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 Accepted manuscript doi:
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Submitted: 27 July 2017
Published online in ‘accepted manuscript’ format: 16 March 2018
Manuscript title: Use of nebulisers to deliver cementation liquid in granular soils to form
biogrout
Authors: Mark Dyer1 and Matteo Viganotti2
Affiliations: 1Faculty of Science and Engineering, University of Waikato, Hamilton,
Waikato, New Zealand and 2Aecom Ltd, Dublin, Ireland
Corresponding author: Mark Dyer, Faculty of Science and Engineering, University of
Waikato, Hamilton, Waikato, New Zealand. Tel.: 0064220447281.
E-mail: mdyer@waikato.ac.nz

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 Accepted manuscript doi:
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Mark Dyer

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 Accepted manuscript doi:
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Abstract

The use of urease enzyme to deposit calcium carbonate for ground treatment of granular soils has typically

relied on injection or percolation of liquid solutions. The approach can result in an uneven distribution of

biogrout and loss of cementation liquid from percolation through the ground into the underlying groundwater

system. As an alternative approach laboratory experiments have demonstrated the potential of using aerosols to

deliver fine micro droplets of reagent liquids into the unsaturated zone to promote calcium carbonate deposition

at soil particle interfaces. SEM images taken of soil samples show markedly different crystal morphology and

particle bond formations for liquid percolation compared with aerosol injection methods. In particular, the bond

formation created from aerosol droplets are shown to replicate the curved surface of liquid menisci between soil

particles leading to a dual bond menisci structure. The accompanying unconfined compression tests are

comparable with similar increase in strength created from percolation and liquid flushing methods. The results

show the potential to use aerosol liquid delivery to promote ground improvement in the vadose zone for granular

soils where possible collapse of residual soils could be addressed or strengthen of granular soils to resist

additional forces such as earthquake loading.

Keywords: aerosols; atomisation; nebuliser; ground improvement; unsaturated soils; vadose zone; carbonate;

precipitation; bacteria; bond formation; bond failure; compressive strength

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 Accepted manuscript doi:
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1.0 INTRODUCTION

The use of urease enzyme to deposit calcium carbonate for formation of biogrout in granular

soils has typically relied on injection or percolation of liquid solutions. The approach can

result in an uneven distribution of biogrout and loss of cementation liquid through percolation

through the ground into the underlying groundwater system. As an alternative approach,

laboratory experiments have demonstrated the potential for nebulisers to deliver fine aerosol

droplets of cementation liquids into the unsaturated zone to promote calcium carbonate

deposition at soil particle interfaces (Dyer et al 2012). This new for the geotechnical sector

but commonplace for other industries. In the case of the chemical and medical sectors,

aerosols and sprays are used to atomise bulk liquids into fine droplets for a wide range of

technologies from administering pharmaceutical drugs to making freeze dried coffee. In

practice, atomisation of bulk liquids depends on liquid, gaseous, mechanical or electrical

energy (Bayvel and Orzechowski 1993). The droplets are commonly termed the dispersed or

discrete phase and the gas is the continuous phase (Lefebvre 1989). A droplet is typically a

small spherical mass of liquid completely bound by free surface and less than 200 microns in

diameter. The ranges of droplet sizes found in nature extending from a drizzle to smoke are

illustrated in Figure 1.

However, there has been negligible work carried out into the transportation of aerosol

droplets in subsurface such as for the treatment of unsaturated soils. Transportation of

chemicals into the ground for engineering or environmental purposes have tended to rely on

the injection of liquids or gases into the subsurface to promote in-situ bioremediation of

polluted land (Grindstaff 2000, USEPA 1998, Baker and Benson 1999, Krauss et al 2003,

Dyer et al 2003) or improve engineering properties (Littlejohn 2002a, Littlejohn 2002b,

Kazemain et al 2010). Studies by Dyer et al (2012) demonstrated the potential to apply

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 Accepted manuscript doi:
10.1680/jgrim.17.00047
aerosol technology from the medial field to transport fine droplets (0.5-5μm diameter

droplets) of bulk liquids through sands and gravel size particles in laboratory column tests.

The results exhibited the potential to deliver bulk liquids in an efficient manner to promote

biological or chemical reactions within the vadose zone for soil stabilisation or soil

remediation; where depending on droplet size the fate of aerosol droplets depended on a)

inertial impaction, b) gravitational sedimentation or c) Brownian diffusion. The researchers

observed that the presence of water menisci acted as filters or traps for the coarser aerosol

droplets that might otherwise be transported through the column. Consequently the menisci

represent the ideal location for deposition of aerosol droplets to promoted inter-particle

bonding.

To explore the potential of using aerosol technology to deliver bulk liquids for ground

treatment, a series of laboratory were carried out using soil columns. For this study droplets

of between 1-10µm were produced using a medical jet nebulisers Cirrus™ manufactured by

Intersurgical (2003). The jet nebulisers operated using a Venturi nozzle and impactor as

shown in Figure 2. High velocity gas was forced through a Venturi nozzle to disintegrate

liquid held in a mixing chamber. The resulting droplets impacted onto a baffle (impactor) to

further reduce droplet size. In this study, droplets from the medical jet nebulisers Cirrus™

were used to deliver the reagents to promote microbial induced calcium carbonate

precipitation (MICP) in sand.

2.0 OLIOGOTROPHIC AND EURTOPHIC TREATMENT BY LIQUID FLUSHING

Ground treatment using microbial induced calcium carbonate precipitation (MICP) has been

gaining tracking in recent years with extensive laboratory and field studies being carried out

by Whiffin et al., 2007, DeJong et al., 2010, Harkes et al., 2010, Van Der Ruyt and Van Der

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 Accepted manuscript doi:
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Zon, 2009, Van Paassen et al., 2010, Filet et al., 2011 and Dejong et al., 2013. The

technology often referred to as biogrout can be classified into two main categories, namely

non-displacement grouting techniques (Karol, 2003) where microbial activities or products

are used to reduce the permeability and/or increase the shear strength of a soil (Chu, 2012).

The new technology has led the filing of several patents (Kucharski et al., 2006, Kucharski et

al., 2008, Cheng and Shen, 2008, Balleur and Girinski, 2008). In practice, the biological

process typically depends on the chemical urea being hydrolysed to ammonium and

carbonate ions in the presence of a high calcium concentration. The resulting increase in pH

and availability of carbonate ions leads to the precipitation of calcium carbonate in the soil

(Stocks-Fischer et al., 1999). Hence, biogrouting or MICP has the potential to transform

loose sediment, such as sand, into a more stable rock-like material (Montoya et al., 2013):

very much like the natural lithogenesis of carbonate rocks.

However recent studies by Dyer and Viganotti (2016) revealed poor differentiation between

different biological protocols used in practice. In essence there is a choice to be made

between growing the necessary bacteria outside of the soil prior to injection or to feed the

bacteria in the soil to promote ongoing population growth in the subsurface. To differentiate

between these two different protocols, the researchers introduced the terms oligotrophic and

eutrophic treatment. Definitions offered for the two new terms are as follows

• Oligotrophic treatment: characterised by the absence of bacterial nutrients in the

cementation liquid and usually includes a fixation phase;

• Eutrophic treatment: characterised by the addition of bacterial nutrients to the

cementation liquid.

In the study by Dyer and Viganotti (2016), MICP biogrout was produced by flushing

saturated soil columns with different solutions of suspended bacteria as well saline fixation

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 Accepted manuscript doi:
10.1680/jgrim.17.00047
and cementation liquids. In particular, for oligotrophic treatment protocol, the columns were

initially flushed from top to bottom at a flow rate of 100 mL h-1 with double deionised water.

Bioaugmentiation was followed by injecting 5 mL of Sporosarcina pasteurii suspension.

Following bioaugmentation, soil samples were treated with saline fixation fluid aseptically

prepared with 7.35 g/L (50 mM) of calcium chloride dehydrate; followed by a series of 30

mL injection rounds (100 mL h-1) to precipitated calcium carbonate. The oligotrophic

cementation medium comprised 60 g/l (1 M) of urea and 147 g/l (1 M) of calcium chloride

dihydrate. In comparison, eutrophic treatment protocols were flushed with a sequence of 30

mL volume of eutrophic cementation liquid prepared comprising 5g/L yeast extract, 10 g/L

(170 mM) urea and 22 g/L (150 mM) of calcium chloride dehydrate, again described in more

detal by Dyer and Viganotti (2016).

Results showed the different treatments to produce markedly different precipitation patterns,

crystal morphology and bond formation and eventual bond failure in silica sand as illustrated

in Figure 3. In the case of oligotrophic treatment the fixation phase resulted in a widespread

pattern of precipitation in silica sand particles with large single rhombohedra crystals formed

on the surface of particles that would have grown continuously with each treatment round

until the soil particles were cemented together. In contrast, eutrophic treatment promoted

preferential precipitation at particle contact points in the form of micritic dome-like structures

of vaterite that acquired a new layer of crystal with each delivery of nutrients. The inter-

particle bonds appeared more complex. The bonding structure was characterised by

coalescing of numerous individual domes into an interlocking structure between the particles.

The bond morphology implied preferential precipitation of vaterite domes during the early

stages of carbonate precipitation. Furthermore, the suture between soil particle and calcium

carbonate deposit was characterised by a jagged edge where the mineral deposit was more

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 Accepted manuscript doi:
10.1680/jgrim.17.00047
evidently crystalline calcite. For this more complex bond structure, diffusion was understood

to be a limiting factor in these tight spaces, which would have shifted the reaction towards the

thermodynamically favoured precipitation of calcite. Not surprisingly, the different crystal

morphologies influenced bond failure mechanism either in the form of a particle-bond

interface mode of failure or internal failure of the carbonate crystal itself.

3.0 OLIOGOTROPHIC AND EURTOPHIC TREATMENT BY ATOMISED

DROPLETS

To date, research into MCIP have tended to concentrate on treatment of saturated soils by

flushing of liquid reagents or percolation of liquid reagent through unsaturated soils. As

witnessed by Van Paassen et al (2010), treatment in the unsaturated zone leads to fingering of

the injected liquid with a resulting highly heterogeneous treatment pattern. To address the gap

in knowledge, a series of unsaturated soil column tests were carried out to explore the effect

of an aerosol delivery method on precipitation patterns, crystal morphology and bond

formation in silica sand. As illustrated in Figure 4, the soil column tests comprised a 140 mm

long perspex tube with an internal diameter of 40 mm (50 mm OD) mount between two

perspex plates and sealed. The top plate provided confinement in order to avoid rupturing of

the samples during injection. A smaller perspex tube (28 mm ID, 32 mm OD) was installed to

connect the nebuliser to the column and a pressure gauge was fitted to the side. The column

was packed with 100 mm of fine silica sand (size range of 150 to 300 µm with estimated

porosity of 46%). A 10 mm thick layer of fine silica gravel (d50 = 2 mm) was employed as

ballast and filter at the top of the column. The gravel also had the purpose of spreading the

confining load at the top. Two scouring pad filters (h = 5mm) were added to prevent the sand

from falling in to the nebuliser unit or exiting the column.

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 Accepted manuscript doi:
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The supplementary unsaturated soil column tests investigated both continuous and pulsed

atomised injection of different concentrations of treatment solutions (urea and calcium ionic).

In each case relying on the atomised injection of a treatment suspension of S. pasteurii (5.04

OD600; UA = 6.49 mM min-1). The microbiological parameters OD600 and UA are defined

as follows.

• Optical Density at 600nm (OD600): the measure of suspended biomass as

determined by spectrophotometric absorbance at 600 nm.

• Urease Activity (UA): the amount of urea hydrolysed in the unit of time by a

bacterial suspension (usually reported in mM∙min-1).

These parameters describe a bacterial culture employed in the process and as such are useful

when characterising the performance and effectiveness of a microbiological species

employed in biogrouting applications. The treatment concentrations along with injection

volumes and rates are shown in Table 1 for continuous injection and Table 3 for pulsed

injection. The atomised liquids were injected using the Cirrus® nebuliser and the same soil

column tests arrangement as described earlier by Viganotti (2014). Based on earlier nebuliser

tests, the aerosol droplets sizes were in the range of 0.5 to 5 μm in diameter (Dyer et al 2012).

From the outset, it was clear that continuous injection of treatment suspensions was

ineffective. These column test involved an initial treatment with 0.1 L (approximately 2 pore

volumes) of S. pasteurii suspension with a 12 hour rest period followed by continuously

injection of aerosol droplets at a rate of 0.25 mL min-1 with a Cirrus® nebuliser in four

cycles as detailed in Table ‎1. Following the final treatment cycle, the samples were left to

react for 24 hours. The treatment resulted in very low unconfined compression strength

(UCS) and carbonate content as reported in Table ‎2. The measured calcium carbonate content

was 0.72% and 0.13% w/w for eutrophic and oligotrophic treatments respectively. The poor

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 Accepted manuscript doi:
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performance was attributed to stripping of reagents from the reaction environment caused by

the atomisation process. To address these issues, a second series of unsaturated tests were

undertaken using pulsed injection.

The second series of soil column tests comprised three unsaturated sand columns prepared

with air-dried fine silica sand (150-300 µm) with an average porosity of 46% and average

coefficient of uniformity of 0.35. The columns were treated with increasing molar

concentrations of reagent liquids as reported in Table 3. Bioaugmentation of each column

was achieved by injecting 90 mL (approximately 2 pore volumes) of nebulised S. pasteurii

suspension at a rate of 0.25 mL min-1. Following bioaugmentation, the columns were rested

for 12 hours to allow for bacterial attachment. Subsequently, the cementation treatment was

delivered by hourly injection periods with hourly pauses, five times daily for a period of 4

days. The volume of cementation liquid delivered daily gradually decreased due to an

increasing backpressures above 150kPa as shown in Table 4. This was attributed to a

reduction in porosity caused by carbonate precipitation at the head of the nebuliser. This

limited the achievable airflow from 8 L min-1 to 6 L min-1. The reaction was stopped 12

hours after the last treatment by flushing 4 pore volumes of deionised water through each

column.

Any liquid accumulating on the top surface of the column was manually removed with a

syringe and the volume recorded. Despite removing the accumulating liquid, however, small

volumes of cementation liquid were seen to percolate from the gravel pack into the sand

column during the reaction period: resulting in increased availability of reagents in the top 20

mm (circa) of the column. This section of the column was therefore carefully removed from

the samples during extraction. The samples were then prepared for UCS testing and

subsequently prepared for calcium carbonate determination and SEM imaging.

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 Accepted manuscript doi:
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The results obtained for unconfined compression tests are reported in Table 5. The highest

measured unconfined compressive strength was 374 kPa for sample E1000. It coincided with

the highest average calcium carbonate content of 3.3% w/w (approximately 50 Kg m-3). In

comparisons, negligible improvement in compressive strength was observed for sample E150

where on average only 1.2% w/w (approximately 18 Kg m-3) of calcium carbonate was

precipitated. The measured increase in compressive strength for sample E500 and E1000

were found to be comparable with published results collated by Al Qabany et al (2013 for

conventional flushing methods in saturated soils as shown in Figure 5. While the data set is

small for three column test employing aerosols injection in unsaturated sands, the results

show a clear increase in strength per density of carbonate deposit at the upper boundary for

increase in compressive strength from conventional flushing or percolation of saturated sands

(Cheng and Cord-Ruwisch, 2012). Furthermore, it was noticed that an increase in

compressive strength with increasing molar concentration of reagents was accompanied by a

change in failure mode as shown in Figure 6. This was most evident for sample E1000 that

failed by axial splitting rather than along inclined shear plane (Zhao, 2008). When

interpreting the test findings, it should be noted that precipitation did not occur

homogeneously along the soil columns column. As illustrated in Figure 7 calcium carbonate

was preferentially precipitated close to the inlet at the base of the column. Measured values

show a 60% to 70% higher carbonate content at the base compared with the top of the

sample. This may explain the localised failure at the top of the test sample E150.

Having commented on unconfined compressive strength results, accompanying SEM images

of representative treated sand samples as shown in Figures 8 and 9. The images provide

additional information to interpret changes in mechanical behaviour. From the outset, the

aerosol injection of treatment reagents into unsaturated sand resulted in noticeably different

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 Accepted manuscript doi:
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precipitation patterns, crystal morphology and bond formation compared with earlier tests

saturated sand samples flushed with liquid reagents as presented earlier in Figure 3. One of

the most noticeable features is the presence of solid curved menisci formed from calcium

carbonate deposits between encapsulated sand particles as illustrated in Figure 8. The curved

menisci imply confinement of aerosol drops between sand particles and in so doing creation

of microenvironments to precipitate carbonate deposits. As shown in Figure 9 and 10, the

microenvironment led to the deposition of copious amounts of spherical shaped vaterite;

which was in marked contrast to the preferential deposition of calcite rhombohedra crystals

observed previously for oligotrophic treated of saturated sands (Zhou et al., 2010). Vaterite in

itself is a metastable phase of calcium carbonate at ambient conditions that is less stable than

calcite with a higher solubility and once exposed to water converts to calcite. A further

noticeable feature from SEM images was the presence of a raised lip of precipitation along

the boundary of the meniscus at the air-liquid-soil (see Figure 10), along with exposed

particle surface at the contact point, which together indicate dual primary and secondary bond

mechanism as explained later in the paper.

4. 0 DISCUSSIONS

Taken together these observations point to a sequence of deposition processes influenced by

differences in ionic strength for reagents combined with differences in surface tension that

result in the formation of a dual bond meniscus structure between soil particles as illustrated

in Figures 10 and 11. The deposition processes firstly depends on droplets of liquid being

deposited at the contact point between soil particles leading to the formation of a meniscus.

The meniscus facilitates the attachment of bacteria to surface of the soil particle at the soil

particle-liquid interface. In turn, the residual bacteria accumulates at the edges of the

meniscus at liquid-air-soil interfaces due to reduced surface tension forces as described by

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 Accepted manuscript doi:
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Zevi et al., 2005. Based on SEM imaging shown in Figure 9, the interpreted distribution of

the bacteria leads to different distribution of calcium carbonate precipitation creating primary

and secondary bonds as follows. The concept of the dual bond model illustrated in Figure 12

as adapted from Cheng and Cord-Ruwisch (2012).

I. Primary Bond. Inner primary bonds are created due to precipitation of calcium

carbonate within the confines of the meniscus at a liquid-soil particle interface closest

to the particle contact point. These primary bonds are characterised by relatively even

distribution of carbonate deposits surrounding untreated particle surface at the contact

point.

II. Secondary Bond. In contrast, curved secondary bonds are created at the outer

boundary of the meniscus at air-liquid-soil and air-liquid interfaces. This peripheral

precipitation process continues until the secondary bond encapsulates the meniscus

resulting in a triple interface between air-liquid-carbonate. The encapsulation process

can in turn confine unprocessed cementation liquid that result in a gap between the

curved secondary bond and inner primary bonds.

Interestingly, the SEM images show the secondary bond often remained unformed for the

heavily treated samples E500 and E1000 as indicated in Figure 11; whereas complete

formation of a secondary bond appears was more widespread for E150, despite the fact that

lower amounts of calcium carbonate were precipitated overall. The scarcity of secondary

bonds in heavily treated samples was interpreted as being a consequence of (a) relatively high

supersaturation occurring in the reaction environment, and (b) higher surface tension of the

cementation liquid. In this case, the higher ionic strength of the cementation liquid would

proportionally lead to a higher surface tension and consequently a larger air-liquid surface

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 Accepted manuscript doi:
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area (Adamson and Gast, 1997). Therefore, larger amounts of calcium carbonate would be

required to complete the secondary bond for the more heavily treated samples.

These variations in crystal morphology as illustrated by SEM images indicated a complex

failure mode that involved fracture of the outer secondary bond together with detachment of

the inner primary bond between carbonate deposit and soil particle. Even though the bond

failure is highly complex and difficult to decipher, the most encouraging feature is the

formation of solid mineral menisci bought about by trapping of aerosol droplets. From an

engineering perspective, the carbonate mineral menisci bond is created at the most optimum

location for ground improvement.

5. 0 CONCLUSIONS

The experiment successfully demonstrated the viability of using aerosol droplets to deliver

bulk liquids to promote in-situ MICP using eutrophic protocols. In particular, a pulsed

injection sequence was shown to be preferable to reduce the risk of stripping liquid reagents

from within the soil matrix. Although SEM images showed the precipitate carbonate mineral

bond morphology and failure modes to be highly complex, the aerosol delivery method in

unsaturated sand had the effect of directing the liquid reagents in droplet form to sand particle

contacts points that in turn lead to formation of solid mineral menisci. From an engineering

perspective, the carbonate mineral menisci bond was located at the most optimum location

for ground improvement. This observation agrees with the hypothesis of Cheng et al. (2012)

which argued that treatment of unsaturated soil should be more effective than treatment of

saturated soils. From a practical outlook however, more research is needed for up scaling of

jet nebuliser technology for large-scale applications and to understand the drained mechanical

behaviour of these lightly cemented ground treatment technique.

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 Accepted manuscript doi:
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Table 1. Test procedure for continuous injection with soil samples left to rest for 24 hours
after each injection round.
Round 1 Round 2 Round 3 Round 4 Injection time
(t0+12h) (t0+36h) (t0+60h) (t0+84h) (continuous)
100 mL 100 mL 100 mL 100 mL 400 min @ 0.25 mL min-1

Table 2. Test results for continuous injection in terms of unconfined compression strength
and calcium carbonate determination
ID Protocol UCS CaCO3 CaCO3 Max CaCO3 Max CaCO3
(kPa) (g)# (w/w) (g)* (w/w)
O1 Oligotrophic 8.52 0.219 0.13% 40 23.4%
E1 Eutrophic 6.97 1.328 0.72% 40 21.7%

Table 3. Pulsed injection test molar concentrations for eutrophic treatment (Column ID
representing micro molar concentration of cementation liquid)
Column ID [Urea] [Ca2+] [Yeast Extract]
E150 0.15 M 0.15 M 5.0 g L-1
E500 0.50 M 0.50 M 5.0 g L-1
E1000 1.00 M 1.00 M 5.0 g L-1

Table 4. Pulsed injection test injection volumes and injection rates for hourly injection
periods with one hour reaction pauses, five times daily (where E150, E500 and E1000 are
column IDs)
E150 E500 E1000
mL mL min-1 mL mL min-1 mL mL min-1
Day 1 75 0.25 75 0.25 75 0.25
Day 2 70 0.23 70 0.23 66 0.22
Day 3 60 0.20 60 0.20 60 0.20
Day 4 60 0.20 60 0.20 60 0.20

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Table ‎5 Pulsed injection tests measured unconfined compressive strength and average
calcium carbonate content for eutrophic treatment
Column ID UCS (kPa) [CaCO3]
E150 6.63 1.2% w/w (18 Kg m-3)
E500 121 2.7% w/w (40 Kg m-3)
E1000 374 3.3% w/w (50 Kg m-3)

Figure 1 Ranges of droplet sizes present in nature extending from a drizzle to smoke
(Lefebvre, 1989).
Figure 2 Illustration of medical jet nebulisers and droplet size distribution (Dyer et al 2012).
Figure 3 SEM images of carbonate precipitation between silica soil particles for Oligotrophic
and Eutrophic treatment protocols for liquid flushing in saturated conditions (with
bond failure visible for eutrophic treated sample).
Figure 4 Schematic representation of the soil column tests injected with aerosol droplets of
cementation reagents from CIRRUS™ nebuliser (Viganotti 2014)
Figure ‎5. Pulsed injection tests results for unconfined compressive strength and calcium
carbonate content compared conventional injection methods in saturated soils by Al
Qabany et al (2012) for 500 µM concentrations eutrophic treatment.
Figure 6 Pulsed injection tests failure surfaces from unconfined compression test for
eutrophic treatment
Figure ‎7 Measured variation of calcium carbonate content along UCS soil samples taken
from column tests E150, E500 and E1000 presented in relation to respective molarity
of cementation liquid.
Figure 8. SEM image of assemblage of sand particles bonded by calcium carbonate deposits
from pulsed injection of atomised eutrophic reagents test E150. The calcium
carbonate precipitation traces the shape of water menisci formed during the
treatment process.
Figure ‎9 SEM image of vaterite spherulites formation from pulsed injection of atomised
eutrophic reagents for test E500 (below) and E1000 (above)
Figure ‎10 SEM image of carbonate lip at the air-liquid-soil interface from pulsed injection of
atomised eutrophic reagents for test E150. Arrows indicate imprint of rod-shaped
bacteria on the surface of the mineral deposit.
Figure ‎11 Sample E500 showing the exposed edge of a carbonate lip (left hand pair of
arrows) where a secondary bond failed to form, and a fully formed secondary bond
(right hand pair of arrows).
Figure 12 Schematic representation of the three-phased process leading to formation of a
dual bond meniscus structure.

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