due to blooming of purple sulfur bacteria, indicating that sulfur cycle is one of the main element cycles in these ponds. In this study, we investigated the microbial diversity of the El Menzeh WSP and focused in particular on the different functional groups of sulfur bacteria. For this purpose, we used denaturing gradient gel electrophoresis of PCR-amplified fragments of the 16S rRNA gene and of different functional genes involved in microbial sulfur metabolism (dsrB, aprA, and pufM). Analyses of the 16S rRNA revealed a relatively high microbial diversity where Proteobacteria, Chlorobi, Bacteroidetes, and Cyanobacteria constitute the major bacterial groups. The dsrB and aprA gene analysis revealed the presence of deltaproteobacterial sulfate-reducing bacteria (i.e., Desulfobacter and Desulfobulbus), while the analysis of 16S rRNA, aprA, and pufM genes assigned the sulfur-oxidizing bacteria community to the photosynthetic representatives belonging to the Chlorobi (green sulfur bacteria) and the Proteobacteria (purple sulfur and non sulfur bacteria) phyla. These results point on the diversity of the metabolic processes within this wastewater plant and/or the availability of sulfate and diverse electron donors.Electronic supplementary materialThe online version of this article (doi:10.1007/s00253-012-3931-5) contains supplementary material, which is available to authorized users.

O2 reduction and denitrification rates in shallow aquifers

O2 reduction and denitrification rates were determined in shallow aquifers of 12 study areas representing a wide range in sedimentary environments and climatic conditions. Zero‐ and first‐order rates were determined by relating reactant or product concentrations to apparent groundwater age. O2 reduction rates varied widely within and between sites, with zero‐order rates ranging from <3 μmol L−1 yr−1 to more than 140 μmol L−1 yr−1 and first‐order rates ranging from 0.02 to 0.27 yr−1. Moderate denitrification rates (10–100 μmol N L−1 yr−1; 0.06–0.30 yr−1) were observed in most areas with O2 concentrations below 60 μmol L−1, while higher rates (>100 μmol N L−1 yr−1; >0.36 yr−1) occur when changes in lithology result in a sharp increase in the supply of electron donors. Denitrification lag times (i.e., groundwater travel times prior to the onset of denitrification) ranged from <20 yr to >80 yr. The availability of electron donors is indicated as the primary factor affecting O2 reduction rates. Concentrations of dissolved organic carbon (DOC) and/or sulfate (an indicator of sulfide oxidation) were positively correlated with groundwater age at sites with high O2 reduction rates and negatively correlated at sites with lower rates. Furthermore, electron donors from recharging DOC are not sufficient to account for appreciable O2 and nitrate reduction. These relations suggest that lithologic sources of DOC and sulfides are important sources of electrons at these sites but surface‐derived sources of DOC are not. A review of published rates suggests that denitrification tends to occur more quickly when linked with sulfide oxidation than with carbon oxidation.

Assessing the mechanisms controlling the mobilization of arsenic in the arsenic contaminated shallow alluvial aquifer in the blackfoot disease endemic area

High levels of arsenic in groundwater and drinking water represent a major health problem worldwide. Drinking arsenic-contaminated groundwater is a likely cause of blackfoot disease (BFD) in Taiwan, but mechanisms controlling the mobilization of arsenic present at elevated concentrations within aquifers remain understudied. Microcosm experiments using sediments from arsenic contaminated shallow alluvial aquifers in the blackfoot disease endemic area showed simultaneous microbial reduction of Fe(III) and As(V). Significant soluble Fe(II) (0.23±0.03mM) in pore waters and mobilization of As(III) (206.7±21.2nM) occurred during the first week. Aqueous Fe(II) and As(III) respectively reached concentrations of 0.27±0.01mM and 571.4±63.3nM after 8 weeks. We also showed that the addition of acetate caused a further increase in aqueous Fe(II) but the dissolved arsenic did not increase. We further isolated an As(V)-reducing bacterium native to aquifer sediments which showed that the direct enzymatic reduction of As(V) to the potentially more-soluble As(III) in pore water is possible in this aquifer. Our results provide evidence that microorganisms can mediate the release of sedimentary arsenic to groundwater in this region and the capacity for arsenic release was not limited by the availability of electron donors in the sediments.

Electron donor availability for microbial reductive processes following thermal treatment

Thermal treatment is capable of removing significant free-phase chlorinated solvent mass while potentially enhancing bioremediation effectiveness by establishing temperature gradients in the perimeter of the source zone and by increasing electron donor availability. The objectives of this study were to determine the potential for enhanced reductive dechlorination activity at the intermediate temperatures that establish in the perimeter of the heated source zone, and to evaluate the effect of electron donor competition on the performance of the microbial reductive dechlorination process. Microcosms, constructed with tetrachloroethene- (PCE-) and trichloroethene- (TCE-) impacted soils from the Great Lakes, IL, and Ft. Lewis, WA, sites were incubated at temperatures of 24, 35, 50, 70, and 95 °C for 4 months. Reductive dechlorination did not occur in microcosms incubated at temperatures above 24 °C even though mesophilic PCE-to- cis-1,2-dichloroethene dechlorinators were present in Ft. Lewis soil suggesting electron donor limitations. Five days after cooling the microcosms to 24 °C and bioaugmentation with the methanogenic, PCE-to-ethene-dechlorinating consortium OW, at least 85% of the initial PCE and TCE were dechlorinated, but dechlorination ceased prior to complete conversion to ethene. Subsequent biostimulation with hydrogen gas mitigated the dechlorination stall, and conversion to ethene resumed. The results of this study demonstrated that temperatures >35 °C inhibit reductive dechlorination activity at the Great Lakes and Ft. Lewis sites, and that the majority of reducing equivalents released from the soil matrix during heat treatment are consumed in methanogenesis rather than reductive dechlorination. These observations suggest that bioaugmenting thermal treatment sites with cultures that do not contain methanogens may allow practitioners to realize enhanced dechlorination activity, a potential benefit of coupling thermal treatment with bioremediation.

Electron donors and acceptors influence anaerobic soil organic matter mineralization in tidal marshes

Anaerobic decomposition in wetland soils is carried out by several interacting microbial processes that influence carbon storage and greenhouse gas emissions. To understand the role of wetlands in the global carbon cycle, it is critical to understand how differences in both electron donor (i.e., organic carbon) and terminal electron acceptor (TEA) availability influence anaerobic mineralization of soil organic matter. In this study we manipulated electron donors and acceptors to examine how these factors influence total rates of carbon mineralization and the pathways of microbial respiration (e.g., sulfate reduction versus methanogenesis). Using a field-based reciprocal transplant of soils from brackish and freshwater tidal marshes, in conjunction with laboratory amendments of TEAs, we examined how rates of organic carbon mineralization changed when soils with different carbon contents were exposed to different TEAs. Total mineralization (the sum of CO2+CH4 produced) on a per gram soil basis was greater in the brackish marsh soils, which had higher soil organic matter content; however, on a per gram carbon basis, mineralization was greater in the freshwater soils, suggesting that the quality of carbon inputs from the freshwater plants was higher. Overall anaerobic metabolism was higher for both soil types incubated at the brackish site where SO42− was the dominant TEA. When soils were amended with TEAs in the laboratory, more thermodynamically favorable respiration pathways typically resulted in greater organic matter mineralization (Fe(III) respiration>SO42− reduction>methanogenesis). These results suggest that both electron donors and acceptors play important roles in regulating anaerobic microbial mineralization of soil organic matter.

Methane emission from rice fields as affected by land use change

The purpose of this study was to evaluate how former upland cultivation history affects CH4 emission from rice paddies. We measured CH4 flux, methanogen population and in situ Fe(III) reduction in the rice paddies following three different lengths of time since upland crop (Soybean) cultivation. Results showed that CH4 emissions from long-term rice paddy (19 year's continuous cultivation) were significantly higher than recently converted ones. Temporal dynamics of methanogens on rice roots also varied among the plots, and showed a good correlation with CH4 emission rates. Cumulative Fe(III) reduction acted as the dominant electron acceptor in all plots, accounting for 68–94% of the total electron consumption. Fe(II) concentration was highest in the 19-year plot and lowest in the 1-year plots, indicating lower electron availability in recently converted paddies necessary for Fe reduction and CH4 production. Anoxic laboratory soil incubation also suggested poor availability of electron donors in the recent paddies. Collectively, our results demonstrate that the conversion of upland to paddy rice cultivation significantly affected CH4 emission through changing availability of electron donors, redox status of soil Fe and activity of methanogens, which ultimately caused low CH4 emissions.

Denitrification at pH 4 by a soil‐derived Rhodanobacter‐dominated community

Soil denitrification is a major source of nitrous oxide emission that causes ozone depletion and global warming. Low soil pH influences the relative amount of N₂O produced and consumed by denitrification. Furthermore, denitrification is strongly inhibited in pure cultures of denitrifying microorganisms below pH 5. Soils, however, have been shown to denitrify at pH values as low as pH 3. Here we used a continuous bioreactor to investigate the possibility of significant denitrification at low pH under controlled conditions with soil microorganisms and naturally available electron donors. Significant NO₃⁻ and N₂O reduction were observed for 3 months without the addition of any external electron donor. Batch incubations with the enriched biomass showed that low pH as well as low electron donor availability promoted the relative abundance of N₂O as denitrification end-product. Molecular analysis of the enriched biomass revealed that a Rhodanobacter-like bacterium dominated the community in 16S rRNA gene libraries as well as in FISH microscopy during the highest denitrification activity in the reactor. We conclude that denitrification at pH 4 with natural electron donors is possible and that a Rhodanobacter species may be one of the microorganisms involved in acidic denitrification in soils.

Plant species traits regulate methane production in freshwater wetland soils

Ecosystem and biogeochemical responses to anthropogenic stressors are the result of complex interactions between plants and microbes. A mechanistic understanding of how plant traits influence microbial processes is needed in order to predict the ecosystem-level effects of natural or anthropogenic change. This is particularly true in wetland ecosystems, where plants alter the availability of both electron donors (e.g., organic carbon) and electron acceptors (e.g., oxygen and ferric iron), thereby regulating the total amount of anaerobic respiration and the production of methane, a highly potent greenhouse gas. In this study, we examined how plant traits associated with plant inputs of carbon (photosynthesis and biomass) and oxygen (root porosity and ferric iron on roots) to mineral soils relate to microbial competition for organic carbon and, ultimately, methane production. Plant productivity was positively correlated with microbial respiration and negatively correlated to methane production. Root porosity was relatively constant across plant species, but belowground biomass, total biomass, and the concentration of oxidized (ferric) iron on roots varied significantly between species. As a result the size of the total root oxidized iron pool varied considerably across plant species, scaling with plant productivity. Large pools of oxidized iron were related to high CO2:CH4 ratios during microbial respiration, indicating that as plant productivity and biomass increased, microbes used non-methanogenic respiration pathways, most likely including the reduction of iron oxides. Taken together these results suggest that increased oxygen input from plants with greater biomass can offset any potential stimulation of methanogenic microbes from additional carbon inputs. Because the species composition of plant communities influences both electron donor and acceptor availability in wetland soils, changes in plant species as a consequence of anthropogenic disturbance have the potential to trigger profound effects on microbial processes, including changes in anaerobic decomposition rates and the proportion of mineralized carbon emitted as the greenhouse gas methane.

Redox evolution in glacial drift aquifers: role of diamicton units in reduction of Fe(III)

High iron concentrations create water quality problems for municipal use in glacial drift aquifer units. The chemical evolution of oxic groundwater in shallow aquifer units to anoxic groundwater in deeper aquifer units, in which soluble Fe(II) is stable, is attributed to coupled reduction of Fe(III) on aquifer solids with oxidation of organic carbon. The objective of this study was to characterize the distribution of organic carbon in aquifer and aquitard sediments to determine the availability of potential electron donors to drive these reactions. To do this, four complete rotasonic cores in a glacial aquifer/aquitard system were sampled at close intervals for analyses of grain-size distribution and organic carbon content. The results indicate significantly higher organic carbon concentrations in diamicton (till) units that function as aquitards, relative to coarse-grained aquifer units. In addition, readily reducible iron content in the diamicton units and lower aquifer unit materials is sufficient to produce far more dissolved iron than is present in the aquifer. Groundwater evolves to the level of iron reduction as a terminal electron-accepting process as it moves downward through aquitard units along flow paths from upland recharge areas to downgradient discharge areas. Deeper aquifer units are therefore unlikely to contain groundwater with low iron concentration.

Characterization of an enrichment culture debrominating tetrabromobisphenol A and optimization of its activity under anaerobic conditions

To study the effects of incubation conditions on the microbial community structure and activity of a TBBPA-debrominating enrichment culture composed of bacterial and archaeal species. The effects of the methanogen inhibitor 2-bromoethanesulfonate (BES), of the antibiotic ampicillin, of substrate (tetrabromobisphenol A, TBBPA) omission and availability of different electron donors on microbial community structure and activity were examined under anaerobic conditions. Debromination of TBBPA was blocked in the presence of ampicillin, while long-term incubation with BES resulted in delayed debromination activity. The results suggest that the bacterial species responsible for the debromination of TBBPA, while archaeal species involved in electron donor metabolism. The enrichment culture lost its debromination activity after cultivation for 9 months without TBBPA, concomitantly with the disappearance of two DNA bands in a denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA gene fragments corresponding to Pelobacter carbinolicus and Sphaerochaeta sp. TQ1 that were present in the original culture. When butyrate was used as an electron donor, TBBPA debromination activity was attenuated. When acetate was used as the electron donor, no debromination was observed and in addition, there was a decrease in the abundance of the mcrA gene. The results indicate that to maintain a high rate of TBBPA debromination activity, it is essential to preserve the microbial community structure (bacterial and archaeal members) of this culture and supply an electron donor that produces high amounts of hydrogen when fermented. The study provides important information for the management of cultures to be used in bioremediation of TBBPA contaminated sites.

Chemolithoautotrophic production mediating the cycling of the greenhouse gases N<sub>2</sub>O and CH<sub>4</sub> in an upwelling ecosystem

Abstract. The high availability of electron donors occurring in coastal upwelling ecosystems with marked oxyclines favours chemoautotrophy, in turn leading to high N2O and CH4 cycling associated with aerobic NH4+ (AAO) and CH4 oxidation (AMO). This is the case of the highly productive coastal upwelling area off central Chile (36° S), where we evaluated the importance of total chemolithoautotrophic vs. photoautotrophic production, the specific contributions of AAO and AMO to chemosynthesis and their role in gas cycling. Chemolithoautotrophy was studied at a time-series station during monthly (2007–2009) and seasonal cruises (January 2008, September 2008, January 2009) and was assessed in terms of the natural C isotopic ratio of particulate organic carbon (δ13POC), total and specific (associated with AAO and AMO) dark carbon assimilation (CA), and N2O and CH4 cycling experiments. At the oxycline, δ13POC averaged −22.2‰; this was significantly lighter compared to the surface (−19.7‰) and bottom layers (−20.7‰). Total integrated dark CA in the whole water column fluctuated between 19.4 and 2.924 mg C m−2 d−1, was higher during active upwelling, and contributed 0.7 to 49.7% of the total integrated autotrophic CA (photo plus chemoautotrophy), which ranged from 135 to 7.626 mg C m−2 d−1, and averaged 20.3% for the whole sampling period. Dark CA was reduced by 27 to 48% after adding a specific AAO inhibitor (ATU) and by 24 to 76% with GC7, a specific archaea inhibitor. This indicates that AAO and AMO microbes (most of them archaea) were performing dark CA through the oxidation of NH4+ and CH4. Net N2O cycling rates varied between 8.88 and 43 nM d−1, whereas net CH4 cycling rates ranged from −0.41 to −26.8 nM d−1. The addition of both ATU and GC7 reduced N2O accumulation and increased CH4 consumption, suggesting that AAO and AMO were responsible, in part, for the cycling of these gases. These findings show that chemically driven chemolithoautotrophy (with NH4+ and CH4 acting as electron donors) could be more important than previously thought in upwelling ecosystems, raising new questions concerning its relevance in the future ocean.

Bioreduction of Nitrobenzene, Natural Organic Matter, and Hematite by Shewanella putrefaciens CN32

We examined the reduction of nitrobenzene by Shewanella putrefaciens CN32 in the presence of natural organic matter (NOM) and hematite. Bioreduction experiments were conducted with combinations and varied concentrations of nitrobenzene, soil humic acid, Georgetown NOM, hematite, and CN32. Abiotic experiments were conducted to quantify nitrobenzene reduction by biogenic Fe(II) and by bioreduced NOMs. We show that S. putrefaciens CN32 can directly reduce nitrobenzene. Both NOMs enhanced nitrobenzene reduction and the degree of enhancement depended on properties of the NOMs (aromaticity, organic radical content). Hematite enhanced nitrobenzene reduction by indirect reaction with biogenic-Fe(II), however, enhancement was dependent on the availability of excess electron donor. Under electron donor-limiting conditions, reducing equivalents diverted to hematite were not all transferred to nitrobenzene. In systems that contained both NOM and hematite we conclude that NOM-mediated reduction of nitrobenzene was more important than Fe(II)-mediated reduction.

Elevated Concentrations of Methyl Mercury in Streams after Forest Clear-Cut: A Consequence of Mobilization from Soil or New Methylation?

Concentrations of inorganic, mercuric mercury (Hg(II)), methyl mercury (MeHg) and ancillary chemistry measured in first-order streams draining 0-4 (N = 20) and 4-10 (N = 27) year-old clear-cuts of former Norway Spruce Picea abies (Karst.) forest stands were compared with concentrations in streams draining >70 year-old Norway Spruce reference stands (N = 10). Concentrations of MeHg, and ratios of MeHg TOC(-1) and Hg(II) TOC(-1), were significantly (p < 0.01) elevated in 0-4 year-old clear-cuts, as compared to references. The only ancillary variable showing a significant elevation for 0-4 year-old clear-cuts was Mn (p < 0.02). The 4-10 year-old clear-cuts showed intermediate concentrations with nonsignificant differences as compared to references. pH, nitrate, sulfate, Ca, Fe, TOC, TON, and the aromaticity of TOC (SUVA(254 nm)) showed nonsignificant differences between clear-cut age classes and references. Assuming that MeHg and Hg(II) are mobilized from soil to stream to a similar relative extent as a consequence of clear-cutting, a calculation showed that (1)/(6) of the elevated MeHg concentration was due to enhanced mobilization from soil and (5)/(6) was due to new methylation of Hg(II) 0-4 years after clear-cut. New methylation after clear-cut is suggested to be stimulated by an increased availability of electron donors for methylating bacteria, as a consequence of degradation of logging residue ("slash") and soil organic matter. A subdivision of sites situated above and below the highest postglacial coastline (HC) revealed a significant elevation of MeHg, MeHg TOC(-1) and Hg(II) TOC(-1) (p < 0.05) beyond their references in 0-4 year-old clear-cuts above (but not below) the HC. This suggests that postglacial deposits of FeS(s) and FeS(2)(s) were not an important factor for elevation of MeHg after clear-cut.

In situ measurements of microbially-catalyzed nitrification and nitrate reduction rates in an ephemeral drainage channel receiving water from coalbed natural gas discharge, Powder River Basin, Wyoming, USA

Nitrification and nitrate reduction were examined in an ephemeral drainage channel receiving discharge from coalbed natural gas (CBNG) production wells in the Powder River Basin, Wyoming. CBNG co-produced water typically contains dissolved inorganic nitrogen (DIN), primarily as ammonium. In this study, a substantial portion of discharged ammonium was oxidized within 50 m of downstream transport, but speciation was markedly influenced by diel fluctuations in dissolved oxygen (> 300 µM). After 300 m of transport, 60% of the initial DIN load had been removed. The effect of benthic nitrogen-cycling processes on stream water chemistry was assessed at 2 locations within the stream channel using acrylic chambers to conduct short-term (2–6 h), in-stream incubations. The highest ambient DIN removal rates (2103 µmol N m − 2 h − 1 ) were found at a location where ammonium concentrations > 350 µM. This occurred during light incubations when oxygen concentrations were highest. Nitrification was occurring at the site, however, net accumulation of nitrate and nitrite accounted for < 12% of the ammonium consumed, indicating that other ammonium-consuming processes were also occurring. In dark incubations, nitrite and nitrate consumption were dominant processes, while ammonium was produced rather than consumed. At a downstream location nitrification was not a factor and changes in DIN removal rates were controlled by nitrate reduction, diel fluctuations in oxygen concentration, and availability of electron donor. This study indicates that short-term adaptation of stream channel processes can be effective for removing CBNG DIN loads given sufficient travel distances, but the long-term potential for nitrogen remobilization and nitrogen saturation remain to be determined.

Systematic evaluation of nitrate and perchlorate bioreduction kinetics in groundwater using a hydrogen-based membrane biofilm reactor

To evaluate the simultaneous reduction kinetics of the oxidized compounds, we treated nitrate-contaminated groundwater (∼9.4 mg-N/L) containing low concentrations of perchlorate (∼12.5 μg/L) and saturated with dissolved oxygen (∼8 mg/L) in a hydrogen-based membrane biofilm reactor (MBfR). We systematically increased the hydrogen availability and simultaneously varied the surface loading of the oxidized compounds on the biofilm in order to provide a comprehensive, quantitative data set with which to evaluate the relationship between electron donor (H 2) availability, surface loading of the electron acceptors (oxidized compounds), and simultaneous bioreduction of the electron acceptors. Increasing the H 2 pressure delivered more H 2 gas, and the total H 2 flux increased linearly from ∼0.04 mg/cm 2-d for 0.5 psig (0.034 atm) to 0.13 mg/cm 2-d for 9.5 psig (0.65 atm). This increased rate of H 2 delivery allowed for continued reduction of the acceptors as their surface loading increased. The electron acceptors had a clear hydrogen-utilization order when the availability of hydrogen was limited: oxygen, nitrate, nitrite, and then perchlorate. Spiking the influent with perchlorate or nitrate allowed us to identify the maximum surface loadings that still achieved more than 99.5% reduction of both oxidized contaminants: 0.21 mg NO 3-N/cm 2-d and 3.4 μg ClO 4/cm 2-d. Both maximum values appear to be controlled by factors other than hydrogen availability.

Chlorobaculum tepidum regulates chlorosome structure and function in response to temperature and electron donor availability

Green sulfur bacteria (GSB) rely on the chlorosome, a light-harvesting apparatus comprised almost entirely of self-organizing arrays of bacteriochlorophyll (BChl) molecules, to harvest light energy and pass it to the reaction center. In Chlorobaculum tepidum, over 97% of the total BChl is made up of a mixture of four BChl c homologs in the chlorosome that differ in the number and identity of alkyl side chains attached to the chlorin ring. C. tepidum has been reported to vary the distribution of BChl c homologs with growth light intensity, with the highest degree of BChl c alkylation observed under low-light conditions. Here, we provide evidence that this functional response at the level of the chlorosome can be induced not only by light intensity, but also by temperature and a mutation that prevents phototrophic thiosulfate oxidation. Furthermore, we show that in conjunction with these functional adjustments, the fraction of cellular volume occupied by chlorosomes was altered in response to environmental conditions that perturb the balance between energy absorbed by the light-harvesting apparatus and energy utilized by downstream metabolic reactions.

Evidence for denitrification regulated by pyrite oxidation in a heterogeneous porous groundwater system

Denitrification is an important natural attenuation process that has been observed in many fissured and porous aquifers. However, an important factor limiting denitrification in aquatic systems is the microbial availability of electron donors. Pyrite as the most abundant sulfide mineral in nature represents one of the potential electron sources for denitrifiers to reduce nitrate, but the reaction mechanisms coupling denitrification processes to pyrite oxidation are still questionable. We utilized hydrochemical data and stable isotopes of nitrate and sulfate in groundwater, isotope ratios of sulfur compounds in aquifer sediments and tritium based groundwater dating for assessing denitrification processes in a pyrite-bearing porous groundwater system. The oxic part of the aquifer with mean water transit times of approximately 60 years was characterized by nitrate concentrations of around 15 mg/l and δ 15N values were similar to those typical for nitrification. In contrast, in the anoxic part with mean water transit times of up to 100 years, low nitrate concentrations accompanied by elevated δ 15N values were observed. Furthermore, isotope data of groundwater sulfate and sulfur compounds in the aquifer sediment suggest that pyrite oxidation is the dominant source of sulfate in the aquifer. The trend of increasing δ 15N values and decreasing nitrate concentrations in concert with depleted δ 34S values of groundwater sulfate similar to δ 34S values of pyrite, FeS 2, suggests that denitrification is coupled to pyrite oxidation, particularly when water mean transit time is elevated.