Microbially mediated fossil concretions and their characterization by the latest methodologies: a review.

The study of well-preserved organic matter (OM) within mineral concretions has provided key insights into depositional and environmental conditions in deep time. Concretions of varied compositions, including carbonate, phosphate, and iron-based minerals, have been found to host exceptionally preserved fossils. Organic geochemical characterization of concretion-encapsulated OM promises valuable new information of fossil preservation, paleoenvironments, and even direct taxonomic information to further illuminate the evolutionary dynamics of our planet and its biota. Full exploitation of this largely untapped geochemical archive, however, requires a sophisticated understanding of the prevalence, formation controls and OM sequestration properties of mineral concretions. Past research has led to the proposal of different models of concretion formation and OM preservation. Nevertheless, the formation mechanisms and controls on OM preservation in concretions remain poorly understood. Here we provide a detailed review of the main types of concretions and formation pathways with a focus on the role of microbes and their metabolic activities. In addition, we provide a comprehensive account of organic geochemical, and complimentary inorganic geochemical, morphological, microbial and paleontological, analytical methods, including recent advancements, relevant to the characterization of concretions and sequestered OM. The application and outcome of several early organic geochemical studies of concretion-impregnated OM are included to demonstrate how this underexploited geo-biological record can provide new insights into the Earth's evolutionary record. This paper also attempts to shed light on the current status of this research and major challenges that lie ahead in the further application of geo-paleo-microbial and organic geochemical research of concretions and their host fossils. Recent efforts to bridge the knowledge and communication gaps in this multidisciplinary research area are also discussed, with particular emphasis on research with significance for interpreting the molecular record in extraordinarily preserved fossils.

Influence of native ureolytic microbial community on biocementation potential of Sporosarcina pasteurii.

Microbially induced calcium carbonate precipitation (MICP)/Biocementation has emerged as a promising technique for soil engineering applications. There are chiefly two methods by which MICP is applied for field applications including biostimulation and bioaugmentation. Although bioaugmentation strategy using efficient ureolytic biocementing culture of Sporosarcina pasteurii is widely practiced, the impact of native ureolytic microbial communities (NUMC) on CaCO3 mineralisation via S. pasteurii has not been explored. In this paper, we investigated the effect of different concentrations of NUMC on MICP kinetics and biomineral properties in the presence and absence of S. pasteurii. Kinetic analysis showed that the biocementation potential of S. pasteurii is sixfold higher than NUMC and is not significantly impacted even when the concentration of the NUMC is eight times higher. Micrographic results revealed a quick rate of CaCO3 precipitation by S. pasteurii leading to generation of smaller CaCO3 crystals (5-40 µm), while slow rate of CaCO3 precipitation by NUMC led to creation of larger CaCO3 crystals (35-100 µm). Mineralogical results showed the predominance of calcite phase in both sets. The outcome of current study is crucial for tailor-made applications of MICP.

Bio-composites treatment for mitigation of current-induced riverbank soil erosion.

Mitigation of erosion along the riverbanks is a global challenge. Stabilisers such as cement can control erosion, but it risks the river ecology. This paper presents the erosion characteristics of riverbank soil treated with two biological stabilisers that alleviate the ecological cost. The riverbank soil of one of the largest river systems, Brahmaputra, is treated by bio-polymeric and bio-cement binders and their composite. Moreover, a novel selective bio-stimulation technique has been employed to achieve bio-mineralisation. The soil stabilisation is assessed by needle penetration tests and CaCO3 contents. The specimens were tested in a flow-controlled hydraulic flume subjected to a critical current profile ranging from 0.06 to 0.62 m/s. Soil samples treated up to four cycles of biocementation have been tested at three different slopes (30°, 45° and 53°). The eroded depth and erosion rate are evaluated with image analysis. Up to four-fold reduction in the erosion rate was observed with biocementation treatment. However, cementation beyond a threshold led to the formation of brittle chunks. A bio-composite was devised through a pre-treatment of low-viscosity biopolymer along with biocementation. The bio-composite was found to effectively mitigate the current-induced erosion with 36% lower ammonia production than the equally erosion resistant biocemented counterpart. The dual characteristics of the bio-composite were confirmed with the microstructural analysis. This study unravels the potential of biopolymer-biocement composite as a sustainable erosion mitigation strategy.

Insights into the influence of cell concentration in design and development of microbially induced calcium carbonate precipitation (MICP) process.

Microbially induced calcium carbonate precipitation (MICP) process utilising the biogeochemical reactions for low energy cementation has recently emerged as a potential technology for numerous engineering applications. The design and development of an efficient MICP process depends upon several physicochemical and biological variables; amongst which the initial bacterial cell concentration is a major factor. The goal of this study is to assess the impact of initial bacterial cell concentration on ureolysis and carbonate precipitation kinetics along with its influence on the calcium carbonate crystal properties; as all these factors determine the efficacy of this process for specific engineering applications. We have also investigated the role of subsequent cell recharge in calcium carbonate precipitation kinetics for the first time. Experimental results showed that the kinetics of ureolysis and calcium carbonate precipitation are well-fitted by an exponential logistic equation for cell concentrations between optical density range of 0.1 OD to 0.4 OD. This equation is highly applicable for designing the optimal processes for microbially cemented soil stabilization applications using native or augmented bacterial cultures. Multiple recharge kinetics study revealed that the addition of fresh bacterial cells is an essential step to keep the fast rate of precipitation, as desirable in certain applications. Our results of calcium carbonate crystal morphology and mineralogy via scanning electron micrography, energy dispersive X-ray spectroscopy and X-ray diffraction analysis exhibited a notable impact of cell number and extracellular urease concentration on the properties of carbonate crystals. Lower cell numbers led to formation of larger crystals compared to high cell numbers and these crystals transform from vaterite phase to the calcite phase over time. This study has demonstrated the significance of kinetic models for designing large-scale MICP applications.

Biocementation mediated by native microbes from Brahmaputra riverbank for mitigation of soil erodibility.

Riverbank erosion is a global problem with significant socio-economic impacts. Microbially induced calcite precipitation (MICP) has recently emerged as a promising technology for improving the mechanical properties of soils. The present study investigates the potential of selectively enriched native calcifying bacterial community and its supplementation into the riverbank soil of the Brahmaputra river for reducing the erodibility of the soil. The ureolytic and calcium carbonate cementation abilities of the enriched cultures were investigated with reference to the standard calcifying culture of Sporosarcina pasteurii (ATCC 11859). 16S rRNA analysis revealed Firmicutes to be the most predominant calcifying class with Sporosarcina pasteurii and Pseudogracilibacillus auburnensis as the prevalent strains. The morphological and mineralogical characterization of carbonate crystals confirmed the calcite precipitation potential of these communities. The erodibility of soil treated with native calcifying communities was examined via needle penetration and lab-scale hydraulic flume test. We found a substantial reduction in soil erosion in the biocemented sample with a calcite content of 7.3% and needle penetration index of 16 N/mm. We report the cementation potential of biostimulated ureolytic cultures for minimum intervention to riparian biodiversity for an environmentally conscious alternative to current erosion mitigation practices.

Understanding and creating biocementing beachrocks via biostimulation of indigenous microbial communities.

Bacterially induced precipitation of minerals leading to cementation of natural geological formations has been well recorded in a variety of environments. A range of microbial pathways and geochemical processes have been found to influence the cementation processes; but detailed formation mechanisms and biogeochemical relationships are still not very clear. There has been a growing demand for the application of bacterially driven biocementation in a number of geotechnical engineering applications recently. Here, we aimed to unpin the mechanisms behind the formation of actively mineralising beachrock sediments at Lucky Bay in Western Australia to understand the natural accretionary processes and potential of indigenous bacterial communities in biocementation. We observed ferruginous, aluminosilicate and carbonate cements along with extensive extra polymeric substances, borings with possible microbial activities in certain sections of native beachrock sediments. Cement precipitation under calcium- and iron-rich microenvironments sourced from seawater and iron creek seems to be driven by both biogenic and abiogenic processes in nature. Native microbial communities with a dominance of the genera Halococcus and Marinobacter were recorded. Enrichment of native bacterial communities under seawater media conditions was conducted which lead to successful biomineralisation of calcitic and ferruginous cements under in vitro conditions although the community composition changed significantly. Nanomechanical properties of natural and laboratory synthesised cement crystals showed that engineered biocement is highly promising. The results of this study clearly demonstrate biological influence in the formation of natural cements and hint significant potential of biostimulation which can be harnessed for different engineering applications including coastal erosion.

Microbial Diversity and Mineralogical-Mechanical Properties of Calcitic Cave Speleothems in Natural and in Vitro Biomineralization Conditions.

Natural mineral formations are a window into important processes leading to carbon storage and mineralized carbonate structures formed through abiotic and biotic processes. In the current study, we made an attempt to undertake a comprehensive approach to characterize the mineralogical, mechanical, and microbial properties of different kinds of speleothems from karstic caves; with an aim to understand the bio-geo-chemical processes in speleothem structures and their impact on nanomechanical properties. We also investigated the biomineralization abilities of speleothem surface associated microbial communities in vitro. Mineralogical profiling using techniques such as X-ray powder Diffraction (XRD) and Tescan Integrated Mineral Analyzer (TIMA) demonstrated that calcite was the dominant mineral in the majority of speleothems with Energy Dispersive X-ray Analysis (EDS) indicating a few variations in the elemental components. Differing proportions of polymorphs of calcium carbonate such as aragonite and vaterite were also recorded. Significant variations in trace metal content were recorded through Inductively Coupled Plasma Mass Spectrometer (ICP-MS). Scanning Electron Microscopy (SEM) analysis revealed differences in morphological features of the crystals which varied from triangular prismatic shapes to etched spiky forms. Microbial imprints and associations were seen in a few sections. Analysis of the associated microbial diversity showed significant differences between various speleothems at Phylum level; although Proteobacteria and Actinobacteria were found to be the predominant groups. Genus level microbial associations showed a relationship with the geochemistry, mineralogical composition, and metal content of the speleothems. The assessment of nanomechanical properties measured by Nanoindentation revealed that the speleothems with a dominance of calcite were stronger than the speleothems with mixed calcium carbonate polymorphs and silica content. The in vitro metabolic activity of the microbial communities associated with the surfaces of the speleothems resulted in calcium carbonate crystal precipitation. Firmicutes and Proteobacteria dominated these populations, in contrast to the populations seen in natural systems. The precipitation of calcium carbonate crystals in vitro indicated that microbial metabolic activity may also play an important role in the synthesis and dissociation of biominerals in the natural environment. Our study provides novel evidence of the close relationship between mineralogy, microbial ecology, geochemistry, and nanomechanical properties of natural formations.

Bacterial Community Dynamics and Biocement Formation during Stimulation and Augmentation: Implications for Soil Consolidation.

Microbially-induced CaCO3 precipitation (MICP) is a naturally occurring process wherein durable carbonates are formed as a result of microbial metabolic activities. In recent years, MICP technology has been widely harnessed for applications in civil engineering wherein synthesis of calcium carbonate crystals occurs at ambient temperature paving way for low energy biocement. MICP using pure urease (UA) and carbonic anhydrase (CA) producing bacteria has been promising in laboratory conditions. In the current study we enriched ureolytic and carbonic anhydrase communities in calcareous soil under biostimulation and bioaugmentation conditions and investigated the effect of microbial dynamics on carbonate precipitation, calcium carbonate polymorph selection and consolidation of biological sand column under nutrient limited and rich conditions. All treatments for stimulation and augmentation led to significant changes in the composition of indigenous bacterial population. Biostimulation as well as augmentation through the UA route was found to be faster and more effective compared to the CA route in terms of extracellular enzyme production and carbonate precipitation. Synergistic role of augmented cultures along with indigenous communities was recorded via both the routes of UA and CA as more effective calcification was seen in case of augmentation compared to stimulation. The survival of supplemented isolates in presence of indigenous bacterial communities was confirmed through sequencing of total diversity and it was seen that both UA and CA isolate had the potential to survive along with native communities under high nutrient conditions. Nutrient conditions played significant role in determining calcium carbonate polymorph fate as calcitic crystals dominated under high carbon supplementation. Finally, the consolidation of sand columns via stimulation and augmentation was successfully achieved through both UA and CA route under high nutrient conditions but higher consolidation in short time period was noticed in UA route. The study reports that based upon the organic carbon content in native soils, stimulation can be favored at sites with high organic carbon content while augmentation with repeated injections of nutrients can be applied on poor nutrient soils via different enrichment routes of microbial metabolism.

Biomineralization of calcium carbonates and their engineered applications: a review.

Microbially induced calcium carbonate precipitation (MICCP) is a naturally occurring biological process in which microbes produce inorganic materials as part of their basic metabolic activities. This technology has been widely explored and promising with potential in various technical applications. In the present review, the detailed mechanism of production of calcium carbonate biominerals by ureolytic bacteria has been discussed along with role of bacteria and the sectors where these biominerals are being used. The applications of bacterially produced carbonate biominerals for improving the durability of buildings, remediation of environment (water and soil), sequestration of atmospheric CO2 filler material in rubbers and plastics etc. are discussed. The study also sheds light on benefits of bacterial biominerals over traditional agents and also the issues that lie in the path of successful commercialization of the technology of microbially induced calcium carbonate precipitation from lab to field scale.