ALBERT

Description: Understanding the natural microbiological mechanismsthat promote iron cycling in iron ore mine environmentsmay provide novel tools for the remediation of the fragile,iron-rich duricrust ecosystems associated with these envir-onmentsaswellasprovide asolutionforthestabilisation ofhillslopes and tailings (waste) dams. A diverse array ofmicrofossils is frequently identified throughout metre-scale duricrusts (canga;〉50 wt.% Fe) that cap iron oredeposits in Brazil, shedding light on the intimate role ofmicroorganisms in the evolution of these crusts. Nanoscalesecondary ion mass spectrometry revealed that carbon andnitrogen biosignatures are occasionally preserved, and typ-ically associated with the cell envelope structures of micro-fossils. The microfossils are 1–5mm in length, withfilamentous and rod-shaped morphologies commonly pre-served1,2. When examined using backscatter electron scan-ning electron microscopy, canga shows a complexmicrostructure from repeated dissolution and re-precipita-tion of iron oxide minerals. Geochronology3, geochemis-try4and microbiology5provide insights into the past andpresent-day role of microorganisms in the evolution ofcanga. These dynamic biogeochemical processes in cangacontribute to the continuous formation of new ironcements, preserving some of world’s longest-lived contin-uously exposed surfaces. Harnessing and accelerating thebiogeochemical cycling of iron may contribute to the de-velopment of novel technologies for mine remediation andwaste stabilisation.

Description: Goethite-cemented duricrusts, also known as canga, commonly occur as a capping rock protecting underlying iron ore deposits. The processes that govern canga formation are still unclear but include recurrent partial dissolution and recrystallisation of goethite through biogeochemical cycling of iron, hypothesised to be catalysed by plants and bacteria. In the present study, the effect of plant exudates on mobilisation of iron in canga was examined using model plants grown on crushed canga in RHIZOtest devices, which separate roots from substrate by a semi-permeable membrane. Moderate plant-induced acidification of the canga was detected, however the primary driver of mineral dissolution was the synergistic effect of reductive and ligand-promoted dissolution, identified by an increase in organic acids concentration and the presence of low concentrations of free ferrous iron. Whilst organic acids exudation lasted, iron cations were stabilised in solution; once the organic acids were degraded by microorganisms, the free cations precipitated as iron oxy-hydroxides. Mineralogical analysis and high-resolution microscopy confirmed our hypothesis that plants that grow in this iron-rich substrate contribute to iron dissolution indirectly (e.g., during phosphate solubilisation), and that the resulting surplus iron not taken up by the plants is redeposited, promoting the cementation of the residual minerals. Understanding the contribution of plants to the iron cycling in canga is crucial when formulating post-mining rehabilitation strategies for iron ore sites.

Description: Canga is a moderately hard iron-rich duricrust primarily composed of goethite as a result of the weathering of banded iron formations. Canga duricrusts lack a well-developed soil profile and consequently form an innate association with rupestrian plants that may become ferruginised, contributing to canga possessing macroscopic biological features. Examination of polished canga using a field emission scanning electron microscope (FE-SEM) revealed the biological textures associated with canga extended to the sub-millimetre scale in petrographic sections and polished blocks. Laminae that formed by abiotic processes and regions where goethite cements were formed in association with microorganisms were observed in canga. Biological cycling of iron within canga has resulted in two distinct forms of microbial fossilisation: permineralisation of multispecies biofilms and mineralisation of cell envelopes. Goethite permineralised biofilms frequently formed around goethite-rich kaolinite grains in close proximity to goethite bands and were composed of micrometre-scale rod-shaped, cocci and filamentous microfossils. In contrast, the cell envelopes immobilised by authigenic iron oxides were primarily of rod-shaped microorganisms, were not permineralised and occurred in pore spaces within canga. Complete mineralisation of intact rod-shaped casts and the absence of permineralisation suggested mineralised cell envelopes may represent fossilised iron-oxidising bacteria in the canga ecosystem. Replication of these iron-oxidising bacteria appeared to infill the porous regions within canga. Synchrotron-based Fourier transform infrared (FTIR) microspectroscopy demonstrated that organic biomarkers were poorly preserved with only weak bands indicative of aliphatic methylene (CH2) associated with permineralised microbial biofilms. High resolution imaging of microbial fossils in canga that had been etched with oxalic acid supported the poor preservation of organic biomarkers within canga, indicating mineralogical replacement of organic biomarkers.

Description: Demonstrating the biogenicity of presumptive microfossils in the geological record often requires supporting chemical signatures, including isotopic signatures. Understanding the mechanisms that promote the preservation of microbial biosignatures associated with microfossils is fundamental to unravelling the palaeomicrobiological history of the material. Organomineralization of microorganisms is likely to represent the first stages of microbial fossilisation and has been hypothesised to prevent the autolytic degradation of microbial cell envelope structures. In the present study, two distinct fossilisation textures (permineralised microfossils and iron oxide encrusted cell envelopes) identified throughout iron-rich rock samples were analysed using nanoscale secondary ion mass spectrometry (NanoSIMS). In this system, aluminium is enriched around the permineralised microfossils, while iron is enriched within the intracellularly, within distinct cell envelopes. Remarkably, while cell wall structures are indicated, carbon and nitrogen biosignatures are not preserved with permineralised microfossils. Therefore, the enrichment of aluminium, delineating these microfossils appears to have been critical to their structural preservation in this iron-rich environment. In contrast, NanoSIMS analysis of mineral encrusted cell envelopes reveals that preserved carbon and nitrogen biosignatures are associated with the cell envelope structures of these microfossils. Interestingly, iron is depleted in regions where carbon and nitrogen are preserved. In contrast aluminium appears to be slightly enriched in regions associated with remnant cell envelope structures. The correlation of aluminium with carbon and nitrogen biosignatures suggests the complexation of aluminium with preserved cell envelope structures before or immediately after cell death may have inactivated autolytic activity preventing the rapid breakdown of these organic, macromolecular structures. Combined, these results highlight that aluminium may play an important role in the preservation of microorganisms within the rock record.

Description: Supergene enriched iron ore deposits in Brazil are typically blanketed by goethite-cemented breccias that form a protective duricrust known as canga. Moderately hard, well consolidated, permeable and resistant to erosion and chemical weathering, the canga blanket protects the relatively friable iron ore below. The protective canga horizons in the Carajás and Quadrilátero Ferrífero mineral provinces represent some of the longest-lived, continuously exposed land surfaces on Earth, and their formation is essential to supergene iron ore enrichment and preservation. Remarkably, the iron-rich duricrusts that have developed in Brazilian tropical rainforest environments, i.e, Carajás, yield geochronological results that indicate that these ancient erosion-resistant surfaces continue to evolve today. Active biogeochemical iron cycling is essential for the ‘self-healing’ cementation/re-cementation occurring in canga, suggesting that recurrent iron reduction and subsequent oxidation are responsible for canga evolution. Macroscopic biological features in canga including ferruginised plant roots and termite tracks have been linked to the biogeochemical cycling of iron. The ‘organic’ textures in canga can be traced to the microscopic scale, preserving fossilised bacterial cell envelopes and permineralised biofilms. At the canga surface, naturally rare and endemic rupestrian plant species carve out an existence, commonly in the absence of soil. Growth of grasses also promotes metal cycling highlighting that the rhizosphere contributes to canga evolution. The fossilisation of microbial biofilms and rhizosphere horizons consolidates canga, affecting its permeability, limiting water transport and enhancing biogeochemical cycling. The development of canga has been essential for the formation, preservation, and discovery of iron ore deposits, and its restoration will ultimately be required for mined land remediation of these unique ecosystems.

Description: Mineralogical and whole rock geochemical analyses for 60 elements on 31 samples of hard ferruginous crust (canga) provide insights into the evolution of the lateritic profile developed on itabirite. Canga can form in two environments: in situ canga that typically caps itabirite and transported canga that covers country rock. Both have similar mineralogical and chemical compositions. Detrital haematite and rare quartz inherited from the itabirite and iron ore comprise the matrix of canga, cemented by goethite, minor gibbsite, and rare manganese oxides and secondary phosphates. Fe2O3 represents more than 91% of its chemical composition and the concentrations of trace elements are low, generally less than 50 ppm. A comparison of the chemical weathering of dolomitic itabirite against the quartz itabirite shows that, although weathering processes are less effective in the former, the geochemical trends of major and trace elements are similar. Negative Ce anomalies (Ce/Ce* = 0.8) and U/Th ratios lower than 1.5 suggest that saprolite formation occurred under slightly anoxic and mildly acidic conditions, allowing rare earth elements (REEs) to remain in the saprolite and also the formation of secondary Al phosphates, instead of Fe phosphates. These conditions became more aggressive during the canga formation process, resulting in further removal of trace elements from the system. The canga formation (pedogenesis) and the chemical weathering of the itabirite (saprolite formation) are independent, but interrelated processes that have been occurring since the Palaeocene.

Description: The surface crust that caps highly weathered banded iron formations (BIFs) supports a unique ecosystem that is a post-mining restoration priority in iron ore areas. Geochemical evidence indicates that biological processes drive the dissolution of iron oxide minerals and contribute to the ongoing evolution of this duricrust. However, limited information is available on present-day biogeochemical processes in these systems, particularly those that contribute to the precipitation of iron oxides and, thus, the cementation and stabilization of duricrusts. Freshly formed iron precipitates in water bodies perched on cangas in Karijini National Park, Western Australia, were sampled for microscopic and molecular analyses to understand currently active microbial contributions to iron precipitation in these areas. Microscopy revealed sheaths and stalks associated with iron-oxidizing bacteria. The iron-oxidizing lineages Sphaerotilus, Sideroxydans, and Pedomicrobium were identified in various samples and Leptothrix was common in four out of five samples. The iron-reducing bacteria Anaeromyxobacter dehalogens and Geobacter lovleyi were identified in the same four samples, with various heterotrophs and diverse cyanobacteria. Given this arid, deeply weathered environment, the driver of contemporary iron cycling in Karijini National Park appears to be iron-reducing bacteria, which may exist in anaerobic niches through associations with aerobic heterotrophs. Overall oxidizing conditions and Leptothrix iron-oxidizers contribute to net iron oxide precipitation in our sampes, rather than a closed biogeochemical cycle, which would result in net iron oxide dissolution as has been suggested for canga caves in Brazil. Enhancements in microbial iron oxide dissolution and subsequent reprecipitation have potential as a surface-crust-ecosystem remediation strategy at mine sites.

Description: Microbial biofilms growing in iron-rich seeps surrounding Lake Violão, Carajás, Brazil serve as a superb natural system to study the role of iron cycling in producing secondary iron cements. These seeps flow across iron duricrusts (referred to as canga in Brazil) into hydraulically restricted lakes in northern Brazil. Canga caps all of the iron ore deposits in Brazil, protecting them from being destroyed by erosion in this active weathering environment. Biofilm samples collected from these seeps demonstrated heightened biogeochemical iron cycling, contributing to the relatively rapid, seasonal formation of iron-rich cements. The seeps support iron-oxidising lineages including Sideroxydans, Gallionella, and an Azoarcus species revealed by 16S rRNA gene sequencing. In contrast, a low relative abundance of putative iron reducers; for example, Geobacter species (〈5% of total sequences in any sample), corresponds to carbon limitation in this canga-associated ecosystem. This carbon limitation is likely to restrict anoxic niches to within biofilms. Examination of a canga rock sample collected from the edge of Lake Violão revealed an array of well- to poorly-preserved microbial fossils in secondary iron cements. These heterogeneous cements preserved bacterial cell envelopes and possibly extracellular polymeric substances within the microfossil iron-rich cements (termed biocements). Bacterial iron reduction initiates the sequence, and intuitively is the rate-limiting step in this broadly aerobic environment. The organic framework of the active- and paleo-biofilms appears to provide a scaffold for the formation of some cements within canga and likely expedites cement formation. The accelerated development of these resilient iron-rich biocements in the lake edge environment compared with surroundings duricrust-associated environments may provide insights into new approaches to remediate mined land, aiding to stabilise slopes, reduce erosion, restore functional hydrogeology and provide a substrate akin to natural canga for revegetation using endemic canga plant species, which have adapted to grow on iron-rich substrates.

Description: Accelerating microbial iron cycling is an innovative environmentally responsible strategy for mine remediation. In the present study, we extend the application of microbial iron cycling in environmental remediation, to include biocementation for the aggregation and stabilization of mine wastes. Microbial iron reduction was promoted monthly for 10 months in crushed canga (a by‐product from iron ore mining, dominated by crystalline iron oxides) in 1 m3 containers. Ferrous iron concentrations reached 445 ppm in treatments and diverse lineages of the candidate phyla radiation dominated pore waters, implicating them in fermentation and/or metal cycling in this system. After a 6‐month evaporation period, iron‐rich cements had formed between grains and 20‐cm aggregates were recoverable from treatments throughout the 1‐m depth profile, while material from untreated and water‐only controls remained unconsolidated. Canga‐adapted plants seeded into one of the treatments germinated and grew well. Therefore, application of this geobiotechnology offers promise for stabilization of mine wastes, as well as re‐formation of surface crusts that underpin unique and threatened plant ecosystems in iron ore regions.