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The fugacity approach in modelling bioventing and enhanced bioremediation of

contaminated soils
M. Cristina Vila1, M. Manuela Carvalho2, Jorge Cardoso1, Manuel Trelles1 and António Fiúza1
1
Faculty of Engineering, University of Porto, Portugal / CERENA
2
Institute of Engineering, Polytechnic Institute of Porto / REQUIMTE

Corresponding author email: mvila@fe.up.pt

ABSTRACT
Enhanced bioremediation and bioventing tests were performed at laboratory scale in order to study the
remediation effectiveness for a residual granitic soil contaminated with benzene. Biodegradation was
promoted by a consortium of heterotrophic microorganisms collected in a refinery area. Microbial
community was extracted from contaminated soil, adapted and developed in a mineral medium enriched
with the benzene. Soil contamination was monitored in discontinuous mode by daily measurements of
benzene concentration in the gas phase using gas chromatography. Bioventing was promoted by phased
ventilation controlled through to a respirometric system which in addition to air supply also monitored
oxygen and carbon dioxide within each column.
Mackay's concept of fugacity was adopted to develop mathematical models able to describe and
reproduce enhanced bioremediation of natural soils contaminated with benzene and biofuels.
Calibration of the developed models was based on experimental data obtained in laboratory tests.
Fugacity based models were successfully applied to lab scale microcosms, opening a very useful path to
access and predict the behaviour of contaminants (mainly their distribution by soil phases) in the
aptitude tests required prior to field application of any remediation technology. These proved robust,
describing and predicting the behaviour and time evolution of the main variables involved in the
processes.
KEYWORDS: Bioremediation technologies; Benzene; Biofuels; Fugacity; Modelling.

1. INTRODUCTION
Soils are one of the most important natural resources that exist since it is the basis sustaining life on the
planet. It is a fragile element of the environment; soil is a non-renewable natural resource because its
rate formation and regeneration is very slow compared to the processes that contribute to its
degradation. Soil contamination is frequently the result of human activities; when contaminants
exceeded the soil’s natural buffering capacity remediation technologies play an important role to reverse
the situation. The selection and application of these technologies depends on the properties of both soils
and contaminants. In this work benzene and biofuels were used as contaminants.
Benzene is present in most fuels, it is an important industrial solvent and it is used as a chemical
intermediate in the production of other chemicals (Morgan et al., 2009). Biofuels are renewable energy
sources that can be used as an alternative to the conventional fuels (Cardoso, 2014), they are liquids or
gaseous fuels that are used for transportation and are usually produced from biomass (EU, 2003). These
contaminants are released to the soil environment through industrial discharges, waste disposal, fuel
leaks or spillages (ATSDR, 2007). Upon release into the soil contaminants will tend to disperse through
the unsaturated zone eventually reaching the saturated zone (Morgan et al., 2009) causing degradation
of the characteristics of soil and groundwater. Enhanced bioremediation and bioventing are cost-
effective remediation technologies that can be used to degrade organic contaminants and remediate
environmental hazards. Both technologies use the degradation capacity of indigenous or adapted
microorganisms to biodegrade organic compounds sorbed to soils, resulting non-toxic by-products (Singh
et al., 2004; Carvalho, 2014).
The contaminants environmental behavior can be evaluated using mathematical models. The concept of
fugacity, which was introduced by G.N. Lewis in 1901 as a more convenient thermodynamic equilibrium

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criterion than chemical potential, has been widely applied to chemical processes. In recent years, mainly
with Mackay's contribution (Pollard et al., 2008; Seth and Mackay, 2002; Hertwich, 2001), the
environmental fate and transport of organic contaminants has been successfully simulated and predicted
by the application of multimedia models based on fugacity, activity and compartment capacity concepts.
In this work, Mackay's concept of fugacity was adopted to develop mathematical models able to describe
and reproduce enhanced bioremediation and bioventing of granitic soil contaminated with benzene and
biofuels. Calibration of the developed models was based on experimental data obtained in laboratory
tests. These models were successfully applied to lab scale microcosms, opening a very useful path to
access and predict the behaviour of contaminants in the aptitude tests required prior to field application
of any remediation technology.

2. METHODOLOGY

2.1 Materials and Analytical Methods

The contaminants used were benzene, n-butanol and soybean biodiesel. Benzene and n-butanol are both
pro-analysis grade with purity ≥ 99.5 % from Panreac Quimica SAU. Biodiesel was synthetized by
homogeneous alkali transesterification (Dias et al., 2008).
The granitic soil used in all remediation tests was collected in the north of Portugal. A sample of sandy-
loam soil contaminated with hydrocarbons was used to extract the microbial consortium to inoculate the
residual granitic soil for bioaugmentation. Standard methodologies were used for the soil
characterization.
The grantic soil was a sandy-loam soil predominantly composed by aluminosilicates with high grade of
kaolinization, large grain size distribution, low electrical conductivity, very low natural organic matter,
and slightly acidic. This soil presented hight water-holding capacity and larger porosity (Carvalho, 2014;
Carvalho et al., 2015; Carvalho et al., 2015a).
During experiments, benzene concentration in the gas phase was monitored by isothermal (150 ºC) gas
chromatography using a GC-Shimadzu-2014 chromatograph equipped with a FID. The carrier gas was N2;
200 μL of gas sample was injected in splitless mode. The operating temperature for both injector and
detector was 200 ºC. Carbon dioxide and oxygen concentrations in the gas phase were determined using
a Servomex - 5200 multipurpose respirometer.

2.2 Methods
Inoculum Preparation
In order to obtain the microbial consortium to inoculate the traials and assess biodegradation, the
selected microbial consortium was developed aerobically in mineral media containing contaminant (only
benzene or benzene and n-butanol or benzene and biodiesel). Resulting cultures were incubated at 28°C
with shaking (150 rpm). The incubation period ended when the benzene concentration in the gas phase
reached 0.5 mg per liter of air (Carvalho, 2014). When the experiments ended the biomass was counted
using spread-plate technique in LB medium agar and incubated at 28 ºC for 3 days (Carvalho, 2014).
These microbial cultures were used in bioremediation tests for soil bioaugmentation.
Remediation Tests
Biodegradation tests were performed at controlled temperature of 25ºC. All bioventing tests were
performed in stainless steel columns with 50 cm high (total volume of 3917 mL), containing 2000 g of
wet soil (25 % of water content). Enhanced bioremediation tests were all performed in stainless steel
columns with 35.5 cm high (total volume of 2788 mL), containing 1420 g of wet soil. The initial
contamination level was obtained by blending 210 mg of benzene per kilogram of wet soil; in some trials
biofuels were also added, 210 mg of n-butanol per kilogram of wet soil or 210 mg of biodiesel per
kilogram of wet soil (Cardoso, 2014; Carvalho, 2014).

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Each inoculated test had its sterilized control. In inoculated trials, enrichment cultures (inoculum) were
used to humidify the previously dryed soils and proceeding to its bioaugmentation and biostimulation; in
the non-inoculated tests, sterilized, distilled and demineralized water was used. In the bioventing tests,
oxygen was supplied through the daily introduction of air into the system, by passing a flow rate of 20 mL
min-1 for 15 minutes.
The concentrations of benzene in the gas phase were daily monitored in all tests. The remediation times
were dictated by the value defined as the residual concentration (0.5 mg of benzene per L of air). In
bioventing tests the concentrations of CO2 and O2 in the gas phase were also monitored daily. At the end
of the experiment, the biomass (CFU) was quantified by previous extraction in sterile saline solution
(0.85% NaCl; w/v) and determined by the method of serial dilution on LB plates (Cardoso, 2014;
Carvalho, 2014; Carvalho et al., 2015).

2.3 Mathematical Modelling

There is a quasi-linear relationship between concentration (c, mol m-3) and fugacity (f, Pa) that can be
expressed in terms of compartmental capacity (Z, mol m-3 Pa-1) by the following equation
c=f·Z (1)
In equilibrium multiphase systems, fugacity is the same in all phases. The compartmental capacity (Z,
mol. m-3. Pa-1) depends on temperature, pressure, contaminant properties and the medium in which it is
present (Thibodeaux and Mackay, 2011). The compartmental capacities of air and water can be
estimated from established thermodynamic equations 2 and 3 (Seth and Mackay, 2002)
Za = 1 / (R · T) (2)
Zw = 1 / H (3)
Where Za is air compartmental capacity (mol m-3 Pa-1), R is the ideal gas constant (8.314 m3 Pa K-1 mol-1),
T is temperature (K), Zw is the compartmental capacity of water (mol m-3 Pa-1), and H is the Henry's law
constant (Pa m3 mol-1 K-1). However, for ill-defined compartments such as soils, sediments, aerosols and
biota it is recommended to determine Z from phase partition coefficients and a known compartmental
capacity such as Za or Zw (Fiúza, 2010).
Zs = (Kw-s · ρs) / H (4)
Being Zs the compartmental capacity of soil (in the present case study it was decomposed into organic
and mineral matter), Kw-s the water-soil partition coefficient (m3 kg-1), and ρs the soil bulk density (kg m-3).
For equilibrium conservative (non-reacting) systems, equilibrium fugacity (f0, Pa) can be obtained from
equation 5.
f0=Mcont / ∑(Vi · Zi ) (5)
Here, Mcont is the total mass of contaminant within the system (mol), Vi and is Zi are the volume (m3) and
the compartment capacity (mol m-3 Pa-1) of the ieth compartment, respectively. In case of a transient
system with contaminant sources and sinks, fugacity becomes a time dependent variable.
In present work, the blank enhanced bioremediation test used a Mackay Level I model, based upon a
closed system, which describes the contaminant phase equilibrium in the media. It served as a first step
in a sensitivity acquiring on the partitioning tendency of the contaminant and gave a magnitude order for
relative concentrations in different compartments. Inoculated tests were described by a Level II model,
corresponding to a steady-state equilibrium system, where contaminant losses occured by advective
transport and/or biodegradation reactions. Only the effective bioremediation period is modelled, being
the microorganism’s adaptation period excluded (a few initial days). Models were implemented in
MatLab® programming routines developed for this purpose.

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3. RESULTS AND DISCUSSION

Benzene distribution in the studied environmental compartments (air, water, soil and organic matter), as
presented in figure 1, shows identical and the higest fractions in solid and liquid phases, in opposite, the
gas phase presented about half of the value, being the benzene fraction in organic matter negligible.
Experimental versus model predicted time evolution of benzene concentration in the gas phase in
inoculated tests are shown in figures 2, 3 and 4. According to the results obtained in enhanced
bioremediation (graphics on the left) the n-butanol presence has virtually no influence on benzene
biodegradation, whereas biodiesel positively influenced the biodegradation. Bioventing results (graphics
on the right) suggest that the addition of biofuels increases the remediation rates. These findings are
corroborated by the kinetic constants obtained from mathematical modelling (Figure 5).

Figure 1. Benzene distribution in the considered environmental compartments: Air, water, soil and
organic matter.

Figure 2. Time evolution of benzene concentration in the gas phase when granitic soil is contaminated
with benzene. Experimental and predicted results for bioremediation (left), and for bioventing (right).

Figure 3. Time evolution of benzene concentration in the gas phase when granitic soil is co-contaminated
with benzene and n-butanol. Experimental and predicted results for bioremediation (left), and for
bioventing (right).

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Figure 4. Time evolution of benzene concentration in the gas phase when granitic soil is co-contaminated
with benzene and biodiesel. Experimental and predicted results for bioremediation (left), and for
bioventing (right).

Figure 5. Kinetics constants of benzene deseapearance obtained by model best fits for the three different
contaminations tested: benzene, benzene plus n-butanol, and benzene plus biodiesel.
a) Enhanced bioremediation; b) Bioventing.

4. CONCLUSIONS
Based on obtained results it is possible to conclude that the presence of biofuels definitively affects the
remediation rates in both studied technologies. In the evaluated situations a positive influence was
observed.
Fugacity based models revealed a good agreement with experimental data for enhanced bioremediation
and for bioventing, as evidenced by the high obtained determination coefficients (R2).
The proposed modelling methodology presents itself as a robust tool to access and predict the behaviour
of contaminants (mainly their distribution by soil phases and concentration time evolution) in the
aptitude tests required prior to field application of any remediation technology.

ACKNOWLEDGEMENTS
This work was supported by FEDER funds through the Operational Program for Competitiveness Factors -
COMPETE and National Funds through FCT - Foundation for Science and Technology via the project
PTDC/AAG-TEC/4403/2012 (ISIS). The authors are gratefull to Joana Dias (LEPABE-FEUP) for her
collaboration in biodiesel supply.

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