Present-day serpentinization generates groundwaters with conditions (pH > 11, Eh < À550 mV) favorable for the microbial and abiotic production of organic compounds from inorganic precursors. Elevated concentrations of methane, C 2 -C 6 alkanes, acetate, and formate have been detected at these sites, but the microbial or abiotic origin of these compounds remains unclear. While geochemical data indicate that methane at most sites of present-day serpentinization is abiogenic, the stable carbon, hydrogen, and clumped isotope data as well as the hydrocarbon gas composition from The Cedars, CA, USA, are consistent with a microbial origin for methane. However, there is no direct evidence of methanogenesis at this site of serpentinization. We report on laboratory experiments in which the microbial communities in fluids and sediments from The Cedars were incubated with 13 C labeled substrates. Increasing methane concentrations and the incorporation of 13 C into methane in live experiments, but not in killed controls, demonstrated that methanogens converted methanol, formate, acetate (methyl group), and bicarbonate to methane. The apparent fractionation between methane and potential substrates (α 13 C CH4-CO2(g) = 1.059 to 1.105, α 13 C CH4-acetate = 1.042 to 1.119) indicated that methanogenesis was dominated by the carbonate reduction pathway. Increasing concentrations of volatile organic acid anions indicated microbial acetogenesis. α 13 C CO2(g)-acetate values (0.999 to 1.000), however, were inconsistent with autotrophic acetogenesis, thus suggesting that acetate was produced through fermentation. This is the first study to show direct evidence of microbial methanogenesis and acetogenesis by the native microbial community at a site of present-day serpentinization.
Previous agglomerate-scale heap bioleaching studies have outlined the variations in cell numbers of the liquid and attached phases during colonisation of sterilised ore by a pure culture. In this study, a mixed mesophilic culture was used in agglomerate-scale columns containing non-sterilised low-grade copper ore. Over a six - month period, columns were harvested at various intervals to provide snapshots of the metal distribution and the quantity, location, and ecological variations of mineral-oxidizing microbes within the ore bed. The initial colonisation period in this experiment was dissimilar to previous work, as the indigenous community was retained within the ore-bed throughout acid agglomeration. The overall colonisation phase lasted for approximately 1,000 hours until cell concentrations stabilised. In each column, less than 0.05% of the total cells were found in the leachate, 15-20% in the interstitial phase and the remaining ~80% were attached to the mineral surface. Once cell numbers had stabilised, interstitial cell concentrations were approximately 2,000× greater than those in the leachate. This difference persisted for the duration of the experiment. Copper concentrations in the two liquid phases generally decreased over time, but were on average 50× higher in the interstitial phase. Iron concentrations were more stable, but again were 30× higher in the interstitial phase. This demonstrates that that the difference in cell concentration between the leachate and interstitial phases cannot be explained through diffusion gradients within the system as it is much greater than those observed for the dissolved metals. It also shows that the specific environmental conditions of the interstitial and attached cells are very different to those inferred through analysis of leachates alone.
An apparatus is described for the continuous monitoring of process water. The phenol contents of six separate flows are recorded. The apparatus monitors water-flows that are normally clean and ultimately dischaxged to a natural watercourse. However, the water is liable to be contaminated by phenolic material ; the apparatus detects such contamination, and the flow is then diverted for treatment a t effluent plants.
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