Microbially Induced Carbonate Precipitation Process Research Articles

Comparison of the efficiency of traditional MICP and two-step MICP method for immobilizing heavy metals in aquatic environments

The application of the microbial-induced carbonate precipitation (MICP) method for remediating heavy metals (i.e., HMs) has recently garnered significant attention. Nevertheless, the inhibition of urease activity by toxic Cd2+, Pb2+, Zn2+, and Cu2+ poses a challenge for MICP-based remediation of HMs contamination. This study: (1) first performs the traditional MICP tests (in which the bacterial solution, urea solution, and HMs are mixed simultaneously), and investigates the toxic effect of HMs on the urease activity and the immobilization efficiency, (2) analyzes the toxicity and immobilization mechanism during the MICP process by combining the simulation and XRD tests, (3) conducts the two-step MICP tests (which initially mix the bacterial solution and urea solution to promote urea hydrolysis, then add the HMs solutions for HMs precipitation) to improve the immobilization efficiency. The tube experiments and simulations were investigated in the HMs concentration range from 1 to 10mM. Indicators including ammonium concentration, HMs concentrations, and pH were measured/ recorded during the tests. The results show that soluble HMs exhibit a concentration-dependent inhibition of urea hydrolysis during the traditional MICP process, resulting in a decreasing immobilization efficiency. The two-step MICP method can effectively immobilize almost the Cd2+ and Zn2+ when the initial urea hydrolysis period exceeds 1~2h. In addition, a high immobilization rate of over 90% can be achieved for Cu-contaminated solutions at the optimal first-step reaction time. Compared with the traditional MICP procedure, the effective two-step MICP method exhibits more promising application prospects for the immobilization of soluble HMs in aquatic environments.

Microbial-induced carbonate precipitation effectively prevents Pb2+ migration through the soil profile: Lab experiment and model simulation

Due to the inappropriate disposal of waste materials containing lead (Pb) and irrigation with sewage containing Pb, the migration of Pb2+ within the soil profile has been extensively investigated. The conventional Pb2+ block method is challenging to implement due to its complex operational procedures and high construction costs. To address this issue, this study introduces the microbial-induced carbonate precipitation (MICP) technique as a novel approach to impede the migration of Pb2+ in the soil profile. Soil acclimatization with urea resulted in an increased proportion of urease-producing microorganisms, including Bacillus, Paenibacillus, and Planococcaceae, along with heightened expression of urea-hydrolyzing genes (UreA, UreB, UreC, and UreG). This indicates that urea-acclimatized soil (Soil-MICP) possesses the potential to induce carbonate precipitation. Batch Pb2+ fixation experiments confirmed that the fixation efficiency of Soil-MICP on Pb2+ exceeded that of soil without MICP, attributed to the MICP process within the Soil-MICP group. Dynamic migration experiments revealed that the MICP reaction transformed exchangeable lead into carbonate-bound Pb, effectively impeding Pb2+ migration in the soil profile. Additionally, the migration rate of Pb2+ in Soil-MICP was influenced by varying urea amounts, pH levels, and pore flow rates, leading to a slowdown in migration. The Two-site sorption model aptly described the Pb2+ migration process in the Soil-MICP column. This study aims to elucidate the MICP biomineralization process, uncover the in-situ blocking mechanism of MICP on lead in soil, investigate the impact of Pb on key genes involved in urease metabolism, enhance the comprehension of the chemical morphology of lead mineralization products, and provide a theoretical foundation for MICP technology in preventing the migration of Pb2+ in soil profiles.

Microbial induced carbonate precipitation for cadmium removal in flue gas from sludge incineration

Flue gas cadmium emission during sludge incineration damages to the health of humans and ecological environment due to its toxic, persistent, and bioaccumulation. Cadmium removal in flue gas based on microbial induced carbonate precipitation (MICP) was investigated using a denitrifying membrane biofilm reactor (DBR). Cadmium removal efficiency obtained 89.8%. Caenispirillum, Halomonas, Stappia, Thauera were the dominant aerobic denitrifiers, which carried twelve cadmium resistance genes (czcA, czcB, czcC, czcD, cadA, cadC, cmtR, cueR, copZ, ctpC, zipB, zntA), and expressed six cadmium resistant proteins (ZntA, CzcA, CtpA, CopZ, CopR and CopB) and ten denitrifying proteins(NarG, NarH, NarI, NapA, NapB, NirK, NirS, NorB, NorC, NosZ), those proteins regulated binding, transport, exportation of cadmium and denitrification. The upregulation of nitrate reductase and nitrite reductase indicated that cadmium could promote the reduction of NO3- to NO, as shown by an integrated metagenomic and metaproteomic sequencing. Cadmium speciation of biofilms, such as cadmium carbonate, increased in proportion, whereas others, such as organic matter-bound cadmium, decreased. The biological adsorption process would gradually yield to MICP process. The MICP stabilized cadmium in flue gas mainly by precipitating Cd through sodium succinate-induced carbonate precipitation. Flue gas cadmium could transform to cadmium carbonate bioprecipitate; carboxyl, hydroxyl and amine groups in cell surface and humic acid could complex cadmium to form Cd-EPS. Denitrifying bacteria could induce MICP process to stabilize cadmium in flue gas. This provides a green and low-cost DBR process as advanced treatment technology for remove cadmium from flue gas.

Study on the mechanism and stability of microbially induced struvite precipitation (MISP) for enhanced immobilization of uranium

The microbial induced carbonate precipitation (MICP) technique has been shown to immobilize a wide range of heavy metals. However, the MICP process for removing uranium-dominated composite heavy metals ignores the formation of substances such as uranyl carbonate, which can easily lead to uranium migration. Furthermore, the by-product ammonia is one of the bottlenecks limiting the application of MICP. In this work, the microbially induced struvite precipitation (MISP) method was used to enhance U immobilization while reducing by-product contamination. The results showed that the optimal uranium removal rate of the MISP process was increased by 82.98% compared to the MICP. Characterization results confirmed the co-precipitation of U with NH4+ in the form of uramphite and struvite, while the participation of U changed the mineral crystal shape. Stability experiments showed that the maximum release of U from struvite was only 1.27%, which is much lower than the minimum release of U from CaCO3. As a result, the MISP process both enhanced uranium immobilization efficiency, decreased the risk of uranium migration-release, and reduced the environmental impact of the by-product ammonia. These results provided a new perspective for studying U contamination remediation and its co-precipitation behavior in microbial mineralization.

Biogenic calcium improved Cd2+ and Pb2+ immobilization in soil using the ureolytic bacteria Bacillus pasteurii

Bioremediation based on microbial-induced carbonate precipitation (MICP) was conducted in cadmium and lead contaminated soil to investigate the effects of MICP on Cd and Pb in soil. In this study, soil indigenous nitrogen was shown to induce MICP to stabilize heavy metals without inputting exogenous urea. The results showed that applying Bacillus pasteurii coupled with CaCl2 reduced Cd and Pb bioavailability, which could be clarified through the proportion of exchangeable Cd and Pb in soil decreasing by 23.65 % and 12.76 %, respectively. Moreover, B. pasteurii was combined separately with hydroxyapatite (HAP), eggshells (ES), and oyster shells (OS) to investigate their effects on soil heavy metals' chemical fractions, toxicity characteristic leaching procedure (TCLP)-extractable Cd and Pb as well as enzymatic activity. Results showed that applying B. pasteurii in soil significantly decreased the heavy metals in the exchangeable fraction and increased them in the carbonate phase fraction. When B. pasteurii was combined with ES and OS, the content of carbonate-bound Cd increased by 114.72 % and 118.81 %, respectively, significantly higher than when B. pasteurii was combined with HAP, wherein the fraction of carbonate-bound Cd increased by 86 %. The combination of B. pasteurii and biogenic calcium effectively reduced the leached contents of Cd and Pb in soil, and the TCLP-extractable Cd and Pb fractions decreased by 43.88 % and 30.66 %, respectively, in the BP + ES group and by 52.60 % and 41.77 %, respectively, in the BP + OS group. This proved that MICP reduced heavy metal bioavailability in the soil. Meanwhile, applying B. pasteurii and calcium materials significantly increased the soil urease enzyme activity. The microstructure and chemical composition of the soil samples were studied, and the results from scanning electron microscope, Fourier transform infra-red spectroscopy, and X-ray diffraction demonstrated the MICP process and identified the formation of CaCO3, Ca0.67Cd0.33CO3, and PbCO3 in heavy metal-contaminated soil.

The Cd immobilization mechanisms in paddy soil through ureolysis-based microbial induced carbonate precipitation: Emphasis on the coexisting cations and metatranscriptome analysis

Microbial induced carbonate precipitation (MICP) can immobilize metals and reduce their bioavailability. However, little is known about the immobilization mechanism of Cd in the presence of soil cations and the triggered gene expression and metabolic pathways in paddy soil. Thus, microcosmic experiments were conducted to study the fractionation transformation of Cd and metatranscriptome analysis. Results showed that bioavailable Cd decreased from 0.62 to 0.29 mg/kg after 330 d due to the MICP immobilization. This was ascribed to the increase in carbonate bound, Fe-Mn oxides bound, and residual Cd. The underlying immobilization mechanisms could be attributed to the formation of insoluble Cd-containing precipitates, the complexation and lattice substitution with carbonate and Fe, Mn and Al (hydr)oxides, and the adsorption on functional group on extracellular polymers of cell. During the MICP immobilization process, up-regulated differential expression urease genes were significantly enriched in the paddy soil, corresponding to the arginine biosynthesis, purine metabolism and atrazine degradation. The metabolic pathway of bacterial chemotaxis, flagellum assembly, and peptidoglycan biosynthesis and the expression of cadA gene related to Cd excretion enhanced Cd resistance of soil microbiome. Therefore, this study provided new insights into the immobilization mechanisms of Cd in paddy soils through ureolysis-based MICP process.

Understanding the formation and structure of bio-mineralization for self-healing of marine concrete: An experimental and thermodynamic approach

In marine engineering, infrastructures are facing serious issues of deterioration resulted largely from concrete cracking due to the hostile seawater environment. Microbial induced carbonate precipitation (MICP) is an effective and environmentally friendly way to achieve self-healing of concrete cracks. However, the formation and structure of MICP products for crack healing in seawater remain unclear, thus hindering the application of MICP in self-healing of marine structures. By means of experimental and thermodynamic approaches, this work first investigated the evolution of aqueous species as well as the phase assemblages and microstructures of bio-minerals in compound solutions simulating concrete cracks in seawater. The coupling effects of Mg2+ and bacteria on bio-mineralization kinetics result in significant discrepancy between experimental results and thermodynamic outputs in terms of phase assemblages in seawater, yet the morphology depends on Mg2+ rather than bacteria. A comparative analysis on healing products collected from real concrete cracks in seawater revealed a fast abiotic precipitation of brucite followed by the MICP process, which is more conducive to attain a high-efficient self-healing in marine environment.

Properties and mechanisms of steel slag strengthening microbial cementation of cyanide tailings

The advantages of microbial induced carbonate precipitation (MICP) as bio-cementation technology for tailings-solidification are under extensive investigation. In order to improve performance of bio-cementation, many strengthening materials were applied to the bio-cementation of tailings. Steel slag (SS) is a kind of industrial solid waste, its chemical composition and mineral composition are similar to cement, and it has a certain application prospect as an auxiliary cementing material. In this study, the properties and mechanism of SS strengthening MICP cementation of cyanide tailings (CT) were investigated. The results showed that Sporosarcina pasteurii growth is not inhibited by SS, and Sporosarcina pasteurii can promote the hydration reaction of SS, providing a suitable alkaline environment and Ca2+, promoting the production of more CaCO3 in the MICP process. When 200 mL of CT leachate was added 1.4 g SS (200–400 mesh), the adsorption of Cu, Pb, Zn, Cd, total cyanide (T-CN), and free cyanide (F–CN) reached 48.05%, 44.28%, 36.25%, 16.67%, 79.05%, and 67.20%, respectively. The maximum unconfined compressive strength（UCS） of the cemented body (with 5%, 150 mesh SS) was 1.97 MPa, which was 3.396 times as high as that without SS. The cemented body with the addition of SS (5%, 150 mesh) contained more carbonate bound Cu (2.75%), Pb (4.89%), Zn (5.37%), and Cd (5.75%), and less exchangeable Cu (3.65%), Pb (6.85%), Zn (2.27%), and Cd (4.42%) than that without SS. In summary, the addition of SS improved the UCS of cemented bodies and the stability of heavy metals and cyanide, reduced the environmental risks existing in the process of CT storage. Meanwhile, it also provides new ideas for resource utilization of industrial solid waste SS and improvement of mine filling materials.

Effect of a harsh circular environment on self-healing microbial-induced calcium carbonate materials for preventing Pb2+ migration

Microbially induced carbonate precipitation (MICP) is increasingly being explored for Pb-contaminated water bodies and soil remediation. However, the Pb-related precipitate resulting from the MICP process can possibly leach acid over time when subjected to harsh environments, causing serious threats to human health. In this study, for the first time, self-healing microbial-induced calcium carbonate (MICC) materials are proposed and applied to prevent Pb2+ migration where the Pb-related precipitate is acid leached after spore germination, spore-vegetative cell transformation, urease secretion, and urea hydrolysis, thereby producing spore-containing precipitation. This process was repeated five times to explore the effect of a harsh circular environment on self-healing MICC materials. Results indicated that Pb immobilization would have deteriorated if the inosine and trace elements had not been intervened during spore germination and spore-vegetative cell transformation, respectively. The spores and vegetative cells provided extra nucleation sites for Pb2+ and minerals to attach. The extracellular polymeric substances (EPSs) combined Pb2+ with functional groups and chemical bonds to prevent their migration to surrounding environments. The scanning electron microscopy–energy-dispersive X-ray spectroscopy (SEM–EDS) images also indicated that the cerussite mineral was precipitated prior to the calcite mineral because Pb2+ had more affinity to combine with CO32- and OH-. An immobilization efficiency of greater than 95% remained nearly the same after five cycles, while it reduced very quickly to less than 10% after three cycles when neglecting the self-healing MICC materials, thus highlighting their relative merits.

Application of electric fields to improve cementation homogeneity of biogrouted sands

AbstractThe inhomogeneity of cementation is a common problem in soil improvement with microbially induced carbonate precipitation process (MICP), which is caused by uneven spatial distribution of bio‐induced calcium carbonate precipitation. Electric field adjustment method was developed and adopted for improving cementation homogeneity of bio‐grouted sands in this research. Horizontal one‐dimensional biogrouting experiments were conducted in sand columns under 0.1 V/cm of direct current (DC) electric field. The column treated with DC electric field showed a significant improvement in homogeneity of calcium carbonate precipitation as compared to the control (without DC electric field). Spatial distribution of calcium carbonate in columns were affected by the properties of bacteria, whether suspended or attached. In general, the attached bacteria with uneven spatial distribution played an important role in the calcium carbonate precipitation when flow rate was regulated at 0.54 cm/min, while the suspended bacteria could also be attached upon times with consumption of cations during biogrouting. With the influence of electric field, suspended bacterial transport in porous media occurred in one direction to the target locations because of the electrophoresis at an average speed of 0.14 cm/h, while sodium ion was found to be moved from anode to cathode by electromigration approach. Electrolysis of water, on the other hand, resulted a slight fluctuation in pH, even though this variation did not show any significant effect on the metabolic activity of bacteria and microbial‐induced carbonate precipitation process. The redistribution of suspended bacteria and sodium ion in porous media provided a possible for attached bacteria to distribute uniformly, which could be considered as the key factor to influence spatial distribution of the bio‐induced calcium carbonate in application of electric field adjustment method.

Reduction of bioavailability and phytotoxicity effect of cadmium in soil by microbial-induced carbonate precipitation using metabolites of ureolytic bacterium Ochrobactrum sp. POC9.

The application of ureolytic bacteria for bioremediation of soil contaminated with heavy metals, including cadmium (Cd), allows for the efficient immobilization of heavy metals by precipitation or coprecipitation with carbonates. Microbially-induced carbonate precipitation process may be useful also in the case of the cultivation of crop plants in various agricultural soils with trace but legally permissible Cd concentrations, which may be still uptaken by plants. This study aimed to investigate the influence of soil supplementation with metabolites containing carbonates (MCC) produced by the ureolytic bacterium Ochrobactrum sp. POC9 on the Cd mobility in the soil as well as on the Cd uptake efficiency and general condition of crop plants (Petroselinum crispum). In the frame of the conducted studies (i) carbonate productivity of the POC9 strain, (ii) the efficiency of Cd immobilization in soil supplemented with MCC, (iii) crystallization of cadmium carbonate in the soil enriched with MCC, (iv) the effect of MCC on the physico-chemical and microbiological properties of soil, and (v) the effect of changes in soil properties on the morphology, growth rate, and Cd-uptake efficiency of crop plants were investigated. The experiments were conducted in soil contaminated with a low concentration of Cd to simulate the natural environmental conditions. Soil supplementation with MCC significantly reduced the bioavailability of Cd in soil with regard to control variants by about 27-65% (depending on the volume of MCC) and reduced the Cd uptake by plants by about 86% and 74% in shoots and roots, respectively. Furthermore, due to the decrease in soil toxicity and improvement of soil nutrition with other metabolites produced during the urea degradation (MCC), some microbiological properties of soil (quantity and activity of soil microorganisms), as well as the general condition of plants, were also significantly improved. Soil supplementation with MCC enabled efficient Cd stabilization and significantly reduced its toxicity for soil microbiota and plants. Thus, MCC produced by POC9 strain may be used not only as an effective Cd immobilizer in soil but also as a microbe and plant stimulators.

Calcium alternative on microbial dust suppressant and the mechanism of action

Calcium alternative is an important factor affecting microbial-induced carbonate precipitation (MICP) process of microbial dust suppressants, however, previous research only focused on a single strain. To analyze the impact of calcium alternative on microbial community, calcium alternative was introduced in the MICP process of treating coal dust with microbial dust suppressants. Results showed that calcium alternative affected the MICP process of the enriched microorganisms. During the growth period of 48 h, calcium alternative improved the species richness of the genera. Urease activity of the microbial community after calcium nitrate treatment could reach 7.48 mM min−1, and the bioavailability during mineralization cycle was 94.57%. The biochemical processes of MICP were more diversified, such as urea hydrolysis, organic oxidation and nitrate dissimilation reduction. In addition, the mechanisms of calcium tolerance were also diverse, such as osmotic adjustment, metabolism, signal transduction, and interspecific cooperation. Quorum sensing strategy was the dominant of defense mechanism. Next, dust suppression experiments showed that calcium nitrate microbial dust suppressant had strong dust suppression ability, with a rain erosion mass loss of 29.25%, and a wind erosion rate 0.92 g m−2·min−1 under the wind speed of 10 m/s. Although further study was still needed, the findings provided a theoretical basis for calcium alternative and exploited great potentials of microorganisms in coal dust suppression.

Inhibited and Retarded Behavior by Ca2+ and Ca2+/OD Loading Rate on Ureolytic Bacteria in MICP Process.

The estimation of optical density (OD) with viable cells is challenging for engineering purposes. In this study, the OD conversion based on previous study was used. The inhibited and retarded behavior of the microbially induced carbonate precipitation (MICP) process was examined. The experimental results showed that high Ca2+ drastically influences the inhibited and retarded behavior on MICP processes. The analysis showed that the inhibition and retardation effects occurred when the Ca2+/OD loading rate exceeded 8.46 M. The critical value was equal to the proportional constant for obtaining carbonate precipitation rate (CPR) from OD. Due to this, the blending design of materials became possible, with no risk of inhibition. In conclusion, the inhibition and retardation of the MICP process are governed by the Ca2+ load and the linear standard line (LSL), which may be attributed to the capacity or tolerance of viable cells, i.e., CPR/OD = 8.46 M or Ca2+/OD = 8.46 M.

Feasible Utilization of Waste Limestone as a Calcium Source for Microbially Induced Carbonate Precipitation (MICP)

Microbial-induced CaCO3 precipitation (MICP) is an innovative and rapidly developing technology for sand solidification. The idea for this research project was built based on the concept of sustainable development and environmental protection. The specific material used for solidification was soluble calcium ions generated by the reaction of limestone waste, a kind of calcium-rich industrial waste from a quarry, and acetic acid. Using Ca(CH3COO)2 (prepared from limestone waste) as a calcium source resulted in a 31.87% lower MICP cost compared to using CaCl2. An unconfined compressive strength (UCS) test was conducted to characterize the macroscopic mechanical properties of bio-cured sand columns. The mineral composition and the microstructure of sand columns were examined by using X-ray diffraction (XRD) and environmental scanning electron microscopy (ESEM). After response surface optimization, the optimal conditions for the reaction of limestone and CH3COOH were determined, and the calcium acetate yield was up to 96.81%. The UCS of sand samples treated with limestone/acetic acid was 10.61% higher than that of samples treated with calcium chloride. This research confirmed the feasibility of cheap limestone waste and soluble calcium ions generated by acetic acid as a calcium source, instead of calcium chloride, for solidifying sand columns in the MICP process.

Investigation of additive-assisted microbial-induced calcium carbonate precipitation in 3D printed cross fractures

Microbially induced carbonate precipitation (MICP) with additives is a promising approach for seepage control in the field of geotechnical engineering. This study designed and fabricated four cross-fracture models using three-dimensional printing. One of the models did not include additives and was treated with a mixed injection strategy. The other three models were prepared using different types of additives such as calcium carbonate, jute fiber, and gravel particles, which were treated with a staged injection strategy to improve our understanding of the optimized MICP process. The increased mass of calcium carbonate in the model was determined by the difference in the model’s weight before and after four-cycle intervals to evaluate the effect of the three additives and two injection strategies on seepage control and precipitation patterns. The results indicated that the masses of the precipitates increased the most for the model without additive owing to gravity during the MICP process; however, the clogging effect was weakest during the seepage experiment. This may be because the smooth inner surfaces of the model and a higher flow rate prevented the floc and cementation solution from settling. The models that included plant fibers and gravel particles provided more surfaces that facilitated calcium carbonate precipitation and contributed to seepage control. The model with calcium carbonate produced a considerable amount of calcium carbonate and interacted positively with the fracture surfaces, leading to the largest decrease in seepage. In the case of calcium-carbonate additives, a layer of precipitates was formed, which increased the inner surface roughness and enhanced the boundary-layer effect. This promoted the precipitation and settling of floc and the formation of more reliable bridges within the cross-fracture model compared with those of other additives.

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.

The Spatial Distribution of Microbially Induced Carbonate Precipitation in Sand Column with Different Grouting Strategies

For ground improvement via microbially induced carbonate precipitation (MICP), the spatial distribution of biocementation considerably affects the mechanical performance of soil. This study systematically assessed the CaCO3 spatial distribution in sand columns with various grouting strategies. CaCO3 dispersion along the radial and vertical directions was quantitatively evaluated. Furthermore, a series of biochemical monitoring experiments was conducted to investigate the MICP process in sand. The results indicated that the reversed and downward chemical solution grouting and high rates of microbial grouting all promoted CaCO3 vertical uniformity. Moreover, the grouting strategy slightly affected the dispersion degree of radially distributed CaCO3. The variation coefficient basically ranged from 0.1 to 0.3. However, the variation coefficient for radially distributed CaCO3 along the vertical direction was significantly affected by the chemical grouting direction. The biochemical monitoring results indicated that 38% Ca2+ was even consumed near the chemical solution outlet in the chemical solution grouting process. The initial Ca2+ concentration during the intermittent period gradually decreased along the chemical solution grouting direction. The CaCO3 vertical distribution was controlled by a combination of the total MICP reaction time, biomass, and Ca2+ availability in the intermittent phase. Different CaCO3 distributions resulted in each grouting strategy exhibiting specific applicable engineering requirements. Furthermore, the carbon footprint was preliminarily evaluated. The results in this study provide references for the selection of grouting strategies for the MICP technique in engineering applications.

Assessment of the bio-cementation effect on shale soil using ultrasound measurement

The assessment of bio-cementation effect using nondestructive testing methods is important for the application of microbial-induced carbonate precipitation (MICP) technique. In this study, a special grouting system was designed to conduct the bio-grouting of shale soils on a testing model. The ultrasound technique was used to evaluate the effectiveness of bio-cementation. A method was derived to estimate the distribution of ultrasonic wave velocity values within the testing model based on the average values measured on the model surfaces. From the changes of isosurfaces of velocity values, the evolution of soil improvement within the model corresponding to the different number of grouting cycles was analyzed numerically and visually. The SEM imaging technique was used to illustrate the mechanism of influence of the calcium carbonate precipitation on the wave velocity of the bio-cemented specimen. The linear relationships were established among the ultrasonic wave velocity, unconfined compressive strength (UCS), and the amount of CaCO3 precipitations formed in the MICP process. In combination with the application of the ultrasound measurement technique as demonstrated in this work, the relationship provides a very useful nondestructive testing tool to assess the effectiveness of bio-cementation within a large soil body.

Morphological Changes of Calcium Carbonate and Mechanical Properties of Samples during Microbially Induced Carbonate Precipitation (MICP)

Recently, microbially induced carbonate precipitation (MICP) has shown potent potential in the field of civil engineering. The calcium carbonate crystals produced by bacteria during the MICP process play a central role in sticking the soil. However, the morphological changes of calcium carbonate crystals in this process and the mechanical performance of soil in the corresponding stages have not been clearly explored. In this paper, the alterations in the morphology of calcium carbonate crystals were continuously observed via scanning electron microscopy during the MICP process in one week, and the mechanical changes of the samples were monitored every day, so as to reveal the relationship between the morphology of calcium carbonate crystals and the mechanical performance of the samples. The results show that the calcium carbonate crystals undergo a gradual change from ellipsoid to rhombic at the 72nd hour. The mechanical properties of both were greatly improved, among which the compressive strength was increased by 2.78 times compared with the previous time point, and the flexure strength was increased by 2.57 times; this time point was also the time when calcite appears. In addition, we found direct evidence on the first day that bacteria act as the nucleation site of calcium carbonate formation. The above findings have certain guiding significance for the in-depth understanding of the internal microscopic changes of MICP and the influence of calcium carbonate morphology on sample mechanics.

Whole cell evaluation and the enzymatic kinetic study of urease from ureolytic bacteria affected by potentially toxic elements

Microbially induced carbonate precipitation (MICP) is a biomineralization process that has various applications in environmental pollution remediation and restoration of a range of building materials. In this study, a ureolytic bacterium, Lysinibacillus sp. GY3, isolated from an E-waste site, was found as a promising catalyst for remediation of heavy metals via the MICP process. This bacterial isolate produced significant amounts of urease and showed a great persistence in immobilization of potentially toxic elements. A reference ureolytic strain, Bacillus megaterium VS1, was selected in order to compare the efficiency of Lysinibacillus sp. GY3. Study on urease localization indicated 80 % more urease activity secreted extracellularly as for Lysinibacillus sp. GY3 compared to B. megaterium VS1. From the investigation on effects of metals on both intra- and extra-cellular urease, it was clear that Lysinibacillus sp. GY3 produced the most stable urease under conditions of metal pressure, especially retaining more than 70 % activity in the presence of 1 g/L Pb2+ and Zn2+. These results suggest that this isolated microorganism could be promisingly introduced in the MICP process to stabilize complex heavy metal pollutions, with reference to the regulating ability under harsh conditions to stabilize urease activity. This species is so important both for its biological features and environmental impacts. In addition, the present study will bring new insight in the field of metal remediation coupled with enzyme engineered biotechnology.