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Ecological Engineering
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Article history: Microbial-induced calcite precipitation (MICP) is a relatively green and sustainable soil improvement
Received 19 June 2015 technique in which by-products of a chemical reaction network managed and controlled by biological
Received in revised form activity alter the engineering properties of the soil. The present study carried out a range of laboratory
20 December 2015
tests to investigate the effect of reducing the injected volume of bacterial suspension and the relative
Accepted 22 January 2016
density (as a soil parameter) on bacterial retention, calcite precipitation, and improvement in the strength
and impermeability of sand specimens. The results show that reducing the volume of injected voids to
Keywords:
up to one third of the pore volume did not significantly affect the improvement performance. This is
Microbial-induced calcite precipitation
Bio-grout
important as it can improve the efficiency of the process and make it economical and more practical for
Biological improvement engineering applications. For similar microbial-induced calcite precipitation conditions and soil type, the
Relative density final strength of the improved columns increased and permeability decreased as the relative density of
the soil increased slightly as precipitated calcite decreased.
© 2016 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.ecoleng.2016.01.010
0925-8574/© 2016 Elsevier B.V. All rights reserved.
50 K. Rowshanbakht et al. / Ecological Engineering 89 (2016) 49–55
Fig. 2. Columns: (a) during treatment; (b) improved; (c) setup schematic.
K. Rowshanbakht et al. / Ecological Engineering 89 (2016) 49–55 51
Fig. 3. Efficiency of bacterial suspension fixation vs. RD and VIP: (a) at OD 600; (b) at urease activity.
Fig. 4. Bacterial concentration remaining in situ in column vs. RD and VIP: (a) at OD 600; (b) at urease activity.
To evaluate the efficiency of the bacterial suspension injection, Changes in the ammonium concentration were investigated for
the OD and urease activity of the effluent were normalized (accord- each cementation fluid injection phase. Changes in the ammonium
ing to volume) and are represented as the percentage of initial value concentration at different densities and VIPs were analyzed, but a
pre-injection in Fig. 3. Both the OD and urease activity showed that significant relation was not found. The average ammonium concen-
increasing the VIP by bacterial suspension, decreased the efficiency tration at each injection steps is compared in Fig. 5. It is evident that
of bacterial retention significantly, although they slightly increased after the second injection stage, the ammonium concentration had
as the RD increased (Fig. 3(a) and (b)). It is clear that increasing the decreased significantly. The results of stoichiometry calculations
injection volume increased the loss of bacteria and urease activity indicate that a major portion of the urea injected in the first and
from the column. At higher RDs, increased compaction, a decrease second step participated in the reaction (85–95%). After the third
in porosity, pore volume, and pore throat size, and filtering of bacte- injection, this decreased to about 25%.
ria between the soil grains increased slightly.
3.4. Uniaxial compressive strength
3.2. Bacteria remaining and enzyme activity in column UCS tests were performed on specimens from the base and top
of each column and the results were analyzed. For both specimen
At the same VIP, the initial net volumes of injection differed for
different soil RDs because of the different volumes of voids. As a 40
basis for comparison, the in situ remaining bacterial concentration
and urease activity in each column was measured. These values
Ammonium concentration (g/L)
30
were then normalized by total injection volume. After normaliza-
tion, the values for the effluent are subtracted from the initial values
(pre-injection) to determine the properties of the retained bacte- 20
rial suspension. Overall, increases in VIP and RD (Fig. 4), increased
bacterial retention, but was not important at values above 2/3 PV. 10
The OD in Fig. 4(a) shows maximum values at 2/3 PV and a RD of
85%. Urease activity increased as the VIP increased, but an injection
0
greater than 2/3 PV had no significant effect on retention. Increas-
1st Inj. 2nd Inj. 3rd Inj.
ing the RD slightly increased the urease activity and OD of the
Injection Steps
remaining fluid (Fig. 4(b) and (d)). Increased adsorption was the
result of smaller voids and throats making filtration easier. Fig. 5. Average ammonium concentration at each injection step.
K. Rowshanbakht et al. / Ecological Engineering 89 (2016) 49–55 53
types, an overall increase in average ultimate strength of the sam- the uncompacted samples and CaCO3 crystals formed over a shorter
ples with an increase in the RD was evident (Fig. 6(a)). The increase distance to bridge the sand particles. These results are in agreement
in strength with the increase in RD was obtained by decreasing the with those of Tsukamoto et al. (2013) and Cheng et al. (2014).
calcite content under the same conditions (Fig. 7(a)). The sand par- The average ultimate strength of samples decreased as the
ticles in the compacted sand columns were closer together than in VIP increased (Fig. 6(b)). The ultimate strength of improved
600
400
UCS (kPa)
200
y = 55.824x - 93.211
R² = 0.2126
0
4 5 6 7 8 9
Calcite Content (%)
(a)
8 8
Specimen from base Specimen from base
Specimen from top Specimen from top
Calcite content (%)
6 6
4 4
2 2
0 0
40% 70% 85% 33% 66% 100%
RD (%) IPV
(b) (c)
Fig. 7. (a) Calcite content vs. UCS; effect of RD (b) and VIP (c) on calcite content.
54 K. Rowshanbakht et al. / Ecological Engineering 89 (2016) 49–55
samples varies from 100 to 500 kPa (Fig. 7a). Maximum strength 3.6. Calcite content
was recorded in samples having a RD of 85%; one sample from
the base of the 1/3 PV injection column and one from the top of Small pieces of the specimens extracted near the entrance, mid-
the 2/3 PV injection column. It was observed that the samples dle, and outlet of the columns were prepared and their calcite
prepared from the top of the columns showed greater strength content determined. The results indicated local irregularities in cal-
than those prepared from the bottom, which were nearer the cite distribution, possibly related to sample size. To account for
injection point. This was true for the majority of samples; how- possible local irregularities, after testing the UCS, the entire sam-
ever, the calcite content of the samples from the base of the ple was powdered and a small portion used for calcimetery and the
column was greater than from those at the top. This could be results used for analysis.
in response to the more homogenous distribution of calcite at The calcite content was first plotted versus UCS as shown in
the top of the column. It appears that calcite forms simulta- Fig. 7(a). The calcite percentage varied from 4.5% to 8%. Although
neously during the injection of cementation fluid. Some of this an increasing trend was recorded for UCS, the R2 value (0.21) was
calcite moves with the pore fluid flow and precipitates away from not favourable. This was caused by the difference in RD, inhomo-
the bottom, is distributed more homogenously and, as a result geneous calcite distribution, and its position in the voids (covering
of filtration, greater precipitation in pore throats increase the the grains or precipitation on contact). The relation between pre-
strength. cipitated calcite and RD was obtained for samples prepared from
UCS test specimens by plotting the precipitated calcite at different
RDs (Fig. 7(b)). The results confirmed that the lower the RD, the
greater the precipitation of CaCO3 , which is in agreement with the
3.5. Permeability tests findings of Tsukamoto et al. (2013). This occurred because the low
RD increased the number of microbes and nutrients absorbed by
The results of the permeability tests are presented in Fig. 6(c) the test specimen as a result of the higher void volume.
and (d). Permeability decreased as RD and VIP increased (Fig. 6(c)). This relation was also be seen for VIP (Fig. 7(c)). In general, the
As RD increased, PV decreased and the pore throats become calcite content at the base of the column was greater than at the
smaller and less permeable. The relation between density and per- top. As the other parameters were held constant, this could be the
meability is in agreement with the results of Ng et al. (2012); result of a greater local concentration of bacteria in proximity to
however, some discrepancy was seen at an RD of 85%. This the injection point at the base of the column.
occurred as the pore throats became smaller, making it more
likely for clogging to occurs; this more easily creates preferen- 3.7. Scanning electron microscopy
tial flow paths during bacterial suspension and cementation fluid
injection. The decrease in permeability with the increasing in The results of SEM and XRF in Fig. 8 show calcite precipitation.
VIP could be the result of more local concentration of bacte- The results in Fig. 8(b) show the silica grains and calcite cement.
ria near the injection point, prompting increased local calcite This image demonstrates the increased tendency for precipitation
precipitation. at particles contacts (Fig. 8(a)). Calcite precipitation was compared
Fig. 8. (a) SEM of calcite concentration at grain contact; (b) XRF analysis; (c), base of column at RD = 40% and VIP = 66; (d) at top.
K. Rowshanbakht et al. / Ecological Engineering 89 (2016) 49–55 55
at different densities and VIPs using SEM samples prepared from DeJong, J.T., Fritzges, M.B., Nüsslein, K., 2006. Microbially induced cementation to
the bottom and top of each column. It is impossible from these control sand response to undrained shear. J. Geotech. Geoenviron. Eng. 132,
1381–1392.
images to clearly quantify the differences between samples, but Dick, J., De Windt, W., De Graef, B., Saveyn, H., Van der Meeren, P., De Belie, N.,
there is an evident difference overall between samples from the top Verstraete, W., 2006. Bio-deposition of a calcium carbonate layer on degraded
and bottom of each column. Differences between samples prepared limestone by Bacillus species. Biodegradation 17, 357–367.
Fujita, Y., et al., 2008. Stimulation of microbial urea hydrolysis in groundwa-
from the bottom and top of the column with a RD of 40% with a VIP ter to enhance calcium carbonate precipitation. Environ. Sci. Technol. 42 (8),
of 33% is shown in Fig. 8(c) and (d). 3025–3032.
Gollapudi, U.K., Knutson, C.L., Bang, S.S., Islam, M.R., 1995. A new method for con-
trolling leaching through permeable channels. Chemosphere 30, 695–705.
4. Conclusion Greenberg, A.E., Clesceri, L.S., Eaton, A.D., 1992. Standard Methods for the Examina-
tion of Waste and Wastewater. American Public Health Society, Washington.
Gurbuz, A., Sari, Y.D., Yuksekdag, Z.N., Cinar, B., 2011. Cementation in a matrix
The present study showed that phase-injection of bacterial sus- of loose sandy soil using biological treatment method. Afr. J. Biotechnol. 10,
pension, fixation, and cementation fluid expanded cementation 7432–7440.
throughout the 20 cm experimental column length by means of Hammes, F., BoonN., de Villiers, J., Verstraete W., Siciliano, S.D., 2003. Strain-specific
ureolytic microbial calcium carbonate precipitation. Appl. Environ. Microbiol.
calcite precipitation. The calcite content varied from 4.5% to 8%.
69, 4901–4909.
The efficiency of the bacterial suspension injection decreased as Harkes, M.P., van Paassen, L.A., Whiffin, V.S., van Loosdrecht, M.C.M., 2010. Fixation
the injection volume of bacterial suspension increased. Increasing and distribution of bacterial activity in sand to induce carbonate precipitation
for ground reinforcement. Ecol. Eng. 36, 112–117.
the RD of the soil at all VIPs slightly increased the efficiency of the
Ivanov, V., Chu, J., 2008. Applications of microorganisms to geotechnical engineering
bacterial suspension injection. Changing the VIP from 1/3 to 1 did for bioclogging and biocementation of soil in situ. Rev. Environ. Sci. Biotechnol.
not improve performance significantly. The permeability of the soil 7, 139–153.
samples decreased as the RD increased; at each RD, permeabil- Jimenez-Lopez, C., Jroundi, F., Pascolini, C., Rodriguez-Navarro, C., Pinar-Larrubia,
G., Rodriguez-Gallego, M., Gonzalez-Munoz, M.T., 2008. Consolidation of quarry
ity decreased as the VIP increased. Increasing the RD, decreased calcarenite by calcium carbonate precipitation induced by bacteria activated
calcite precipitation and increased UCS. The decrease in calcite among the microbiota inhabiting the stone. Int. Biodeterior. Biodegrad. 62,
precipitation is related to the reduction in void volume and the 352–363.
Jonkers, H.M., Thijssen, A., Muyzer, G., Copuroglu, O., Schlangen, E., 2010. Application
resulting decrease in bacterial and cementation fluid. Sand particles of bacteria as self-healing agent for the development of sustainable concrete.
in the compacted columns were closer together than in the uncom- Ecol. Eng. 36, 230–235.
pacted columns and CaCO3 crystals formed over a shorter distance Kaltenbacher, Z., Mate, M., Kopenetz, L., Farcas, V., Molnar, C., 2014. Synthesis of
research on biogrout soil improvement method. Construcţii 2, 89–95.
to bridge the sand particles and increase the strength. Increasing Kim, D., Park, K., Kim, D., 2014. Effects of ground conditions on microbial cementation
the volume of bacterial fluid injection to more than 1/3 PV did in soils. Materials 7, 143–156.
not increase calcite precipitation, but decreased it. Furthermore, it Martinez, B.C., DeJong, J.T., Ginn, T.R., Montoya, B.M., Barkouki, T.H., Hunt, C., Tanyu,
B., Major, D., 2013. Experimental optimization of microbial-induced carbon-
decreased the UCS. Overall, the calcite content of the samples from
ate precipitation for soil improvement. J. Geotech. Geoenviron. Eng. 139 (4),
the bottom of the column was greater than those from the top of the 587–598.
column, although the strength decreased. This could be the result Mortensen, B.M., Haber, M.J., DeJong, J.T., Caslake, L.F., Nelson, D.C., 2011. Effects of
environmental factors on microbial induced calcium carbonate precipitation. J.
of higher local and nonhomogeneous concentrations of calcite at
Appl. Microbiol. 111, 338–349.
the bottom of the column. Despite the local irregularities, electron Nelson, S.J., Launt, P.D., 1991. Stripper well production increased with MEOR treat-
microscopy showed differences in calcite precipitation between the ment. Oil Gas J. 89, 114–118.
top and bottom of the column. Decreasing the volume of bacterial Nemati, M., Voordouw, G., 2003. Modification of porous media permeability, using
calcium carbonate produced enzymatically in situ. Enzym. Microb. Technol. 33,
suspension made the process more practical for field applications 635–642.
and more economical. Nemati, M., Greene, E.A., Voordouw, G., 2005. Permeability profile modification
using bacterially formed calcium carbonate: comparison with enzymic option.
Process Biochem. 40, 925–933.
References Ng, W.S., Lee, M.L., Hii, S.L., 2012. An overview of the factors affecting microbial-
induced calcite precipitation and its potential application in soil improvement.
ASTM D2487-11, 2006. Standard Practice for Classification of Soils for Engineering World Acad. Sci. Eng. Technol. 62, 723–729.
Purposes (Unified Soil Classification System). Annual book of ASTM Standards, Ramachandran, S.K., Ramakrishnan, V., Bang, S.S., 2001. Remediation of concrete
West Conshohocken. using microorganisms. ACI Mater. J. 98, 3–9.
ASTM D2166, 2006. Standard Test Method for Unconfined Compressive Strength of D’Aquino Henriques, R.A., 2011. Estudio Relativo al Hormigón Bacteriano: Fab-
Cohesive Soil. Annual book of ASTM Standards, West Conshohocken. ricacióny Potenciales Campos de Aplicación. Tesis de Master. Universitat
ASTM D421-87, 2007. Standard Practice for Dry Preparation of Soil Samples for Politecnica de Catalunya.
Particle-Size Analysis and Determination of Soil Constants. Annual book of ASTM Sarda, D., Choonia, H., Sarode, D., Lele, S., 2009. Biocalcification by Bacillus pasteurii
Standards, West Conshohocken. urease: a novel application. J. Ind. Microbiol. Biotechnol. 36, 1111–1115.
ASTM D422-63, 2007. Standard Test Method for Particle-Size Analysis of Soils. Sarmast, M., Farpoor, M.H., Sarcheshmehpoor, M., Eghbal, M.K., 2014. Micromor-
Annual book of ASTM Standards, West Conshohocken. phological and biocalcification effects of Sporosarcina pasteurii and Sporosarcina
ASTM D4253-00, 2006. Standard Test Methods for Maximum Index Density and Unit ureae in sandy soil columns. J. Agric. Sci. Technol. 16, 681–693.
Weight of Soils Using a Vibratory Table. Annual book of ASTM Standards, West Shirakawa, M.A., Kaminishikawahara, K., Vanderley, M.J., Henrique, K., Marcos, M.F.,
Conshohocken. 2011. Sand bioconsolidation through the precipitation of calcium carbonate by
ASTM D4254-00, 2006. Standard Test Methods for Minimum Index Density and two ureolytic bacteria. Mater. Lett. 65, 1730–1733.
Unit Weight of Soils and Calculation of Relative Density. Annual book of ASTM Tagliaferri, F., Waller, J., Ando, E., Hall, S.A., Viggiani, G., Besuelle, P., DeJong, J.T.,
Standards, West Conshohocken. 2011. Observing strain localization processes in bio-cemented sand using X-ray
Achal, V., Pan, X., Özyurt, N., 2011. Improved strength and durability of fly ash- imaging. Granul. Matter 13, 247–250.
amended concrete bymicrobial calcite precipitation. Ecol. Eng. 37, 554–559. Tsukamoto, M., Inagaki, T., Sasaki, Y., Oda, K., 2013. Influence of relative den-
Cheng, L., Shahin, M.A., Cord-Ruwisch, R., Addis, M., Hartanto, T., Elms, C., 2014. Soil sity on microbial carbonate precipitation and mechanical properties of sand.
stabilisation by microbial induced calcium carbonate precipitation: investiga- In: Proceedings of the 18th International Conference on Soil Mechanics and
tion of some important physical and environmental aspects. In: 7th International Geotechnical Engineering, Paris.
Congress on Environmental Geotechnics, Australia. Van Paassen, L.A., 2009. Ground Improvement by Microbially Induced Carbonate
Chou, Chiung-Wen, 2007. Bioimprovement of Geotechnical Properties of Sandy Precipitation. Ph.D. Thesis. Delft University of Technology.
Soils. Master of Science. Faculty of the Graduate School of the University of van Paassen, L.A., Ghose, R., van der Linden, T.J.M., van der Star, W.R.L., van
Maryland. Loosdrecht, M.C.M., 2010. Quantifying biomediated ground improvement by
De Belie, N., De Muynck, W., 2009. Crack repair in concrete using biodeposition. In: ureolysis: large-scale biogrout experiment. J. Geotech. Geoenviron. Eng. 136
Concrete Repair, Rehabilitation and Retrofitting II., pp. 777–781. (12), 1721–1728.
De Muynck, W., Debrouwer, D., De Belie, N., Verstraete, W., 2008. Bacterial carbonate van Wijngaarden, W.K., Vermolen, F.J., van Meurs, G.A.M., Vuik, C., 2012. A math-
precipitate on improves the durability of cementitious materials. Cement Concr. ematical model and analytical solution for the fixation of bacteria in Biogrout.
Res. 38, 1005–1014. Transp. Porous Med. 92, 847–866.
DeJong, J.T., Mortensen, B.M., Martinez, B.C., Nelson, D.C., 2010. Biomediated soil Whiffin, V.S., van Paassen, L.A., Harkes, M.P., 2007. Microbial carbonate precipitation
improvement. Ecol. Eng. 36, 197–210. as a soil improvement technique. Geomicrobiol. J. 25 (5), 417–423.