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Journal of Arid Land  2023, Vol. 15 Issue (7): 797-811    DOI: 10.1007/s40333-023-0018-3
Research article     
Optimization of growth medium for microbially induced calcium carbonate precipitation (MICP) treatment of desert sand
Monika DAGLIYA1, Neelima SATYAM1,*(), Ankit GARG2
1Department of Civil Engineering, Indian Institute of Technology, Indore 453552, India
2Guangdong Engineering Center for Structure Safety and Health Monitoring, Shantou University, Shantou 515063, China
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Wind-induced sand erosion is a natural process, and can have several negative impacts on human health, environment, and economy. To mitigate the wind-induced sand erosion, an environmental friendly technique that helps to bind soil particles is desirable. The microbially induced calcium carbonate precipitation (MICP) treatment has lately become renowned and a viable alternative to enhance the binding of sand particles (especially against wind erosion). The efficiency of Sporosarcina pasteurii bacteria in inducing calcite formation can be influenced by various factors, including the type of growth media used for bacterial culture. Most of the studies have mainly validated the efficiency of S. pasteurii bacteria usually under single growth media for the MICP treatment. However, the efficiency of S. pasteurii under different growth media on calcite formation is rarely explored. The current study explores the effect of S. pasteurii bacteria on calcite formation under the presence of three different growth media, namely, molasses (MS), tryptic soy broth (TB), and nutrient broth (NB). The three growth media have been applied in the laboratory with and without bacterial solution (control samples). Altered cementation media concentrations (0.5 and 1.0 M) with different pore volumes (PVs), namely, 0.25, 0.50, and 1.00 PV were used in sand-filled tubes for 7 and 14 treatment cycles (1 cycle=24 h). The pH and EC were measured for 12-h period in every 2 h interval, to monitor values at the time of treatment at room temperature. The calcite precipitation was confirmed using SEM (scanning electron microscope), PXRD (powder X-ray diffraction), and calcimeter tests. It was observed that MS generates lower calcite precipitation as compared with NB and TB. However, MS has the advantage of being more economical and abundant (waste product from sugar mills and refineries) as compared with other growth media (NB and TB). It was observed that the minimum and the maximum calcite precipitation using MS is 5% and 12%, respectively. The findings using MS in the present study was compared with the literature and found that precipitation of calcite using MS is effective to stabilize soil against wind erosion.

Key wordsgrowth media      molasses      tryptic soy broth      nutrient broth      S. pasteurii      calcium carbonate     
Received: 06 December 2022      Published: 31 July 2023
Corresponding Authors: *Neelima SATYAM (E-mail:
Cite this article:

Monika DAGLIYA, Neelima SATYAM, Ankit GARG. Optimization of growth medium for microbially induced calcium carbonate precipitation (MICP) treatment of desert sand. Journal of Arid Land, 2023, 15(7): 797-811.

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Fig. 1 Mechanism of Sporosarcina pasteurii precipitation in desert sand. MICP, microbially induced calcium carbonate precipitation.
Fig. 2 Particle size distribution curve for sand sample (Dagliya et al., 2022b)
Cu (mm) Cc (mm) D10 (mm) D30 (mm) D60 (mm) D50 (mm) OMC
G emax emin
1.83 1.09 0.13 0.18 0.23 0.21 12.70 1.65 2.57 0.90 0.62
Table 1 Basic properties of desert sand
Fig. 3 Systematic diagram for the preparation of bacterial solution, treatment process, and testing kit. (a), bacterial culture MS (molasses); (b), bacterial culture NB (nutrient broth); (c), bacterial culture TB (tryptic soy broth); (d), orbital shaking incubator; (e), spectrophotometer; (f), treatment setup; (g), tubes with cementation solution; (h), drain the cementation solution.
No. Ingredient 0.5 M 1.0 M
1 Nutrient broth (NB; g/L) 3.00 3.00
2 Tryptic soy broth (TB; g/L) 2.50 2.50
3 Molasses (MS; mL/L) 100.00 100.00
4 Urea (mL/L) 30.03 30.03
5 Calcium chloride dehydrate (mL/L) 36.75 73.50
6 Sodium bicarbonate (mL/L) 2.12 2.12
7 Ammonium chloride (mL/L) 10.00 10.00
Table 2 Typical ingredients for cementation solution in 0.5 and 1.0 M concentrations
Sample designation Cementation solution (M) Growth media Pore volume Treatment days
0.5 1.0 TB NB MS 0.25 0.50 1.00 7 14
Table S1 Testing plan summarizing treatment combinations with different growth mediums, cementation solutions, pore volumes, and duration
Fig. 4 Electrical conductivity (EC) for three different growth media and dual cementation media concentration (1.0 M and 0.5 M). (a) bacteria; (b), without bacteria. S, Sporosarcina pasteurii; MS, molasses; NB, nutrient broth; TB, tryptic soy broth.
Fig. 5 pH for three different growth media and dual cementation media concentration (1.0 M and 0.5 M). (a) bacteria; (b), without bacteria. S, Sporosarcina pasteurii; MS, molasses; NB, nutrient broth; TB, tryptic soy broth.
Fig. 6 Variation in CaCO3 percentage for different growth media with and without bacteria solution, different pore volumes, and treatment days. (a), 0.5 M cementation solution; (b), 1.0 M cementation solution. S, Sporosarcina pasteurii; MS, molasses; NB, nutrient broth; TB, tryptic soy broth; PV, pore volume; D, treatment days.
Fig. 7 Scanning electron microscope (SEM) images at different magnifications of soil samples treated with different pore volumes and growth medias after 7 d of curing. (a), untreated sand; (b1-b4), NB with 0.5 M and 0.50 PV or 1.0 M and 1.00 PV; (c1-c4), TB with 0.5 M and 0.50 PV or 1.0 M and 1.00 PV; (d1-d4), MS with 0.5 M and 0.50 PV or 1.0 M and 1.00 PV; (b1-d1, b3-d3), ×5000 magnification; (b2-d2, b4-d4), ×20,000 or 25,000 magnification; S, Sporosarcina pasteurii; NB, nutrient broth; TB, tryptic soy broth; MS, molasses; PV, pore volume; D, treatment days.
Fig. 8 Powder X-ray diffraction (PXRD) analysis under different pore volumes (PVs) and growth medias for 7 and 14 d. (a), untreated sand; (b1-b3), 0.5 M, 0.50 PV, and 7 d with bacteria; (c1-c3), 0.5 M, 0.50 PV, and 14 d with bacteria; (d1-d3), 1.0 M, 1.00 PV, and 7 d with bacteria; (e1-e3), 1.0 M, 1.00 PV, and 14 d with bacteria; (f1-f3), 0.5 M, 0.50 PV, and 7 d without bacteria; (g1-g3), 0.5 M, 0.50 PV, and 14 d without bacteria; S, Sporosarcina pasteurii; MS, molasses; NB, nutrient broth; TB, tryptic soy broth; PV, pore volume; D, treatment days; C, calcium carbonate; Q; quartz.
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