Carbonate polymorphism controlled by microbial iron redox dynamics at a natural CO2 leakage site (Crystal Geyser, Utah)

Crystal Geyser (Utah, USA) is a CO2-rich low-temperature geyser that is studied as a natural analog for CO2 leakage from carbon capture and storage (CCS) sites. In order to better constrain the biogeochemical processes influencing CaCO3 precipitation at geological CO2 escape sites, we characterized fast-forming iron-rich calcium carbonate pisoids and travertines precipitating from the fluids expelled by the geyser. The pisoids, located within a few meters from the vent, are composed of concentric layers of aragonite and calcite. Calcite layers contain abundant ferrihydrite shrubs in which iron is encasing bacterial forms. The aragonite layers contain less abundant and finely dispersed iron, present either as iron-oxide microspherules or iron adsorbed to organic matter dispersed within the carbonate matrix. We propose that carbonate polymorphism in the pisoids is mostly controlled by local fluctuations of the iron redox state of the fluids from which they form, caused by episodic blooms of iron-oxidizing bacteria. Indeed, the waters expelled by Crystal Geyser contain >200 µM dissolved iron (Fe2+), a known inhibitor of calcite growth. The calcite layers of the pisoids may record episodes of intense microbial iron oxidation, consistent with observations of iron-oxide rich biofilms thriving in the rimstone pools around the geyser and previous metagenomic analyses showing abundant neutrophilic, microaerophilic iron-oxidizing bacteria in vent water. In turn, aragonite layers of the pisoids likely precipitate from Fe2+-rich waters, registering periods of less intense iron oxidation. Separately, CaCO3 polymorphism in the travertines, where calcite and aragonite precipitate concurrently, is not controlled by iron dynamics, but may be locally influenced by the presence of microbial biofilms. This study documents for the first time an influence of microbial iron oxidation on CaCO3 polymorphism in the environment, and informs our understanding of carbonate formation at CO2 leakage sites and in CCS contexts.

Approaches for Eliminating Bacteria Introduced during In Situ Bioleaching of Fractured Sulfidic Ores in Deep Subsurface

The major objective of the EU Horizon 2020 project “BioMOre” is the technical realization of indirect in situ leaching of Kupferschiefer sandstone and black shale ore by a ferric iron lixiviant generated by a mixed culture of autotrophic, acidophilic, iron-oxidizing bacteria and archaea in a ferric iron-generating bioreactor (FIGB). These organisms could colonize the deeply buried geological formations even under anaerobic conditions as most are able to grow by coupling the reduction of ferric iron to the oxidation of reduced sulfur compounds in the absence of oxygen. Development of an inhibition protocol to eliminate these allochthonous microbial bioreactor populations subsequent to the completion of in situ bioleaching was therefore investigated. Column bioleaching experiments using a laboratory-scale FIGB confirmed not only that metals were solubilised from both the sandstone and shale ores, but also that significant numbers of bacteria were released from the FIGB. The efficacy of 13 different chemical compounds in inhibiting microbial iron oxidation has been tested at different concentrations in shake flask and FIGB-coupled columns. Iron-oxidation activity, microcalorimetrically-determined activity and ATP measurements, in combination with microscopic cell counts and biomolecular analysis (T-RFLP, qPCR), plate counts and most-probable-number (MPN), were used to monitor the inhibiting effects on the acidophiles. Complete inhibition of metabolic activity of iron-oxidizing acidophiles was achieved in the presence of 0.4 mM formate, 300 mM chloride, 100 mM nitrate, 10 mM of primary C6 to C8 alcohols, 100 mM 1-butanol, 100 mM 1-pentanol, 0.1 mM SDS or 0.35 mM benzoic acid. No inhibition was found for 0.6 mM acetic acid and 200 mM methanol. Based on these results a recipe for the chemical composition of the “decommissioning solution” is proposed.

Microbial Iron Oxidation in the Arctic Tundra and Its Implications for Biogeochemical Cycling

ABSTRACTThe role that neutrophilic iron-oxidizing bacteria play in the Arctic tundra is unknown. This study surveyed chemosynthetic iron-oxidizing communities at the North Slope of Alaska near Toolik Field Station (TFS) at Toolik Lake (lat 68.63, long −149.60). Microbial iron mats were common in submerged habitats with stationary or slowly flowing water, and their greatest areal extent is in coating plant stems and sediments in wet sedge meadows. Some Fe-oxidizing bacteria (FeOB) produce easily recognized sheath or stalk morphotypes that were present and dominant in all the mats we observed. The cool water temperatures (9 to 11°C) and reduced pH (5.0 to 6.6) at all sites kinetically favor microbial iron oxidation. A microbial survey of five sites based on 16S rRNA genes found a predominance ofProteobacteria, withBetaproteobacteriaand members of the familyComamonadaceaebeing the most prevalent operational taxonomic units (OTUs). In relative abundance, clades of lithotrophic FeOB composed 5 to 10% of the communities. OTUs related to cyanobacteria and chloroplasts accounted for 3 to 25% of the communities. Oxygen profiles showed evidence for oxygenic photosynthesis at the surface of some mats, indicating the coexistence of photosynthetic and FeOB populations. The relative abundance of OTUs belonging to putative Fe-reducing bacteria (FeRB) averaged around 11% in the sampled iron mats. Mats incubated anaerobically with 10 mM acetate rapidly initiated Fe reduction, indicating that active iron cycling is likely. The prevalence of iron mats on the tundra might impact the carbon cycle through lithoautotrophic chemosynthesis, anaerobic respiration of organic carbon coupled to iron reduction, and the suppression of methanogenesis, and it potentially influences phosphorus dynamics through the adsorption of phosphorus to iron oxides.

New Insight into Microbial Iron Oxidation as Revealed by the Proteomic Profile of an Obligate Iron-Oxidizing Chemolithoautotroph

ABSTRACTMicroaerophilic, neutrophilic, iron-oxidizing bacteria (FeOB) grow via the oxidation of reduced Fe(II) at or near neutral pH, in the presence of oxygen, making them relevant in numerous environments with elevated Fe(II) concentrations. However, the biochemical mechanisms for Fe(II) oxidation by these neutrophilic FeOB are unknown, and genetic markers for this process are unavailable. In the ocean, microaerophilic microorganisms in the genusMariprofundusof the classZetaproteobacteriaare the only organisms known to chemolithoautotrophically oxidize Fe and concurrently biomineralize it in the form of twisted stalks of iron oxyhydroxides. The aim of this study was to identify highly expressed proteins associated with the electron transport chain of microaerophilic, neutrophilic FeOB. To this end,Mariprofundus ferrooxydansPV-1 was cultivated, and its proteins were extracted, assayed for redox activity, and analyzed via liquid chromatography-tandem mass spectrometry for identification of peptides. The results indicate that a cytochromec4,cbb3-type cytochrome oxidase subunits, and an outer membrane cytochromecwere among the most highly expressed proteins and suggest an involvement in the process of aerobic, neutrophilic bacterial Fe oxidation. Proteins associated with alternative complex III, phosphate transport, carbon fixation, and biofilm formation were abundant, consistent with the lifestyle ofMariprofundus.

Seasonal and spatial variations in microbial activity at various phylogenetic resolutions at a groundwater – surface water interface

We investigated the seasonal and spatial variation in activity and density of the metabolically active in situ microbial community (AIMC) at a landfill leachate-impacted groundwater – surface water interface (GSI). A series of AIMC traps were designed and implemented for AIMC sampling and microbial activity and density examinations. Measurements were made not only at the level of bacterial domain but also at the levels of alphaproteobacterial Rhizobiales order and gammaproteobacterial Pseudomonas genus, both of which included a large number of iron-oxidizing bacteria as revealed from previous analysis. Consistently higher microbial activities with less variation in depth were measured in the AIMC traps than in the ambient sediments. Flood disturbance appeared to control AIMC activity distributions at the gradually elevated GSI. The highest AIMC activities were generally obtained from locations closest to the free surface water boundary except during the dry season when microbial activities were similar across the entire GSI. A clone library of AIMC 16S rRNA genes was constructed, and it confirmed the predominant role of the targeted alphaproteobacterial group in AIMC activity and composition. This taxon constituted 2%–14% of all bacteria with similar activity distribution profiles. The Pseudomonas group occupied only 0.1‰–0.5‰ of the total bacterial density, but its activity was 27 times higher than the bacterial average. Of the 16S rRNA sequences in the AIMC clone library, 7.5% were phylogenetically related to putative IOB, supporting the occurrence and persistence of active microbial iron oxidation across the studied iron-rich GSI ecosystem.

Evidence for microbial dissolution of pyrite from the Lower Cambrian oolitic limestone, South China

The oxidative dissolution of the sulphide mineral pyrite (FeS2) has been of significant interest since it affects global geochemical cycles, generates acid mine drainage, and is used in industrial metal extraction. Several different groups of prokaryotes are known to catalyze the dissolution of pyrite and use the free energy generated from the oxidation, which may result in the dissolution of the mineral and the precipitation of the secondary ferric iron minerals either on the cell surface or is separated from the cells. However, straightforward evidence for such metabolic process in the ancient sediments is rare. Here we report pyrite crystals from the Lower Cambrian oolitic limestones that show indications of microbial erosion in various degrees. Erosion pits and tubular micro-tunnels with characteristic shapes and sizes in our samples are generally similar to those obtained from the laboratory studies on the oxidative dissolution of pyrite by iron-oxidizing bacteria. Diagenetic examination demonstrates that the bioerosion predates the consolidation of the limestone. In addition, bacillus-sized and -shaped microfossils encrusted with iron oxides are present in our samples, which are very likely to be fossilized sheaths produced by iron-oxidizing bacteria. Our findings indicate that the microbial oxidative dissolution of pyrite existed in the Cambrian shallow marine carbonate sediments. Furthermore, we suggest that characteristic pitting patterns on the pyrite crystals from ancient sediments are an important clue to trace the evolution of life, in particular, the evolution of metabolism like microbial iron oxidation in the remote past on our planet, independent of biomarkers, isotopic signals and body fossils as well.

Distribution and Diversity ofGallionella-Like Neutrophilic Iron Oxidizers in a Tidal Freshwater Marsh

ABSTRACTMicrobial iron oxidation is an integral part of the iron redox cycle in wetlands. Nonetheless, relatively little is known about the composition and ecology of iron-oxidizing communities in the soils and sediments of wetlands. In this study, sediment cores were collected across a freshwater tidal marsh in order to characterize the iron-oxidizing bacteria (FeOB) and to link their distributions to the geochemical properties of the sediments. We applied recently designed 16S rRNA primers targetingGallionella-related FeOB by using a nested PCR-denaturing gradient gel electrophoresis (DGGE) approach combined with a novel quantitative PCR (qPCR) assay.Gallionella-related FeOB were detected in most of the samples. The diversity and abundance of the putative FeOB were generally higher in the upper 5 to 12 cm of sediment than in deeper sediment and higher in samples collected in April than in those collected in July and October. Oxygen supply by macrofauna appears to be a major force in controlling the spatial and temporal variations in FeOB communities. The higher abundance ofGallionella-related FeOB in April coincided with elevated concentrations of extractable Fe(III) in the sediments. Despite this coincidence, the distributions of FeOB did not exhibit a simple relationship to the redox zonation inferred from the geochemical depth profiles.