Bioresource Technology 101 (2010) 5168–5174

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Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Biological treatment of waste gas containing mixture of monochlorobenzene

(MCB) and benzene in a bench scale biofilter
R.A. Pandey *, P.R. Joshi, S.N. Mudliar, S.C. Deshmukh
National Environmental Engineering Research Institute, Nehru Marg, Nagpur 440020, India

a r t i c l e i n f o a b s t r a c t

Article history: The paper outlines treatment of waste gas containing monochlorobenzene (MCB) and benzene in a mix-
Received 25 November 2009 ture using biofilter packed with compost and woodchips seeded with Acinetobacter calcoaceticus. The bio-
Received in revised form 30 January 2010 filter could treat waste gas containing MCB and benzene effectively with an efficiency of (99 ± 5%) and
Accepted 3 February 2010
(97 ± 6%) at optimal empty bed contact time (EBCT) of 3 min with a loading of 57 g/m3/h of MCB and
Available online 3 March 2010
2 g/m3/h of benzene. At optimum loading of MCB and benzene, the biofilter showed total bacterial count
of 13  105 CFU/g of compost, while the MCB and benzene degrading bacterial count was 71  104 CFU/g
Keywords:
and 5  104 CFU/g compost respectively. The experimental removal efficiency of MCB and benzene were
Monochlorobenzene
Benzene
in good agreement with the model predicted value.
Biofilter Ó 2010 Elsevier Ltd. All rights reserved.
Treatment
Waste gas

1. Introduction
Monochlorobenzene (MCB) is manufactured worldwide to the
tune of approximately 700,000 metric tonnes (Othmer, 2000).
MCB is used as an intermediate in dye and pesticide synthesis;
as solvent, insect repellents, degreasers, deodorants, heat transfer
agents etc. The MCB is manufactured via chlorination of benzene
and the process produces waste gases containing MCB and ben-
zene, which are obnoxious odour emitting pollutants (Shareefdeen
and Singh, 2005). Similarly industries where MCB and benzene are
used as raw materials also generate waste gases containing these
compounds. These constituents on release into the air give obnox-
ious odour and remain for a longer period into the environment as
a volatile organic compound (VOCs). The estimated release of MCB
and benzene into air is above between 0.2% and 0.3% respectively
of the total production and use (Shareefdeen and Singh, 2005).
Dow Chemical Company has reported that about 30–50% of the an-
nual MCB produced is released to air (Joshi, 2009). MCB and ben-
zene have been identified as priority pollutants by the United
States Environmental Protection Agency (USEPA, http://www.epa.-
gov/waterscience/methods/pollutants.htm) and have several
health and environmental implications (Joshi, 2009; ATSDR, 2005).
The conventional physical or chemical processes are employed
for the treatment of volatile organic compounds (VOCs) viz.
adsorption, chemical oxidation etc. (Joshi, 2009). Chemical pro-
cesses include typically thermal or catalytic oxidation (Joshi,
* Corresponding author. Tel.: +91 712 2240097; fax: +91 712 2249961.
E-mail address: ra_pandey@neeri.res.in (R.A. Pandey).
2009). All of these options are more expensive than biological pro-
cesses for treatment of low concentration of VOCs (Mathur et al.,
2006; Bhatt et al., 2006, 2007; Braeckevelt et al., 2007; Zhang
et al., 2007; Haines and Glanda, 2008). A number of microorgan-
isms such as Pseudomonas, Alcaligenes, Rhodococcus, Azotobacter
and Acinetobacter sp. have been reported for degradation of chlori-
nated pollutants (Bhatt et al., 2007). Similarly, other cited litera-
ture (Mathur et al., 2006; Bhatt et al., 2007) have reported the
mineralization of MCB and its derivatives in liquid broth under
appropriate conditions in the laboratory by the bacteria isolated
from solid and water. The treatment of waste gas containing
MCB has also been cited using biotrickling filter (Young-Sook and
Richard, 1994; Mathur et al., 2006). However, no literature has
been sighted for treatment of waste gas containing MCB and ben-
zene in a mixture. A potential culture for biodegradation of MCB
and benzene was isolated and identified as Acinetobacter calcoace-
ticus. This culture was used for the treatment of waste gas contain-
ing MCB and benzene in mixture. The paper outlines the treatment
of waste gas containing MCB and benzene in a mixture using bio-
filter packed with compost and wood chips augmented with
seeded culture A. calcoaceticus.
2. Methods
2.1. Materials
2.1.1. Bench scale biofilter unit
The schematic of the bench scale biofiltration unit is presented
in Fig. 1. The laboratory scale biofilter was made-up of PVC

0960-8524/$ - see front matter Ó 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2010.02.017
 R.A. Pandey et al. / Bioresource Technology 101 (2010) 5168–5174 5169

material. The biofilter unit consisted of a packing section, blower, Table 1

an odour-generating unit, a rotameter (1–10 LPM), temperature Dimensional details and performance of biofilter at optimal condition for treatment of
waste gas containing MCB and benzene in a mixture.
indicator, and manometer for pressure reading as well as sample
collection ports. A leachate collection system with water seal was Parameter Average value
also provided at the bottom of the biofilter. The packing material Height of the packed bed (m) 0.90
consisted of compost (% w/w carbon 40, nitrogen 1.2, phosphorous Diameter column (OD/ID) (m) 0.20/0.195
0.02, potassium 0.80, calcium 1.50, magnesium 0.12, and sodium Total volume (l) 27
Volume of packing (l) 12.1
0.02; other metal elements mg/kg: copper 44, manganese 360, Porosity of packing 0.30
and zinc 70) and wood chips in alternate layers. Dimensional de- Relative humidity of inlet air (%) 80
tails of the bioreactor are given in Table 1. Moisture content of packing medium 60–65
EBCT (min) 3
Pressure drop across packing medium (mmwc) 1–2
2.1.2. Chemicals and basal medium Operating temperature (°C) 25–30
All the chemicals used for analysis were of analytical grade. The pH 6.7–7
chemicals/solvents used for gas chromatographic analysis were of MCB loading (g/m3/h) 57
Removal efficiency of MCB (%) 99 ± 5
chromatographic grade. The MCB and benzene used in the investi-
Removal efficiency of benzene (%) 97 ± 6
gation was procured from M/s. Aldrich Co., Germany. The nutrient Benzene loading (g/m3/h) 2
media used in the present investigation (liquid and solid medium) Total count 1.1  106
were prepared as per Standard Method of Water and Wastewater Specific count on MCB (CFU/g of compost) 17  105
Analysis (APHA, AWWA, WEF, 2005). The basal medium for the Specific count of benzene (CFU/g of compost) 5  105
Chloride concentration (mg/L) 500–600
growth of microorganisms consisted of dipotassium hydrogen
phosphate (K2HPO4, 0.625 g/L), potassium dihydrogen phosphate
(KH2PO4, 0.375 g/L), ammonium sulphate [(NH4)2SO4, 0.5 g/L].
Magnesium sulphate (MgSO47H2O) calcium chloride (CaCl2), cop- Turbochrome Navigator Software attached with the system. The
per sulphate (CuSO45H2O) and ferrous sulphate (FeSO4). The basal GC method for estimation of MCB and benzene was standardized
agar medium was supplemented with yeast extract (0.1% w/v) for and method is reproducible for the estimation of these constituents
the evaluation of total count of the microorganisms of the biofilter (MCB: 20 lg/L and benzene: 15 lg/L) (Joshi, 2009).pH meter (Ther-
unit, while for the specific count, the medium yeast extract was re- mo make), incubator, hot air oven, centrifuge (IEC Model PR-2), ro-
placed with MCB (50 mg/L) and benzene (50 mg/L) separately. tary shaker (EMENVEE make), suction pump (Rotovac FP2F Model
PRM3), autoclave (P-95K, wates No. 191), electronic weighing ma-
chine (Schimadzu AX200), muffle furnace (Tempo make) and air
2.1.3. Analytical equipments
compressor (ELGI make) were used in the experiments.
MCB and benzene in the waste gas before and after treatment was
analysed by GC fitted with flame ionization detector (FID) and Supe-
lco equity five capillary column having length 30 m, ID- 2.2. Methods
0.25 mm  0.5 lm film thicknesses. The oven temperature was
maintained at 100 °C for one minute and then programme at the rate 2.2.1. Isolation and identification bacterial cultures degrading MCB
25 °C per minute to 200 °C. The temperature of the injector and and benzene
detector was kept at 250 °C and 300 °C respectively with nitrogen Three cultures degrading MCB were isolated form the soil by
as the carrier gas. The data was acquired, stored and analysed with enrichment technique (Giri et al., 2010) and identified by colony

Clean Gas
Legend
R1, R2 --- Rotameter
B --- Blower
P --- Pump
Biofilter SP1 M --- Packed
bed Mixer
I --- Impinger
SP2
SP ---- Sampling port
M
SP3

SP4
Nutrients
R1 R2 P

B Leachate

Fig. 1. Schematic diagram of air phase biofilter for treatment of waste gas containing MCB.
5170 R.A. Pandey et al. / Bioresource Technology 101 (2010) 5168–5174
morphology followed by biochemical test and 16S-rRNA matching
with NCB1 library database. Out of these three cultures A. calcoace-
ticus was potential to degrade MCB in the range of 94–99%. Further,
this culture has also potential to degrade benzene in the range of
50–60% with an input concentration of 50 mg/L. The other two cul-
tures namely Bacillus fusiformis and Bacillus sphaericus have only
potential to degrade monochlorobenzene in the range of 20–30%
with an input concentration of 50 mg/L (Joshi, 2009). Further, the
kinetics of biodegradation of MCB and benzene was carried out
using flasks culture experiments. The value of microbial growth
rate of MCB: lmax(m), half saturation constant of MCB: Ks(m), micro-
bial growth rate of benzene: lmax(b) and half saturation constant of
benzene Ks(b) has been evaluated. Based on the kinetics of biodeg-
radation of MCB and benzene, A. calcoaceticus was selected a seed
for the biofilter for the treatment of waste gas containing MCB and
benzene in mixture.
2.2.2. Seed inoculum and biofilter operation
The isolated culture A. calcoaceticus was grown in 100 mL basal
medium containing benzene (50 mg/L) and MCB (50 mg/L) individ-
ually as well as in a mixture (25 mg/L each) in separate flasks. The
biomass was harvested from all the experimental flasks and an
inoculum having biomass with a tune of 1% w/v biomass dry
weight was prepared. Further, 200 ml of broth containing biomass
(1% w/v) was mixed with compost (2 kg) prior to packing into the
biofilter. Further, this mixed biomass in a 100 ml nutrient medium
was also sprinkled each time thrice a day to keep the packing med-
ium of the biofilter in moist condition (60–65%). The waste gas
containing the benzene and MCB generated in the odorant-gener-
ating unit was passed into the biofilter using a blower.
The operation of the biofilter resulted in the growth of microor-
ganism and the removal of MCB and benzene from the waste gas.
Continuous operation of the biofilter was carried out for evaluation
of different parameters for degradation of MCB and benzene. The
biofilter was operated for more than 365 days on a bench scale
on a continuous feed basis, and the parameters, viz. Empty bed
contact time (EBCT), MCB and benzene loadings at optimal EBCT,
requirement of moisture content and pH of the packing medium
were evaluated. The flow rates of waste gas to the biofilter were
varied in the range of 2–10 LPM in order to achieve different EBCT
(1.2–6.05 min) in the biofilter. Similarly by changing the input con-
centration of MCB and benzene in the waste gas at optimal EBCT
different loadings of MCB and benzene was achieved in the biofil-
ter. The MCB and benzene loading was varied in the range of 11–
90 g/m3/h at constant load of benzene 2 g/m3/h. The benzene load-
ing in the waste gas was kept lower as compared to MCB loading
Table 2
Details of model parameters.
Symbol Parameter
As Biofilm surface area
H Biofilter bed height
d Biofilm thickness
Dm Effective diffusivity of MCB in the biofilm
Db Effective diffusivity of benzene in the
biofilm
mm Air/biofilm partition coefficient of MCB
mb Air/biofilm partition coefficient of benzene
lmax(m) Microbial growth rate of MCB
lmax(b) Microbial growth rate of benzene
Ks(m) Half saturation constant of MCB
Ks(b) Half saturation constant of benzene
Model predication MCB % removal efficiency
error in comparison Benzene % removal efficiency
with experimental
observation
since the industries manufacturing MCB emits chlorobenzene
and benzene in a mixture with a higher loading of chlorobenzene
(Joshi, 2009). In the second set of investigation, the load of benzene
was varied in the range of 1–10 g/m3/h keeping the load of MCB
constant.
During assessment of the moisture content of the packing med-
ium of the biofilter, the volume of sprinkled nutrient medium was
varied in order to achieve different moisture content in the system.
All the experimental observations and assessment of the different
parameters were made during pseudo steady state of performance
of the biofilter.
Model parameters pertaining to kinetics of microbial degrading
MCB and benzene (based on Monod kinetics) were estimated
through batch experiments. The significant biodegradation kinetic
parameters such as maximum growth rate (lmax), half saturation
constant (Ks) was obtained from the weighted average in the
hyperbolic regression. The other model parameters evaluated
through experiments (Table 2).
3. Results and discussion
3.1. Biofilter start-up
The bench scale biofilter packed with compost containing MCB
and benzene degrading microorganism A. calcoaceticus, was ex-
posed to the waste gas containing mixture of MCB (0.68 g/Nm3)
and benzene (0.3 g/Nm3) at the waste gas flow rate of 2 litres per
minute (LPM). At the initial stage of the operation, there was no
major change in the magnitude of benzene in the inlet and outlet
of the biofilter. However, MCB could be degraded in the range of
94–99%. This may be attributed to the presence of MCB degrading
microorganisms in the biofilter. At the same time the microbial
population, which could degrade benzene, were not available to
the extent required for effective degradation of benzene and it took
7 days for the biofilter to get acclimatised to benzene with limited
degradation of benzene (50–60%). Therefore, biofilter was further
seeded with A. calcoaceticus as it has potential to degrade MCB
and benzene. The start-up of biofilter took total 15 days at the
end of which the benzene specific count and MCB specific counts
were raised from initial 21  101 to 31  102 CFU/g and 62  104
to 70  104 CFU/g respectively. Kim and Lee (2002) reported
start-up period of 15 days for biodegradation of benzene after inoc-
ulation of Pseudomonas sp. in a biofilter packed with activated
charcoal. Similar, observations were reported in the literature dur-
ing studies on conventional VOCs removal in biofilter (Delhomenie
and Michele, 2003).
Value Source
150 m2/m3 Experiment
0.94 m Experiment
0.0001 m Experiment
4.916  106 m2/h Mathur et al. (2006) and adjusted by simulation
3.708  106 m2/h Lyman et al. (1990) and adjusted by simulation
0.095 Staudinger and Roberts (1996) and adjusted by simulation
0.167 Staudinger and Roberts (1996) and adjusted by simulation
0.17 1/h Experiment
0.23 1/h Experiment
12.8 g/m3 Experiment
15.3 g/m3 Experiment
<15%
<10%
 R.A. Pandey et al. / Bioresource Technology 101 (2010) 5168–5174 5171

120 %RE (MCB) % RE (Benzene)

100

% Removal efficiency
80

60

40

20

0
1.21 1.51 2.02 3.03 6.05
EBCT (min)
Fig. 2. Performance of biofilter for the treatment of waste gas containing MCB and benzene in mixture at different empty bed contact times.

3.2. Performance evaluation of the biofilter for the treatment of waste
gas containing MCB and benzene in mixture
The empty bed contact time (EBCT) is an important parameter
for designing a biofilter system. The MCB and benzene load in
the biofilter was kept constant to an average magnitude of 28 g/
m3/h and 2 g/m3/h respectively. At the EBCT of 3 min, the maxi-
mum removal efficiency of MCB could be achieved to 96 ± 8% while
benzene could get removed with the maximum efficiency of
78.4 ± 5% from the waste gas (Fig. 2). The observed value of rela-
tively high EBCT for effective treatment of MCB and benzene in
the biofilter can be attributed to the poor water solubility of the
MCB and benzene along with their limited biodegradability (Bhatt
et al., 2007). Further, the bacteriological status of the biofilter also
supports this observation, as the specific count with reference to
benzene degrading microorganisms (2  104 – 5  104 CFU/g of
the compost) is less than that of MCB (50  104 – 70  104 CFU/g
of the compost).
The effect of MCB and benzene loading on the respective sub-
strate removal efficiency is presented in Figs. 3 and 4. The maxi-
mum removal efficiencies of MCB and benzene was found to be
more than 99 ± 5% and 97 ± 6% respectively at a load of 57 g/m3/
h of MCB and 2 g/m3/h of benzene.
Moisture content of 65% was found to be optimum at which
maximum removal of MCB and benzene was achieved. At higher
moisture content in the biofilter results in carrying away the waste
gas containing MCB and benzene along with the nutrient medium,
which found its way into the leachate of the biofilter.
At the optimal performance conditions, the biofilter showed a
total bacterial count of 13  105 CFU/g of compost, while the
MCB and benzene degrading bacterial count was 71  104 CFU/g
and 5  104 CFU/g of compost respectively.
During biodegradation of MCB, the chlorine atom in the MCB
ring is released as chloride ion (Bhatt et al., 2007). After the estab-
lishment of MCB and benzene degrading microorganisms in the
biofilter, aerobic mineralization of both the constituents was
achieved. Thus, the MCB and benzene present in the waste gas bio-
degraded with the release of chloride from the MCB. Aerobic min-
eralization has been well documented for chlorobenzenes with up
to four chlorine substituents in microcosms and by pure cultures
(Bhatt et al., 2007). Aerobic microorganisms usually initiate degra-
dation by activation of the aromatic nucleus through oxygenation

120

100
% Removal Efficiency

80

60

40

20

0
0 20 40 60 80 100
-3 -1
Loading (gm h )
Fig. 3. Assessment of MCB load at constant load of benzene (2 g/Nm3/h) in the biofilter.
5172 R.A. Pandey et al. / Bioresource Technology 101 (2010) 5168–5174
120
100
% Removal efficiency
80
60
40
20
0
1.01 1.19
Fig. 4. Assessment of benzene load in presence of optimal load of MCB (57 g/Nm3/h) in the biofilter.
reactions. The introduction of hydroxyl-group, usually in ortho-po-
sition to one another, results in a few central intermediates such as
catechols, protocatechuate, gentisate and hydroxyhydroquinones.
These intermediates are subjected to oxygenolytic ring-cleavage
followed by channelling of the ring-cleavage products into the cen-
tral metabolism. Chlorocatechol is one of the central intermediate
in the degradation of various chlorinated pollutants (Bhatt et al.,
2007). Various alternative pathways have also been reported in
the literatures viz. oxidative pathway or meta-cleavage pathway
for such chloro constituents (Pieper and Reineke, 2004).
The chloride concentration is often monitored as a measure of
MCB biodegradation. In the present investigation on bench scale
biofilter for the treatment of MCB, the formation of chloride as
a by-product was observed in the range of 50–600 mg/L as chlo-
ride ion in the leachate of the biofilter during the operational
period.
The biofilter for treatment of waste gas was operated continu-
ously at optimal EBCT (3 min) and MCB (57 g/m3/h) and benzene
(2 g/m3/h) loadings for a period of 150 days and the results are pre-
sented in Table 1. The inlet and outlet concentrations of MCB and
benzene in the biofilter have also been depicted in Figs. 5 and 6.
Fig. 5. Profile of benzene in inlet and outlet of the biofilter during the treatment of waste gas containing MCB and benzene in a mixture.
1.62 2.22 6.32 10.41 13.13 17.87 20.75 26.88
Load (gm-3h-1)
The overall removal of MCB was found to be 99 ± 5% while the ben-
zene removal efficiency was 97 ± 6%.
3.3. Mathematical model, simulation and comparison with
experimental results
The mathematical model describing the biofiltration of mixture
of MCB and benzene is based on the diffusion of the compounds
through the biofilm and degradation of the compound along the
biofilter column. The details on these aspects through model equa-
tions are presented as Appendix A. The approach is to solve the
one-dimensional mass balance equations over the microscopic
cross section of a biofilm assuming simplified reaction kinetics
(Mohseni and Allen, 1999). The kinetic model parameters obtained
through flask experiments in the present investigation and other
optimized through simulation are presented in Table 2. The MCB
and benzene degradation along the biofilter followed Monod kinet-
ics and oxygen was not a limiting factor for the range of MCB and
benzene concentrations studied in present investigation, as con-
firmed through batch experiments during determination of the
model parameters (data not shown). The model predictions were
 R.A. Pandey et al. / Bioresource Technology 101 (2010) 5168–5174 5173

Fig. 6. Profile of MCB in inlet and outlet of the biofilter during the treatment of waste gas containing MCB and benzene in a mixture.

in good agreement with experimental values with prediction error X and Y (Mohseni and Allen, 1999). The volumetric kinetic expres-
of <15% for MCB and <10% for benzene (Table 2). sion is generally used in biofilter modeling due to the difficulty to
determinate the biomass density in the system with a non-destruc-
4. Conclusions tive technique and without modifying experimental conditions. For
the case where the compound fully penetrates into the biofilm or
The mixture of volatile organic compounds such as MCB and reaction limiting case, the boundary conditions for Eq. (1) and (2)
benzene are released during their manufacture and process opera- are:
tion in which they are used. The studies indicate that it is possible Cm Cb
to effectively treat waste gas containing mixture of MCB and ben- Sm ¼ and Sb ¼ ð3Þ
Mm Mb
zene in a conventional biofilter seeded with A. calcoaceticus. MCB
and benzene could be removed with a maximum efficiency of dSm dSb
99 ± 5% and 97 ± 6% respectively at loading of 57 g/m3/h MCB ¼ ¼0 ð4Þ
dx dx
and at benzene loading of 2 g/m3/h at an EBCT of 3 min.
Cm and Cb is the MCB and benzene concentration in bulk gas
phase. The boundary condition as represented by Eq. (3) assumes
Acknowledgements
that the gas and liquid phase at the interface are at equilibrium,
a condition similar to Henry’s law and represented by a dimension-
The authors express gratitude to Dr. T. Chakrabarti, Acting
less partition coefficient Mm and Mb. The other boundary condition
Director, NEERI for his kind permission to publish this article.
represented by Eq. (4) assumes that the flux in solid phase is zero,
The authors also acknowledge Mitsui Environmental Engineering
which as an interpretation that flux into the solid phase due to
Trust, New Delhi for providing financial support for this
adsorption is equal to the flux out of the solid phase due to
investigation.
desorption.
The gas phase concentrations of MCB and benzene, along the
Appendix A
biofilter column, can be described by:
 
A.1. Governing equations dC m dSm
Ug ¼ As Dem ð5Þ
dh dx x¼0
The differential equations describing the removal of MCB and
 
benzene in a differential section of the biofilm at steady state can dC b dSb
Ug ¼ As Deb ð6Þ
be represented as: dh dx x¼0
2
d Sm X lmaxðmÞ Sm where Ug is superficial gas velocity, As is the biofilm surface area
Dem ¼ ð1Þ
dx
2 Y x=s K m þ Sm and h is position along the biofilter.
The corresponding boundary conditions are:
2
d Sb X lmaxðbÞ Sb C m ¼ C mi and C b ¼ C bi ð7Þ
Deb 2
¼ ð2Þ
dx Y x=s K b þ Sb
where the subscript ‘i’ represents conditions at the inlet of the
where lmax(m) and lmax(b) is the maximum specific growth rate, Km biofilter.
and Kb is the half saturation constant of MCB and benzene respec-
tively, Dem and Deb is the effective diffusion coefficient of the MCB A.2. Model solution
and benzene in the biofilm, Sm and Sb represent the liquid phase
concentration of MCB and benzene in biofilm and x is the position There were four differential equations (Eqs. (1), (2), (5), and (6)),
in the biofilm. X is the dry cell density in g/m3 and Yx/s is the bio- that needed to be solved using numerical technique in MATLAB
mass to substrate yield coefficient. Since both these values were platform. The code is based on the use of the Shooting method
not known single parameter mmax (=lmax X/Yx/s) was used in the for the biofilm, and finite difference method for the gas phase along
model in order to avoid further adjustments and assumptions for the column (Chapra and Canale, 2003; Cuptil and Shacham, 2007).
5174 R.A. Pandey et al. / Bioresource Technology 101 (2010) 5168–5174

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