Monitoring of biocement- and biogrout- producing bacteria in desert habitats of Iran
Subject Areas : BacteriologyMehdi Kargar 1 , Mohammad Kargar 2
1 - M.Sc., Young Researchers and Elite Club, Shiraz Branch, Islamic Azad University, Shiraz, Iran.
2 - Professor, Department of Microbiology, Jahrom Branch, Islamic Azad University, Jahrom, Iran
Keywords: 16S rRNA gene, Urease, Biocement, Sporosarcina pasteurii,
Abstract :
Background & Objectives: Microbially induced carbonate precipitation (MICP) is one of the proposed methods for soil stabilization and strengthening. The aim of this study was to isolate and characterize biocement-producing ureolytic bacteria from arid soils. Materials & Methods: In this cross-sectional descriptive study, 280 soil samples were collected from different areas of Iran. Samples were cultured using urea broth medium. Urease-positive bacteria were assessed for the level of urease activity. Identification of active strains was performed based on conventional biochemical tests and 16S rRNA gene sequencing. Optimum temperature and pH conditions for biocement production were evaluated in urease-positive bacteria and was compared with sporosaicina pasteurii (ATCC 11589) strain. Finally, calcium carbonate crystals were assessed byX-ray diffraction (XRD) crystallography test at the temperature of 25 °C and pH of 8.5 as optimal conditions. Results: 304 strains were isolated from 5 different bacterial genera. Bacillus and Sporosarcina were the major ureolytic isolates of arid regions. The most amount of calcium carbonate was produced at the temperature of 25 °C and a pH of 8.5. Ten most effective isolates in the production of urease enzymes and biocement were identified by molecular analysis and subsequently registered in NCBI Genbank. In addition, biocement production by selected isolates was confirmed by XRD test. Conclusion: Our results showed that the amount of urea and calcium chloride plays an important role in calcium carbonate crystals production. S. pasteurii was one of the most important biocement-producing bacteria. Therefore, optimization of growth conditions in higher scale is recommended for further applications in stabilizing the sand, ground strength and bridging.
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2. Rajesh KV, Leena Ch, Vishakha B, Manisha Th. Bio-mineralization and bacterial carbonate
precipitation in mortar and concrete. Biosci Bioengin. 2015; 1: 5-11.
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biocement formation during stimulation and augmentation: implications for soil consolidation.
Front Microbiol. 2017; 8(1267): 17.
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by bacteria and its multiple applications. Springer Plus. 2016; 2 (250): 1-26.
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for the production of inorganic nano- and microstructures. Angew Chem Int. 2003; 42: 614-641.
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of iron oxides. Soil Sci Soc America J. 1998; 62: 930-937.
8. Torkzaban S, Tazehkand SS, Walker SL, Bradford SA. Transport and fate of bacteria in porous
media: coupled effects of chemical conditions and pore space geometry. Water Resources Res.
2008; 44: 1-12.
9. Al-Thawadi SM. High strength in situ biocementation of soil by calcite precipitating locally
isolated ureolytic bacteria. Ph.D. thesis. Perth Western Australia. Mudroch University. 2008;
264.
10. Qian C, Wang R, Cheng L, Wang J. Theory of microbial carbonate precipitation and its
application in restoration of cement-based materials defects. Chin J Chem. 2010; 28: 847-857.
11. Rodriguez-Navarro C, Rodriguez-Gallego M, Ben Chekroun K, Gonzalez-Munoz MT.
Conservation of ornamental stone by Myxococcus xanthus induced carbonate biomineralization.
Appl Env Microbiol. 2003; 69: 2182-2193.
12. Wilson DC, Greenfield EM, Crenshaw MA. Ionotropic nucleation of calcium carbonate by
molluscan matrix. Amer Zool. 1984; 24: 925-932.
13. Worrell E, Price L, Martin N, Hendriks C, Ozawa Meida L. Carbon dioxide emissions from the
global cement industry. Ann Rev Energy Environ. 2001; 26: 303-329.
14. Rahmani S, Forozandeh M, Mosavp M, Rezaeej A. Detection of bacteria by amplifying the
16S rRNA gene with universal primers and RFLP. Med J Islam Repub Iran. 2006; 19(4):
333-338.
15. Whiffin VS, van Paassen L, Harkes MP. Microbial carbonate precipitation as a soil improvement technique. Geomicrobiol J. 2007; 24: 417-423.
16. Laxmana R, Sri R, Manjusha A, Arun Kumar M. Bio cement: An eco friendly construction
material. Int J Curr Eng Technol. 2015; 5(4): 2880-2883.
17. C i C r RC a g Z . C rr c i “ ai a i c m
production via microbially induced calcium carbonate precipitation: Use of limestone and acetic
aci ri r m p r i ig c ic i ma ”. AC ai a C m g. 17; 5
(6): 5183-5190.
18. Maleki M, Ebrahimi S, Asadzadeh F, Emami Tabrizi M. Performance of microbial-induced
carbonate precipitation on wind erosion control of sandy soil. Int JEnviron Sci Technol. 2016;
13(3): 937-944.
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with bacteria. Geostrata-Geo Institute for ASCE. 2005; 36: 13-16.
2. Rajesh KV, Leena Ch, Vishakha B, Manisha Th. Bio-mineralization and bacterial carbonate
precipitation in mortar and concrete. Biosci Bioengin. 2015; 1: 5-11.
3. Dhami NK, Alsubhi WR, Watkin E, Mukherjee A. Bacterial community dynamics and
biocement formation during stimulation and augmentation: implications for soil consolidation.
Front Microbiol. 2017; 8(1267): 17.
4. Periasamy A, Chang-Ho K, Yu Jin Sh, Jae Seong So. Formations of calcium carbonate minerals
by bacteria and its multiple applications. Springer Plus. 2016; 2 (250): 1-26.
5. Bnuerlein E. Biomineralization of unicellular organisms: An unusual membrane biochemistry
for the production of inorganic nano- and microstructures. Angew Chem Int. 2003; 42: 614-641.
6. Van Paassen LA. Biogrout-ground improvement by microbially induced carbonate precipitation.
Delft, the Netherlands, Delft University of Technology. 2009: 195.
7. Brennan EW, Lindsay WL. Reduction and oxidation effect on the solubility and transformation
of iron oxides. Soil Sci Soc America J. 1998; 62: 930-937.
8. Torkzaban S, Tazehkand SS, Walker SL, Bradford SA. Transport and fate of bacteria in porous
media: coupled effects of chemical conditions and pore space geometry. Water Resources Res.
2008; 44: 1-12.
9. Al-Thawadi SM. High strength in situ biocementation of soil by calcite precipitating locally
isolated ureolytic bacteria. Ph.D. thesis. Perth Western Australia. Mudroch University. 2008;
264.
10. Qian C, Wang R, Cheng L, Wang J. Theory of microbial carbonate precipitation and its
application in restoration of cement-based materials defects. Chin J Chem. 2010; 28: 847-857.
11. Rodriguez-Navarro C, Rodriguez-Gallego M, Ben Chekroun K, Gonzalez-Munoz MT.
Conservation of ornamental stone by Myxococcus xanthus induced carbonate biomineralization.
Appl Env Microbiol. 2003; 69: 2182-2193.
12. Wilson DC, Greenfield EM, Crenshaw MA. Ionotropic nucleation of calcium carbonate by
molluscan matrix. Amer Zool. 1984; 24: 925-932.
13. Worrell E, Price L, Martin N, Hendriks C, Ozawa Meida L. Carbon dioxide emissions from the
global cement industry. Ann Rev Energy Environ. 2001; 26: 303-329.
14. Rahmani S, Forozandeh M, Mosavp M, Rezaeej A. Detection of bacteria by amplifying the
16S rRNA gene with universal primers and RFLP. Med J Islam Repub Iran. 2006; 19(4):
333-338.
15. Whiffin VS, van Paassen L, Harkes MP. Microbial carbonate precipitation as a soil improvement technique. Geomicrobiol J. 2007; 24: 417-423.
16. Laxmana R, Sri R, Manjusha A, Arun Kumar M. Bio cement: An eco friendly construction
material. Int J Curr Eng Technol. 2015; 5(4): 2880-2883.
17. C i C r RC a g Z . C rr c i “ ai a i c m
production via microbially induced calcium carbonate precipitation: Use of limestone and acetic
aci ri r m p r i ig c ic i ma ”. AC ai a C m g. 17; 5
(6): 5183-5190.
18. Maleki M, Ebrahimi S, Asadzadeh F, Emami Tabrizi M. Performance of microbial-induced
carbonate precipitation on wind erosion control of sandy soil. Int JEnviron Sci Technol. 2016;
13(3): 937-944.