Process Biochemistry 40 (2005) 1501–1508

www.elsevier.com/locate/procbio

Treatment of complex chemical wastewater in a sequencing batch

reactor (SBR) with an aerobic suspended growth configuration$
S. Venkata Mohan, N. Chandrashekara Rao, K. Krishna Prasad,
B.T.V. Madhavi, P.N. Sharma*
Biochemical and Environmental Engineering Centre, Indian Institute of Chemical Technology, Hyderabad 500007, India

Received 11 September 2002; accepted 20 February 2003

Abstract

A sequencing batch reactor (SBR) has certain advantages over conventional activated sludge processes (ASP) for the treatment of
complex wastewater. The performance of a sequencing batch reactor in treating complex chemical effluents was investigated with a
suspended biomass configuration and operating under aerobic conditions. A total sequence of 24 h (15 min: filling phase; 23 h: reaction
phase (aeration with recirculation); 30 min: setting; and 15 min: withdrawal) was employed and studied with various organic loading rates
(1.0 kg COD/m3/day: 1.7 kg COD/m3/day and 3.5 kg COD/m3/day). The SBR performance was assessed by means of carbon removal and
operational parameters such as pH, oxidation–reduction potential (ORP), sludge volume (SV), sludge volume index (SVI), suspended solids
(SS) and volatile suspended solids (VSS), which were monitored during reactor operation. The SBR showed relatively more efficient
performance over conventional suspended growth systems. About 8% sulphate was removed under anoxic conditions at all the studied
organic loading rates. Enhanced performance with SBR over conventional ASP may be due to enforced short term unsteady state conditions
coupled with periodic exposure of the microorganisms (physiological state) to defined process conditions which facilitate the required
metabolic conditions for treating complex chemical effluents.
# 2004 Published by Elsevier Ltd.

Keywords: Sequencing batch reactor; Complex chemical wastewater; Suspended growth; Oxygen consumption rate

1. Introduction serve as microbial substrates. Alternative approaches like

Industrial wastewater, in particular, originating from
pharmaceutical and chemical industries, normally contains
toxic organic compounds [1]. The variability of the
wastewater on both flow and composition (change of
manufacturing product, transitory operation of the plant,
washing, etc.) inherently inhibits the treatment process and
makes it difficult to treat using conventional biological
processes [2,3]. Traditional continuous flow systems such as
activated sludge process (ASP) have serious difficulties to
meet the stipulated standards. Biological treatment of
complex chemicals is particularly challenging owing to
the inhibition and/or toxicity of these compounds when they
$
IICT communication number: 020902.
* Corresponding author.
E-mail address: kousik@iict.ap.nic.in (P.N. Sharma).
discontinuous processes (sequencing batch reactor (SBR)
technology), which promote the mineralization of the
industrial wastewater containing toxic compounds seem
to be promising.
Sequencing batch reactor technology has been developed
on the basic scientific assumption that periodic exposure of
the microorganisms to defined process conditions is
effectively achieved in a fed batch system in which exposure
time, frequency of exposure and amplitude of the respective
concentration can be set independently of any inflow
condition [4]. SBR technology differs in various ways from
conventional technologies used in biological treatment of
wastewater. The most obvious difference is that the reactor
volume varies with time, whereas it remains constant in the
traditional continuous flow system. From the process
engineering point of view, the SBR system is distinguished
by the enforcement of controlled short term unsteady state

0032-9592/$ – see front matter # 2004 Published by Elsevier Ltd.

doi:10.1016/j.procbio.2003.02.001
1502 S.V. Mohan et al. / Process Biochemistry 40 (2005) 1501–1508
conditions leading in the long run to a stable steady state
with respect to composition and metabolic properties of the
microbial population growing in the reactor by controlling
the distribution and physiological state of the microorgan-
isms. The success of SBR technology depends upon the
great potential provided by the possibilities of influencing
the microbial system in the SBR and also upon the fact that
SBRs are comparatively easy to operate and are cost
efficient. SBR processes are known to save more than 60%
of expenses required for conventional activated sludge
process in operating cost [5]. Interest has been growing
worldwide both in scientific research and in practical
application of SBR technology. SBR with various reactor
configurations for nutrient removal have been studied
extensively [4–10]. One of the main advantages of SBR
is that enhanced phosphorus removal was due to phosphate
accumulation during the anaerobic process and utilization in
the subsequent aerobic process of the sequence [9].
SBR has been successfully applied for the treatment of
domestic wastewater, medium and lower strength land fill
leachates, simulated dye wastewaters and contaminated soils
[11–21]. However, a through literature search showed that
SBR technology has not been investigated with complex
chemical wastewaters such as pharmaceutical, drug and
chemical manufacturing units so far. The wastewater
generated from such industries constitute various organic
substances used in the process, inorganic salts, organic
solvents, etc. which result in high COD, low BOD, high salt
content (TDS), toxic and inhibitory substances in waste-
water which inhibit the biological process [3].
The purpose of the present study was to investigate the
application of sequencing batch reactor technology for
treating complex chemical effluents by adopting a
suspended growth configuration. SBR is operated with
varying organic loading rates and process performance is
assessed by monitoring COD during the cycle operation. To
understand the ongoing biochemical process, BOD, pH,
oxidation–reduction potential (ORP), volatile suspended
solids (VSS), suspended solids (SS), oxygen consumption
rate (OCR) and sulphates are also monitored during the
cycle operation. The results obtained are presented and
discussed in this communication.
2. Material and methods
2.1. Chemicals
All chemicals used during the experiments were of analy-
tical reagent (AR) grade and purchased from various sources.
Glass doubled distilled water was used wherever necessary.
2.2. Wastewater characteristics
The wastewater used in this study was collected from a
common effluent treatment plant in Hyderabad, where
Table 1
Characteristics of wastewater used as feed to SBR
Parameters Concentrations
pH 7.83
ORP (mV) 92
TDS (g/l) 11
Suspended solids (mg/l) 900
Oil and grease (mg/l) 14
Total residual chlorine (mg/l) 0.21
Total nitrogen (TKN) (mg/l) 125
Ammonical nitrogen (mg/l) 35
COD (mg/l) 6000
BOD (mg/l) 2600
Chlorides (mg/l) 5000
Phosphates (mg/l) 360
Sulphates (mg/l) 1750
Sulphides (mg/l) 35
Phenol (mg/l) 7.8
wastewater received from various chemical industries was
being treated. The wastewater was a composite one from
about 100 industries producing a variety of chemicals, drugs,
pharmaceuticals, pesticides and various chemical inter-
mediates. The detailed characteristics of the wastewater
used in this study as feed are given in Table 1. The complex
chemical characteristics of the effluent could be assessed by
the presence of a low BOD/COD ratio (<0.3), high sulphate
content (>1.75 g/l) and high TDS concentration (>11 g/l).
2.3. SBR configuration and operation
A sequencing batch reactor was fabricated in the
laboratory from perplex glass material having a total
working volume of 1.7 l capacity. The reactor had an
internal diameter of 0.07 m and 0.22 m length with L/D ratio
of
3. The outlet of the reactor used for wastewater
withdrawal was present at 0.045 m length from bottom of
the reactor. This outlet arrangement prevents loss of biomass
in the reactor after the settling phase is over. About 0.34 l of
mixed liquor will be present in the reactor after the
withdrawal phase is completed resulting in a total liquid
volume during the reactor phase of 1.34 l. The reactor was
fabricated with proper inlet and outlet arrangements. A
schematic representation of the SBR is given in Fig. 1.
The reactor was operated in suspended growth config-
uration in sequencing batch mode at a constant temperature
of 26  2 8C. The total cycle period of 24 h (HRT)
consisting of 15 min of filling phase, 23 h of reaction
(aerobic) phase with recycling, 30 m of settling phase and
15 min of withdrawal phase was employed throughout
(Table 2). The sequence of the SBR operation was controlled
by pre-programmed timers (feeding, aeration, recycling and
withdrawal). At the beginning of each cycle, immediately
after withdrawal (earlier sequence), a pre-defined feed
volume (1 l) was pumped into the system and the reactor
volume was recirculated with aeration during the reaction
phase. At the end of the cycle, suspended biomass (VSS)
 S.V. Mohan et al. / Process Biochemistry 40 (2005) 1501–1508
Fig. 1. Schematic representation of the sequencing batch reactor (SBR).
settled and effluent was withdrawn from the reactor. The
sequence of various operations was done with pre-
programmed timers (ETTS, Germany). Feeding, wastewater
withdrawal and recirculation operations were done with the
help of peristaltic pumps (Watson Marlow 101 U/R). Air
supply was by means of diffused aerators connected to a
sparger arrangement. The sequence of various operations
was done with the pre-programmed timers (ETTS,
Germany). During the reaction phase, aqueous phase
dissolved oxygen was maintained in the range of 3.0–
4.5 mg/l. The pH of the influent was adjusted to 7.1  0.2
before wastewater feeding.
Recirculation at a rate of 4 l/day was maintained
throughout the investigation to achieve a homogeneous
distribution of substrate as well as uniform distribution of
suspended biomass along the reactor depth. Recirculation
Table 2
Cycle period and phase details of reactor during sequence
Phase Cycle period Air supply
Filling (min) 15 Off
Reaction with recirculation (h) 23 On
Settling (min) 30 Off
Withdrawal (min) 15 Off
1503
also facilitates linear velocity, which restricts the
existence of a concentration gradient during the reaction
phase of the SBR operation. The reactor can be considered
as completely mixed during the reaction phase of the
sequence.
2.4. Start up of the SBR
The SBR was inoculated with biomass (aerobic) acquired
from an operating laboratory scale activated sludge process
unit, which had been operated continuously for 3 years for
the treatment of complex chemical effluents. The mixed
liquor (VSS: 3.5 g/l) from the aerobic chamber of the ASP
unit was acquired and was fed to the SBR reactor at a ratio of
1:5 with reactor volume as inoculum. After inoculation, the
reactor was operated with synthetic feed (glucose: 1 g/l;
sodium acetate: 1 g/l; and Na2HPO4: 0.3 g/l) to build up the
biomass. When the SBR was built up with 3 g/l of VSS, it
was fed with the designated effluent at an organic loading
rate of 0.8 kg COD/m3/day and subsequently after stable
Condition performance was achieved, the reactor was operated at
Anoxic higher organic loading rates. The reactor was operated
Aerobic continuously for a period of three months with varying
Anoxic organic loading rates (0.8 kg COD/m3/day; 1.7 kg COD/m3/
Anoxic
day; and 3.5 kg COD/m3/day).
1504 S.V. Mohan et al. / Process Biochemistry 40 (2005) 1501–1508
2.5. Analytical procedures
The performance of SBR with complex chemical
effluents was assessed by monitoring carbon removal
(COD) throughout the reactor operation and during the
cycle period. In addition, pH, oxidation–reduction potential,
BOD, sulphates, volatile suspended solids, suspended solids,
oxygen consumption rate (OCR) and dissolved oxygen (DO)
were also determined during sequence operation to assess
the performance of the SBR. The analytical procedures for
monitoring the above parameters were adopted from the
procedure outline in Standard Methods [22].
The suspended biofilm in the mixed liquor of the SBR
was subjected to scanning electron microscopy (SEM) to
observe morphological details after carefully dehydrating
(30 8C, hot air oven for 24 h) the biomass without disturbing
the actual morphology of the biofilm.
3. Results and discussion
3.1. SBR performance
SBR was operated in sequencing batch mode with a total
24 h cycle period with an organic loading rate of 0.8 kg
COD/m3/day to assess the suitability of the reactor for
treating the complex chemical wastewater under study.
Initially after the start up of the reactor (15 days), the reactor
was operated with an organic loading rate of 0.8 kg COD/
m3/day and the reactor performance was assessed by
monitoring carbon removal (COD and BOD) during the
sequence (cycle) operation and also throughout the reactor
operation. The variation of COD and BOD removal with the
function of the cycle period is depicted in Fig. 2. Sixty four
percent COD removal was observed at an organic loading
rate of 0.8 kg COD/m3/day. The COD removal rate was slow
(23%) during the initial phase of sequence operation (up to
Fig. 2. COD and BOD variation during SBR cycle operation.
Fig. 3. COD and BOD variation during SBR operation.
10 h). With an increase of sequence time a relatively rapid
removal was noticed after 10 h and approached 64% at the
end of the reaction phase (23.3 h). The initial low COD
removal may be due to the relatively high concentration
gradient of the substrate. With an increase in sequence time,
the native suspended biofilm might have become acclima-
tized to the new substrate (system) conditions facilitating
rapid removal of the organic substrate through mineraliza-
tion. The BOD profile during the sequence operation
(reaction phase) showed comparably the same pattern as the
COD profile. BOD removal of 69% was observed after the
reactor attained stability. It can be concluded from the
reactor performance data obtained that SBR showed
relatively better performance with respect to COD removal
when compared to the conventional ASP system.
The SBR was operated at various organic loading rates
(0.8 kg COD/m3/day; 1.7 kg COD/m3/day; and 3.5 kg COD/
m3/day) (Fig. 3). With continued operation, the reactor
showed enhanced performance with respect to COD and
BOD removal and attained stable conditions within 3 days
after feeding and remained more or less constant thereafter.
About 66% of COD removal and 92% of BOD removal was
observed during stabilized operation of the reactor. On day
21 after of startup, the reactor was fed with an organic
loading rate of 1.7 kg COD/m3/day. Immediately after
increase in the organic loading rate, the reactor showed an
increase in the outlet COD and BOD levels and approached
47% COD removal and 72% BOD removal within 4 days.
On day 28 after startup, the reactor was fed at an organic
loading rate of 3.5 kg COD/m3/day and reactor had a
performance with 35% BOD removal and 57% COD
removal. Consolidated data of SBR performance at various
organic loading rates are presented in Table 3. It is evident
from the data, that with increase in organic load, the COD
removal rate was reduced. COD removal of 47% was
 S.V. Mohan et al. / Process Biochemistry 40 (2005) 1501–1508 1505

Table 3
Performance of SBR at various organic loading rate
Organic loading rate Percent COD Percent BOD Time to achieve stable Percent sulphate F/M ratio
(kg COD/m3/day) removal removal performance (days) removal (as BOD) range
0.8 66.4 92.2 4 7.8 0.24–0.29
1.7 47.1 72.7 5 8.3 0.44–0.38
3.5 25.4 57.0 7 8.8 0.94–1.10

Table 4
Comparative performance of conventional ASP and SBR
Reactors Configuration Organic loading rate HRT Percent COD Percent BOD Sludge age VSS in mixed
(kg COD/m3/day) (day) removal removal (days) liquor (mg/l)
ASP Suspended growth (aerobic) 1.1 5 55.0 67 12 2000–2250
SBR Suspended growth (aerobic and anoxic) 0.8 1 66.4 92 10 2000–2250

achieved at 1.7 kg COD/m3/day. On increasing the organic network (internal). The biofilm was formed on well define
loading rate to 3.5 kg COD/m3/day, the COD and BOD mats of filamentous base.
removal rates were inhibited markedly. To achieve stable
performance (with respect to carbon removal), the reactor 3.2. Sulphate removal
required 4 days at 0.8 kg COD/m3/day and 5 days for 1.7 kg
COD/m3/day. About 7 days were required to achieve stable The reactor was fed with 1.75 g/l sulphates, which is
performance at an organic loading rate of 3.5 kg COD/m3/ considered to be highly inhibitory and not suitable for
day. Sulphate reduction of about 8% was recorded at all the anaerobic process. A conventional aerobic system cannot
studied organic loading rates. The relatively poor perfor-
mance of the SBR at higher organic load can be attributed to
the presence of high substrate gradients with relatively high
concentration of the toxic and inhibitory substances in the
wastewater.
From the results obtained, it can be concluded that SBR
(with suspended growth system) showed a relative effective
performance compared to conventional ASP system with
same wastewater in terms of carbon removal (Table 4). The
performance of SBR is superior compared to conventional
ASP operated with the same complex chemical wastewater
[23]. Conventional ASP was operated at 1.1 kg COD/m3/day
continuously for a period of 6 months with 5 days HRT and
showed shown 55% COD removal and 67% of BOD
removal. In comparison, the SBR operated at 0.8 kg COD/
m3/day showed a comparatively effective performance
(COD removal of 66% and BOD removal of 92%) with
only 24 h of HRT. Conventional ASP operated with this
effluent required 35 days of operation to achieve stable
performance. However, in the case of SBR, 5 days were
found to be sufficient to achieve stable performance.
Periodic processes that induce controlled unsteady state
conditions have organisms that have high substrate uptake
and storage capabilities are generally more robust and can
withstand shock loads [4,24].
A scanning electron micrograph (SEM) of the suspended
biofilm, shown in Fig. 4 indicates the suspended biofilm
acquired during the SBR operation at 1.7 kg COD/m3/day
loading rate, where inhibition to the process and disintegra-
tion of biofilm was not observed. From the micrograph, the
morphology of the suspended biofilm was heterogeneous
with irregular (uneven) surface texture having porous Fig. 4. Scanning electron micrograph (SEM) of suspended biofilm.
1506 S.V. Mohan et al. / Process Biochemistry 40 (2005) 1501–1508

Fig. 6. Variation of pH and ORP during SBR cycle operation.

ORP during the sequence were also monitored and presented

Fig. 5. Removal of sulphate during the operation of the SBR cycle at in Fig. 6. The influent pH of 7.9 was introduced at the
various organic loading rates.
starting of the reaction phase. The aqueous phase pH
gradually decreased with time and approached 7.5 at the end
reduce sulphates, as it requires an anoxic/anaerobic en- of the reaction phase. ORP (mV) profile visualized a mirror
vironment to bring about conversion. In this study, about 8% image to pH and with increase of sequence time the ORP
of sulphate reduction was achieved at all the organic loading approached zero (0 mV).
rates studied (Table 3 and Fig. 5). Sulphate transformation in Sludge volume index (SVI) is one of the important
the SBR reactor may be attributed to the prevailing anoxic parameters to assess the stability of sludge in any aerobic
zone in the internal layers of the suspended biofilm and the suspended growth system. SVI was monitored during the
induced anoxic conditions during the sequence operation. end of each cycle before withdrawal. Variation in SVI
The internal biofilm layer where anoxic conditions facilitate including sludge volume (SV) in the reactor is shown in
sulphate transformation as shown are: Fig. 7. The SV was relatively high (20 cm) initially at 0.8 kg
COD/m3/day. With increase in organic loading rate, SV
anoxic
SO2
4 þ organic substrate ! S
2 decreased. This may be attributed to the fact that increase in
condition
organic loading rate increased the toxic load in the influent,
The size of the biofilm floc had a significant influence on the thereby causing inhibition to the native biomass growth and
extent and presence of the anoxic zone. Normally, the biofilm its metabolic activities. The SVI was good (50–100) during
particle size in the ASP plant was in the range 10–110 mm. In
the present study, the suspended biofilm size encountered
during experiments were in the size range of 50–770 mm
(average size 317 mm). The biofilm floc of 200 mm size and
above will have an anoxic microniches in the internal part of
the thick flocs [25]. The suspended biofilm particle
(>200 mm) in the present study have induced anoxic phase,
which lead to sulphate transformation. The persistent induced
anoxic environment (15 min) during the feeding phase and
the subsequent settling and withdrawal phase (45 min) of the
cycle operation facilitated a suitable environment for native
biomass for sulphate conversion.

3.3. Process monitoring

To understand the on going biochemical process during

sequence operation, the process was monitored by deter-
mining pH, ORP, DO, SVI and VSS. The variation of pH and Fig. 7. Variation of SVI and SV during SBR operation.
 S.V. Mohan et al. / Process Biochemistry 40 (2005) 1501–1508
Fig. 8. Variation of VSS and SS during SBR operation.
SBR operation at 1.0 kg COD/m3/day and 1.7 kg COD/m3/
day, which is considerably less than 100, commonly used as
the limit between good and poor settling sludge. Sludge
normally settles well in SBR as per earlier reported work and
at high concentration of DO, SBR suspended growth
systems produce sludge with good settling and thickening
properties (floc high compactness) [26]. In this study,
aqueous phase DO was maintained in the range of 2.5–
4.0 mg/l and the results of sludge showed good settling
properties.
Volatile suspended solids and suspended solids were
monitored throughout the study to assess the viability of the
suspended biomass during SBR operation with complex
chemical effluents (Fig. 8). The SS was in the range 2250–
2000 mg/l during reactor operation at 0.8 kg COD/m3/day
and 1.7 kg COD/m3/day organic loading rates, respectively.
With increase in organic loading rate (3.5 kg COD/m3/day),
the SS content rapidly decreased, indicating the toxic nature
of the influent feed. Variation of VSS during SBR operation
was also comparable with the reactor SS. The variation of
F/M ratio during reactor operation is presented in Table 3.
The F/M ratio was in the range of 0.24–0.29 during steady
state conditions of reactor operation with 1.2 kg COD/m3/
day organic loading rate. At 1.7 kg COD/m3/day organic
loading rate, the F/M ratio varied around 0.4 (optimum for
good system) showing a relatively lower microorganism
to substrate ratio as compared to 1.0 kg COD/m3/day.
However, the F/M ratio at 3.0 kg COD/m3/day was quite
high (0.9–1.1), which indicates process inhibition due to
high substrate loading containing toxic substances.
Oxygen consumption/transfer capacity is often one of the
important factors that limit the capacity of suspended growth
biological systems. Oxygen consumption rate (OCR) is
monitored to assess the ability of suspended biomass to
degrade complex substrate in aerobic environments. Oxygen
1507
Fig. 9. OCR profile during SBR cycle operation.
consumption capacity indicates the ongoing biochemical
process. The oxygen consumption rate up to 4 h of cycle
operation was 0.122 mg O2/min and subsequently increased
to 0.129 mg O2/min and remained more or less constant
thereafter up to 23.3 h (Fig. 9). The oxygen consumption
rate was expected to increase with sludge retention time
due to higher endogenous respiration requirements. The
relative low performance of the SBR at high organic loading
rate (3.5 kg COD/m3/day) was further indicated by the
reduction in SS and VSS contents of the reactor as well as
reduction in the sludge volume.
It can be concluded from the data obtained that the SBR
was efficient when compared to a conventional ASP system,
while treating complex wastewater in that the startup (within
15 days) and the stabilization of reactor (within 2 days after
loading) were rapid in addition to relatively lower HRT.
Organic loading (shock loads) had relatively small effects on
performance and at higher loads stabilization of the reactor
was also rapid. The varying of individual components of
incoming wastewater with time in each process step place
microorganisms under nutritional changes from feast to
famine. This maintains a wide distribution in the population
of microorganisms. Periodic operations with altering feast
and famine conditions results in high uptake of substrate and
also better settalibility of the biomass [27]. Grau et al. [28] in
their accumulation regeneration theory stated that micro-
organisms are able to store substrate during an imposed
transient phase. Subsequent reuse for growth during
substrate-limited conditions provides competitive advan-
tage. Recent research indicated that sequential feast and
famine conditions will enhance the overall performance of
the biological system [29]. Enforced short term unsteady
state conditions coupled with periodic exposure of the
microorganisms to defined process conditions by controlling
their physiological state (incorporating required metabolic
1508 S.V. Mohan et al. / Process Biochemistry 40 (2005) 1501–1508
conditions) in SBR showed comparatively efficient perfor-
mance over conventional suspended growth systems treating
complex chemical wastewater.
4. Conclusions
The SBR showed relatively efficient performance
compared to a conventional ASP system in treating complex
chemical effluent. It is concluded that SBR needs a relatively
short period for start up and stabilization of reactor was
achieved within 2–5 days compared to an ASP system. The
performance of SBR is dependent on organic loading rate
and the system can withstand its performance up to 1.7 kg
COD/m3/day loading rate. With increase in organic loading
rate (3.5 kg COD/m3/day) the performance becomes
inhibited. Enforced short term unsteady state conditions
coupled with periodic exposure of the microorganisms to
defined process conditions which can control the physio-
logical state in SBR have resulted in comparatively efficient
performance over the conventional suspended growth
systems (ASP) for the treatment of complex chemical
effluents.
Acknowledgement
One of the authors (Dr. S. Venkata Mohan) gratefully
acknowledges Alexander von Humboldt (AvH) Foundation
for giving the fellowship and Prof. Dr.-Ing P.A.Wilderer for
exposing him to the field of sequencing batch reactor (SBR)
technology. Mr. N. Chandrashekara Rao is thankful to CSIR
for providing him Senior Research Fellowship.
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