Seeding improves the strength of bio-tiles grown with microbially induced calcium carbonate precipitation.

Bio-tiles are a biobased alternative to conventional tiles that utilise a promising technology called microbially induced calcium carbonate (CaCO3) precipitation (MICP). This technology has low energy requirements and also sequesters carbon. Bio-tiles have been made in previous work using a submersion method, however, the process required additives such as 0.3 M magnesium chloride to achieve bio-tiles that meet international standards. The current study aimed to improve the bio-tile strength properties with CaCO3 crystal seeding and a pumping method instead of the use of magnesium that also increases ionic strength. With this technique, cementation solution containing the required calcium and urea for the MICP reaction was pumped through a sealed mould in a series of programmed treatments. The highest concentration of ureolytic Sporosarcina pasteurii with an effective urease activity of 40 mmol NH4-N/L·min was found to be most beneficial to the breaking strength of the bio-tiles, as were the shortest retention times of 1 h between treatments. Seeding with CaCO3 crystals offered significant benefit to the MICP process. Pre-seeding of the geotextiles was explored and the mass of seeds initially present on the geotextiles was found to have a direct improvement on the breaking strength of 21-82 %, increasing with seed loading. The highest CaCO3 seed loading tested of 0.072 g seeds/cm2 geotextile resulted in bio-tiles with a breaking strength of 940 ± 92 N and a modulus of rupture of 16.4 ± 1.7 N/mm2, meeting international targets for extruded tiles with 6-10 % water absorption. When a seed loading of 0.021 g/cm2 was used instead, bio-tiles meeting targets for tiles with a water absorption of >10 % were produced at 628 ± 18 N and 10.5 ± 1.1 N/mm2.

Synergistic enhancement of cadmium immobilization and soil fertility through biochar and artificial humic acid-assisted microbial-induced calcium carbonate precipitation.

Microbially induced carbonate precipitation (MICP) is emerging as a favorable alternative to traditional soil remediation techniques for heavy metals, primarily due to its environmental friendliness. However, a significant challenge in using MICP for farmland is not only to immobilize heavy metals but also to concurrently enhance soil fertility. This study explores the innovative combination of artificial humic acid (A-HA), biochar (BC), and Sporosarcina pasteurii (S. pasteurii) to mitigate the bioavailability of cadmium (Cd) in contaminated agricultural soils through MICP. X-ray diffraction (XRD) and scanning electron microscope (SEM) analyses revealed that the integration of BC and A-HA significantly enhances Cd immobilization efficiency by co-precipitating with CaCO3. Moreover, this treatment also improved soil fertility and ecological functions, as evidenced by increases in total nitrogen (TN, 9.0-78.2 %), alkaline hydrolysis nitrogen (AN, 259.7-635.5 %), soil organic matter (SOM, 18.1-27.9 %), total organic carbon (TOC, 43.8-48.8 %), dissolved organic carbon (DOC, 36.0-88.4 %) and available potassium (AK, 176.2-193.3 %). Additionally, the relative abundance of dominant phyla such as Proteobacteria and Firmicutes significantly increased with the introduction of BC and A-HA in MICP. Consequently, the integration of BC and A-HA with MICP offers a promising solution for remediating Cd-contaminated agricultural soil and synergistically enhancing soil fertility.

The Effect of Bacteria-to-Calcium Ratio on Microbial-Induced Carbonate Precipitation (MICP) under Different Sequences of Calcium-Source Introduction.

To explore the effects of the introduction order of calcium sources and the bacteria-to-calcium ratio on the microbially induced calcium carbonate precipitation (MICP) product CaCO3 and to achieve the regulation of CaCO3 crystal morphology, the mineralisation products of MICP were compared after combining bacteria and calcium at ratios of 1/9, 2/9, 3/9, 4/9, 5/9, and 6/9. A bacterial solution was combined with a urea solution in two calcium addition modes: calcium-first and calcium-later modes. Finally, under the calcium-first addition method, the output of high-purity vaterite-type CaCO3 was achieved at bacteria-to-calcium ratios of 2/9 and 3/9; under the calcium-later addition method, the output of calcite-type CaCO3 could be stabilised, and the change in the bacteria-to-calcium ratio did not have much effect on its crystalline shape.

Feeding strategies for Sporosarcina pasteurii cultivation unlock more efficient production of ureolytic biomass for MICP.

The bacterium Sporosarcina pasteurii is the most commonly used microorganism for Microbial Induced Calcite Precipitation (MICP) due to its high urease activity. To date, no proper fed-batch cultivation protocol for S. pasteurii has been published, even though this cultivation method has a high potential for reducing costs of producing microbial ureolytic biomass. This study focusses on fed-batch cultivation of S. pasteurii DSM33. The study distinguishes between limited fed-batch cultivation and extended batch cultivation. Simply feeding glucose to a S. pasteurii culture does not seem beneficial. However, it was exploited that S. pasteurii is auxotrophic for two vitamins and amino acids. Limited fed-batch cultivation was accomplished by feeding the necessary vitamins or amino acids to a culture lacking them. Feeding nicotinic acid to a nicotinic acid deprived culture resulted in a 24% increase of the specific urease activity compared to a fed culture without nicotinic acid limitation. Also, extended batch cultivation was explored. Feeding a mixture of glucose and yeast extract results in OD600 of ≈70 at the end of cultivation, which is the highest value published in literature so far. These results have the potential to make MICP applications economically viable.

Biotrapping Ureolytic Bacteria on Sand to Improve the Efficiency of Biocementation.

Microbially induced calcium carbonate precipitation (MICP) has emerged as a novel technology with the potential to produce building materials through lower-temperature processes. The formation of calcium carbonate bridges in MICP allows the biocementation of aggregate particles to produce biobricks. Current approaches require several pulses of microbes and mineralization media to increase the quantity of calcium carbonate minerals and improve the strength of the material, thus leading to a reduction in sustainability. One potential technique to improve the efficiency of strength development involves trapping the bacteria on the aggregate surfaces using silane coupling agents such as positively charged 3-aminopropyl-methyl-diethoxysilane (APMDES). This treatment traps bacteria on sand through electrostatic interactions that attract negatively charged walls of bacteria to positively charged amine groups. The APMDES treatment promoted an abundant and immediate association of bacteria with sand, increasing the spatial density of ureolytic microbes on sand and promoting efficient initial calcium carbonate precipitation. Though microbial viability was compromised by treatment, urea hydrolysis was minimally affected. Strength was gained much more rapidly for the APMDES-treated sand than for the untreated sand. Three injections of bacteria and biomineralization media using APMDES-treated sand led to the same strength gain as seven injections using untreated sand. The higher strength with APMDES treatment was not explained by increased calcium carbonate accrual in the structure and may be influenced by additional factors such as differences in the microstructure of calcium carbonate bridges between sand particles. Overall, incorporating pretreatment methods, such as amine silane coupling agents, opens a new avenue in biomineralization research by producing materials with an improved efficiency and sustainability.

Effect of (in)organic additives on microbially induced calcium carbonate precipitation.

AIM: This study aimed to understand the morphological effects of (in)organic additives on microbially induced calcium carbonate precipitation (MICP). METHODS AND RESULTS: MICP was monitored in real time in the presence of (in)organic additives: bovine serum albumin (BSA), biofilm surface layer protein A (BslA), magnesium chloride (MgCl2), and poly-l-lysine. This monitoring was carried out using confocal microscopy to observe the formation of CaCO3 from the point of nucleation, in comparison to conditions without additives. Complementary methodologies, namely scanning electron microscopy, energy-dispersive X-ray spectroscopy and X-ray diffraction, were employed to assess the visual morphology, elemental composition, and crystalline structures of CaCO3, respectively, following the crystals' formation. The results demonstrated that in the presence of additives, more CaCO3 crystals were produced at 100 min compared to the reaction without additives. The inclusion of BslA resulted in larger crystals than reactions containing other additives, including MgCl2. BSA induced a significant number of crystals from the early stages of the reaction (20 min) but did not have a substantial impact on crystal size compared to conditions without additives. All additives led to a higher content of calcite compared to vaterite after a 24-h reaction, with the exception of MgCl2, which produced a substantial quantity of magnesium calcite. CONCLUSIONS: The work demonstrates the effect of several (in)organic additives on MICP and sets the stage for further research to understand additive effects on MICP to achieve controlled CaCO3 precipitation.

Steerable artificial magnetic bacteria with target delivery ability of calcium carbonate for soil improvement.

The microbial-induced carbonate precipitation (MICP) has acquired significant attention due to its immense potential in sustainable engineering applications, particularly in soil improvement. However, the precise control of microbial-induced calcium carbonate precipitation remains a formidable challenge in engineering practices, owing to the uncertain movement paths of bacteria and the nonuniform distribution of soil pores. Taking inspiration from targeted therapy in medicine, this paper presents novel research on the development and validation of magnetically responsive bacteria. These bacteria demonstrate the ability to target calcium carbonate precipitation in a microfluidic chip, thereby promoting an environmentally friendly and ecologically sustainable biomineralization paradigm. The study focuses on investigating the migration of magnetite nanoparticles (MNPs) in aqueous solutions and enhancing the stability of MNP culture liquids. A specially designed microfluidic chip is utilized to simulate natural sand particles and their pores, while an external magnetic field is applied to precisely control the movement path of the artificial magnetic bacteria, enabling targeted precipitation of calcium carbonate at the micron-scale. Verification of the engineered artificial magnetic bacteria and their ability to induce calcium carbonate precipitation is conducted through SEM-EDS analysis, microfluidic chip observations, and the application of the K-means algorithm and ImageJ software to analyze calcium carbonate formation. The influence of the concentration of magnetic nanoparticles on the calcium carbonate production rate was also studied. The results confirm the potential of the artificial magnetic bacteria for future engineering applications. KEY POINTS: • Sporosarcina pasteurii is first time successfully engineered into artificial magnetic bacteria. • The artificial magnetic bacteria show excellent performance of targeted transportation and directional deposition of CaCO3 in microfluidic chip. • The emergence of artificial magnetic bacteria promotes paradigm shift of next generation environmentally friendly biomineralization.

Microbially Induced Calcium Carbonate Precipitation by Sporosarcina pasteurii: a Case Study in Optimizing Biological CaCO3 Precipitation.

Current production of traditional concrete requires enormous energy investment that accounts for approximately 5 to 8% of the world's annual CO2 production. Biocement is a building material that is already in industrial use and has the potential to rival traditional concrete as a more convenient and more environmentally friendly alternative. Biocement relies on biological structures (enzymes, cells, and/or cellular superstructures) to mineralize and bind particles in aggregate materials (e.g., sand and soil particles). Sporosarcina pasteurii is a workhorse organism for biocementation, but most research to date has focused on S. pasteurii as a building material rather than a biological system. In this review, we synthesize available materials science, microbiology, biochemistry, and cell biology evidence regarding biological CaCO3 precipitation and the role of microbes in microbially induced calcium carbonate precipitation (MICP) with a focus on S. pasteurii. Based on the available information, we provide a model that describes the molecular and cellular processes involved in converting feedstock material (urea and Ca2+) into cement. The model provides a foundational framework that we use to highlight particular targets for researchers as they proceed into optimizing the biology of MICP for biocement production.

Growing bio-tiles using microbially induced calcium carbonate precipitation.

Using the biomimetic process known as microbially induced calcium carbonate precipitation (MICP), the growth of bio-tiles was investigated as an alternative to conventionally fired ceramic tiles which require operating temperatures above 1000 °C, therefore adding to global carbon emissions. The ureolytic activity of Sporosarcina pasteurii was controlled by centrifuging and dilution with fresh yeast extract media. The bio-tiles were grown using a novel submersion method in which custom moulds were placed in exact positions within the bio-reactor and each was mixed individually from beneath. Five parameters were optimised to achieve bio-tiles (dimensions of 100 × 100 × 10 mm) of breaking strength comparable to conventional tiles of equivalent thickness. By optimising ureolytic activity (4.0 mmol/L·min), the cementation solution concentration (0.3 M), the particle size distribution (D10 = 312 µm; D50 = 469 µm), the volume of cementation solution, as well as the addition of supplemental magnesium (0.3 M), bio-tiles with a breaking strength 637 N ± 60 N and a modulus of rupture of 13.0 N/mm2 ± 2.3 N were produced. These parameters exceed the conventional standards of breaking strength and modulus of rupture of 600 N and 8 N/mm2, respectively, the standards set for tiles with a water absorption above 10 %. This is also the first time that an optimum CaCO3 precipitation rate constant has been identified (0.11-0.18 day-1) for producing bio-tiles that meet the strength properties of conventional extruded ceramic tiles. The tile manufacturing technique described in this study is easy to operate and scale since multiple bio-tiles can be produced in larger cementation tanks. This natural tile making process also benefits the environment by operating at room temperature.

Biologically-induced synthetic manganese carbonate precipitate (BISMCP) for potential applications in heavy metal removal.

Heavy metal pollution of water is a burning issue of today's world. Among several strategies involved for heavy metal remediation purpose, biomineralization has shown great potential. Of late, research has been focused on developing effective mineral adsorbents with reduced time and cost consumption. In this present paper, the Biologically-Induced Synthetic Manganese Carbonate Precipitate (BISMCP) was produced based on the biologically-induced mineralization method, employing Sporosarcina pasteurii in aqueous solutions containing urea and MnCl2. The prepared adsorbent was characterized using Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), SEM-energy dispersive X-ray spectroscopy (SEM-EDX), X-ray diffraction (XRD) and BET surface area analyzer. EDX analysis showed the elements in the crystal BISMCP were Mn, C, and O. XRD result of BISMCP determined the crystal structure, which is close to rhodochrosite (MnCO3). Spectral peaks of FTIR at 1641.79 cm-1 confirmed the appearance of C[bond, double bond]O binding, with strong stretching of CO32- in Amide I. From the six kinds of BISMCP produced, sample MCP-6 has the higher specific surface area by BET analysis at 109.01 m2/g, with pore size at 8.76 nm and higher pore volume at 0.178 cm3/g. These specifications will be suitable as an adsorbent for heavy metal removal by adsorption process. This study presents a preliminary analysis of the possibility of BISMCP for heavy metals adsorption using ICP multi-element standard solution XIII (As, Cr, Cd, Cu, Ni, and Zn). BISMCP formed from 0.1 MnCl2 and 30 ml of bacteria volume (MCP-6) produced a better adsorbent material than others concentrations, with the adsorption efficiency of total As at 98.9%, Cr at 97.0%, Cu at 94.7%, Cd at 88.3%, Zn at 48.6%, and Ni at 29.5%. Future work could be examined its efficiency adsorbing individual heavy metals.

Improvement of cadmium immobilization in contaminated paddy soil by using ureolytic bacteria and rice straw.

Cadmium (Cd) in paddy soil can be immobilized via microbially induced carbonate precipitation (MICP), but it poses a risk to the properties and eco-function of the soil. In this study, rice straw coupled with Sporosarcina pasteurii (S. pasteurii) was used to treat Cd-contaminated paddy soil with minimizing the detrimental effects of MICP. Results showed that the application of rice straw coupled with S. pasteurii reduced Cd bioavailability. The X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS) confirmed that Cd immobilization efficiency was increased in the rice straw coupled with S. pasteurii treatment via co-precipitating with CaCO3. Moreover, rice straw coupled with S. pasteurii enhanced soil fertility and ecological functions as reflected by the high amount of alkaline hydrolysis nitrogen (AN) (14.9 %), available phosphorus (AP) (13.6 %), available potassium (AK) (60.0 %), catalase (9.95 %), dehydrogenase (736 %), and phosphatase (214 %). Further, the relative abundance of dominant phyla such as Proteobacteria and Firmicutes significantly increased when applying both rice straw coupled with S. pasteurii. The most significant environmental factors that affected the composition of the bacterial community were AP (41.2 %), phosphatase (34.2 %), and AK (8.60 %). In conclusion, using rice straw mixed with S. pasteurii is a promising application to treat Cd-contaminated paddy soil due to its positive effects on treating soil Cd as well as its ability to reduce the detrimental effects of the MICP process.

Experimental Study on Different Improvement Schemes of EICP-Lignin Solidified Silt.

In practical engineering applications, silt is prone to liquefaction and quicksand. This paper mainly studies the improvement effects of urease, lignin and their mixture on the strength and liquefaction resistance of silt. Based on the results and phenomena of an unconfined compressive strength and dynamic triaxial test, the improvement effects of the compressive strength, deformation resistance and liquefaction resistance of silt under different improvement schemes are analyzed, and the optimal values of the cement or lignin when enzyme-induced calcium carbonate precipitation (EICP) technology, lignin alone or EICP and lignin are obtained. The results show that the optimum concentration of the constant temperature and humidity sample (referred to as the constant humidity sample) and the constant temperature immersion sample (referred to as the soaking sample) of urease in the unconfined compressive strength test is 1.0 mol/L, and the compressive strength of the soaking sample is 4.9 MPa, which is 1.56 times that of the constant humidity sample; the optimum addition ratio of the lignin-improved constant humidity sample is 3%, and its compressive strength is 2.07 Mpa; the optimum addition ratio of the samples immersed at constant temperature is 4%, and the compressive strength is 3.05 MPa; when urease combines with lignin to improve silt, 4% is the best lignin addition ratio, the compressive strength of the constant humidity sample reaches 1.57 Mpa and the compressive strength of the soaking sample reaches 3.75 MPa; in the dynamic triaxial multi-stage cyclic load test, all samples were cured at constant humidity sample, and in the urease modified silt scheme, 1.0 mol/L was the optimal cement concentration; in the scheme of improving silt with lignin, 3% is the optimal addition ratio; when 1.25 mol/L cementation solution plus urease crude extract is combined with different ratios of lignin in the experimental scheme, 3% is the best lignin addition ratio.

Treatment of engineering waste slurries by microbially induced struvite precipitation mechanisms.

With societal development, the growing scale of engineering construction, and the increase in environmental protection requirements, the necessity of engineering waste mud disposal is becoming increasingly prominent. In this study, microbially induced struvite precipitation (MISP) was introduced to treat engineering waste mud. The study mainly focused on: i) the optimal mineralization scheme for microbially induced struvite precipitation, ii) the feasibility of the process and the effect of reaction parameters on treating engineering waste mud with microbially induced struvite precipitation, and iii) the mechanism of microbially induced struvite precipitation in treating engineering waste mud. The results showed that the waste mud could be well treated with 8.36 × 10 6   c e l l ⋅ m L - 1 bacteria, 10 mM urea, 20 mM phosphate buffer, and 25 mM M g C l 2 at pH 7. The kaolin suspension could be effectively flocculated. The flocculation rate reached approximately 87.2% under the optimum mineralization conditions. The flocculation effect was mainly affected by the concentrations of reactants and heavy metals and the suspension pH. The X-ray diffraction (XRD) patterns showed a strong struvite (MAP) diffraction peak. Scanning electron microscopy (SEM) images indicated that under the optimal mineralization conditions, the crystals were large and showed prismatic shapes tilted at both ends with adhered kaolin particles. In summary, this manuscript provides an effective way to treat engineering waste mud, and the findings should have a positive effect on enhancing soil fertility and preventing secondary pollution.

Improvement of microstructure of cementitious composites by microbially-induced calcite precipitation.

In this study, microstructural improvement of cementitious composites was achieved by bacterial CaCO3 precipitation using three bacterial species, namely Sporosarcina pasteurii, Bacillus cereus, and Actinobacteria sp. M135-3, respectively. The final product was comparatively investigated regarding the physical effects of urease activity of different cells on the mortar in the long term.Microstructural improvement was determined by evaluating the pore structure by determining the increase in strength, decrease in water absorption, and capillary water absorption rate of the cement mortars having different microorganism concentrations (106-109 bacteria/ml). These measurements were taken on bacteria-containing and control samples on the 2nd, 7th, 28th, and 56th days, respectively. In addition, calcite and vaterite as calcium carbonate polymorphs formed by the precipitation of calcium carbonate by three types of bacteria were identified by Scanning electron microscopy and energy dispersive X-ray spectroscopy (SEM/EDS), X-ray diffraction (XRD) and Thermogravimetric analysis - Differential scanning calorimetry (TGA-DSC) analyzes.The bacteria-containing mortar samples showed that bacterial species and concentrations directly affect cementitious composites' mechanical and physical properties. Composite samples containing bacteria resulted in statistically significant microstructural improvements measured by higher mechanical strength, lower water absorption value, and capillary water absorption rate compared to control samples, especially at early ages. However, the effect of microbial calcite formation diminishes at later ages, especially at 56-days, attributed to the bacteria cells losing their vitality and integrity and forming spaces inside the mortars.

Sporosarcina pasteurii can be used to print a layer of calcium carbonate.

When using microbiologically induced calcium carbonate precipitation (MICP) to produce calcium carbonate crystals in the cavities between mineral particles to consolidate them, the inhomogeneous distribution of the precipitated calcium carbonate poses a problem for the production of construction materials with consistent parameters. Various approaches have been investigated in the literature to increase the homogeneity of consolidated samples. One approach can be the targeted application of ureolytic organisms by 3D printing. However, to date, this possibility has been little explored in the literature. In this study, the potential to use MICP to print calcium carbonate layers on mineral particles will be investigated. For this purpose, a dispensing unit was modified to apply both a suspension of Sporosarcina pasteurii and a calcination solution containing urea and calcium chloride onto quartz sand. The study showed that after passing through the nozzle, S. pasteurii preserved consistent cell vitality and therefore its potential of MICP. Applying cell suspension and calcination solution through a printing nozzle resulted in a layer of calcium carbonate crystals on quartz sand. This observation demonstrated the proof of concept of printing calcium carbonate by MICP through the nozzle of a dispensing unit. Furthermore, it was shown that cell suspensions of S. pasteurii can be stored at 4°C for a period of 17 days while maintaining its optical density, urease activity and cell vitality and therefore the potential for MICP. This initial concept could be extended in further research to printing three-dimensional (3D) objects to solve the problem of homogeneity in consolidated mineral particles.

Developing a fluorometric urease activity microplate assay suitable for automated microbioreactor experiments.

Quantifying urease activity is an important task for Microbial Induced Calcite Precipitation research. A new urease activity microplate assay using a fluorescent pH indicator is presented. The method is also suitable for automated measurements during microbioreactor experiments. The assay reagent consists of the green fluorescent pH-indicator fluorescein, urea and a phosphate buffer. After sample addition, the microbial urease hydrolyses urea, which results in a pH and hence fluorescence increase. The fluorescence signal can be measured with a microplate reader or with the microbioreactor system BioLector, allowing for automated urease activity measurements during cultivation experiments. In both measurement systems, the fluorescence signal slope highly correlates with the urease activity measured offline with standard methods. Automated measurement is possible, as no sample preparation such as centrifugation or adjusting of the optical density is required. The assay was developed so that the culture samples turbidity, salinity or buffer concentration does not have a negative impact on the fluorescence signal. The assay allows for straightforward, non-hazardous, parallelized, cheap and reliable measurements, making research on ureolytic bacteria for Microbial Induced Calcite Precipitation more efficient. The assay could be adapted to other enzymes, which have a strong impact on the pH value.

Effect of cell density on decrease in hydraulic conductivity by microbial calcite precipitation.

The effect of number of cells deposited on decrease in hydraulic conductivity of porous media using CaCO3 precipitation induced by Sporosarcina pasteurii (ATCC 11,859) was examined in columns packed with glass beads in the range of 0.25 mm and 3 mm in diameter. After resting Sporosarcina pasteurii cells were introduced into the columns, a precipitation solution, which consisted of 500 mM CaCl2 and 500 mM urea, was introduced under continuous flow conditions. It was shown that hydraulic conductivity was decreased by formation of microbially induced CaCO3 precipitation from between 8.37 * 10-1 and 6.73 * 10-2 cm/s to between 3.69 * 10-1 and 1.01 * 10-2 cm/s. The lowest hydraulic conductivity was achieved in porous medium consisting of the smallest glass beads (0.25 mm in diameter) using the highest density of cell suspension (OD600 2.25). The number of the deposited cells differed depending on the glass bead size of the columns. According to the experiments, 7 * 10-9 g CaCO3 was produced by a single resting cell. The urease activity, which led CaCO3 precipitation, depended on presence of high number of cells deposited in the column because the nutrients were not included in the precipitation solution and consequently, the amount of CaCO3 precipitated was proportional with the cell number in the column. A mathematical model was also developed to investigate the experimental results, and statistical analysis was also performed.

Microdroplet-Based In Situ Characterization Of The Dynamic Evolution Of Amorphous Calcium Carbonate during Microbially Induced Calcium Carbonate Precipitation.

Amorphous calcium carbonate (ACC) plays an important role in microbially induced calcium carbonate precipitation (MICP), which has great potential in broad applications such as building restoration, CO2 sequestration, and bioremediation of heavy metals, etc. However, our understanding of ACC is still limited. By combining microscopy of cell-laden microdroplets with confocal Raman microspectroscopy, we investigated the ACC dynamics during MICP. The results show that MICP inside droplets can be divided into three stages: liquid, gel-like ACC, and precipitated CaCO3 stages. In the liquid stage, the droplets are transparent. As the MICP process continues into the gel-like stage, the ACC structure appears and the droplets become opaque. Subsequently, dissolution of the gel-like structure is accompanied by growth of precipitated CaCO3 crystals. The size, morphology, and lifetime of the gel-like structures depend on the Ca2+ concentration. Using polystyrene colloids as tracers, we find that the colloids exhibit diffusive behavior in both the liquid and precipitated CaCO3 stages, while their motion becomes arrested in the gel-like ACC stage. These results provide direct evidence for the formation-dissolution process of the ACC-formed structure and its gel-like mechanical properties. Our work provides a detailed view of the time evolution of ACC and its mechanical properties at the microscale level, which has been lacking in previous studies.

Effects of Bacterial Culture and Calcium Source Addition on Lead and Copper Remediation Using Bioinspired Calcium Carbonate Precipitation.

Lead and copper ions from wastewater induced by metallurgical processes are accumulated in soils, threatening plant and human health. The bioinspired calcium carbonate precipitation is proven effective in improving the cementation between soil particles. However, studies on capsulizing heavy metal ions using the bioinspired calcium carbonate precipitation are remarkably limited. The present study conducted a series of test tube experiments to investigate the effects of bacterial culture and calcium source addition on the remediation efficiency against lead and copper ions. The calcium carbonate precipitation was reproduced using the Visual MINTEQ software package to reveal the mechanism affecting the remediation efficiency. The degradation in the remediation efficiency against lead ions relies mainly upon the degree of urea hydrolysis. However, higher degrees of urea hydrolysis cause remediation efficiency against copper ions to reduce to zero. Such high degree of urea hydrolysis turns pH surrounding conditions into highly alkaline environments. Therefore, pursuing higher degrees of urea hydrolysis might not be the most crucial factor while remedying copper ions. The findings shed light on the importance of modifying pH surrounding conditions in capsulizing copper ions using the bioinspired calcium carbonate precipitation.

Advances in Enzyme Induced Carbonate Precipitation and Application to Soil Improvement: A Review.

Climate change and global warming have prompted a notable shift towards sustainable geotechnics and construction materials within the geotechnical engineer's community. Earthen construction materials, in particular, are considered sustainable due to their inherent characteristics of having low embodied and operational energies, fire resistance, and ease of recyclability. Despite these attributes, they have not been part of the mainstream construction due to their susceptibility to water-induced deterioration. Conventional soil improvement techniques are generally expensive, energy-intensive, and environmentally harmful. Recently, biostabilization has emerged as a sustainable alternative that can overcome some of the limitations of existing soil improvement methods. Enzyme-induced carbonate precipitation (EICP) is a particularly promising technique due to its ease of application and compatibility with different soil types. EICP exploits the urease enzyme as a catalyst to promote the hydrolysis of urea inside the pore water, which, in the presence of calcium ions, results in the precipitation of calcium carbonate. The purpose of this paper is to provide a state-of-the-art review of EICP stabilization, highlighting the potential application of this technique to field problems and identifying current research gaps. The paper discusses recent progress, focusing on the most important factors that govern the efficiency of the chemical reactions and the precipitation of a spatially homogenous carbonate phase. The paper also discusses other aspects of EICP stabilization, including the degree of ground improvement, the prediction of the pore structure of the treated soil by numerical simulations, and the remediation of potentially toxic EICP by-products.