Methanogenic Sediments Research Articles

Archaeal polar lipids in subseafloor sediments from the Nankai Trough: Implications for the distribution of methanogens in the deep marine subsurface

Although the methane in marine methane hydrates is mainly of microbial origin, information about the distribution of methanogens in subseafloor sediments is limited. To address this issue, we analyzed sediment core samples from two sites in the Nankai Trough, off the Pacific coast of central Japan, including those bearing methane hydrates from depths >100m below the seafloor (mbsf), for isopranyl ether-linked polar lipids (i.e. with polar head groups of phosphate, sugar, or both) as biomarkers of archaea, including methanogens. In most samples, including the deepest (381mbsf), archaeol, and sn-2- and sn-3-hydroxyarchaeols were detected as their hydrolyzed derivatives. Concentrations of these three archaeal lipids correlated strongly with each other, suggesting a common biological source. The δ13C values of phytane derived from the phytanyl groups in the archaeal lipids were distinctly higher than those of methane, indicating that methanogens rather than anaerobic methanotrophic archaea were the major biological source. Depth profiles of polar sn-2-hydroxyarchaeol concentration were consistent with those of the potential methane production activity previously estimated from incubation of core sediments from the same sites. This observation, together with results of previous studies showing the presence of sn-2-hydroxyarchaeol mainly in shallow young sediments, strongly suggests that this polar lipid is a valid biomarker for in situ methanogens in sediments. There was a strong correlation between the concentration of polar sn-2-hydroxyarchaeol and that of total organic carbon, suggesting that bulk organic matter concentration is a primary control on the distribution of methanogens in sediments.

Gas hydrate distribution and carbon sequestration through coupled microbial methanogenesis and silicate weathering in the Krishna–Godavari Basin, offshore India

The National Gas Hydrate Program Expedition 01 cored ten sites in the Krishna–Godavari basin, located on the southeastern margin of India. A comprehensive suite of pore water solute concentrations and isotope ratios were analyzed to investigate the distribution and concentration of gas hydrate along the margin, in situ diagenetic and metabolic reactions, and fluid migration and flow pathways. Gas hydrate was present at all of the sites cored, and in situ microbial methanogenesis leads to estimates of depth-integrated average gas hydrate saturations that are typically <5%. Deep-sourced fluid and gas migration produces gas hydrate saturations up to 68% along an isolated coarser-grained stratigraphic horizon at Site 15 and up to 41% within a fractured clay-dominated system at Site 10. Our results show that the CO2 produced through net microbial methanogenesis is effectively neutralized by silicate weathering throughout the sediment column drilled at each site (∼100–300 m), buffering the pore water pH and generating excess alkalinity via the same reaction sequence as continental silicate weathering. Most of the excess alkalinity produced by silicate weathering in the Krishna–Godavari basin is sequestered in Ca- and Fe-carbonates as a result of ubiquitous calcium release from weathering detrital silicates and dissolved Fe production within the methanogenic sediments. Formation of secondary hydrous silicates (e.g. smectite) related to incongruent primary silicate dissolution acts as a significant sink for pore water Mg, K, Li, Rb, and B. The consumption of methane through anaerobic oxidation of methane, sequestration of methane in gas hydrate, and sequestration of dissolved inorganic carbon in authigenic carbonates keeps methanogenesis as a thermodynamically feasible catabolic pathway. Our results combined with previous indications of silicate weathering in anoxic sediments in the Sea of Okhotsk, suggest that silicate weathering coupled to microbial methanogenesis should be occurring in continental margins worldwide, providing a net sink of atmospheric CO2.

Extreme organic carbon burial fuels intense methane bubbling in a temperate reservoir

Organic carbon (OC) burial and greenhouse gas emission of inland waters plays an increasingly evident role in the carbon balance of the continents, and particularly young reservoirs in the tropics emit methane (CH4) at high rates. Here we show that an old, temperate reservoir acts simultaneously as a strong OC sink and CH4 source, because the high sedimentation rate supplies reactive organic matter to deep, anoxic sediment strata, fuelling methanogenesis and gas bubble emission (ebullition) of CH4from the sediment. Damming of the river has resulted in the build‐up of highly methanogenic sediments under a shallow water column, facilitating the transformation of fixed CO2 to atmospheric CH4. Similar high OC burial and CH4 ebullition is expected in other reservoirs and natural river deltas.

The effect of diagenesis and fluid migration on rare earth element distribution in pore fluids of the northern Cascadia accretionary margin

Analytical challenges in obtaining high quality measurements of rare earth elements (REEs) from small pore fluid volumes have limited the application of REEs as deep fluid geochemical tracers. Using a recently developed analytical technique, we analyzed REEs from pore fluids collected from Sites U1325 and U1329, drilled on the northern Cascadia margin during the Integrated Ocean Drilling Program (IODP) Expedition 311, to investigate the REE behavior during diagenesis and their utility as tracers of deep fluid migration. These sites were selected because they represent contrasting settings on an accretionary margin: a ponded basin at the toe of the margin, and the landward Tofino Basin near the shelf's edge. REE concentrations of pore fluid in the methanogenic zone at Sites U1325 and U1329 correlate positively with concentrations of dissolved organic carbon (DOC) and alkalinity. Fractionations across the REE series are driven by preferential complexation of the heavy REEs. Simultaneous enrichment of diagenetic indicators (DOC and alkalinity) and of REEs (in particular the heavy elements Ho to Lu), suggests that the heavy REEs are released during particulate organic carbon (POC) degradation and are subsequently chelated by DOC. REE concentrations are greater at Site U1325, a site where shorter residence times of POC in sulfate-bearing redox zones may enhance REE burial efficiency within sulfidic and methanogenic sediment zones where REE release ensues. Cross-plots of La concentrations versus Cl, Li and Sr delineate a distinct field for the deep fluids (z > 75 mbsf) at Site U1329, and indicate the presence of a fluid not observed at the other sites drilled on the Cascadia margin. Changes in REE patterns, the presence of a positive Eu anomaly, and other available geochemical data for this site suggest a complex hydrology and possible interaction with the igneous Crescent Terrane, located east of the drilled transect.

Environmental evidence for net methane production and oxidation in putative ANaerobic MEthanotrophic (ANME) archaea

Uncultured ANaerobic MEthanotrophic (ANME) archaea are often assumed to be obligate methanotrophs that are incapable of net methanogenesis, and are therefore used as proxies for anaerobic methane oxidation in many environments in spite of uncertainty regarding their metabolic capabilities. Anaerobic methane oxidation regulates methane emissions in marine sediments and appears to occur through a reversal of a methane-producing metabolism. We tested the assumption that ANME are obligate methanotrophs by detecting and quantifying gene transcription of ANME-1 across zones of methane oxidation versus methane production in sediments from the White Oak River estuary, North Carolina. ANME-1 consistently transcribe 16S rRNA and mRNA of methyl coenzyme M reductase (mcrA), the key gene for methanogenesis, up to 45 cm into methanogenic sediments. CARD-FISH shows that ANME-1 exist as single rod-shaped cells or pairs of cells. Integrating normalized depth distributions of 16S rDNA and rRNA (measured with qPCR and RT-qPCR respectively) shows that 26-77% of the rDNA (a proxy for ANME-1 cell numbers), and 18-76% of the rRNA (a proxy for ANME-1 activity) occurs within methane-producing sediments. These results, along with a re-assessment of the published Iiterature, change the perspective to ANME-1 as methanogens that are also capable of methane oxidation.

The biogeochemistry of sorbed methane in marine sediments

Sorption of volatile hydrocarbon gases (VHCs) to marine sediments is a recognized phenomenon that has been investigated in the context of petroleum exploration. However, little is known about the biogeochemistry of sorbed methane and higher VHCs in environments that are not influenced by thermogenic processes. This study evaluated two different extraction protocols for sorbed VHCs, used high pressure equipment to investigate the sorption of methane to pure clay mineral phases, and conducted a geochemical and mineralogical survey of sediment samples from different oceanographic settings and geochemical regimes that are not significantly influenced by thermogenic gas. Extraction of sediments under alkaline conditions yielded higher concentrations of sorbed methane than the established protocol for acidic extraction. Application of alkaline extraction in the environmental survey revealed the presence of substantial amounts of sorbed methane in 374 out of 411 samples (91%). Particularly high amounts, up to 2.1 mmol kg −1 dry sediment, were recovered from methanogenic sediments. Carbon isotopic compositions of sorbed methane suggested substantial contributions from biogenic sources, both in sulfate-depleted and sulfate-reducing sediments. Carbon isotopic relationships between sorbed and dissolved methane indicate a coupling of the two pools. While our sorption experiments and extraction conditions point to an important role for clay minerals as sorbents, mineralogical analyses of marine sediments suggest that variations in mineral composition are not controlling variations in quantities of sorbed methane. We conclude that the distribution of sorbed methane in sediments is strongly influenced by in situ production.

Silicate weathering in anoxic marine sediments

Two sediment cores retrieved at the northern slope of Sakhalin Island, Sea of Okhotsk, were analyzed for biogenic opal, organic carbon, carbonate, sulfur, major element concentrations, mineral contents, and dissolved substances including nutrients, sulfate, methane, major cations, humic substances, and total alkalinity. Down-core trends in mineral abundance suggest that plagioclase feldspars and other reactive silicate phases (olivine, pyroxene, volcanic ash) are transformed into smectite in the methanogenic sediment sections. The element ratios Na/Al, Mg/Al, and Ca/Al in the solid phase decrease with sediment depth indicating a loss of mobile cations with depth and producing a significant down-core increase in the chemical index of alteration. Pore waters separated from the sediment cores are highly enriched in dissolved magnesium, total alkalinity, humic substances, and boron. The high contents of dissolved organic carbon in the deeper methanogenic sediment sections (50–150mgdm−3) may promote the dissolution of silicate phases through complexation of Al3+ and other structure-building cations. A non-steady state transport-reaction model was developed and applied to evaluate the down-core trends observed in the solid and dissolved phases. Dissolved Mg and total alkalinity were used to track the in-situ rates of marine silicate weathering since thermodynamic equilibrium calculations showed that these tracers are not affected by ion exchange processes with sediment surfaces. The modeling showed that silicate weathering is limited to the deeper methanogenic sediment section whereas reverse weathering was the dominant process in the overlying surface sediments. Depth-integrated rates of marine silicate weathering in methanogenic sediments derived from the model (81.4–99.2mmol CO2m−2year−1) are lower than the marine weathering rates calculated from the solid phase data (198–245mmol CO2m−2year−1) suggesting a decrease in marine weathering over time. The production of CO2 through reverse weathering in surface sediments (4.22–15.0mmol CO2m−2year−1) is about one order of magnitude smaller than the weathering-induced CO2 consumption in the underlying sediments. The evaluation of pore water data from other continental margin sites shows that silicate weathering is a common process in methanogenic sediments. The global rate of CO2 consumption through marine silicate weathering estimated here as 5–20Tmol CO2year−1 is as high as the global rate of continental silicate weathering.

Response of fermentation and sulfate reduction to experimental temperature changes in temperate and Arctic marine sediments

Anaerobic degradation of organic material generally proceeds through a sequence of steps, including polymer hydrolysis, fermentation and respiration or methanogenesis. The intermediates, such as volatile fatty acids (VFA) or H(2), are generally maintained at low concentration, showing a close coupling of the terminal oxidation to fermentation. We exposed marine sediments to extreme temperature perturbations to study the nature and robustness of this coupling. Bacterial sulfate reduction and its dependence on fermentation were studied experimentally over a broad temperature range of -0.3 to 40 degrees C in sediments from temperate and permanently cold environments. In an Arctic sediment from Svalbard, the apparent optimum temperature for sulfate reduction decreased with prolonged incubation, whereas sulfate reduction rates increased. In a temperate sediment from the North Sea, the apparent optimum temperature was higher and did not change with incubation time. Up to a critical temperature, the concentrations of VFA remained low, <3 microM for acetate and <1 microM for the other VFA, the H(2) concentration showed thermodynamic control by sulfate-reducing bacteria, revealing a close coupling of fermentation and sulfate reduction. Above the critical temperature, the concentrations of VFA and H(2) increased transiently by 100-1000-fold. According to the different in situ temperatures of the samples, the critical temperature was lower for sediments from the Arctic than from the North Sea. The H(2) concentrations decreased again upon prolonged incubation to values typical for sulfate-depleted methanogenic sediments. This suggests that fermentative bacteria and methanogenic archaea in both sediments tolerated higher temperatures than the sulfate-reducing community.

Silicate weathering in anoxic marine sediments

Two sediment cores retrieved at the northern slope of Sakhalin Island, Sea of Okhotsk, were analysed for biogenic opal, organic carbon, carbonate, sulphur, major element concentrations, mineral contents and dissolved substances including nutrients, sulphate, methane, major cations, humic substances and total alkalinity. Down-core trends in mineral abundance suggest that plagioclase feldspars and other reactive silicate phases (olivine, pyroxene, volcanic ash) are transformed into smectite in the methanogenic sediment sections. The element ratios Na/Al, Mg/Al and Ca/Al in the solid phase decrease with sediment depth indicating a loss of mobile cations with depth and producing a significant down-core increase in the chemical index of alteration.

Enantioselective Microbial Transformation of the Phenylpyrazole Insecticide Fipronil in Anoxic Sediments

Fipronil, a chiral insecticide, was biotransformed initially to fipronil sulfide in anoxic sediment slurries following a short lag period. Sulfidogenic or methanogenic sediments transformed fipronil with half-lives of approximately 35 and 40 days, respectively. In all microbially active sediment slurries tested, the transformation of fipronil to fipronil sulfide was enantioselective. In the sulfidogenic sediment slurry, the enantiomeric fraction (EF) of fipronil decreased from an initial racemic EF value of 0.46 to a value of 0.22 during the incubation period of active fipronil transformation, indicating preferential transformation of the S-(+)-enantiomer. A previously unidentified product, 5-amino1-[2,6-dichloro-4-(trifluoromethyl)-phenyl]-4-(trifluoromethylthio)-1-H-pyrazole-3-carboxyamide, or fipronil sulfide-amide, was detected in the sulfidogenic slurries and coincided with the loss of fipronil sulfide. Biota from methanogenic freshwater sediment slurries also transformed fipronil enantioselectively but with a preference for the R-(-)-enantiomer. In all microbially inhibited (autoclaved) sediment slurries tested, no changes in the enantiomeric fractions of fipronil were observed and only low levels (< 5% of the added fipronil) of the fipronil sulfide metabolite were detected. In defined (model) chemical experiments, solutions of pyrite (FeS2) and iron sulfide (FeS) non-enantioselectively transformed fipronil primarily to either 2,6-dichloro-4-(trifluoromethyl)-aniline or to fipronil sulfide and fipronil amide, respectively. This report provides the first experimental evidence of enantioselective microbial transformation of fipronil in a natural environment (soil, water, and sediment) as well as identification of a novel fipronil biotransformation product.

Accumulation of dechlorination daughter products: A valid metric of chloroethene biodegradation

AbstractIn situ reductive dechlorination of perchloroethene (PCE) and trichloroethene (TCE) generates characteristic chlorinated (cis‐dichloroethene [cis‐DCE] and vinyl chloride [VC]) and nonchlorinated (ethene and ethane) products. The accumulation of these daughter products is commonly used as a metric for ongoing biodegradation at field sites. However, this interpretation assumes that reductive dechlorination is the only chloroethene degradation process of any significance in situ and that the characteristic daughter products of chloroethene reductive dechlorination persist in the environment. Laboratory microcosms, prepared with aquifer and surface‐water sediments from hydrologically diverse sites throughout the United States and amended with [1,2‐14C] TCE, [1,2‐14C] DCE, [1,2‐14C] DCA, or [1,2‐14C] VC, demonstrated widely variable patterns of intermediate and final product accumulation. In predominantly methanogenic sediment treatments, accumulation of 14C‐DCE, 14C‐VC, 14C‐ethene, and 14C‐ethane predominated. Treatments characterized by significant Fe(III) and/or Mn(IV) reduction, on the other hand, demonstrated substantial, and in some cases exclusive, accumulation of 14CO2 and 14CH4. These results suggest that relying on the accumulation of cis‐DCE, VC, ethene, and ethane may substantially underestimate overall chloroethene biodegradation at many sites. © 2007 Wiley Periodicals, Inc.*This article is a U.S. government work and, as such, is in the public domain in the United States of America.

Community structure of Archaea and Bacteria in a profundal lake sediment Lake Kinneret (Israel)

The microbial community structure of an anoxic profundal lake sediment, i.e., subtropical Lake Kinneret, was analysed with respect to its composition by culture-independent molecular methods including terminal restriction fragment length polymorphism (T-RFLP) analysis, comparative sequence analysis, and quantitative real-time PCR. In particular we were interested in the structure, species composition, and relative abundance of the overall microbial community in the methanogenic sediment layer (0–10 cm depth). Pairwise comparison of archaeal and bacterial 16S rRNA gene T-RFLP profiles obtained from three independent samplings indicated stability of the microbial community. The numbers of Archaea and Bacteria, quantified by real-time PCR, amounted to about 10 8 and 10 10 16S rRNA gene copies cm −3 sediment, respectively, suggesting that Archaea may account for only a minor fraction (approximately 1%) of the total prokaryotic community. Hydrogenotrophic Methanomicrobiales and acetoclastic Methanosaeta spp. dominated T-RFLP profiles of the archaeal community. T-RFLP profiles of the bacterial community were dominated by Deltaproteobacteria, sulphate reducers and syntrophs in particular. The second most abundant group was assigned to the Bacteroidetes–Chlorobi-group. Only one bacterial group, which was affiliated with halorespiring bacteria of subphylum II of the Chloroflexi, showed variation in abundance within the sediment samples investigated. Our study gives a comprehensive insight into the structure of the bacterial and archaeal community of a profundal lake sediment, indicating that sulphate reducers, syntrophs, bacteroidetes, halorespirers and methanogens are of particular importance in Lake Kinneret sediment.

Nitroaromatic Reduction Kinetics as a Function of Dominant Terminal Electron Acceptor Processes in Natural Sediments

The reductive transformation of p-cyanonitrobenzene (pCNB) was investigated in laboratory batch slurries exhibiting dominant terminal electron accepting processes (TEAPs). Pseudo-first-order rate constants (k(obs)) were measured for the reduction of pCNB in nitrate-reducing, iron-reducing, sulfate-reducing, and methanogenic sediment slurries. Reduction was extremely slow in nitrate-reducing slurries but increased in slurries exhibiting TEAPs with significant concentrations of solution phase Fe(ll). As the reduction of pCNB progressed in the Fe(ll) rich systems, significant but nonstoichiometric decreases in aqueous Fe(ll) concentration were measured. Normalization of k(obs) to initial aqueous Fe(ll) concentrations (k(obs)/[Fe(ll)]t=0) gave values ranging from 0.0040 to 0.0052 d(-1) microM(-1) for nitrate-reducing, iron-reducing, and methanogenic sediment slurries as well as sulfate-reducing sediment slurries in which lactate served as a source of organic carbon. The k(obs)/ [Fe(ll)]t=0 ratios were 1-fold greater for sulfate-reducing batch slurries amended with acetate and iron-reducing slurries equilibrated with a 3% H2 atmosphere indicating that the electron source and system parameters such as pH play a determinant role in the reaction kinetics. Although these data demonstrate that aqueous phase Fe(ll) must be present for significant reduction to occur, a limited role for aqueous phase Fe(ll) as a quantitative indicator of reactivity is suggested.

Sediment porewater exchange and solute release during ebullition

The ebullitive exchange of porewater between low-permeability methanogenic sediment and overlying water was measured in the laboratory using fluoride as a tracer. Sediment was equilibrated with fluoride at 3.1 mM, followed by purging with fluoride-free water for 4.5 h to decrease the porewater concentration gradient and diffusive flux. Bubbles with an average volume of about 0.23 ml were injected 4.6 cm below the sediment–water interface at a constant rate of 42 ml/m 2/min for 3 h in the test chambers, while no bubbles were injected in the control chambers. Fluoride concentrations in the overlying water of both test and control chambers increased linearly with time, due to a combination of ebullition and molecular diffusion in the test chambers and molecular diffusion only in the control chambers. The fluoride fluxes in test chambers were about 50% greater than in the controls and the differences are attributed to ebullitive exchange of porewater. The flux in the test chambers correlated strongly ( r 2 = 0.99) with the volumetric rate of bubble release. On a bubble volume basis, the volume of porewater corresponding to the fluoride released was 0.069 ± 0.006 ml porewater/ml bubbles. The volume released per bubble was 0.016 ± 0.003 ml. When ebullition was stopped, flux in the test chamber decreased to a rate that was slightly less than in the control chamber. After accounting for slight differences in concentration gradients, the post-ebullition fluxes in the test and control chambers were nearly identical, indicating that the bubble tube and vent structures do not enhance diffusion in the absence of ebullition. Calculations suggest that molecular diffusive fluxes for near surface solutes are greater than ebullitive fluxes in all but highly methanogenic and ebullitive sediments.

Salt marsh rhizosphere affects microbial biotransformation of the widespread halogenated contaminant tetrabromobisphenol-A (TBBPA)

Estuarine sediments are the repository for a wide range of contaminants. Anthropogenic impacts and variations in the belowground biomass of salt marsh plants potentially select for different sediment microbial communities with different functional capabilities, including the ability to biotransform anthropogenic contaminants. There are large differences in both root morphology and the amount of fine root biomass of Spartina alterniflora and Phragmites australis; Spartina is the species commonly used to replace Phragmites in northeastern US salt marsh restoration projects. Our study compared the effect of these two macrophyte species on sediment microbial communities responsible for the biotransformation of the halogenated flame retardant tetrabromobisphenol A (TBBPA). Sediments were obtained from contaminated and uncontaminated salt marsh field sites in New Jersey. Anaerobic methanogenic sediment microcosms were established and incubated for up to 130 days. TBBPA was reductively dehalogenated resulting in the transient formation of two intermediates, identified as tribromobisphenol A and dibromobisphenol A, and the formation and accumulation of bisphenol A (BPA) as the end product. Spartina sediments from both sites were found to dehalogenate TBBPA more rapidly than the Phragmites or unvegetated sediments, resulting in greater production of BPA. Microbial community diversity as measured by in situ sediment phospholipid fatty acid (PLFA) composition prior to TBBPA exposure, was found to be higher in the uncontaminated sediments; differences in microbial PLFA diversity were not seen in contaminated sediments associated with either the different plant species or unvegetated sediment. The results of this study demonstrate that these two plant species affected sediment microbial community function with respect to dehalogenation capabilities, even though the disturbed and undisturbed sediments varied in microbial community composition.

Carbon Isotopic Fractionation during Anaerobic Biotransformation of Methyl tert-Butyl Ether and tert-Amyl Methyl Ether

The fuel oxygenate methyl tert-butyl ether (MTBE) has been frequently detected in groundwater and surface water. Since contaminated sites are often subsurface, anaerobic degradation of MTBE will likely be significant for remediation. As traditional approaches to evaluate biodegradation generally involve laboratory microcosm studies which require time and resources, innovative approaches are needed to demonstrate active in situ biodegradation of MTBE. This study was conducted to gather information at the laboratory level to evaluate the potential of applying carbon isotope fractionation as an indicator for in situ biodegradation of the fuel oxygenates MTBE and tert-amyl methyl ether (TAME). In this study, MTBE utilization was observed in a methanogenic sediment microcosm after a lengthy lag period of about 400 days. MTBE utilization was sustained upon refeeding and subculturing. tert-Butyl alcohol (TBA) was found to accumulate after propagation of cultures. The MTBE-grown cultures also utilized TAME and produced tert-amyl alcohol (TAA). The detection of TBA and TAA indicated that ether bond cleavage was the initial step in degradation for both compounds. Carbon isotope fractionation during anaerobic MTBE and TAME degradation was studied, and isotopic enrichment factors (epsilon) with 95% confidence intervals of -15.6 +/-4.1% and -13.7+/-4.5% were estimated for anaerobic MTBE and TAME degradation, respectively. Addition of 2-bromoethanesulfonic acid, an inhibitor of methanogenesis, substantially prolonged the lag period before transformation, but did not influence carbon isotope fractionation. Our experiment provided strong evidence of significant carbon isotope fractionation during anaerobic MTBE and TAME degradation, demonstrating that this technique can be used as an indicator for in situ MTBE and TAME degradation.

Importance of clay size minerals for Fe(III) respiration in a petroleum‐contaminated aquifer

ABSTRACTThe availability of Fe(III)‐bearing minerals for dissimilatory Fe(III) reduction was evaluated in sediments from a petroleum‐contaminated sandy aquifer near Bemidji, Minnesota (USA). First, the sediments from a contaminated area of the aquifer, in which Fe(III) reduction was the predominant terminal electron accepting process, were compared with sediments from a nearby, uncontaminated site. Data from 0.5 m HCl extraction of different size fractions of the sediments revealed that the clay size fraction contributed a significant portion of the ‘bio‐available’ Fe(III) in the background sediment and was the most depleted in ‘bio‐available’ Fe(III) in the iron‐reducing sediment. Analytical transmission electron microscopy (TEM) revealed the disappearance of thermodynamically unstable Fe(III) and Mn(IV) hydroxides (ferrihydrite and Fe vernadite), as well as a decrease in the abundance of goethite and lepidocrocite in the clay size fraction from the contaminated sediment. TEM observations and X‐ray diffraction examination did not provide strong evidence of Fe(III)‐reduction‐related changes within another potential source of ‘bio‐available’ Fe(III) in the clay size fraction – ferruginous phyllosilicates. However, further testing in the laboratory with sediments from the methanogenic portion of the aquifer that were depleted in microbially reducible Fe(III) revealed the potential for microbial reduction of Fe(III) associated with phyllosilicates. Addition of a clay size fraction from the uncontaminated sediment, as well as Fe(III)‐coated kaolin and ferruginous nontronite SWa‐1, as sources of poorly crystalline Fe(III) hydroxides and structural iron of phyllosilicates respectively, lowered steady‐state hydrogen concentrations consistent with a stimulation of Fe(III) reduction in laboratory incubations of methanogenic sediments. There was no change in hydrogen concentration when non‐ferruginous clays or no minerals were added. This demonstrated that Fe(III)‐bearing clay size minerals were essential for microbial Fe(III) reduction and suggested that both potential sources of ‘bio‐available’ Fe(III) in the clay size fraction, poorly crystalline Fe(III) hydroxides and structural Fe(III) of phyllosilicates, were important sources of electron acceptor for indigenous iron‐reducing microorganisms in this aquifer.

Evidence for the removal of CFC-11, CFC-12, and CFC-113 at the groundwater–surface water interface in the Everglades

Poor agreement between 3H/ 3He ages and CFC-11 and CFC-12 ages suggests that CFCs may not be conservative tracers in the Everglades National Park. 3H/ 3He ages were used to calculate the expected concentration of CFC-11 and CFC-12 in groundwater from wells 2 to 73 m deep. The expected concentrations of CFCs were compared to the measured concentrations and plots of the % CFC-12 and CFC-11 remaining offered no evidence that significant CFC removal was occurring in the groundwater at depths ≥2 m, suggesting that CFC removal occurs at shallower depths. Except where CFC contamination was suspected, CFC-11, CFC-12 and CFC-113 concentrations in fresh surface water were nearly always below solubility equilibrium with the atmosphere. Measurements of CFC-11, CFC-12 and CFC-113 in pore water indicate a 50–90% decrease in concentration 5 cm below the groundwater–surface water (GW–SW) interface. In the same 5 cm interval CH 4 concentrations increased by 300–1000%. This suggested that CFCs were removed at the GW–SW interface, possibly by methane-producing bacteria. CFC derived recharge ages should therefore be viewed with caution when recharging water percolates through anoxic methanogenic sediments.

H2 consumption during the microbial reductive dehalogenation of chlorinated phenols and tetrachloroethene.

Competition for molecular hydrogen exists among hydrogen-utilizing microorganisms in anoxic environments, and evidence suggests that lower hydrogen concentrations are observed with more energetically favorable electron-accepting processes. The transfer of electrons to organochlorines via reductive dehalogenation reactions plays an important role in hydrogen dynamics in impacted systems. We studied the flux of aqueous hydrogen concentrations in methanogenic sediment microcosms prior to and during reductive dehalogenation of a variety of substituted chlorophenols (CP) and tetrachloroethene (perchloroethylene, PCE). Mean hydrogen concentrations during reductive dehalogenation of 2,4-CP, 2,3,4-CP, and PCP were 3.6 nM, 4.1 nM, and 0.34 nM, respectively. Sediment microcosms that were not dosed with chlorophenols yet were actively methanogenic maintained a significantly higher mean hydrogen concentration of 9.8 nM. During active PCE dehalogenation, sediment microcosms maintained a mean hydrogen concentration of 0.82 nM. These data indicate that during limiting hydrogen production, the threshold ecosystem hydrogen concentration is controlled by microbial populations that couple hydrogen oxidation to thermodynamically favorable electron accepting reactions, including reductive dehalogenation of chloroaromatic and chloroaliphatic compounds. We also present revised estimates for the Gibbs free energy available from the reductive dehalogenation of a variety of substituted chlorophenols based on recently published values of vapor pressure, solubility, and pKa for these compounds.

Reductive dechlorination of 2,4-dichlorophenol and related microbial processes under limiting and non-limiting sulfate concentration in anaerobic mid-Chesapeake Bay sediments

The effects of sulfate, added electron donors, and metabolic inhibitors on reductive dechlorination of 2,4-dichlorophenol (DCP) in anaerobic sediment from a relatively unpolluted site in the mesohaline region of Chesapeake Bay were investigated. The onset of ortho reductive dechlorination of DCP was often rapid in sulfate-depleted sediments and the metabolite, 4-chlorophenol, persisted for >44-60 days at nearly stoichiometric levels, relative to the DCP lost, with no further degradation or dechlorination observed. Dechlorination rates increased by a factor of 6-10 after re-feeding DCP, suggesting enrichment of dechlorinators. Sulfate inhibited reductive dechlorination greatly in fresh sediments, but only partially in aged, methanogenic or DCP-acclimated sediments. Exogenous H2 stimulated rates of reductive dechlorination in the absence, but not presence of sulfate. Molybdate severely inhibited reductive dechlorination and also significantly inhibited methanogenesis in sulfate-depleted sediments. Direct inhibition of methanogenesis with bromoethane sulfonic acid did not negatively affect dechlorination rates, suggesting that molybdate may have inhibited production of electron donors for methanogenesis and reductive dechlorination. These results demonstrate a potential for rapid dechlorination of DCP in relatively uncontaminated estuarine sediments and reveal confounding effects of a putative 'specific' metabolic inhibitor, molybdate.