International Biodeterioration & Biodegradation 85 (2013) 451e457

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International Biodeterioration & Biodegradation

journal homepage: www.elsevier.com/locate/ibiod

Biological treatment of polychlorinated biphenyls (PCBs)

contaminated transformer oil by anaerobiceaerobic sequencing
batch biofilm reactors
B.F. Nabavi, M. Nikaeen*, M.M. Amin, H. Farrokhzadeh
Department of Environmental Health Engineering, Environment Research Center, Isfahan University of Medical Science, Hezar Jerib Avenue, Isfahan, Iran

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

Article history: Concern about the environmental fate of PCBs contaminated transformer oils has increased recently.
Received 2 August 2012 Therefore, there is a real need to develop and improve treatment methods of transformer oils to mini-
Received in revised form mize their environmental side effects. This study was carried out to evaluate the efficiency of sequential
19 June 2013
anaerobiceaerobic batch biofilm reactors for the biological treatment of transformer oil containing PCBs.
Accepted 12 September 2013
The process performance was studied by increasing the organic loading rate (OLR) in the range of 0.9
Available online 8 October 2013
e32.2 g COD/L.d. Average chemical oxygen demand (COD) removal efficiencies of 99.8% were achieved in
the system at OLR of 21.5 g COD/L.d. Also, high PCBs biodegradation percentages 96.5 were observed.
Keywords:
Transformer oil
Analysis of common indicators for the monitoring of anaerobic and aerobic processes confirmed the high
Biodegradation ability of the anaerobiceaerobic process for treatment of PCBs contaminated transformer oil, charac-
Polychlorinated biphenyls (PCBs) terized by high COD concentration and by highly chlorinated biphenyls content. The StovereKincannon
Sequencing batch biofilm reactors model found to be the most appropriate model for predicting transformer oil biodegradation in anaer-
obiceaerobic SBBR. This biological system as an environmentally friendly and cost effective method has
proved to be a suitable technology for treatment of PCB contaminated transformer oil with high effi-
ciency in organic matter and PCBs removal.
Ó 2013 Elsevier Ltd. All rights reserved.

1. Introduction insulating properties (Pieper, 2005; Furukawa and Fujihara, 2008).

In recent decades, there has been an increasing concern about
the fate of industrial chemicals and wastes such as transformer
oils (Haus et al., 2001). Transformer oil is used in transformers as a
cooling and insulating medium (Lucas et al., 2001). Mineral oils
have traditionally used as the major compounds of transformer oil
due to their superior dielectric properties. Mineral oils, complex
hydrocarbon mixtures with three main chemical groups including
parafins, naphthenes and aromatics, are produced from crude
petroleum by various processing steps. These chemical groups are
known to differ in their microbial degradation with range varies
from 15 to 75% (Haus et al., 2001). Aromatic hydrocarbons and
polar compounds with high resistance to biodegradation (Haus
et al., 2001) are a major concern for environment and human
health. Furthermore, since 1929, polychlorinated biphenyls (PCBs)
have been widely used for transformer oils formulation due to
their excellent chemical and physical stability and electrical
* Corresponding author. Tel.: þ98 3117922660; fax: þ98 3116682509.
E-mail address: Nikaeen@hlth.mui.ac.ir (M. Nikaeen).
PCBs, a family of synthetic organic chemicals with toxigenic,
carcinogenic and reproduce effects pose a great risk to human
health and the environment safety (Hatamian-Zarmi et al., 2009).
The high chemical stability and superhydrophobicity of PCBs cause
them to bioaccumulate in cells and pass by the food chain (Borja
et al., 2005; Pieper and Seeger, 2008). At present, there are large
amounts of end-of-life transformer oils all over the world, which
actual or potential release of them to the environment seriously
threats human and ecosystem (Rojas-Avelizapa et al., 1999).
Therefore, it is urgent to develop and improve treatment methods
of transformer oils to minimize their environmental side effects.
Different physical and chemical methods such as incineration and
direct dechlorination for the disposal of transformer oils and
destruction of PCBs have been proposed (Sobiecka et al., 2009).
However, incineration as a conventional disposal method with
high efficiency; could emit toxic compounds and is extremely
costly (Kastánek and Kastánek, 2005). Since, bacteria play a
fundamental role in the removal of waste chemical compounds
(Pieper and Seeger, 2008); recently, biological methods of PCBs
degradation as an environmentally friendly solution were applied
(Sobiecka et al., 2009). However, the number of chlorine atoms per

0964-8305/$ e see front matter Ó 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.ibiod.2013.09.007
452 B.F. Nabavi et al. / International Biodeterioration & Biodegradation 85 (2013) 451e457

molecule is a critical feature for biodegradation of PCBs (Furukawa, Table 1

2000; Field and Sierra-Alvarez, 2008). Highly chlorinated bi- Operational parameters of the anaerobiceaerobic sequencing batch biofilm reactors.

phenyls are persistent under aerobic condition, but anaerobic Anaerobic-SBBR Aerobic-SBBR
dechlorination of highly chlorinated congeners generates lower Influent OLR HRT Concentration Influent OLR HRT (d)a
chlorinated congeners which are easily degraded under aerobic COD (g/L) (g COD/L.d) (d)a of influent COD (g/L) (g COD/L.d)
conditions (Abraham et al., 2002). Since, the main mixture of PCBs PCBs (mg/L)
in transformer oils is high chlorinated compounds including 4.7 0.9 5 0.034 0.12 0.01 10
Aroclor 1260 and 1254 (Rojas-Avelizapa et al., 1999; Martínez 7.8 1.6 5 0.041 0.25 0.03 10
et al., 2005), sequential anaerobiceaerobic treatment is expected 10.7 2.1 5 0.055 0.24 0.02 10
13.5 2.7 5 0.068 1.09 0.11 10
to provide complete biodegradation of PCBs contaminated trans-
14.5 2.9 5 0.082 1.18 0.12 10
former oils (Tartakovsky et al., 2001). There are several study 18.9 3.8 5 0.102 1.55 0.15 10
about the biodegradation and bioremediation of PCBs (Fedi et al., 23.7 4.7 5 0.136 1.66 0.17 10
2001; Borja et al., 2006; Adebusoye et al., 2008; Dercová et al., 28.4 5.7 5 0.205 1.09 0.11 10
2008; Hatamian-Zarmi et al., 2009; Bertin et al., 2011). Most of 31.1 6.2 5 0.273 1.10 0.11 10
36.2 7.2 5 0.341 0.84 0.08 10
the studies focused on batch cultures involving pure cultures or
39.4 7.9 5 0.478 1.14 0.11 10
mixture of bacteria in laboratory media containing a single PCBs 53.7 10.7 5 1.854 0.98 0.10 10
congener or an individual Aroclor. Sobiecka et al. (2009) reported 53.7 21.5 2.5 3.709 2.09 0.42 5
the ability of commercial mixtures of microorganisms in biological 53.7 32.2 1.7 5.563 12.26 3.7 3.3
treatment of transformer oil using glass tubes in batch mode for a
HRT ¼ VReactor =VFeed tC , tC: cycle period (Siman et al., 2004).
several days. They observed PCB congeners biodegradation in the
ranges of 0e99%, 2e97% and 40e94% under anoxic, oxic and
anoxic/oxic treatments. Tartakovsky et al. (2001) evaluated 14 L aerobic sequencing batch biofilm reactor (SBBR) as shown in
degradation of Aroclor 1242 by coupled anaerobiceaerobic bio- Fig. 1. The reactors were made of Plexiglas and placed into water
reactors and a near complete mineralization of Aroclor 1242 was bathes heated by aquarium heaters for keeping the operational
observed in this study. Hatamian-Zarmi et al. (2009) reported temperature at 37  1  C. The anaerobic reactor was fed with
biodegradation of highly chlorinated biphenyl and Aroclor 1242 by synthetic wastewater from a storage tank using an aquarium
Pseudomonas aeruginosa TMU56. Gas chromatograph analysis of pump. A reciprocal pump and a peristaltic pump were also used to
Aroclor 1242 following 4 days incubation showed about 73% transfer effluent from anaerobic to aerobic reactor and to
degradation of PCBs. In the other study, Borja et al. (2006) discharge final effluent, respectively. The SBBR reactors were filled
investigated biodegradation of polychlorinated biphenyls using with 1 cm polyurethane foam cubes (initial porosity of 85%) as
biofilm grown in a fluidized bed reactor. Based on our knowledge, immobilizing support. These cubes were contained in glass bas-
however, there is no study about the biological treatment of PCBs kets inside the reactors. In order to minimize the air pollution
contaminated transformer oils in biological reactors. In this study, from PCBs, granular activated carbon was used on the top of aer-
for the first time, we evaluated the efficiency of sequential obic reactor.
anaerobiceaerobic batch biofilm reactors for the biological treat-
ment of transformer oil containing PCBs. 2.2. Experimental setup

2. Material and methods Both reactors were inoculated with a mixture of wastewater
sludge and transformer contaminated soil microbial consortium
2.1. SBBR reactors have already been acclimatized to PCB contaminated transformer
oil in a batch experiment .The anaerobic reactor was fed with
The coupled anaerobiceaerobic bioreactors consisted of 7 L synthetic wastewater containing PCB-contaminated transformer
anaerobic sequencing batch biofilm reactor (ASBBR) connected to oil at a rate of 1 L/cycle. The synthetic wastewater was supplied

Fig. 1. Scheme of anaerobiceaerobic sequencing batch biofilm reactors containing immobilized biomass: (1) substrate tank, (2) ASBBR, (3) SBBR, (4) feed pump, (5) water bath, (6)
wet gas meter, (7) gas bag, (8) reciprocal pump, (9) discharge pump, (10) air pump, (11) treated effluent, (12) Programmable logic controller (PLC) and timers.
 B.F. Nabavi et al. / International Biodeterioration & Biodegradation 85 (2013) 451e457
to the bioreactor in cycles of 24, 12 and 8 h corresponding to
hydraulic retention times (HRT) of 5, 2.5 and 1.7 days. It was
made up freshly every day with tap water, different volumes of
transformer oil in acetone and mineral medium. The composition
of mineral medium was as follow (in mg L-1 of tap water): NH4Cl,
76.45; KH2PO4, 10; K2HPO4, 25.32; FeCl3, 1.021; CaCl2$2H2O, 2.06;
MgSO4$7H2O, 2.14; MnCl2$2H2O, 0.34; COCl2$6H2O, 0.092;
NiSO4$6H2O, 0.0763; ZnSO4, 0.0592; Na2MoO4$2H2O, 0.0822;
CuCl2$2H2O, 0.016; H3BO3, 0.02. The pH was adjusted 7e7.5 with
the NaOH solution. The waste transformer oil (Askarel) was ob-
tained from power plant of Isfahan, Iran; with an initial PCBs
concentration of 200 mg/L. Aroclors 1242, 1254 and 1260 were
the major PCBs compounds of transformer oil with the concen-
tration of 22.42 mg/L, 65.89 mg/L, and 97.13 mg/L, respectively.
The reactors were run for a start-up period of 85 days with an
organic loading rate (OLR) of 0.4 g COD/L.d which gradually
increased to 0.9 g COD/L.d, to allow biofilm formation. Then, the
system was operated for about 14 months with step-wise in-
crease of the organic loading rate (OLR) from 0.9 g COD/L.d to
32.2 g COD/L.d. Each step was run for several weeks to ensure
that steady state conditions were achieved. Kinetic analysis was
carried out based on values obtained from stable performance
stage. The operational parameters of reactors are presented in
Table 1.
2.3. Analytical methods
The performance of reactors was evaluated by monitoring
physicalechemical variables in the influent and effluent of the re-
actors at different steps. Chemical oxygen demand (COD), volatile
suspended solids (VSS), volatile fatty acids (VFA), total alkalinity
and pH were measured according to the Standard Methods (APHA,
2005). The biogas production was measured by wet gas meter
(ELSTER, AMCO, Germany) and determination of methane pro-
duced was carried out by gas chromatography. pH and DO were
monitored daily with pH-meter (SCHOTT, model GC 824) and DO-
meter (YSI, model 55, American).
2.4. PCBs analysis
PCBs analysis (Aroclors 1242, 1254, 1260) was performed with
a gas chromatography (AGILENT 6890N), equipped with an
electron capture detector (ECD) and a capillary column
(30m  0.32 mm) with HP-5 phase after extraction. Purification
and separation of PCBs compounds in transformer oils and
effluent samples were conducted by liquideliquid extraction
using n-hexane. Extraction procedure was performed in 250 ml
separatory funnels. 20 ml of hexane was added to 100 ml of the
sample and PCBs were extracted by vigorous shaking and
collection of the hexane layer. The procedure was repeated
several times. The hexane layer, then, was rehydrated with so-
dium sulfate and evaporated to a final volume of 2 ml using a
rotary vacuum evaporator. The extracts were injected into the
capillary column. The injector and detector were kept at 270  C
and 300  C, respectively. Nitrogen was used as carrier gas
(0.9 ml/min). The temperature program was set at an initial
temperature of 120  C (0 min). This was increased by 15  C/min
to 210  C (1 min) and then by 2  C/min to 245  C (1 min) and
finally by 30  C/min to 290  C (3 min).
2.5. Scanning electron microscopy
The presence and structure of the biofilm attached to the
polyurethane foam carriers was examined by scanning electron
microscopy (SEM, LEO, England).
453
Fig. 2. Influence of OLR on the COD removal efficiency.
3. Result and discussion
3.1. Reactor performance
The anaerobic reactor was operated at OLRs of 0.9e32.2 g COD/
L.d, with an increasing COD concentration from 4.7 g/L to 53.7 g/L
and HRT decreasing from 5 to 1.7 days. The performance of the
reactors with regard to the COD removal efficiency is shown in
Fig. 2. The average COD removal efficiency of the anaerobic-SBBR
and aerobic-SBBR was 94.3% and 85.3%, respectively. As shown in
Fig. 2, the most of COD was removed in the anaerobic stage. These
results would be in consistent with those reported by Işık and
Sponza (2004) and Hosseini koupaie et al. (2012). Isik and Sponza
reported 77% of COD removal in anaerobic stage of a sequential
anaerobiceaerobic system treated azo dye in a synthetic waste-
water. Hosseini Koupaie et al. also achieved anaerobic COD removal
of 81e83% in a sequential ASBR/MB-SBBR system treated waste-
water containing Acid Red 18. Total COD removal efficiency in the
range of 99e99.8% was achieved in the anaerobiceaerobic reactors
at OLR of between 0.9 g COD/L.d and 21.5 g COD/L.d. The results
confirm the capability of anaerobiceaerobic process in treatment of
high strength industrial wastewaters (Chan et al., 2009).
Recently, anaerobiceaerobic systems have been widely used for
treatment of high organic strength wastewaters such as textile.
Application of anaerobiceaerobic SBR systems in the treatment of
textile wastewaters has proved to be a suitable technology as high
COD removal of more than 85% is achieved (Chan et al., 2009). It is
also noticeable that the use of immobilized microorganisms as
biofilm in sequential anaerobiceaerobic biological systems offers
some advantages over suspended growth for biodegradation of
organic matters (Chan et al., 2009). The enhanced treatment ca-
pacity and operational stability caused by a large active biomass
concentration is one of the most significant advantages of the
biofilm process. Long sludge retention time (SRT) due to the
microorganism immobilization makes the biofilm reactors also
suitable for slow-growing (Elenter et al., 2007). Calculation of the
SRT in the present study showed a long SRT as 122 days, which
confirms the ability of biofilm processes in providing of high
biomass concentration.
As shown in Fig. 2, the COD removal efficiency was 98.2% and
96.1% for anaerobic and 99.8% for anaerobiceaerobic reactors at the
OLR of 10.7 g COD/L.d and 21.5 g COD/L.d corresponding to a HRT of
5 and 2.5 days, respectively. The average COD concentration of the
effluent was 1.02 g/L and 0.1 g/L for anaerobic and aerobic reactor,
respectively. However, in higher organic loading rate (32.2 g COD/
L.d and HRT of 1.7 days), the system presented reduced organic
removal efficiency down to 78% and 97.1% for anaerobic and
anaerobiceaerobic reactors, respectively. At the highest OLR, there
454 B.F. Nabavi et al. / International Biodeterioration & Biodegradation 85 (2013) 451e457

Fig. 3. Influence of OLR on the VFA concentration and biogas production as anaerobic
indicator parameters.

was a marked increase in the effluent COD concentration to about

12.26 g/L and 1.6 g/L for anaerobic and aerobic reactor, respectively
.In overall, the results indicated that very high efficiencies of COD
removal (99.8%) is attained even at high organic loading rate
(21.5 g COD/L.d) for anaerobiceaerobic SBBR. The result was com-
parable to the study of Majone et al. (2010). They investigated
anaerobic treatment of an industrial wastewater from a Fischere
Tropsch process in a continuous-flow packed-bed biofilm reactor.
The reactor was operated at an OLR of 20 g COD/L.d and a hydraulic
retention time of 1.4 days with a COD removal of 96% with nearly
complete conversion of the removed COD into methane.

3.2. Process monitoring

Process was monitored continually to understand the

biochemical process during reactors operation, by determining pH, Fig. 4. StovereKincannon model plot: (A) ASBBR, (B) SBBR.

DO, total alkalinity, volatile fatty acids, and volatile suspended

solids (VSS) and biogas production. Values of pH varied from 6.8 to
The VFA/Alkalinity ratio can be also used as a measure of
7.05 and 6.9 to 8.1 for anaerobic and aerobic bioreactor, respec-
anaerobic biological treatment stability, which a ratio of less than
tively. The DO concentration in aerobic bioreactor varied from 2.1 to
0.3e0.4, the process is considered to be operating favorably
3.1 mg/L, except for the OLR of 21.5 g COD/L.d and 32.2 g COD/L.d
without acidification risk (Borja et al., 2004). In our study the VFA/
which DO content was reduced to 1.7 and 0.04 mg/L, respectively.
alkalinity ratio values were in the suggested limit value or a little
With OLR increasing to 32.2 g COD/L.d, effluent VFA sharply
higher. However, a considerable increase of the VFA/Alkalinity ratio
increased to a maximum value indicating that VFA accumulation
was observed at OLR of 32.2 g COD/L.d as a result of VFA accumu-
occurred (Fig. 3). High level concentration of VFA could be related to
lation during overload.
the inability of methanogens to cope with the high organic loading
rate (Majone et al., 2010) which leads to the accumulation of in-
3.3. Kinetic models
termediate products such as VFA (Ghaniyari-Benis et al., 2009).
Despite of increasing level of VFA at high OLR, the total alkalinity
Many models have been used to describe the overall kinetics of
(expressed as equivalent CaCO3) did not decreased. This can be
biological organic removal in reactors (Borghei et al., 2008). First-
attributed to carbonate/bicarbonate buffering capacity of the
order, StovereKincannon and Grau-second order (Grau) model
reactor. The carbonate/bicarbonate buffering capacity is related to
are widely used for the study of biodegradation kinetic. The kinetic
the presence of CO2 in the reactor that is not completely reduced to
models were plotted for anaerobic and aerobic SBBR reactors. The
CH4 as biogas (Borja et al., 2004).
linear correlation coefficient (R2) was selected for choosing the
The biogas production rate is a one of the most important in-
most suitable model to represent organic matter removal kinetic in
dicators for the monitoring of anaerobic processes. The biogas
the study (Padilla-Gasca and López, 2010). According to the highest
production rate as a function of OLR is shown in Fig. 3. The biogas
R2 values from the assessed models, the StovereKincannon model
production increased with increasing OLR up to 21.5 g COD/L.d.
with a value of 0.999 and 0.99 for anaerobic and aerobic reactor,
However, the VFA accumulation during organic overload
respectively found to be the most appropriate model. The Stovere
(32.2 g COD/L.d) responded very quickly by biogas production rate.
Kincannon model (Eq. (1)) is the most widely accepted method in
This marked change in the biogas production might be attributed to
modeling a variety of different types of biofilm reactors.
an inhibition of the methanogens activity because of VFA accu-
mulation at highest OLR (Borja et al., 2004). The analysis of biogas  
V KB V 1
at the end of experiment showed 81.4% of methane, which corre- ¼ þ (1)
Q ðS0  SÞ Umax QS0 Umax
sponded to the yield coefficient of methane production about 0.39 L
of methane per gram COD removed. The result confirms the pres- In the StovereKincannon model, [V/Q (S0  S)] was plotted
ence and good activity of methanogens in the anaerobic reactor. versus V/(QS0) (Fig. 4), where V is the reactor volume (L), Q is the
 B.F. Nabavi et al. / International Biodeterioration & Biodegradation 85 (2013) 451e457 455

Fig. 5. The removal percentage of PCBs in anaerobic and anaerobiceaerobic processes.

inflow rate (L/d), S0 and S are the substrate concentration in the

feed and effluent (g/L), respectively.
Linear regression was used to calculate the slope (KB/Umax) and
the intercept point of the straight line (1/Umax). KB (saturation value
constant) and Umax (maximum substrate utilization rate) were
calculated, as 171.2 and 166.7 g/L.d for anaerobic and 0.37 and
0.23 g/L.d for aerobic bioreactor, respectively. The linear regression
with high R2 for both of the reactors showed that the removal ef-
ficiencies were a function of organic loading rate and transformer
oil biodegradation increased with increasing its concentration in
the feed solution.
Compared with the maximum OLR of 32.2 g COD/L.d obtained
in this study, the predicted Umax is significantly higher in
anaerobic reactor, which indicates potentially higher capacity of
the ASBBR in coping with high organic strength transformer oil
(Wang et al., 2009). In the results reported by Wang et al. (2009)
and Ahn and Forster (2000) there was also remarked difference
between the maximum OLR and the Umax predicted by Stovere
Kincannon model. However, in comparison to the values of Umax
and KB obtained by the same StovereKincannon model for
various substrates, the highest values were obtained in the pre-
sent study for biodegradation of transformer oil by ASBBR (Wang
et al., 2009).

3.4. PCBs biodegradation

The biodegradation PCBs-contaminated transformer oil was

investigated under anaerobiceaerobic treatment. No carbon
sources other than the transformer oil were provided, because of
the presence of other hydrocarbons different from PCBs in
transformer oil (Rojas-Avelizapa et al., 1999). Overall, PCBs
biodegradation percentages of 94.2 and 96.5 were observed un-
der anaerobic and anaerobiceaerobic treatment (Fig. 5), respec-
tively. Sequential anaerobiceaerobic treatment of PCBs especially
highly chlorinated compounds has been successfully tested in
batch cultures for several weeks to several months. Master et al.
(2002) reported a sequential anaerobiceaerobic laboratory scale
treatment of soil contaminated with Aroclor 1260 with an initial
concentration of 59 mg/L in a period of 4 months (Borja et al.,
2005). Based on several studies, however, the rate of PCBs
biodegradation differ from organism to organism and depends
on the initial concentration as well as the composition of the
PCBs mixture or types of Aroclor (Adebusoye et al., 2008). As
seen in Fig. 5, considerable values of PCBs mixture have been
removed in anaerobic stage. Calculation of total biomass con-
centration based on the VSS analysis showed that about 4.3 mg
PCBs/gVSS were removed in anaerobic reactor. It has been
Fig. 6. SEM observation of microorganisms immobilized in polyurethane foam: (A)
polyurethane foam, (B) SBBR biofilm and (A) ASBBR biofilm.
demonstrated that biodegradation of highly chlorinated bi-
phenyls (i.e. Aroclors 1242, 1254, 1260) under aerobic conditions
is limited (Borja et al., 2005). In general, the persistence of PCBs
increases with an increase in the degree of chlorination (Sobiecka
et al., 2009). It was proposed that anaerobic dechlorination and
subsequent aerobic mineralization of dechlorinated compounds
by oxidative attack of the aerobic bacteria is the main mecha-
nisms for anaerobiceaerobic treatment of PCBs (Tartakovsky
et al., 2001). Nevertheless, some aerobic bacteria have the ca-
pacity to degrade highly chlorinated PCB congeners using an
initial oxygenase reaction (Sobiecka et al., 2009). The results of
the study showed the average removal percentage of PCB
456 B.F. Nabavi et al. / International Biodeterioration & Biodegradation 85 (2013) 451e457
compounds under aerobic conditions was about 50%. Adebusoye
et al. (2008) reported 37% degradation of Aroclor 1242 from
transformer oil in aerobic condition. Comparison of degradation
rate of Aroclors showed the highest average removal percentage
for Aroclor 1242 with a value of 85.4% in anaerobic reactor.
Quesen et al. also reported extensive dechlorination of Aroclor
1242 and 1248 in comparison with Aroclors 1254 and 1260 (Borja
et al., 2005). Generally, in accordance with the previous studies
our results demonstrated that microorganisms were more cable
of metabolizing high-chlorinated Aroclors under anaerobic con-
ditions than aerobic conditions. Although, only few studies
report the use of biofilm to degrade PCBs (Mukerjee-Dhar et al.,
1998; Kuncova et al., 2002; Borja et al., 2006), application of
biofilms as an alternative in the degradation of PCBs offers some
advantages such as the presence of large microbial diversity in a
stable form (Borja et al., 2006). In overall, sequential anaerobice
aerobic SBBR achieved successfully the biodegradation of
highly chlorinated biphenyls more than 99.9% at high OLR of
21.5 g COD/L.d.
Due to low solubility of PCBs in water, potential accumulation
or adsorption on biofilm and media must be considered. Extrac-
tion of PCBs from media and attached biofilm at the end of
experiment showed a content of 3.68 mg PCBs/g biofilm. This
meant that only 1.9% of the total content of applied PCBs mixture
to the reactor was adsorbed, and decrease in PCB concentration
can really be attributed to biotransformation and not to any other
mechanisms such as adsorption. In the other word, biodegradation
was the predominant mechanisms of PCBs removal. The study of
Tartakovsky et al. (2001) also showed only 4e9% of the total
amount of Aroclor fed to the reactor was accumulated in the
sludge.
3.5. Scanning electron microscopy
The structure of the biofilm attached to the polyurethane foam
carriers is shown for aerobic and anaerobic biofilm in Fig. 6.
4. Conclusion
The results of this study suggest that anaerobiceaerobic SBBR
can be used successfully as an alternative for treatment of PCBs
contaminated transformer oil, characterized by high COD con-
centration and by highly chlorinated biphenyls content. This bio-
logical system as an environmentally friendly and cost effective
method has proved to be a suitable technology for treatment of
PCBs contaminated transformer oil with high efficiency of 99.8%
for COD as well as 99.9% for PCBs removal at high OLR of
21.5 g COD/L.d.
Acknowledgment
This research was funded by the vice chancellery for research
of Isfahan University of Medical Sciences (Grant No. 389268). The
authors wish to thank the Mr. Jafarpoor from the Power Plant of
Isfahan for him kind help and providing the transformer oil
sample. We also thankful for the initial financial support of this
project provided by R & D section of Iron-Foundry Company of
Isfahan.
References
Abraham, W.R., Nogales, B., Golyshin, P.N., Pieper, D.H., Timmis, K.N., 2002. Poly-
chlorinated biphenyl-degrading microbial communities in soils and sediments.
Curr. Opin. Microbiol. 5, 246e253.
Adebusoye, S.A., Ilori, M.O., Picardal, F.W., Amund, O.O., 2008. Extensive biodegra-
dation of polychlorinated biphenyls in Aroclor 1242 and electrical transformer
fluid (Askarel) by natural strains of microorganisms indigenous to contami-
nated African systems. Chemosphere 73, 126e132.
Ahn, J.H., Forster, C.F., 2000. Kinetic analyses of the operation of mesophilic and
thermophilic anaerobic filters treating a simulated starch wastewater. Process
Biochem. 36, 19e23.
APHA, AWWA, WEF, 2005. Standard Methods for the Examination of Water &
Wastewater, 21 ed. American Public Health Association, New York.
Bertin, L., Capodicasa, S., Fedi, S., Zannoni, D., Marchetti, L., Fava, F., 2011.
Biotransformation of a highly chlorinated PCB mixture in an activated sludge
collected from a Membrane Biological Reactor (MBR) subjected to anaerobic
digestion. J. Hazard. Mater. 186, 2060e2067.
Borghei, S.M., Sharbatmaleki, M., Pourrezaie, P., Borghei, G., 2008. Kinetics of
organic removal in fixed-bed aerobic biological reactor. Bioresour. Technol. 99,
1118e1124.
Borja, J., Taleon, D.M., Auresenia, J., Gallardo, S., 2005. Polychlorinated biphenyls and
their biodegradation. Process. Biochem. 40, 1999e2013.
Borja, J.Q., Auresenia, J.L., Gallardo, S.M., 2006. Biodegradation of polychlorinated
biphenyls using biofilm grown with biphenyl as carbon source in fluidized bed
reactor. Chemosphere 64, 555e559.
Borja, R., Rincón, B., Raposo, F., Dominguez, J.R., Millán, F., Martin, A., 2004. Meso-
philic anaerobic digestion in a fluidised-bed reactor of wastewater from the
production of protein isolates from chickpea flour. Process. Biochem. 39, 1913e
1921.
Chan, Y.J., Chong, M.F., Law, C.L., Hassell, D.G., 2009. A review on anaerobic-aerobic
treatment of industrial and municipal wastewater. Chem. Eng. J. 155, 1e18.
Dercová, K., Cicmanová, J., Lovecká, P., Demnerová, K., Macková, M., Hucko, P.,
Kusnír, P., 2008. Isolation and identification of PCB-degrading microor-
ganisms from contaminated sediments. Int. Biodeterior. Biodegrad. 62,
219e225.
Elenter, D., Milferstedt, K., Zhang, W., Hausner, M., Morgenroth, E., 2007. Influence
of detachment on substrate removal and microbial ecology in a heterotrophic/
autotrophic biofilm. Water Res. 41, 4657e4671.
Fedi, S., Carnevali, M., Fava, F., Andracchio, A., Zappoli, S., Zannoni, D., 2001. Poly-
chlorinated biphenyl degradation activities and hybridization analyses of
fifteen aerobic strains isolated from a PCB-contaminated site. Res. Microbiol.
152, 583e592.
Field, J.A., Sierra-Alvarez, R., 2008. Microbial transformation and degradation of
polychlorinated biphenyls. Environ. Pollut. 155, 1e12.
Furukawa, K., 2000. Biochemical and genetic bases of microbial degradation of
polychlorinated biphenyls (PCBs). J. Gen. Appl. Microbiol. 46, 283e296.
Furukawa, K., Fujihara, H., 2008. Microbial degradation of polychlorinated bi-
phenyls: biochemical and molecular features. J. Biosci. Bioeng. 105, 433e449.
Ghaniyari-Benis, S., Borja, R., Monemian, S.A., Goodarzi, V., 2009. Anaerobic treat-
ment of synthetic medium-strength wastewater using a multistage biofilm
reactor. Bioresour. Technol. 100, 1740e1745.
́ ́
Hatamian-Zarmi, A., Shojaosadati, S.A., Vasheghani-Farahani, E., Hosseinkhani, S.,
Emamzadeh, A., 2009. Extensive biodegradation of highly chlorinated biphenyl
and Aroclor 1242 by Pseudomonas aeruginosa TMU56 isolated from contami-
nated soils. Int. Biodeterior. Biodegrad. 63, 788e794.
Haus, F., German, J., Junter, G.A., 2001. Primary biodegradability of mineral base oils
in relation to their chemical and physical characteristics. Chemosphere 45,
983e990.
Hosseini koupaie, E., Alavi moghaddam, M.R., Hashemi, S.H., 2012. Investigation of
decolorization kinetics and biodegradation of azo dye Acid Red 18 using
sequential process of anaerobic sequencing batch reactor/moving bed
sequencing batch biofilm reactor. Int. Biodeterior. Biodegrad. 71, 43e49.
Işık, M., Sponza, D.T., 2004. Monitoring of toxicity and intermediates of C.I. Direct
Black 38 azo dye through decolorization in an anaerobic/aerobic sequential
reactor system. J. Hazard. Mater. 114, 29e39.
Kastánek, F., Kastánek, P., 2005. Combined decontamination processes for wastes
containing PCBs. J. Hazard. Mater. 117, 185e205.
Kuncova, G., Triska, J., Vrchotova, N., Podrazky, O., 2002. The influence of immobi-
lization of Pseudomonas sp. 2 on optical detection of polychlorinated biphenyls.
Mater. Sci. Eng. 21, 195e201.
Lucas, J.R., Abeysundara, D.C., Weerakoon, C., Perera, K., Obadage, K.C.,
Gunatunga, K.A.I., 2001. Coconut oil insulated distribution transformer. In:
Presented at the 8 th Annual Conference of the IEE Sri Lanka on 22 September
2001.
Majone, M., Aulenta, F., Dionisi, D., D’addario, E.N., Sbardellati, R., Bolzonella, D.,
Beccari, M., 2010. High-rate anaerobic treatment of FischereTropsch waste-
water in a packed-bed biofilm reactor. Water Res. 44, 2745e2752.
Martínez, H.A.R., Rodríguez, G.C., Castillo, D.H., 2005. Determination of PCBs in
transformers oil using gas chromatography with mass spectroscopy and Aro-
clors (A1254: A1260). J. Mex. Chem. Soc. 49, 263e270.
Mukerjee-Dhar, G., Shimura, M., Kimbara, K., 1998. Degradation of polychlorinated
biphenyl by cells of Rhodococcus opacus strain TSP203 immobilized in alginate
and in solution. Enzyme Microb. Technol. 23, 34e41.
Master, E.R., Lai, V.V., Kuipers, B., Cullen, W.R., Mohn, W., 2002. Sequential anaer-
obiceaerobic treatment of soil contaminated with weathered Aroclor 1260.
Environ. Sci. Technol. 36, 100e103.
Padilla-Gasca, E., López, A.L., 2010. Kinetics of organic matter degradation in an
upflow anaerobic filter using slaughterhouse wastewater. J. Bioremed. Bio-
degrad. 1, 106e112.
Pieper, D.H., 2005. Aerobic degradation of polychlorinated biphenyls. Appl. Micro-
biol. Biotechnol. 67, 170e191.
 B.F. Nabavi et al. / International Biodeterioration & Biodegradation 85 (2013) 451e457
Pieper, D.H., Seeger, M., 2008. Bacterial metabolism of polychlorinated biphenyls.
J. Mol. Microbiol. Biotechnol. 15, 121e138.
Rojas-Avelizapa, N.G., Rodriguez-Vázquez, R., Enriquez-Villanueva, F., Martinez-
Cruz, J., Poggi-Varaldo, H.M., 1999. Transformer oil degradation by an indigenous
microflora isolated from a contaminated soil. Resourc. Conserv. Recycl. 27, 15e26.
Siman, R.R., Borges, A.C., Ratusznei, S.M., Rodrigues, J.A.D., Zaiat, M., Foresti, E.,
Borzani, W., 2004. Influence of organic loading on an anaerobic sequencing
biofilm batch reactor (ASBBR) as a function of cycle period and wastewater
concentration. J. Environ. Manage. 72, 241e247.
́ ́ ́
457
Sobiecka, E., Cedzynska, K., Bielski, C., Antizar-Ladislao, B., 2009. Biological treat-
ment of transformer oil using commercial mixtures of microorganisms. Int.
Biodeterior. Biodegrad. 63, 328e333.
Tartakovsky, B., Michotte, A., Cadieux, J.C.A., Lau, P.C.K., Hawari, J., Guiot, S.R., 2001.
Degradation of Aroclor 1242 in a single-stage coupled anaerobic/aerobic
bioreactor. Water Res. 35, 4323e4330.
Wang, S., Chandrasekhara Rao, N., Qiu, R., Moletta, R., 2009. Performance and ki-
netic evaluation of anaerobic moving bed biofilm reactor for treating milk
permeate from dairy industry. Bioresour. Technol. 100, 5641e5647.