Thomas M. McCollom

Isolation and Characterization of Novel Psychrophilic, Neutrophilic, Fe-Oxidizing, Chemolithoautotrophic α- and γ-Proteobacteria from the Deep Sea

ABSTRACT We report the isolation and physiological characterization of novel, psychrophilic, iron-oxidizing bacteria (FeOB) from low-temperature weathering habitats in the vicinity of the Juan de Fuca deep-sea hydrothermal area. The FeOB were cultured from the surfaces of weathered rock and metalliferous sediments. They are capable of growth on a variety of natural and synthetic solid rock and mineral substrates, such as pyrite (FeS2), basalt glass (∼10 wt% FeO), and siderite (FeCO3), as their sole energy source, as well as numerous aqueous Fe substrates. Growth temperature characteristics correspond to the in situ environmental conditions of sample origin; the FeOB grow optimally at 3 to 10°C and at generation times ranging from 57 to 74 h. They are obligate chemolithoautotrophs and grow optimally under microaerobic conditions in the presence of an oxygen gradient or anaerobically in the presence of nitrate. None of the strains are capable of using any organic or alternate inorganic substrates tested. The bacteria are phylogenetically diverse and have no close Fe-oxidizing or autotrophic relatives represented in pure culture. One group of isolates are γ-Proteobacteria most closely related to the heterotrophic bacterium Marinobacter aquaeolei (87 to 94% sequence similarity). A second group of isolates are α-Proteobacteria most closely related to the deep-sea heterotrophic bacterium Hyphomonas jannaschiana (81 to 89% sequence similarity). This study provides further evidence for the evolutionarily widespread capacity for Fe oxidation among bacteria and suggests that FeOB may play an unrecognized geomicrobiological role in rock weathering in the deep sea.

A reassessment of the potential for reduction of dissolved CO 2 to hydrocarbons during serpentinization of olivine

The concept that aqueous CO2 can be reduced to hydrocarbons abiotically during serpentinization of olivine has become widespread in the earth and planetary sciences. This process has been invoked to explain the occurrence of hydrocarbons in crystalline igneous rocks and proposed as a source of prebiotic organic compounds for the origin of life. We reevaluate this scenario through an experimental study of the reaction of dissolved CO2 in the presence of olivine under hydrothermal conditions (300°C, 350 bar). Reduction of CO2 to formate (HCOO−) was found to proceed rapidly, with H2 generated from hydrothermal alteration of olivine serving as the reductant. The reverse reaction, decomposition of formic acid to CO2 and H2, was also found to proceed rapidly. Although dissolved hydrocarbon concentrations increased throughout the experiments, isotopic labeling of dissolved CO2 with 13C indicated that these compounds were primarily generated from reduced carbon compounds already present in olivine at the beginning of the experiment rather than by reduction of CO2. The only hydrocarbon product from reduction of CO2 observed in the experiments was a small amount of methane (<0.04% conversion of dissolved CO2 in more than 2500 h of heating). Comparison of the reaction products with thermodynamic data indicates that reactions between dissolved CO2 and formate rapidly achieved metastable equilibrium at the experimental conditions, suggesting that similar reactions could control the concentration of formate in geologic fluids. The results indicate that the potential for abiotic formation of hydrocarbons during serpentinization may be much more limited than previously believed, and other mineral catalysts or vapor phase reactions may be required to explain many occurrences of abiotic hydrocarbons in serpentinites and igneous rocks.

Methanogenesis as a potential source of chemical energy for primary biomass production by autotrophic organisms in hydrothermal systems on Europa

Geochemical models are used to explore the possibility that lithoautotrophic methanogenesis (the conversion of CO2 plus H2 to methane) could be a source of metabolically useful chemical energy for the production of biomass at putative Europan hydrothermal systems. Two cases are explored: a relatively reduced methane-rich ocean and a relatively oxidized sulfate- and bicarbonate-rich ocean. In the case of a methane-rich ocean, a source of CO2 for methanogenesis is provided by conversion of dissolved methane to CO2 during reaction of ocean water with igneous rocks at high temperatures in the subsurface. Fluid-rock reactions also provide a source of dissolved H2 in the hydrothermal fluid. When this fluid circulates back to the ocean floor and mixes with seawater, conversion of the dissolved CO2 and H2 to methane provides a potential source of chemical energy that can be used to drive metabolic processes. For the case of a sulfate- and carbonate-rich ocean, reaction with reduced igneous rocks at high temperatures will also produce hydrothermal fluids with high H2 concentrations (as occurs in hydrothermal systems on Earth). Mixing of the resulting hydrothermal fluid with seawater in a relatively oxidized ocean could supply energy from either methanogenesis or sulfate reduction. For plausible compositions of a Europan ocean, methanogenesis can supply similar amounts of energy to that which supports the prolific ecosystems surrounding submarine hydrothermal vents on Earth. Even in the most optimistic case, however, the total amount of biomass that could be supported globally by lithoautotrophic microbes on Europa is extremely small compared to the biomass produced photosynthetically on Earth. Nevertheless, sufficient metabolic energy could apparently be available at hydrothermal systems on Europa to support an origin of life and localized ecosystems.

Neutrophilic Iron-Oxidizing Bacteria in the Ocean: Their Habitats, Diversity, and Roles in Mineral Deposition, Rock Alteration, and Biomass Production in the Deep-Sea

The importance of metals to life has long been appreciated. Iron (Fe) is the fourth most abundant element overall, and the second most abundant element that is redox-active in near-surface aqueous habitats, rendering it the most important environmental metal. While it has long been recognized that microorganisms participate in the global iron cycle, appreciation for the pivotal role that redox cycling of iron plays in energy conservation among diverse prokaryotes has grown substantially in the past decade. In addition, redox reactions involving Fe are linked to several other biogeochemical cycles (e.g., carbon), with significant ecological ramifications. The increasing appreciation for the role of microbes in redox transformations of Fe is reflected in a recent surge in biological and environmental studies of microorganisms that conserve energy for growth from redox cycling of Fe compounds, particularly in the deep ocean. Here we highlight some of the key habitats where microbial Fe-oxidation plays significant ecological and biogeochemical roles in the oceanic regime, and provide a synthesis of recent studies concerning this important physiological group. We also provide the first evidence that microbial Fe-oxidizing bacteria are a critical factor in the kinetics of mineral dissolution at the seafloor, by accelerating dissolution by 6–8 times over abiotic rates. We assert that these recent studies, which indicate that microbial Fe-oxidation is widespread in the deep-sea, combined with the apparent role that this group play in promoting rock and mineral weathering, indicate that a great deal more attention to these microorganisms is warranted in order to elucidate the full physiological and phylogenetic diversity and activity of the neutrophilic Fe-oxidizing bacteria in the oceans.

Experimental constraints on the hydrothermal reactivity of organic acids and acid anions: I. Formic acid and formate

A series of hydrothermal experiments covering a range of temperatures from 175 to 260°C examined the decomposition of formic acid and formate and also investigated the production of formate from reduction of CO2. Decomposition rates measured in this study, which were conducted in gold-TiO2 reactors, were several orders of magnitude slower than those reported in previous studies conducted in steel and Ti-metal reactors, indicating the previous studies substantially overestimated the rate of the reaction owing to reactor catalysis. Although experiments were conducted with several different minerals present (hematite, magnetite, serpentinized olivine, NiFe-alloy), the decomposition rates were similar in each experiment once the effects of fluid pH were accounted for, suggesting that the minerals had no effect on the stability of formic acid or formate. At higher temperatures (>225°C), the rates of both the decomposition of formate and the reduction of CO2 to formate were sufficiently rapid that reactions between dissolved CO2 and formate rapidly attained a state of metastable thermodynamic equilibrium. The results suggest that the amount of formate in many subsurface and hydrothermal fluids is likely to be controlled by equilibrium with dissolved CO2 at the prevailing oxidation state and pH of the fluid. This may account for the high concentrations of formate observed in strongly reducing environments such as serpentinites, as well as the low concentrations relative to other organic acid anions in mildly reducing environments such as oil-filed brines and formation waters in sedimentary basins. Although formate has been suggested to be a reaction intermediate in the formation of abiotic hydrocarbons from reduction of aqueous CO2, production of hydrocarbons was not observed in any of the experiments, except for trace amounts of methane, despite high concentrations of formate and strongly reducing conditions.

A volcanic environment for bedrock diagenesis at Meridiani Planum on Mars

Exposed bedrocks at Meridiani Planum on Mars display chemical and mineralogical evidence suggesting interaction with liquid water. On the basis of morphological observations as well as high abundances of haematite and sulphate minerals, the rocks have been interpreted as sediments that were deposited in a shallow body of briny water with subsequent evaporation leaving behind the sulphate minerals. The iron-sulphur mineralization at Meridiani has also been inferred to be analogous to that produced during oxidative weathering of metal sulphide minerals, such as occurs at acid mine drainage sites. Neither of these interpretations, however, is consistent with the chemical composition of the rocks. Here we propose an alternative model for diagenesis of Meridiani bedrock that involves deposition of volcanic ash followed by reaction with condensed sulphur dioxide- and water-bearing vapours emitted from fumaroles. This scenario does not require prolonged interaction with a standing body of surface water and may have occurred at high temperatures. Consequently, the model invokes an environment considerably less favourable for biological activity on Mars than previously proposed interpretations.

Seafloor bioalteration of sulfide minerals: Results from in situ incubation studies

We present results of incubation studies conducted at low temperatures (∼4°C) in the vicinity of a seafloor hydrothermal vent system. We reacted Fe-, S-, Cu-, and Zn-bearing minerals including pyrite, marcasite, chalcopyrite, sphalerite, elemental sulfur, and a portion of a natural chimney sulfide structure for 2 months at the Main Endeavour Segment of the Juan de Fuca Ridge in the Pacific Ocean. Our study utilizes Fluorescent In Situ Hybridizations (FISH), Scanning and Transmission Electron Microscopy (SEM, TEM), and light microscopic analysis. The surfaces of these minerals are solely colonized by Bacteria and not by Archaea. Colonization densities vary over an order of magnitude with the following sequence: elemental sulfur > chimney sulfide > marcasite > pyrite > sphalerite > chalcopyrite, and correspond well with the abiotic oxidation kinetics of these materials, excepting elemental sulfur, which is both the least reactive to oxidizing species and the most heavily colonized. Colonization densities also correspond with apparent degree of reaction (dissolution pitting + accumulation of secondary alteration products). Heavy accumulations of secondary Fe oxides on Fe-bearing minerals, most notably on the chimney sulfide, form in situ as the result of mineral dissolution and the activity of neutrophilic Fe-oxidizing bacteria. Results suggest that mineral-oxidizing bacteria play a prominent role in weathering of seafloor sulfide deposits, and that microbial utilization of mineral substrates contributes to biomass production in seafloor hydrothermal environments.

Fluid-rock interactions in the lower oceanic crust: Thermodynamic models of hydrothermal alteration

Rocks from the lower oceanic crust show both petrologic and isotopic evidence for extensive alteration by circulating hydrothermal fluids derived from seawater. Deeply circulating fluids may have a substantial, but as yet largely undetermined, impact on the thermal regime of spreading centers, magmatic processes at mid-ocean ridges, and elemental fluxes at ridge hydrothermal systems. In the present study, the consequences of water-rock interactions during the penetration of seawater into hot lower oceanic crust were investigated using thermodynamic reaction path models. Alteration assemblages predicted for reaction of an evolving seawater with olivine gabbro, gabbronorite, and troctolite during heating from 300° to 900°C closely resemble alteration assemblages observed in rock samples from the lower oceanic crust. These assemblages include sodic plagioclase + actinolite ± diopsidic clinopyroxene ± epidote ± chlorite ± prehnite ± analcime ± quartz at lower temperatures (300°–500°C) and plagioclase + hornblende ± diopside ± chlorite ± trace magnetite at intermediate temperatures (500–700°C). The relative abundances of these alteration products show much greater variation with temperature than with water/rock (W/R) ratio, indicating that variations in the amounts of hydrated minerals such as amphiboles may not be an accurate indication of the W/R ratio experienced by rocks in the lower crust since the same amount of fluid may produce widely differing amounts of amphibole at slightly different temperatures. Depending on the temperature of interaction, some rocks in the lower oceanic crust may have experienced much higher W/R ratios than has previously been proposed based on the amount of amphibole present. At higher temperatures (750°–900°C),. the models predict that very little alteration occurs before the fluid equilibrates with the rock, producing only trace amounts of amphibole as an alteration phase, with the remainder of the rock retaining its igneous mineral composition essentially unchanged. This result suggests that extensive penetration of rocks by aqueous fluids may occur in the lower ocean crust at temperatures >∼700°C and leave very little conspicuous petrologic evidence for their presence. Such cryptic alteration is consistent with isotopic evidence for hydrothermal alteration of oxygen and Sr isotopes by seawater in lower ocean crust rock samples which otherwise retain their igneous compositions and textures. Taken together, the model results allow for much more extensive circulation of fluids in the lower oceanic crust and at much higher temperatures than has been proposed by previous studies based on petrologic interpretations.

Geochemical Constraints on Sources of Metabolic Energy for Chemolithoautotrophy in Ultramafic-Hosted Deep-Sea Hydrothermal Systems

Numerical models are employed to investigate sources of chemical energy for autotrophic microbial metabolism that develop during mixing of oxidized seawater with strongly reduced fluids discharged from ultramafic-hosted hydrothermal systems on the seafloor. Hydrothermal fluids in these systems are highly enriched in H(2) and CH(4) as a result of alteration of ultramafic rocks (serpentinization) in the subsurface. Based on the availability of chemical energy sources, inferences are made about the likely metabolic diversity, relative abundance, and spatial distribution of microorganisms within ultramafic-hosted systems. Metabolic reactions involving H(2) and CH(4), particularly hydrogen oxidation, methanotrophy, sulfate reduction, and methanogenesis, represent the predominant sources of chemical energy during fluid mixing. Owing to chemical gradients that develop from fluid mixing, aerobic metabolisms are likely to predominate in low-temperature environments (<20-30 degrees C), while anaerobes will dominate higher-temperature environments. Overall, aerobic metabolic reactions can supply up to approximately 7 kJ of energy per kilogram of hydrothermal fluid, while anaerobic metabolic reactions can supply about 1 kJ, which is sufficient to support a maximum of approximately 120 mg (dry weight) of primary biomass production by aerobic organisms and approximately 20-30 mg biomass by anaerobes. The results indicate that ultramafic-hosted systems are capable of supplying about twice as much chemical energy as analogous deep-sea hydrothermal systems hosted in basaltic rocks.

Formation of meteorite hydrocarbons from thermal decomposition of siderite (FeCO3)

Thermal decomposition of siderite has been proposed as a source of magnetite in martian meteorites. Laboratory experiments were conducted to evaluate the possibility that this process might also result in abiotic synthesis of organic compounds. Siderite decomposition in the presence of water vapor at 300°C generated a variety of organic products dominated by alkylated and hydroxylated aromatic compounds. The results suggest that formation of magnetite by thermal decomposition of siderite on the precursor rock of the martian meteorite ALH84001 would have been accompanied by formation of organic compounds and may represent a source of extraterrestrial organic matter in the meteorite and on Mars. The results also suggest that thermal decomposition of siderite during metamorphism could account for some of the reduced carbon observed in metasedimentary rocks from the early Earth.