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Abstract Ureolysis-induced calcium carbonate precipitation (UICP) is a biomineral solution where the urease enzyme converts urea and calcium into calcium carbonate. The resulting biomineral can bridge gaps in fractured shale, reduce undesired fluid flow, limit fracture propagation, better store carbon dioxide, and potentially enhance well efficiency. The mechanical properties of shale cores were investigated using a modified Brazilian indirect tensile strength test. An investigation of intact shale using Eagle Ford and Wolfcamp cores was conducted at varying temperatures. Results show no significant difference between shale types (average tensile strength = 6.19 MPa). Eagle Ford displayed higher strength at elevated temperature, but temperature did not influence Wolfcamp. Comparatively, cores with a single, lengthwise heterogeneous fracture were sealed with UICP and further tested for tensile strength. UICP was delivered via a flow-through method which injected 20–30 sequential patterns of ureolytic microorganisms and UICP-promoting fluids into the fracture until permeability reduced by three orders of magnitude or with an immersion method which placed cores treated with guar gum and UICP-promoting fluids into a batch reactor, demonstrating that guar gum is a suitable inclusion and may reduce the number of flow-through injections required. Tensile results for both delivery methods were variable (0.15–8 MPa), and in some cores the biomineralized fracture split apart, possibly due to insufficient sealing and/or heterogeneity in the composite UICP-shale cores. Notably in other cores the biomineralized fracture remained intact, demonstrating more cohesion than the surrounding shale, indicating that UICP may produce a strong seal for subsurface application. 

Abstract Microbially-induced calcium carbonate precipitation (MICP) is a biological process in which microbially-produced urease enzymes convert urea and calcium into solid calcium carbonate (CaCO₃) deposits. MICP has been demonstrated to reduce permeability in shale fractures under elevated pressures, raising the possibility of applying this technology to enhance shale reservoir storage safety. For this and other applications to become a reality, non-invasive tools are needed to determine how effectively MICP seals shale fractures at subsurface temperatures. In this study, two different MICP strategies were tested on 2.54 cm diameter and 5.08 cm long shale cores with a single fracture at 60 ℃. Flow-through, pulsed-flow MICP-treatment was repeatedly applied to Marcellus shale fractures with and without sand (“proppant”) until reaching approximately four orders of magnitude reduction in apparent permeability, while a single application of polymer-based “immersion” MICP-treatment was applied to an Eagle Ford shale fracture with proppant. Low-field nuclear magnetic resonance (LF-NMR) and X-Ray computed microtomography (micro-CT) techniques were used to assess the degree of biomineralization. With the flow-through approach, these tools revealed that while CaCO₃precipitation occurred throughout the fracture, there was preferential precipitation around proppant. Without proppant, the same approach led to premature sealing at the inlet side of the core. In contrast, immersion MICP-treatment sealed off the fracture edges and showed less mineral precipitation overall. This study highlights the use of LF-NMR relaxometry in characterizing fracture sealing and can help guide NMR logging tools in subsurface remediation efforts. 

Abstract Environmentally relevant metagenomes and BONCAT-FACS derived translationally active metagenomes from Powder River Basin coal seams were investigated to elucidate potential genes and functional groups involved in hydrocarbon degradation to methane in coal seams with high- and low-sulfate levels. An advanced subsurface environmental sampler allowed the establishment of coal-associated microbial communities under in situ conditions for metagenomic analyses from environmental and translationally active populations. Metagenomic sequencing demonstrated that biosurfactants, aerobic dioxygenases, and anaerobic phenol degradation pathways were present in active populations across the sampled coal seams. In particular, results suggested the importance of anaerobic degradation pathways under high-sulfate conditions with an emphasis on fumarate addition. Under low-sulfate conditions, a mixture of both aerobic and anaerobic pathways was observed but with a predominance of aerobic dioxygenases. The putative low-molecular-weight biosurfactant, lichysein, appeared to play a more important role compared to rhamnolipids. The methods used in this study—subsurface environmental samplers in combination with metagenomic sequencing of both total and translationally active metagenomes—offer a deeper and environmentally relevant perspective on community genetic potential from coal seams poised at different redox conditions broadening the understanding of degradation strategies for subsurface carbon. 

Abstract. The kinetics of urea hydrolysis (ureolysis) and induced calcium carbonate(CaCO3) precipitation for engineering use in the subsurface wasinvestigated under aerobic conditions using Sporosarcina pasteurii(ATCC strain 11859) as well as Bacillus sphaericus strains 21776and 21787. All bacterial strains showed ureolytic activity inducingCaCO3 precipitation aerobically. Rate constants not normalized tobiomass demonstrated slightly higher-rate coefficients for both ureolysis(kurea) and CaCO3 precipitation (kprecip)for B. sphaericus 21776 (kurea=0.10±0.03 h−1, kprecip=0.60±0.34 h−1) compared toS. pasteurii (kurea=0.07±0.02 h−1,kprecip=0.25±0.02 h−1), though these differences werenot statistically significantly different. B. sphaericus 21787showed little ureolytic activity but was still capable of inducing someCaCO3 precipitation. Cell growth appeared to be inhibited duringthe period of CaCO3 precipitation. Transmission electron microscopy (TEM) images suggest this is dueto the encasement of cells and was reflected in lower kureavalues observed in the presence of dissolved Ca. However, biomass regrowthcould be observed after CaCO3 precipitation ceased, which suggeststhat ureolysis-induced CaCO3 precipitation is not necessarilylethal for the entire population. The kinetics of ureolysis andCaCO3 precipitation with S. pasteurii was furtheranalyzed under anaerobic conditions. Rate coefficients obtained in anaerobicenvironments were comparable to those under aerobic conditions; however, nocell growth was observed under anaerobic conditions with NO3-,SO42- or Fe3+ as potential terminal electronacceptors. These data suggest that the initial rates of ureolysis andureolysis-induced CaCO3 precipitation are not significantlyaffected by the absence of oxygen but that long-term ureolytic activity mightrequire the addition of suitable electron acceptors. Variations in theureolytic capabilities and associated rates of CaCO3 precipitationbetween strains must be fully considered in subsurface engineering strategiesthat utilize microbial amendments.