Abstract
Biocementation or microbially induced calcium carbonate precipitation (MICP) is a feasible biochemical process in enhancing the behavior of geomaterial, that is, soil strengthening and/or remediation. This biochemical process encounters several biotic and abiotic challenges while implementing it in real field conditions. With this, the present study aims to investigate the efficiency and suitability of different ex situ and in situ ureolytic microbes in enhancing the geotechnical properties of the sand in different environmental conditions. The studies of MICP revealed a hindrance of microbial growth and ureolytic enzyme activity of one ex situ strain of S. pasteurii in the prevailing soil anoxic (air restrict) or anaerobic condition. The hindrance was the major reason for the minimal amount of precipitation and no strength gain in the biomodified sample. In contrast, high compressive strength was achieved with an abundant amount of precipitation for the sample catalyzed by another strain of S. pasteurii and isolated Proteus species. The results showed that the specific urease activity varies substantially pertaining to the type of microbes in similar chemical and environmental conditions which directly impact the biomineral precipitation and the rate of strength enhancement. The whole study recommends two major tools, that is, the value of specific urease activity and the ureolysis rate in the prevailing soil condition to compute the suitability of the ureolytic microbe for a successful MICP implementation.
Practical Applications
Microbially induced calcium carbonate precipitation (MICP) via ureolysis is a cost-effective, ecofriendly biochemical process that helps strengthen or remediate the geomaterial. In this process, the ureolytic microbes precipitate the carbonate biomineral in the soil pore and bridge the soil particles which helps in soil strengthening. Isolating and stimulating the native ureolytic microbes is preferable to successfully implement the MICP process in real field conditions. In addition, it is essential to analyze the suitability or efficiency of ureolytic microbes in the MICP process since the type of ureolytic microbes affects the biomineral precipitation activity and also the soil enhancement process. This study isolated four unique native ureolytic microbes from the soil and investigated their efficiency for soil strengthening. The investigation suggests using the isolated facultative ureolytic microbes during MICP field implementation since most of the soil strengthening or remediation is in the subsurface with low or no oxygen availability. For this, feasible stepwise isolation and selection of the ureolytic microbes process are provided to further use it either in soil strengthening or remediation. The study also provides two major parameters to choose the suitability of microbes, that is, the specific enzyme/ureolytic activity unit and the enzyme activity in the prevailing soil environmental conditions for MICP implementation.
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Data Availability Statement
All data, models, or codes that support the findings of this study are available from the corresponding author upon reasonable request.
Acknowledgments
The author is thankful to Prof Dali Naidu Arnepalli of the Department of Civil Engineering at IIT Madras for his supervision and guidance.
Notation
The following symbols are used in this paper:
- %
- percentage;
- (v/v)
- volume/volume;
- (w/v)
- weight/volume;
- µmole
- micromole;
- ATCC
- American Type Culture Collection;
- Cc
- coefficient of gradation;
- Cu
- coefficient of uniformity;
- D50
- average particle diameter;
- EDX
- energy dispersive X-ray;
- ES1
- Ennore fine-sand;
- FTIR
- Fourier-transform infrared spectroscopy analysis;
- g
- gram;
- IMViC
- indole test, methyl red test, Voges–Proskauer test, and citrate utilization test;
- KSP
- solubility product;
- L
- liter;
- M
- Molar;
- MICP
- microbially induced calcium carbonate;
- min
- minute;
- mL
- milliliter;
- mM
- millimolar;
- mm
- millimeter;
- NBU
- nutrient broth urea;
- NCIM
- National Collection of Industrial Microorganisms;
- O/F
- oxidation/fermentation test;
- OD600
- optical density at 600 nm;
- PS1
- Proteus species;
- rpm
- revolution per minute;
- SEM
- scanning electron microscopy;
- SP
- poorly graded sand;
- SP1
- S. pasteurii (NCIM);
- SP2
- S. pasteurii (ATCC);
- TGA
- thermogravimetric analysis;
- U
- unit;
- UCS
- unconfined compressive strength; and
- XRD
- X-ray diffraction.
References
Achal, V., A. Mukherjee, P. C. Basu, and M. S. Reddy. 2009. “Lactose mother liquor as an alternative nutrient source for microbial concrete production by Sporosarcina pasteurii.” J. Ind. Microbiol. Biotechnol. 36 (3): 433–438. https://doi.org/10.1007/s10295-008-0514-7.
Achal, V., and X. Pan. 2011. “Characterization of urease and carbonic anhydrase producing bacteria and their role in calcite precipitation.” Curr. Microbiol. 62 (3): 894–902. https://doi.org/10.1007/s00284-010-9801-4.
Ahmad, J., M. A. Khan, and S. Ahmad. 2023. “State of the art on factors affecting the performance of MICP treated fine aggregates.” Mater. Today: Proc. https://doi.org/10.1016/j.matpr.2023.04.087.
Aksornchu, P., P. Prasertsan, and V. Sobhon. 2008. “Isolation of arsenic tolerant bacteria from arsenic-contaminated soil.” Songklanakarin J. Sci. Technol. 30 (1): 95–102.
Anbu, P., C.-H. Kang, Y.-J. Shin, and J.-S. So. 2016. “Formations of calcium carbonate minerals by bacteria and its multiple applications.” SpringerPlus 5 (1): 250–276. https://doi.org/10.1186/s40064-016-1869-2.
ASTM. 2007. Standard test method for particle-size analysis of soils. ASTM D422. West Conshohocken, PA: ASTM.
ASTM. 2011. Standard practice for classification of soils for engineering purposes (unified soil classification system). ASTM D2487. West Conshohocken, PA: ASTM.
ASTM. 2013. Standard test method for unconfined compressive strength of cohesive soil. ASTM D2166. West Conshohocken, PA: ASTM.
ASTM. 2014. Standard test method for ammonia nitrogen in water. ASTM D1426. West Conshohocken, PA: ASTM.
Baskar, S., R. Baskar, L. Mauclaire, and J. A. Mckenzie. 2006. “Microbially induced calcite precipitation in culture experiments: Possible origin for stalactites in Sahastradhara caves, Dehradun, India.” Curr. Sci. 90 (1): 58–64.
Cappuccino, J. G., and N. Sherman. 2008. “Microbiology: A laboratory manual.” In Benjamin cummings. 8th ed., edited by K. Churchman, 133. San Francisco, CA: Pearson.
Chen, L., Y. Shen, A. Xie, B. Huang, R. Jia, R. Guo, and W. Tang. 2009. “Bacteria-mediated synthesis of metal carbonate minerals with unusual morphologies and structures.” Cryst. Growth Des. 9 (2): 743–754. https://doi.org/10.1021/cg800224s.
Cheng, L., and R. Cord-Ruwisch. 2012. “In situ soil cementation with ureolytic bacteria by surface percolation.” Ecol. Eng. 42 (5): 64–72. https://doi.org/10.1016/j.ecoleng.2012.01.013.
Cheng, L., R. Cord-Ruwisch, and M. A. Shahin. 2013. “Cementation of sand soil by microbially induced calcite precipitation at various degrees of saturation.” Can. Geotech. J. 50 (1): 81–90. https://doi.org/10.1139/cgj-2012-0023.
Costin, S., and S. Ionut. 2007–2013. “ABIS online-bacterial identification software.” Accessed January 4, 2023. http://www.tgw1916.net/bacteria_logare.html.
Dejong, J. T., M. B. Fritzges, and K. Nusslein. 2006. “Microbially induced cementation to control sand response to undrained shear.” J. Geotech. Geoenviron. Eng. 132 (11): 1381–1392. https://doi.org/10.1061/(ASCE)1090-0241(2006)132:11(1381).
Dhami, N. K., M. S. Reddy, and M. S. Mukherjee. 2013. “Biomineralization of calcium carbonates and their engineered applications: A review.” Front. Microbiol. 4 (314): 1–13.
Genge, M. J., A. P. Jones, and G. D. Price. 1995. “An infrared and Raman study of carbonate glasses: Implications for the structure of carbonatite magmas.” Geochim. Cosmochim. Acta 59 (5): 927–937. https://doi.org/10.1016/0016-7037(95)00010-0.
Gomez, M. G., C. M. Anderson, C. M. R. Graddy, J. T. DeJong, D. C. Nelson, and T. R. Ginn. 2017. “Large-Scale comparison of bioaugmentation and biostimulation approaches for biocementation of sands.” J. Geotech. Geoenviron. Eng. 143 (5): 1–13. https://doi.org/10.1061/(ASCE)GT.1943-5606.0001640.
Harkes, M. P., L. A. van Paassen, J. L. Booster, V. S. Whiffin, and M. C. M. van Loosdrecht. 2010. “Fixation and distribution of bacterial activity in sand to induce carbonate precipitation for ground reinforcement.” Ecol. Eng. 36 (2): 112–117. https://doi.org/10.1016/j.ecoleng.2009.01.004.
Ivanov, V., and J. Chu. 2008. “Applications of microorganisms to geotechnical engineering for bioclogging and biocementation of soil in situ.” Rev. Environ. Sci. Bio/Technol. 7 (2): 139–153. https://doi.org/10.1007/s11157-007-9126-3.
Jain, S. 2021. “An overview of factors influencing microbially induced carbonate precipitation for its field implementation.” In Building materials for sustainable and ecological environment, edited by V. Achal, and C. S. Chin, 73–99. Singapore: Springer.
Jain, S., and D. N. Arnepalli. 2019. “Biochemically induced carbonate precipitation in aerobic and anaerobic environments by Sporosarcina pasteurii.” Geomicrobiol. J. 36 (5): 443–451. https://doi.org/10.1080/01490451.2019.1569180.
Jain, S., and D. N. Arnepalli. 2020. “Adhesion and deadhesion of ureolytic bacteria on sand under variable pore fluid chemistry.” J. Environ. Eng. 146 (6): 04020038. https://doi.org/10.1061/(ASCE)EE.1943-7870.0001708.
Jiang, N.-J., et al. 2022. “Bio-mediated soil improvement: An introspection into processes, materials, characterization and applications.” Soil Use Manage. 38 (1): 68–93. https://doi.org/10.1111/sum.12736.
Jin, D., S. Zhao, N. Zheng, D. Bu, Y. Beckers, S. E. Denman, C. S. McSweeney, and J. Wang. 2017. “Differences in ureolytic bacterial composition between the rumen digesta and rumen wall based on ureC gene classification.” Front. Microbiol. 8: 385.
Kang, C.-H., J.-H. Choi, J.-G. Noh, D.-Y. Kwak, S.-H. Han, and J.-S. So. 2014. “Microbially induced calcite precipitation-based sequestration of strontium by Sporosarcina pasteurii WJ-2.” Appl. Biochem. Biotechnol. 174 (7): 2482–2491. https://doi.org/10.1007/s12010-014-1196-4.
Kang, C.-H., Y.-J. Kwon, and J.-S. So. 2016. “Soil bioconsolidation through microbially induced calcite precipitation by Lysinibacillus sphaericus WJ-8.” Geomicrobiol. J. 33 (6): 473–478. https://doi.org/10.1080/01490451.2015.1053581.
Kumar, A., H.-W. Song, S. Mishra, W. Zhang, Y.-L. Zhang, Q.-R. Zhang, and Z.-G. Yu. 2023. “Application of microbial-induced carbonate precipitation (MICP) techniques to remove heavy metal in the natural environment: A critical review.” Chemosphere 318: 137894. https://doi.org/10.1016/j.chemosphere.2023.137894.
Knorst, M. T., R. Neubert, and W. Wohlrab. 1997. “Analytical methods for measuring urea in pharmaceutical formulations.” J. Pharm. Biomed. Anal. 15 (11): 1627–1632. https://doi.org/10.1016/S0731-7085(96)01978-4.
Krieg, N. R., and J. G. Holt. 1984. “Bergey’s manual of systematic bacteriology.” In Vol. 1 of Williams and Wilkins. 1st ed., edited by D. R. Boone, R. W. Castenholz, and G. M. Garrit, 27–57. Baltimore, MD: Springer.
Li, M., K. Wen, Y. Li, and L. Zhu. 2018. “Impact of oxygen availability on microbially induced calcite precipitation (MICP) treatment.” Geomicrobiol. J. 35 (1): 15–22. https://doi.org/10.1080/01490451.2017.1303553.
Lin, W., Y. Gao, W. Lin, Z. Zhuo, W. Wu, and X. Cheng. 2023. “Seawater-based bio-cementation of natural sea sand via microbially induced carbonate precipitation.” Environ. Technol. Innovation 29: 103010. https://doi.org/10.1016/j.eti.2023.103010.
Liu, Y., A. Ali, J. F. Su, K. Li, R. Z. Hu, and Z. Wang. 2023. “Microbial-induced calcium carbonate precipitation: Influencing factors, nucleation pathways, and application in waste water remediation.” Sci. Total Environ. 860: 160439. https://doi.org/10.1016/j.scitotenv.2022.160439.
Mitchell, A. C., E. J. Espinosa-Ortiz, S. L. Parks, A. Phillips, A. B. Cunningham, and R. Gerlach. 2018. “Kinetics of calcite precipitation by ureolytic bacteria under aerobic and anaerobic conditions.” Biogeosciences 16 (4): 1–26.
Mortensen, B. M., M. J. Haber, J. T. Dejong, L. F. Caslake, and D. C. Nelson. 2011. “Effects of environmental factors on microbial induced calcium carbonate precipitation.” J. Appl. Microbiol. 111 (2): 338–349. https://doi.org/10.1111/j.1365-2672.2011.05065.x.
Mujah, D., M. A. Shahin, and L. Cheng. 2017. “State-of-the-art review of bio-cementation by microbially induced calcite precipitation (MICP) for soil stabilization.” Geomicrobiol. J. 34 (6): 524–537. https://doi.org/10.1080/01490451.2016.1225866.
Naveed, M., J. Duan, S. Uddin, M. Suleman, Y. Hui, and H. Li. 2020. “Application of microbially induced calcium carbonate precipitation with urea hydrolysis to improve the mechanical properties of soil.” Ecol. Eng. 153: 105885. https://doi.org/10.1016/j.ecoleng.2020.105885.
Nemati, M., E. A. Greene, and G. Voordouw. 2005. “Permeability profile modification using bacterially formed calcium carbonate: Comparison with enzymic option.” Process Biochem. 40 (2): 925–933. https://doi.org/10.1016/j.procbio.2004.02.019.
Ng, W., M. Lee, and S. Hii. 2012. “An overview of the factors affecting microbial-induced calcite precipitation and its potential application in soil improvement.” World Acad. Eng. 62 (2): 723–729.
Omoregie, A. I., G. Khoshdelnezamiha, N. Senian, D. E. L. Ong, and P. M. Nissom. 2017. “Experimental optimisation of various cultural conditions on urease activity for isolated Sporosarcina pasteurii strains and evaluation of their biocement potentials.” Ecol. Eng. 109: 65–75. https://doi.org/10.1016/j.ecoleng.2017.09.012.
Parks, S. L. 2009. “Kinetics of calcite precipitation by ureolytic bacteria under aerobic and anaerobic conditions.” Master in Science thesis, Dept. of Chemical and Biological Engineering, Montana State Univ.
Patil, U. K., and K. Muskan. 2009. Essentials of biotechnology. New Delhi, India: I. K. International Publishing House.
Rahman, M. M., R. N. Hora, I. Ahenkorah, S. Beecham, M. R. Karim, and A. Iqbal. 2020. “State-of-the-Art review of microbial-induced calcite precipitation and its sustainability in engineering applications.” Sustainability 12 (15): 6281. https://doi.org/10.3390/su12156281.
Rajasekar, A., S. Wilkinson, and C. K. S. Moy. 2021. “MICP as a potential sustainable technique to treat or entrap contaminants in the natural environment: A review.” Environ. Sci. Technol. 6: 100096.
Reig, F. B., J. V. Adelantado, and M. C. Moya Moreno. 2002. “FTIR quantitative analysis of calcium carbonate (calcite) and silica (quartz) mixtures using the constant ratio method. Application to geological samples.” Talanta 58 (4): 811–821. https://doi.org/10.1016/S0039-9140(02)00372-7.
Sarda, D., H. S. Choonia, D. D. Sarode, and S. S. Lele. 2009. “Biocalcification by Bacillus pasteurii urease: A novel application.” J. Ind. Microbiol. Biotechnol. 36 (8): 1111–1115. https://doi.org/10.1007/s10295-009-0581-4.
Stocks-Fischer, S., J. K. Galinat, and S. S. Bang. 1999. “Microbiological precipitation of CaCO3.” Soil Biol. Biochem. 31 (11): 1563–1571. https://doi.org/10.1016/S0038-0717(99)00082-6.
Song, C., C. Wang, D. Elsworth, and S. Zhi. 2022. “Compressive strength of MICP-treated silica sand with different particle morphologies and gradings.” Geomicrobiol. J. 39 (2): 148–154. https://doi.org/10.1080/01490451.2021.2020936.
Tang, C. S., L. Yin, N. Jiang, C. Zhu, H. Zeng, H. Li, and B. Shi. 2020. “Factors affecting the performance of microbial-induced carbonate precipitation (MICP) treated soil: A review.” Environ. Earth Sci. 79 (5): 94. https://doi.org/10.1016/S0039-9140(02)00372-7.
Tobler, D. J., M. O. Cuthbert, R. B. Greswell, M. S. Riley, J. C. Renshaw, S. Handley-sidhu, and V. R. Phoenix. 2011. “Comparison of rates of ureolysis between Sporosarcina pasteurii and an indigenous groundwater community under conditions required to precipitate large volumes of calcite.” Geochim. Cosmochim. Acta 75 (11): 3290–3301. https://doi.org/10.1016/j.gca.2011.03.023.
Wang, Z., X. Zhao, X. Chen, P. Cao, L. Cao, and W. Chen. 2023. “Mechanical properties and constitutive model of calcareous sand strengthened by MICP.” J. Mar. Sci. Eng. 11 (4): 819. https://doi.org/10.3390/jmse11040819.
Wei, S., H. Cui, Z. Jiang, H. Liu, H. He, and N. Fang. 2015. “Biomineralization processes of calcite induced by bacteria isolated from marine sediments.” Braz. J. Microbiol. 46 (2): 455–464. https://doi.org/10.1590/S1517-838246220140533.
Whiffin, V. S., L. A. van Paassen, and M. P. Harkes. 2007. “Microbial carbonate precipitation as a soil improvement technique.” Geomicrobiol. J. 24 (5): 417–423. https://doi.org/10.1080/01490450701436505.
Yan, Y., Y. Tang, G. Xu, J. Lian, and D. Fu. 2019. “Study on the relationship between mechanical properties and mesostructure of microbial cemented sand bodies.” Adv. Mater. Sci. Eng. 2019: 1–13. https://doi.org/10.1155/2019/3684645.
Zhao, Q., L. Li, C. Li, M. Li, F. Amini, and H. Zhang. 2014. “Factors affecting improvement of engineering properties of MICP-treated soil catalyzed by bacteria and urease.” J. Mater. Civ. Eng. 26 (12): 1–10. https://doi.org/10.1061/(ASCE)MT.1943-5533.0001013.
Zhu, T., and M. Dittrich. 2016. “Carbonate precipitation through microbial activities in natural environment, and their potential in biotechnology: A review.” Front. Microbiol. 4 (4): 1–21.
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Published In
Journal of Hazardous, Toxic, and Radioactive Waste
Volume 28 • Issue 1 • January 2024
Copyright
© 2023 American Society of Civil Engineers.
History
Received: Jan 4, 2023
Accepted: Jun 29, 2023
Published online: Aug 17, 2023
Published in print: Jan 1, 2024
Discussion open until: Jan 17, 2024
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