Microbially Induced CaCO3 Precipitation Research Articles

Coupled effect of microbiologically induced calcium carbonate and biofilms in leachate

Fouling and clogging are persistent challenges to the collection and treatment of leachate. The main components of fouling and clogging are calcium carbonate (CaCO3) and biofilms. However, the relationships between CaCO3 and biofilms remain to be clarified. In this study, the interaction between microbially induced CaCO3 precipitation (MICP) and biofilms was investigated using Luria–Bertani (LB) or urea media. Results showed that the bacteria promoted the precipitation of CaCO3 and the formation of a complex mixture of biofilms. The amount of formed CaCO3 in the urea medium was 12.9 times of that in the LB medium. The high MICP potential in the urea medium was associated with increased pH and alkalinity. In addition, the clogging materials exhibited a layered structure and uneven distribution over the clogging width and depth profile. These results indicated the presence of nucleation sites of CaCO3 on the surface of and inside the bacteria. This research provides insights into the regulation of MICP and biofilms through dynamic control of clogging and fouling.

Calcifying Bacteria Flexibility in Induction of CaCO3 Mineralization.

Microbially induced CaCO3 precipitation (MICP) is considered as an alternative green technology for cement self-healing and a basis for the development of new biomaterials. However, some issues about the role of bacteria in the induction of biogenic CaCO3 crystal nucleation, growth and aggregation are still debatable. Our aims were to screen for ureolytic calcifying microorganisms and analyze their MICP abilities during their growth in urea-supplemented and urea-deficient media. Nine candidates showed a high level of urease specific activity, and a sharp increase in the urea-containing medium pH resulted in efficient CaCO3 biomineralization. In the urea-deficient medium, all ureolytic bacteria also induced CaCO3 precipitation although at lower pH values. Five strains (B. licheniformis DSMZ 8782, B. cereus 4b, S. epidermidis 4a, M. luteus BS52, M. luteus 6) were found to completely repair micro-cracks in the cement samples. Detailed studies of the most promising strain B. licheniformis DSMZ 8782 revealed a slower rate of the polymorph transformation in the urea-deficient medium than in urea-containing one. We suppose that a ureolytic microorganism retains its ability to induce CaCO3 biomineralization regardless the origin of carbonate ions in a cell environment by switching between mechanisms of urea-degradation and metabolism of calcium organic salts.

Bio-precipitation of CaCO3 for soil improvement: A Review

The bio-mineralization of calcium carbonates is an innovative and eco-friendly technique for improving soil, which has recently become increasingly popular in the field of geotechnical engineering. Different bio-cementation methods are employed for improving granular soils, including microbially-induced CaCO3 precipitation (MICP) and enzymatic-induced carbonate precipitation (EICP). These methods offer innovativeness, effectiveness and sustainability when applied in geotechnical engineering, and have been suggested as possible solutions for various different geotechnical issues. More research has been conducted into MICP compared to EICP. Nevertheless, urease sourced from plants likely offers many benefits over the use of bacteria urease in bio-cementation processes. The present study reviews the mechanisms, possible areas for implementation, related benefits and drawbacks of the bio-cementations techniques. Additionally, it will concentrate on the techniques used in the precipitation of CaCO3 involving the urease hydrolysis of urea. The study findings will enhance the comprehension of biotechnical mechanisms and ensure that geotechnical specialists are suitably informed with up-to-date knowledge on this subject.

Constraints on CaCO3 precipitation in superabsorbent polymer by aerobic bacteria.

Microbially induced CaCO3 precipitation (MICP) can give concrete self-healing properties. MICP agents are typically bacterial endospores which are coated into shelled granules, infused into expanded clay, or embedded into superabsorbent polymer (SAP). When small cracks appear in the cured concrete, the encapsulation is broken and the metabolic CO2 production from the germinated bacteria causes healing of the cracks by precipitation of CaCO3. Such systems are being tested empirically at large scales, but survival of endospores through preparation and application, as well as germination and growth kinetics of the germinated vegetative cells, remains poorly resolved. We encapsulated endospores of Bacillus subtilis and Bacillus alkalinitrilicus in crosslinked acrylamide-based SAP and quantified their germination, growth, and, in the case of B. alkalinitrilicus, CaCO3 precipitation potential. The endospores survived crosslinking and desiccation inside the polymer matrix. Microcalorimetry and microscopy showed that ~ 80% of the encapsulated endospores of both strains readily germinated after rehydration of freeze-dried SAP. Germinated cells grew into dense colonies of cells inside the SAP, and those of B. alkalinitrilicus calcified with up to 0.3g CaCO3 produced per g desiccated SAP when incubated aerobically. Measurements by planar optodes indicated that the precipitation rates were inherently oxygen limited due to diffusional constraints, rather than limited by electron donor or Ca2+ availability. Such oxygen limitation will limit MICP in all water-saturated and oxygen-dependent systems, and MICP agents based on anaerobic bacteria, e.g., nitrate reducers, should be developed to broaden the applicability of bioactive self-healing concretes to wet and waterlogged environments.

Aragonite formation induced by open cultures of microbial consortia to heal cracks in concrete: Insights into healing mechanisms and crystal polymorphs

Microbially induced CaCO3 precipitation (MICP), mainly in the form of calcite, has been reported to be an efficient approach for self-healing cracks in concrete. However, little is known about the implementation of aragonite, the other common crystalline form of CaCO3, in bioconcrete. Therefore, a systematic investigation of the crystal polymorphs and the healing efficiency of different cultures, i.e., two microbial consortia under anaerobic (MC-Aa) and anoxic (MC-Ao) conditions and nonureolytic pure-culture bacteria (Bacillus cohnii), was conducted in this study. The results showed that the MC-Ao agent exhibited the maximum values of completely healed crack widths (1.22 mm) after 28 d of healing, which is larger than the values of 0.79 and 0.73 mm for B. cohnii and MC-Aa, respectively. Field emission scanning electron microscopy (FESEM) and X-ray diffraction (XRD) analyses confirmed that the biominerals induced by MC-Aa and B. cohnii are calcite, while those of MC-Ao were 82% aragonite and 18% calcite. It is noteworthy that this proportion of aragonite is the highest reported level for the MICP system to date. In addition, economic evaluation verified that the microbial consortia resulted in a 61% decrease in production costs compared to pure cultures. The intriguing findings obtained in this study may provide a scientific basis for the potential implementation of microbial consortia under anoxic conditions, such as a new self-healing agent, in bioconcrete.

Biochemical process of ureolysis-based microbial CaCO3 precipitation and its application in self-healing concrete.

Urea hydrolysis has already been considered as the most effective pathway for microbially induced CaCO3 precipitation (MICP). The present work first studied the combination of several key factors including initial pH, temperature, and dosage of urea, which contribute to the biochemical process of MICP. Under an amiable condition of pH and temperature, the dosage of urea has a significant impact on the rate of urea degradation and CaCO3 precipitation. A bacteria-based self-healing system was developed by loading healing agents on ceramsite carriers. The self-healing efficiency was evaluated by visual inspection on crack closure, compressive strength regain, and capillary water absorption. A preferable healing effectiveness was obtained when the bacteria and organic nutrients were co-immobilized in carriers. Image analysis showed that cracks up to 273μm could be healed with a crack closure ratio of 86% in 28days. The compressive strength regain increased 24% and the water absorption coefficient decreased 27% compared to the reference. The findings indicated a promising application of ureolysis-based MICP in restoring the mechanical properties and enhancing the durability of concrete.

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.

Long-term sustainability of microbial-induced CaCO3 precipitation in aqueous media

Microbially induced CaCO3 precipitation (MICP) via urea hydrolysis is an emerging technique for soil amelioration, building materials rehabilitation and pollutants sequestration amongst other various environmental applications. The successful application of MICP requires the sustainability of the precipitated CaCO3; to which the fate of ammonia, the main by-product of ureolysis, is potentially significante. Ammonia volatilization and biological ammonia oxidation both induce a pH decrease, which, in turn, might cause CaCO3 dissolution. To examine the potential effect of accumulated ammonia on precipitated CaCO3, we conducted a long-term MICP batch experiment, using environmental enrichment cultures of ureolytic bacteria. Here we show that CaCO3 precipitation was completed within 15–27 days, along with a rise in ammonium concentration. Following completion of ureolysis and precipitation, ammonium concentrations decreased, leading to a pH decrease. About 30 days after precipitation was completed, as much as 30% CaCO3 dissolution, was observed. A two-step model, describing urea hydrolysis followed by the removal of ammonia from the precipitation solution, predicted CaCO3 dissolution due to ammonia volatilization. We suggest that ureolytic MICP might result in ammonia volatilization, leading to significant CaCO3 dissolution. These results provide basic insights into the sustainability of ureolytic MICP and should further encourage removal of the accumulated ammonia from the treated site.

Microbially induced CaCO3 precipitation through denitrification: An optimization study in minimal nutrient environment

So far, researchers investigated microbially induced CaCO3 precipitation (MICP) for soil reinforcement, self-repairing concrete and Ca2+ removal from industrial waste streams. Reported MICP yields were mainly achieved under nutrient-rich conditions. However, creating the tested nutrient-rich conditions in intended applications is both an economical and a practical issue. Therefore, investigation of MICP in more realistic conditions is necessary. This study presents optimization of MICP through denitrification in minimal nutrient conditions. To optimize their MICP performances, we isolated two strains, Pseudomonas aeruginosa and Diaphorobacter nitroreducens, by following an application oriented selection procedure. Upon performance optimization, in 2 days, D. nitroreducens and P. aeruginosa precipitated 14.1 and 18.9gCaCO3/gNO3-N, respectively. Repetitive CaCO3 precipitation was also achieved from a single inoculum in both 2 days and 3 weeks intervals. Selected strains and the process were further evaluated for three MICP applications: (1) Ca2+ removal from paper mill wastewater (2) soil reinforcement, (3) crack repair in concrete. Overall, denitrification was found to be an effective process to remove Ca2+ from paper mill wastewater. P. aeruginosa and D. nitroreducens could be introduced as potential candidates for soil and concrete applications due to their enhanced precipitation yields, resilience and performance under minimal nutrient conditions.