Microfluidic study in a meter-long reactive path reveals how the medium's structural heterogeneity shapes MICP-induced biocementation.

Microbially induced calcium carbonate (CaCO3) precipitation (MICP) is one of the major sustainable alternatives to the artificial cementation of granular media. MICP consists of injecting the soil with bacterial- and calcium-rich solutions sequentially to form calcite bonds among the soil particles that improve the strength and stiffness of soils. The performance of MICP is governed by the underlying microscale processes of bacterial growth, reactive transport of solutes, reaction rates, crystal nucleation and growth. However, the impact of pore-scale heterogeneity on these processes during MICP is not well understood. This paper sheds light on the effect of pore-scale heterogeneity on the spatiotemporal evolution of MICP, overall chemical reaction efficiency and permeability evolution by combining two meter-long microfluidic devices of identical dimensions and porosity with homogeneous and heterogeneous porous networks and real-time monitoring. The two chips received, in triplicate, MICP treatment with an imposed flow and the same initial conditions, while the inlet and outlet pressures were periodically monitored. This paper proposes a comprehensive workflow destined to detect bacteria and crystals from time-lapse microscopy data at multiple positions along a microfluidic replica of porous media treated with MICP. CaCO3 crystals were formed 1 h after the introduction of the cementation solution (CS), and crystal growth was completed 12 h later. The average crystal growth rate was overall higher in the heterogeneous porous medium, while it became slower after the first 3 h of cementation injection. It was found that the average chemical reaction efficiency presented a peak of 34% at the middle of the chip and remained above 20% before the last 90 mm of the reactive path for the heterogeneous porous network. The homogeneous porous medium presented an overall lower average reaction efficiency, which peaked at 27% 420 mm downstream of the inlet and remained lower than 12% for the rest of the microfluidic channel. These different trends of chemical efficiency in the two networks are due to a higher number of crystals of higher average diameter in the heterogeneous medium than in the homogeneous porous medium. In the interval between 480 and 900 mm, the number of crystals in the heterogeneous porous medium is more than double the number of crystals in the homogeneous porous medium. The average diameters of the crystals were 23-46 µm in the heterogeneous porous medium, compared to 17-40 µm in the homogeneous porous medium across the whole chip. The permeability of the heterogeneous porous medium was more affected than that of the homogeneous system, while the pressure sensors effectively captured a higher decrease in the permeability during the first two hours when crystals were formed and a less prominent decrease during the subsequent seeded growth of the existing crystals, as well as the nucleation and growth of new crystals.

Controlling the calcium carbonate microstructure of engineered living building materials.

The fabrication of responsive soft materials that enable the controlled release of microbial induced calcium carbonate (CaCO3) precipitation (MICP) would be highly desirable for the creation of living materials that can be used, for example, as self-healing construction materials. To obtain a tight control over the mechanical properties of these materials, needed for civil engineering applications, the amount, location, and structure of the forming minerals must be precisely tuned; this requires good control over the dynamic functionality of bacteria. Despite recent advances in the self-healing of concrete cracks and the understanding of the role of synthesis conditions on the CaCO3 polymorphic regulation, the degree of control over the CaCO3 remains insufficient to meet these requirements. We demonstrate that the amount and location of CaCO3 produced within a matrix, can be controlled through the concentration and location of bacteria; these parameters can be precisely tuned if bacteria are encapsulated, as we demonstrate with the soil-dwelling bacterium Sporosarcina pasteurii that is deposited within biocompatible alginate and carboxymethyl cellulose (CMC) hydrogels. Using a competitive ligand exchange mechanism that relies on the presence of yeast extract, we control the timing of the release of calcium ions that crosslink the alginate or CMC without compromising bacterial viability. With this novel use of hydrogel encapsulation of bacteria for on-demand release of MICP, we achieve control over the amount and structure of CaCO3-based composites and demonstrate that S. pasteurii can be stored for up to 3 months at an accessible storage temperature of 4 °C, which are two important factors that currently limit the applicability of MICP for the reinforcement of construction materials. These composites thus have the potential to sense, respond, and heal without the need for external intervention.

Direct currents stimulate carbonate mineralization for soil improvement under various chemical conditions.

The present study integrates direct electric currents into traditional calcium carbonate mineralization to investigate electrochemical interactions and the subsequent crystalline growth of CaCO3 bonds in sand. A specific line of focus refers to the effect of three chemical reactive species involved in the stimulated geo-chemo-electric system, namely CaCl2, Ca(CH3COO)2 and Ca(CH3CH2(OH)COO)2. By altering treatment conditions and the applied electric field, we capture distinctive trends related to the: (i) overall reaction efficiencies and distribution of CaCO3 crystals is sand samples; (ii) promotion of CaCO3 mineralization due to DC (iii) crystallographic and textural properties of mineralized bonds. The study introduces the concept of EA-MICP which stands for Electrically Assisted Microbially Induced Carbonate Precipitation as a means of improving the efficiency of soil bio-cementation compared to traditional MICP-based works. Results reveal both the detrimental and highly beneficial effects that electric currents can hold in the complex, reactive and transport processes involved. An interesting observation refers to the "doped" morphology of CaCO3 crystals, which precipitate under electric fields, validated by crystallographic analyses and microstructural observations.

3-D micro-architecture and mechanical response of soil cemented via microbial-induced calcite precipitation.

We introduce the application of microbial-induced calcite precipitation via the ureolytic soil bacterium Sporosarcina Pasteurii in freeze-dried form, as a means of enhancing overall MICP efficiency and reproducibility for geotechnical engineering applications. We show that the execution of urea hydrolysis and CaCO3 precipitation persist as a "cell-free" mechanism upon the complete breakdown of rehydrated cell clusters. Further, strength and stiffness parameters of bio-cemented sands are determined. Medium-grained bio-cemented sand yields compressive strengths up to 12 MPa while, surprisingly, fine-grained sand yields up to 2.5 MPa for similar bond contents. To understand the observed discrepancies, we undertake a systematic study of the bio-cemented material's microstructure, by combining a series of microstructural inspection tools. The study extends beyond conventional qualitative and textural characterization and provides with new insight into the material's peculiar 3D micro-architecture. We apply a new methodology towards quantifying crucial microscopic characteristics such as the particle sizes of the crystalline bond lattice, the bond-grain contacts and particle orientations. Bonds are found to exhibit distinctive geometries and morphologies when MICP applies to different base materials. We thus contribute to the debate on the importance of factors affecting: (i) MICP efficiency, (ii) the mechanical response and (iii) peculiar micro-architecture of bio-improved geo-materials.