Tori M. Hoehler

Field and laboratory studies of methane oxidation in an anoxic marine sediment: Evidence for a methanogen‐sulfate reducer consortium

Field and laboratory studies of anoxic sediments from Cape Lookout Bight, North Carolina, suggest that anaerobic methane oxidation is mediated by a consortium of methanogenic and sulfate-reducing bacteria. A seasonal survey of methane oxidation and CO2 reduction rates indicates that methane production was confined to sulfate-depleted sediments at all times of year, while methane oxidation occurred in two modes. In the summer, methane oxidation was confined to sulfate-depleted sediments and occurred at rates lower than those of CO2 reduction. In the winter, net methane oxidation occurred in an interval at the base of the sulfate-containing zone. Sediment incubation experiments suggest both methanogens and sulfate reducers were responsible for the observed methane oxidation. In one incubation experiment both modes of oxidation were partially inhibited by 2-bromoethanesulfonic acid (a specific inhibitor of methanogens). This evidence, along with the apparent confinement of methane oxidation to sulfate-depleted sediments in the summer, indicates that methanogenic bacteria are involved in methane oxidation. In a second incubation experiment, net methane oxidation was induced by adding sulfate to homogenized methanogenic sediments, suggesting that sulfate reducers also play a role in the process. We hypothesize that methanogens oxidize methane and produce hydrogen via a reversal of CO2 reduction. The hydrogen is efficiently removed and maintained at low concentrations by sulfate reducers. Pore water H2 concentrations in the sediment incubation experiments (while net methane oxidation was occurring) were low enough that methanogenic bacteria could derive sufficient energy for growth from the oxidation of methane. The methanogen-sulfate reducer consortium is consistent not only with the results of this study, but may also be a feasible mechanism for previously documented anaerobic methane oxidation in both freshwater and marine environments.

Thermodynamic control on hydrogen concentrations in anoxic sediments

Abstract Molecular hydrogen plays a central role in bacterially mediated anoxic sediment chemistry, as both an important electron transfer agent and a key thermodynamic control. We studied the response of hydrogen concentrations to changes in temperature, specific electron acceptor, sulfate concentration, and pH in a series of laboratory experiments using sediments from Cape Lookout Bight, North Carolina. Hydrogen concentrations were found to depend significantly on each of these factors in a fashion that suggests thermodynamic control. In general, the change in hydrogen concentrations was apparently driven by a necessity to maintain a constant ΔG for the predominant terminal electron-accepting process. We hypothesize this situation derives from highly competetive conditions that force terminal metabolic bacteria to operate right at their thermodynamic limits. The response of hydrogen to individual controls in the laboratory experiments was reflected in relatively quantitiative fashion in down-core, seasonal, and inter-environmental variations observed in sediment cores from Cape Lookout Bight and the White Oak River, NC.

Factors that control the stable carbon isotopic composition of methane produced in an anoxic marine sediment

The carbon isotopic composition of methane produced in anoxic marine sediment is controlled by four factors: (1) the pathway of methane formation, (2) the isotopic composition of the methanogenic precursors, (3) the isotope fractionation factors for methane production, and (4) the isotope fractionation associated with methane oxidation. The importance of each factor was evaluated by monitoring stable carbon isotope ratios in methane produced by a sediment microcosm. Methane did not accumulate during the initial 42-day period when sediment contained sulfate, indicating little methane production from “noncompetitive” substrates. Following sulfate depletion, methane accumulation proceeded in three distinct phases. First, CO2 reduction was the dominant methanogenic pathway and the isotopic composition of the methane produced ranged from −80 to −94‰. The acetate concentration increased during this phase, suggesting that acetoclastic methanogenic bacteria were unable to keep pace with acetate production. Second, acetate fermentation became the dominant methanogenic pathway as bacteria responded to elevated acetate concentrations. The methane produced during this phase was progressively enriched in 13C, reaching a maximum δ13C value of −42‰. Third, the acetate pool experienced a precipitous decline from >5 mM to <20 μM and methane production was again dominated by CO2 reduction. The δ13C of methane produced during this final phase ranged from −46 to −58‰. Methane oxidation concurrent with methane production was detected throughout the period of methane accumulation, at rates equivalent to 1 to 8% of the gross methane production rate. Thus methane oxidation was too slow to have significantly modified the isotopic signature of methane. A comparison of microcosm and field data suggests that similar microbial interactions may control seasonal variability in the isotopic composition of methane emitted from undisturbed Cape Lookout Bight sediment.

The Carbon Isotope Biogeochemistry of Methane Production in Anoxic Sediments: 2. A Laboratory Experiment

The factors that contribute to variability in the isotopic composition of methane produced by anoxic sediment were evaluated by monitoring stable carbon isotope ratios in methane produced by a sediment microcosm. Anoxic marine sediment from Cape Lookout Bight, North Carolina, was placed in a 2-L pyrex incubation syringe for 114 d. Sediment samples were removed at weekly intervals and analyzed for sulfate, acetate, and methane concentrations as well as13C/12C ratios in the dissolved CO2 and methane pools. In addition, rates of sulfate reduction, methane production from acetate, and methane production from CO2 were measured by radiotracer techniques. The isotopic composition of the methane pool varied by more than 40%0 over the course of the experiment. The changes appear to be the result of at least two factors: (1) changes in the isotopic composition of the dissolved CO2 pool and (2) changes in the fraction of the methane production derived from acetate. A comparison of the laboratory experiment with field data suggests that similar biogeochemical processes operate to control carbon isotope ratios of methane in both undisturbed and laboratory-incubated sediment.