Journal of Water Process Engineering 19 (2017) 60–66

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Journal of Water Process Engineering

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Short communication

Treatment of Adsorbable Organic Halide (AOX) from pulp and paper MARK
industry wastewater using aerobic granules in pilot scale SBR
⁎ ⁎
I.H. Farooqi , Farrukh Basheer
Department of Civil Engineering, Z.H. College of Engineering and Technology, A.M.U., Aligarh 202002, India

A R T I C L E I N F O A B S T R A C T

Keywords: Aerobic granulation of activated sludge was achieved in a pilot scale column type sequencing batch reactor
AOX (SBR) employed for the treatment of pulp and paper industry wastewater containing Adsorbable Organic Halide
SBR (AOX) in the range of 15–20 mg/L. Throughout the 780 days of operation the biomass trends to remain in
Aerobic granules between 7 and 8 g/L MLVSS and SVI reduced to 60–80 mL/g MLSS. After applying different selection pressure
Pulp and paper industry wastewater
the average size of aerobic granule in the SBR was in the range of 2–4 mm. After achieving steady state con-
ditions in the pilot plant, at an organic loading rateof 4.5 kg COD m−3day−1 at 6 h HRT the degradation of AOX
was significant and the effluent AOX concentration of below 5 mg/L was achieved. The results achieved in the
study exhibit the viability of formation of aerobic granules in pilot scale SBR and maintenance of long-term
strength of aerobic granular sludge with a high reduction of AOX in pulp and paper industry wastewater.

1. Introduction
The pulp and paper industry is a water demanding production and
ranks only third in the world, after the primary metals and the chemical
industries, in terms of freshwater consumption [1]. Around 500 diverse
chlorinated organic compounds have been identified including chloro-
form, chlorate, resin acids, chlorinated hydrocarbons, phenols, ca-
techols, guaiacols, furans, dioxins, syringols, vanillins, etc [2,3]. The
discharge of the Halogenated Organic Compounds in the environment is
increasingly becoming a matter of concern due to their carcinogenic/
toxic effects. Most of these compounds are adsorbable and in waste-
water, these compounds are estimated in a group as “Adsorbable Or-
ganic Halides” (AOX).
The formation of these compounds is due to the use of chlorine and
chlorinated compounds in the various industrial processes including the
bleaching process in the pulp and paper industries. The pulp and paper
industries also being one of the major consumers of chlorine and its
compounds need main concern in the perspective of removal of AOX
discharges in their effluents. The toxic effects of AOX (e.g. chlor-
ophenols, guaiacols, furans and dioxins) range from carcinogenic and
mutagenic effects to acute and chronic toxicity and can act as inhibitors
of microorganism growth, [3–5]. Conventional wastewater treatment
systems such as activated sludge process have been used for treatment
of pulp and paper mill wastewater. The process has disadvantage of
excess sludge production and fluctuations in removal efficiency for
recalcitrant compounds [6]. Activated sludge process for treatment of
pulp and paper mill wastewater have bulking problem, which in result
causes poor settling of sludge and effluent have more suspended solids
[7,8]. The use of chemicals such as chlorine, hydrogen peroxide, metal
ions (Fe, Ca, Mg) are effective in getting better settling properties of
sludge in paper mill wastewater treatment [9,10]. Moreover these
chemical addition methods are not cost effective and they do not pro-
vide long term solutions for bulking problem. Conventional adsorption
technique, post-treatment using a granular activated carbon (GAC) has
been widely used for removal of AOX from pulp and paper mill was-
tewater. However GAC have to be regularly regenerated, because ad-
sorption site are exhausted by pollutants after some time [11]. Osman
et al. [5]. have used GAC in sequencing batch biofilm reactor (GAC-
SBBR) to treat recycle paper mill wastewater and results showed high
AOX removal efficiency by maintaining longest hydraulic retention
time (HRT). The advantage of the biofilms in these systems save energy
and less area is required, efficient to handle toxicity and can withheld
high organic loading rate [12].
These days a new technology of aerobic granulation in column type
Sequential Batch Reactors (SBR) have been widely explored based on
modified activated sludge process. Due to the excellent settling capacity
of aerobic granules, large area for a settling tank will not be required,
which in turn reduces the wastewater treatment plant area by 80%
[13]. Aerobic granules have unique structure and microorganisms that
can withstand high loading rate as well as high toxicant concentration.
Aerobic granulation can be considered as a special type of self-ag-
gregation of microorganism, without any carrier medium for growth of

⁎
Corresponding authors.
E-mail addresses: farooqi_izhar@yahoo.com (I.H. Farooqi), farrukhbasheer@yahoo.com, farrukhbasheer.cv@amu.ac.in (F. Basheer).

http://dx.doi.org/10.1016/j.jwpe.2017.07.005
Received 28 March 2017; Received in revised form 21 June 2017; Accepted 9 July 2017
2214-7144/ © 2017 Elsevier Ltd. All rights reserved.
I.H. Farooqi, F. Basheer
biofilms. The immobilization of microbial cells into granule formation
generally improved the ability of cells inside the granules to tolerate
high toxicity concentration and achieve good structural properties and
biodegradation performance. Sequential batch reactors have ad-
vantages of low operating costs, no secondary clarifier required, no
recycling pumps required, and good control over filamentous microbial
growth, tolerance to shock loads and denitrification during anoxic fill
and settle phase [14]. Most of the lab scale and pilot scale studies were
carried out for municipal wastewater treatment using aerobic granu-
lation [15–18] and for synthetic wastes [13,19]. Very few studies have
been carried out for development of aerobic granules on actual in-
dustrial wastewater like petrochemical wastewater [20], palm oil mill
effluent (POME) [21], Brewery wastewater [22]. However, till date no
work has been reported for the treatment of AOX compounds from pulp
and paper industry using aerobic granulation technology. The present
work carried out to evaluate the feasibility of a pilot scale SBR for the
development of aerobic granules and to assess its performance for the
biodegradation of AOX presents in the pulp and paper industry waste-
water
2. Materials and methods
2.1. Reactor setup and operation
The Naini Tissue industry, Uttarakhand, India has been chosen for
the study as it utilises agro based material and chlorine for bleaching
purpose. The formation of AOX compounds occur when agro based
materials are used as raw materials and chlorine is used for bleaching.
The diameter of the pilot scale SBR was 600 mm and the effective
height was 3000 mm resulting in total volume of 3.394 m3 and height
to diameter (H/D) ratio of 5:1 (Fig. 1). The Pilot Plant was seeded with
the aerobic sludge of the aeration tank of the existing effluent treatment
plant of the industry. Initially a 24 h cycle was maintained and was
strictly followed. Later on the cycle of operation was reduced to 12 h,
8 h and 6 h. Programmable Logical Control (PLC) arrangements for the
automation of the Pilot Plant SBR were used. Effluent was withdrawn
from a port located at 40% height from the bottom resulting in volume
exchange ratio of 60% (2.04 m3).At maximum organic loading rate at
HRT of 6 h the working capacity of pilot SBR was 8.16 m3/day of
wastewater.
2.2. Acclimatization and development process of aerobic granules
The pilot plant was seeded with the aerobic sludge of the aeration
tank of the effluent treatment plant of the industry. The reactor was fed
with the wastewater of the equalization tank. Diffused aeration was
provided with the help of a blower and special types of diffusers were
fitted at the bottom of the reactor which prevented the choking with
sludge. Initially a 24 h cycle of operation was maintained which in-
cluded fill react settle and decant timings of respectively. The samples
were collected daily and were analysed for pH, alkalinity, Chemical
Oxygen Demand (COD), Biochemical Oxygen Demand (BOD),
Adsorbable Organic Halides (AOX). Provision of online measurement of
dissolved oxygen was made using online HACH D.O. monitoring
system. The sludge of the reactor was also analysed for its volatile
content on a routine basis.
In the initial phase the reactor was fed with diluted pulp and paper
industry wastewater with sewage (1 in 10) for proper acclimatization
and growth of the microorganism. Thereafter the dilution was reduced
to 1in 5, 1in 4 and 1in 3.sucessfully operating the pilot plant at reduced
dilution for 220 days of operation wastewater having characteristics
shown in Table 1 was fed to the reactor. The reactor was run on a 24 h
cycle till a substantial removal of AOX, BOD and COD removal took
place. The cycle of operation was reduced to 12, 8 and 6 h (on day 150,
day 300 and day 475) once pseudo steady state conditions were
achieved in each stage as shown in Table 2. The duration of each phase
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Journal of Water Process Engineering 19 (2017) 60–66
(fill, react, settle and decant) in individual cyclic operation is shown in
Table 3. Initially settling time was kept at 30 min thereafter selection
load was applied on day 150 as the settling time was reduced from
30 min to 5 min. This was done to retain the heavier aerobic sludge in
the reactor which helps in formation of dense and compact aerobic
granules. The reactor run was completed at 6 h of cycle after achieving
a significant reduction in the values of AOX and COD and was operated
for a total period of 780 days.
2.3. Analytical methods
Measurement of pH, biomass concentration, COD, BOD, Sludge
Volume Index SVI were conducted in accordance with Standard
Methods [23]. COD was measured using Closed Reflux method/Hach
Reagents (DR5000 UV/Vis spectrophotometer, HACH, USA),pH was
measured using HQ30d portable meter coupled with pH probe, HACH,
USA. Continuous DO was measured using online HACH probe attached
with PLC. Dissolved oxygen concentration (DO) was measured using
HACH, USA HQ30d portable meter coupled with LBOD10101 probe.
Samples were taken from the reactor at a predetermined time intervals
for analysis.
Aerobic Granule was analysed using Cenisco binocular petrological
microscope with SANYO digital camera. The photographs were ana-
lysed using image analysis (IA) system (Averz Software).The mor-
phology of aerobic granules in stipulation to aspect ratio was analysed
by using IA technique. Aspect ratio is the ratio between minor axis and
major axis of ellipse corresponding to the granule (0 = line,
1 = circle). Granule strength is defined as an integrity coefficient (IC)
(%) which is the residual volatile suspended solids (VSS) after sample
has been agitated for 5 min at 200 rpm on a platform shaker to total
VSS of the intact granules prior to agitation [24]. The stronger granules
have higher ICs.
2.4. Adsorbable Organic Halide
The AOX measurements were done by Multi X 2500 halide analyzer
(Jena, Germany). Nie et al. [25] described the basic method and process
as follows: Effluent first passed through the activated carbon column,
the organic chloride of the bleaching effluent then adsorbed by the
activated carbon column. Sodium nitrate was used in washing the ad-
sorbed inorganic chloride of the activated carbon column, and then the
activated carbon column burned in the combustion furnace. Content of
AOX calculated though the micro coulomb titration method [25,26].
2.5. Gas chromatography–mass spectrometry
GC (Gas chromatography) and GC/MS (Gas chromatography–mass
spectrometry) analysis of influent and effluent samples were done in
order to identify the number of products formed during aerobic bio-
degradation of pulp and paper mill wastewater. GC/MS equipped with
a split/splitless injector and a Perkin Elmer Clarus 680, USA gas chro-
matograph interfaced with a Turbomass spectrometric (Perkin Elmer
600T, USA) mass spectrometric mass selective detector system were
used. The samples were prepared and analysed under conditions as
described previously [13]. The metabolic intermediates were derived
from degradation of AOX identified by comparing their retention time
(minutes) and mass spectra with that of the National Institute of
Standard and Technology (NIST) library available.
2.6. Extracellular polymeric substances
The extracellular polymeric substances (EPS) of the flocculent
sludge and aerobic granules were extracted using a heat extraction
method as described earlier [27]. The carbohydrate content of the ex-
tract was analysed by the phenol-sulphuric method and expressed as
the glucose equivalent, and its protein substance content were
I.H. Farooqi, F. Basheer Journal of Water Process Engineering 19 (2017) 60–66

Fig. 1. (a) Schematic diagram of the pilot-scale SBR. (b) Pilot Scale SBR reactor for cultivating aerobic granules with pulp and paper mill wastewater in Naini Tissue industry,
Uttarakhand, India.

Table 1 Table 3
Charecteristics of Pulp and Paper industry wastewater. Duration of each phase during cyclic operation in SBR.

Parameter Units Range Cycl (hours) Fill (hours) React(Aeration) Settle Decant
(hours) (hours) (hours)
Chemical Oxygen Demand (COD) mg/L 2000–3000
Biochemical Oxygen Demand (BOD) mg/L 600–1350 24 0.25 23 0.5 0.25
Adsorbable Organic Halide (AOX) mg/L 10–22 12 0.25 11.5 0.083 0.167
Total Suspended Solids (TSS) mg/L 2500–4000 8 0.25 7.5 0.083 0.167
pH – 7.5–8.5 6 0.25 5.5 0.083 0.167

Table 2 measured according to the modified Lowry method [27].

Operational parameters and results of the sequencing batch reactor (average value).

Parameters Units Operational parameters 3. Results and discussion

Phase 1st 2nd 3rd 4th 3.1. Pilot plant SBR performance
Period. Days 0–150 150–300 300–475 475–800
Cycle Time/HRT Hours 24 12 8 6 Mounteer et al. [28] studied the removal of recalcitrant COD of
COD loading rate kg COD 0.3 2.25 3.45 4.5 bleached Kraft pulp effluents in SBR. Their results suggested that the
m−3day−1 microorganisms in activated sludge were capable of degrading re-
AOX Removal% 60 70 75 79
COD Removal Removal% 80 75 85 88
fractory COD, but were not metabolically active during activated sludge
SVI ml g−1 MLSS−1 120–130 110 75–85 60–70 process treatment of pulp and paper mill effluent. They also suggested
MLVSS g L−1 3–5 5–6.5 6.5–7.5 7–8 the use of biological selector in the treatment process that can enhance
Size of Granules mm – 0.5–1 1–2 2–4 the efficacy of microorganisms to remove refractory compounds. The
Integrity % – – 90 95
Sequencing Batch Reactor operation consisted of cycles of feed, aera-
Coefficient
tion, settling, and decant. The aeration phase is of two stages: biode-
gradation stage in which substrate was degraded to lowest level, fol-
lowed by the second stage of food scarcity in which no food was
available. The feast and scarcity periods in one cycle are the typical

62
I.H. Farooqi, F. Basheer
Fig. 2. (a) Variation of influent and effluent chemical oxygen demand (COD) and (b)
biochemical oxygen demand (BOD) with time.
characteristics of SBR operation. This alternate feast and famine si-
tuations play important role in microbial growth process which lead to
formation of strong and dense granules. The aerobic granules have been
reported to be very efficient for the removal of organics and toxic
compounds. In the present study a column type SBR based pilot plant of
total volume 3.394 m3 run on a 24 h cycle till substantial removal of
AOX, BOD and COD took place. The cycle of operation was reduced to
12, 8 and 6 h (on day 150, day 300 and day 475) once pseudo steady
state conditions were achieved in each stage. The reactor run was
completed at 6 h of cycle after achieving a significant reduction in the
value of AOX and COD and total run of pilot plant was 780 days.
Fig. 2 shows the variation of influent and effluent chemical oxygen
demand and biochemical oxygen demand with time. Initially diluted
pulp and paper industry wastewater with sewage (1 in 10) was fed to
the reactor for proper acclimatization and growth of the microorgan-
isms. This ratio was maintained for 30 days. The COD was gradually
increased by reducing the dilution ratio. Thereafter, the dilution ratio
was reduced to 1 in 5 and the feeding was continued till 100 day of
operation. The dilution of wastewater was further reduced to 1 in 4 and
was maintained till day 140 of reactor operation. After achieving sig-
nificant COD reduction the dilution ratio was further decreased and
diluted wastewater (1 in 3) was fed to the Pilot plant for another
80 days i.e. 220th day of reactor operation. The effluent COD was
monitored continuously. It was observed that the effluent COD was
nearly constant. After operating the Pilot Plant for two hundred and
twenty days at HRT of 24 h raw wastewater was fed to the Pilot Plant.
There was a sudden increase in the concentration of influent COD.
However, there was not much effect on the effluent COD. This might be
attributed to the fact that the microorganisms have been acclimatised
for a longer time and hence were able to biodegrade higher loads of
organic matter. The reactor was then continued to operate on the raw
influent till 300th day of operation. After this the organic loading was
increased to 3.45 kg COD m3d-1 by reducing the cycle to 8 h and the 8 h
cycle was maintained till reactor attained steady state conditions. The
values of BOD and COD in the treated effluent were around of
250 mgL−1 and 30 mgL−1 respectively. After 475 days of reactor
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Journal of Water Process Engineering 19 (2017) 60–66
Fig. 3. Biomass concentration and sludge volume index (SVI).
operation the cycle of operation was reduced to 6 h resulted in organic
loading rate of 4.5 kg COD m3d−1 and the cycle was maintained till the
end of reactor operation. The influent and effluent COD, BOD profiles
show (Fig. 2) the proper acclimatization of Pilot Plant SBR for de-
gradation of organic compounds present in pulp and paper wastewater.
By operating the reactor in such a way it was possible for industry to
attain 250 mgL−1 of effluent COD discharge standard as per Central
Pollution Control Board (CPCB), India [29].
The sludge volume index was also found out and showed excellent
settling properties of the aerobic granules and sludge (Fig. 3).It is evi-
dent from Fig. 4 that there was a substantial built up of the biomass in
the reactor as the MLVSS concentration at the bottom reached around
8 g/L. Fig. 4 shows the sludge profile at different heights of the reactor
on day 400. The results shows that still lower settling time may be
provided to reduce the timings of the cycle of operation.
3.2. Adsorbable Organic Halide (AOX) removal
The main aim of the research work was to evaluate the performance
of column type sequencing batch reactor for the biodegradation of AOX
compounds. The monitoring of the influent and effluent was carried out
through AOX analyser (Analytic Jena, Germany). Fig. 5 shows the
variation of influent and effluent AOX for the entire tenure of operation
of the Pilot Plant. It is seen that the influent value of AOX varied a lot
from 8 to 22 mgL−1. However, once the reactor had achieved steady
state conditions at organic loading rate 4.5 kg COD m−3day−1 the
values of AOX remained almost constant and were below 5 mgL−1. At
the start the AOX removal efficiency was less however, over a period of
time the microorganisms have developed necessary enzymes in aerobic
granules for the biodegradation of AOX. The results of AOX degradation
have been confirmed through GC MS analysis. Fig. 6 shows GC–MS
anlaysis of influent and effluent. Chromatographic analysis was done in
order to determine the number of intermediates formed during biode-
gradation. About 90% of the compounds (Table 4.) that were present in
influent were completely degraded and were not found in effluent
Fig. 6. A similar study on the accumulation of AOX in aerobic granules
has also been conducted by Wang et al. [30]. They stated that small
sized aerobic granules grew rapidly, while flocculent sludge tends to be
Fig. 4. Sludge profile at different heights of the reactor on day 400.
I.H. Farooqi, F. Basheer Journal of Water Process Engineering 19 (2017) 60–66

Table 4
Different compounds identified in the pulp and paper mill wastewater in GCe MS
Analysis.

S.No Identification from Mass spectra

1. Phenol 2 methoxy
2. 4h-pyran-4-one, 2,3-dihydro-3,5 dihydroxy-6 methyl
3. 2 methoxy phenol
4. Benzene, 1,4-dimethoxy
5. Phenol 2 methoxy-4 methyl
6. 3 methyl phenol
7. 4 vinylguaiacol
8. 3 methoxy 5 methyl phenol
9. Phenol 3,4-dimethoxy
Fig. 5. The variation of influent and effluent AOX with time. 10. Phenol 2,6 dimethoxy
11. 4 chloro, 2,5 xylenol
12. Benzene, methyl (1-methyl ethyl)
washed out in the initial period of acclimatization indicating the ac- 13. Pentanoic acid
cumulation of granules, and 94 and 95% of the total 2, 4-Di- 14. Phenol 2,6-dimethoxy-4-(2propenyl)
chlorophenol and COD removal efficiencies were achieved respectively. 15. Pentalchlorophenol
16. 2,2-bis(4Hydroxyphenyl) propane
17. Cedrol
3.3. Development of aerobic granules 18. 2,6 Diisopropylphenol

The initial biomass concentration and SVI in the reactor were

granules and thus were shielded from the direct contact to toxicants
3.5 gL−1 and 130 mLg−1 MLSS respectively (Fig. 3). After the selection
[13]. The biomass concentration generally showed an upward trend
load on day 150 as the settling time was reduced from 30 min to 5 min,
and it remained in between 7 and 8 gL−1 MLVSS and SVI reduced to
all the biomass having lesser settling velocities were washed out from
60–80 mLg−1MLSS. The process of development of granules was ob-
the reactor. Owing to this selection pressure the formation of aerobic
served in the pilot plant SBR. The granules were taken out and their size
granules was improved as the thick and heavy biomass was retained in
was determined. The process of granulation continued and the size of
the reactor. In next 30 days formation of aerobic granules took place
the granules increased. After 475 days of operation the average size of
having size around 5 mm. However, after operation for a period of
the granule were maximum and was in the range 2–4 mm (Fig. 7).
around 200 days the microorganisms were found to agglomerate to
Proteins (PN) and carbohydrates (PS) were found to be
form thick and larger size granules. There is a food concentration gra-
7.34 mg g−1VSS and 2.98 mg g−1VSS respectively in aerobic granules
dient inside the granules, which made the aerobic granules less sus-
on day 350. In this work the protein/carbohydrate ratio was approxi-
ceptible to the toxicity of refractory compounds present in wastewater.
mately 2.46 for pulp and paper mill wastewater fed aerobic granules.
Fewer numbers of microorganisms are present on the surface of aerobic
Previous findings have shown that there is high proportion of proteins
granules and were directly exposed to the toxicants. Majority of the
over the carbohydrates (polysaccharides) for aerobic granules in SBRs
microorganisms were present in the interior arrangement of the aerobic

Fig. 6. Chromatographic analysis of pulp and paper industry wastewater. (a) influent and (b) effluent.

64
I.H. Farooqi, F. Basheer
[31,13]. McSwain et al. [31] reported the protein/carbohydrate ratio
for aerobic granules as high as 7.5. The presence of protein in extra
cellular polymeric substance (EPS) suggested its important role in
aerobic granule formation. Because protein has a high content of ne-
gatively charged amino acids, protein is more involved than sugars in
electrostatic bonds with multivalent cations, an important factor in
stabilizing aggregate structure [31]. EPS could improve the polymeric
interface, encourage the hydrophobicity, link within cells and produces
clusters to trap microbes to form more dense granular structure
[13,31,32].
4. Conclusion
Aerobic granulation of activated sludge was achieved in pilot scale
column type sequencing batch reactor (SBR) for the treatment of pulp
and paper industry wastewater having Adsorbable Organic Halide
(AOX) in the range of 15–20 mgL−1. After applying different selection
pressure the average size of aerobic granule in the SBR was in the range
of 2–4 mm having very good settling properties. The granules were
found to have high proteins in extra cellular polymeric substances
which helped in the agglomeration of bacteria to form dense aerobic
granules. The SBR based pilot plant was successfully operated at an
organic loading rate of 4.5 kg COD m3d−1. The effluent COD and AOX
concentration were below 250 mg/L and 5 mg/L respectively. The re-
actor was operated at minimum HRT of 6 h. The results achieved in the
study exhibit the feasibility of formation of aerobic granules in pilot
scale SBR and maintenance of long-term strength of granular sludge
with a high reduction of AOX in pulp and paper industry wastewater.
The outcome of the study would benefit pulp and Paper industry sector
a lot as column based SBR requiring less space, power and may be re-
placed with conventional activated sludge process for achieving the
desired effluent standards.
Acknowledgement
Authors are thankful to Ministry of Environment, Forest and Climate
Change, Government of India for providing research grant to conduct
this research.
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