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The 9th AUN/SEED-Net Regional Conference on Natural Disaster (RCND 2021) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1091 (2022) 012059 doi:10.1088/1755-1315/1091/1/012059

To Optimize Sequencing Batch Reactor (SBR) and Monitor

Aerobic Granular Sludge (AGS) Development for Domestic
Wastewater Treatment

Norazah Mohd Kassim1*, Aznah Nor Anuar 1*

, Hakim Halim1*, Noor Atiqah Sharani2,
Alijah Md Aris2, Khor Bee Chin2
1
Department of Chemical and Environmental Engineering, Malaysia-Japan International
Institute of Technology, Universiti Teknologi Malaysia, Kuala Lumpur, Malaysia
2
Indah Water Research Centre, Kuala Lumpur, Malaysia

*Corresponding Author: [email protected], [email protected], [email protected]

Abstract. The aim of this study is to determine design and process parameters of the aerobic
granular sludge system to treat actual domestic wastewater. The aerobic granular sludge was
developed in a 2.5 L lab-scale bioreactor which uses the operational system based on the
sequencing batch reactor (SBR). The bioreactor was fed with influent from local sewerage
treatment plant and seeding with fresh inoculums (seed sludge) from activated sludge tank. The
operation of the reactor was based on the SBR system with a complete cycle time of 4 hours and
specifically designed to be operated for 24 hours continuously. Two sets of experiments were
carried out in a single-walled cylindrical glass column bioreactor. These two sets of experiments
are referred to as SBRA and SBRB with different time setting. SBR systems have four stages
which is feeding, aeration, settling and decant. It was carried out in sequence: (1) ‘feeding with
aeration’ for SBRA (feed and aerate pumps operated simultaneously); ‘feeding without aeration’
for SBRB (feed and aerate pumps operated separately), (2) ‘aeration only’, (3) ‘settling’, and (4)
‘discharge’. Removal performance of the SBR such as BOD, COD, AMN, NO3-, TSS, VSS and
TP along with the other several parameters of the SBR such as DO, SOUR, ORP and settling
profile are monitored daily. Starting from day 1 until day 26 the average DO values were around
1 - 2 mg/l but after 1.5 hours the DO values increased drastically. It shows that the microbe did
not consumed air during the process. Day 13 onwards SOUR profiles show that the microbe
consumed oxygen slowly. These results suggest that low amount of readily oxidized organic
available in the bioreactor. The SVI reading shows good settling performance which is less than
100 mL/g. The settling profile shows the sludge has been further compact day by day. From the
finding, it is suggested that it would be better for feed and aerate pumps to be operated separately.

1. Introduction
The dramatic increase of human population and industrialization has resulted in excess scale of
wastewater to be handled before it becomes reusable and enters into ecosystems, which means new
development of sewage technique are needed [1]. Aerobic granulation is described as a self-aggregation
process that transform loose biomass into dense and compact granules under control conditions [2].
AGS is well known for its physical characteristics including regular in shape and compact structure [3].
As a dense microbial aggregate, AGS has great settling ability which can enhance the effectiveness of
sludge separation from treated effluent. It also has high biomass levels that would ensure faster and

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The 9th AUN/SEED-Net Regional Conference on Natural Disaster (RCND 2021) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1091 (2022) 012059 doi:10.1088/1755-1315/1091/1/012059

efficient removal of pollutants [4]. The high surface area and porosity of AGS also provide excellent
resistance towards toxic substances. Besides, other notable features of AGS are the ability to tolerate
high organic loads and carry out the simultaneous removal of organic substances and nutrients [5]. AGS
technology demonstrates a 20-25% reduction in operation costs, 23-40% less electricity requirement
and a 50-75% reduction in space requirements compared to the conventional activated sludge treatment
system [6].
In an existing activated sludge system, a mixed culture of suspended biomass is growing and
removing organic carbon and nutrients from the influent. In such a process the biomass (the activated
sludge), which are usually present as flocs, are mixed with the wastewater in a large aerated basin.
Supply of fresh wastewater to the bioreactor and discharged of treated wastewater from the bioreactor
occur continuously. The discharged wastewater is led to a settling tank. In the settler, separation of
activated sludge from the treated wastewater is carried out by means of gravitational forces. The treated
wastewater can then be discharged into surface waters, or is used for further treatment. Existing
activated sludge plants produce surplus sludge. Part of the settled activated sludge is recycled to the
bioreactor. The remainder of the sludge is usually treated anaerobically and later disposed in landfills,
or is used as fertilizer in agriculture.
Performance of activated sludge system deteriorates due to sludge separation problem caused by
sludge bulking. Bulking activated sludge which settles slowly and compacts poorly is still one of the
most common operational problems plaguing the wastewater treatment plant. Moreover However,
existing activated sludge plants take up a substantial footprint. In order to treat large amounts of
wastewater completely, large aeration tanks are needed. The settling tanks cover a large area, because
the settling velocity of the activated sludge flocs is very low, normally <1 mh–1.
AGS cultivation has been preferentially carried out in sequencing batch reactors (SBRs). In this
system, all phases (filling, reaction, settling, decanting, idling) take place in the same tank. Therefore,
a secondary clarifier is not needed as in activated sludge system. AGS has a small footprint and a low
energy demand compared to conventional municipal wastewater treatment technologies [7]. Since the
available ground area to build the treatment plant is usually limited, especially in Malaysia, there is a
need for a more compact and high-performance reactor. This need for more compact reactors and short
hydraulic retention time (HRT) directed the study towards the development of systems with high
biomass concentrations. Furthermore, it demonstrated unique features including granules without
carrier material, excellent settling properties, great biomass enrichment, simple single-tank concept with
simultaneously biochemical oxygen demand (BOD), chemical oxygen demand (COD), nitrate (NO3-),
ammoniacal nitrogen (AMN) and total phosphate (TP) removal and low costs compared with
conventional activated sludge processes.
The objectives of this study are (i) to optimize sequencing batch reactor (SBR) and (ii) to monitor
aerobic granular sludge (AGS) development for domestic wastewater treatment whereby this study will
utilize fully hundred percent actual domestic wastewater to comply with the existing wastewater
treatment operation. This could be a benchmark for local sewerage services to start-up their full-scale
aerobic granulated sludge system.

2. Materials and Methods

2.1. Designed and Setting Up Bioreactor
Cyclic Aerobic Granular Sludge Bioreactor (CAgABio); a 2500 mL laboratory-scale reactor have been
used to develop the granules as shown in Figure 1. The basis of this reactor set-up was developed
according to the reactor configuration built in the Kluyver Laboratory for Biotechnology, Delft
University of Technology, the Netherlands [8]-[10]. The schematic diagram of the reactor set-up is
shown in Figure 2. The operation of the reactor was based on the normal SBR system with a complete
cycle of 4 hours and specially designed to be operated for 24 hours continuously. Two sets of
experiments were carried out in a single-walled cylindrical glass column bioreactor with a working
volume of 2.5 L. The two sets of experiments are referred to as SBRA and SBRB. The process flow
diagram of the CAgABio system is shown in Figure 3.

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The 9th AUN/SEED-Net Regional Conference on Natural Disaster (RCND 2021) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1091 (2022) 012059 doi:10.1088/1755-1315/1091/1/012059

Figure 1. 2500 mL laboratory-scale SBR.

2.2. Seeding Sludge

A fresh activated sludge taken from a Sequencing Batch Reactor Treatment Plant (Bunus STP – KLR
403, Kuala Lumpur) was used as seed sludge in the reactor. The seed sludge characteristics used in this
experiment is given in Table 1. The characterisation study is essential to provide physical and
morphological information for the initialisation of the granulation process where at the end of the study,
the final characteristics of granulated seed sludge could be figured out. The physical characteristics
were determined in terms of sludge volume index (SVI), size and settling velocity (v).

2.3. Influent Feeding

The influent for the experiments with actual domestic wastewater were collected from wastewater
treatment plant (BUNUS STP – KLR 403 Kuala Lumpur). For every cycle, 1000 mL of actual domestic
wastewater was dosed to the reactor. The composition of the influent dosed to the laboratory-scale is
given in Table 2.

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The 9th AUN/SEED-Net Regional Conference on Natural Disaster (RCND 2021) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1091 (2022) 012059 doi:10.1088/1755-1315/1091/1/012059

2.4. Analytical Procedures

All analytical measurements performed in this study were conducted according to Standard Methods for
the examination of water and wastewater [11].

Figure 2. Schematic diagram of reactor.

Figure 3. Process flow diagram.

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The 9th AUN/SEED-Net Regional Conference on Natural Disaster (RCND 2021) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1091 (2022) 012059 doi:10.1088/1755-1315/1091/1/012059

Table 1. Physical characteristic of activated sludge (seed sludge).

(BUNUS STP – KLR 403 Kuala Lumpur).

No Set Value Descriptions

1. pH 7.0
2. DO (mg/L) 2.0
3. SV30 (mL/g) 150
4. Size (mm) <0.2
5. Settling velocity (m/h) 0.6

Table 2. Composition of influent from BUNUS STP – KLR 403 Kuala Lumpur.

No Set Value Descriptions

1. pH 6.7
2. COD (mg/L) 304
3. BOD5 (mg/L) 123
4. Ammoniacal Nitrogen (AMN), NH3-N 20
(mg/L)
5. Nitrate, NO3- (mg/L) 0.5
6. Total Phosphate (TP), PO₄³⁻ (mg/L) 5

3. Results and Discussion

3.1. Bioreactor time setting
SBR systems have four processes and it was carried out in sequence feeding, aeration, settling and
decant: (1) ‘feeding with aeration’ (feed and aerate pumps operated simultaneously), (2) ‘aeration only
and without feeding’, (3) ‘settling’, and (4) ‘discharge’. Starting from day 1 until day 26 the average
Dissolved Oxygen (DO) values were around 1 - 2 mg/l but after 1.5 hours the DO values increased
drastically. It shows that the microbe did not consumed air during the process. Day 13 onwards SOUR
profiles show that the microbe consumed oxygen slowly. These results suggest that low amount of
readily oxidized organic available in the bioreactor. Oxygen Reduction Potential (ORP) reading shows
that no denitrification happened along the cycle. The lowest value of ORP monitored was 111 mV
whereby the normal (ORP) value for denitrification should be ranges between -50 mV to +50 mV.
Oxidation-reduction potential or ORP has been used for many years in facilities that process wastewater
generated by metal finishing plants, but only recently has it become prominent in municipal wastewater
treatment plants [12]. To meet the ORP corresponding values, setting time for aeration has changed by
separated operating of the aeration pump during feeding (feeding without aerate).

3.2. Formation and settling velocity of Granules

Day 32 first batch of AGS was harvested using 0.3mm, 0.6mm and 1.0mm sieve. The diameter of the
granules was obtained under microscope observation with an average diameter of 1.64mm (irregular
shapes of granules were developed). The developed granules at 32 days are shown in Figure 4.
Morphology characteristic are shown in Figure 5 (micro colonies Cocci-shaped bacteria acted as a
supporting consortia during the formation of granular), Figure 6 (extracellular polymeric substances
EPS acts as a sticky glue between the microbial cells and strengthen the structure of the granules) and

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The 9th AUN/SEED-Net Regional Conference on Natural Disaster (RCND 2021) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1091 (2022) 012059 doi:10.1088/1755-1315/1091/1/012059

Figure 7 (cavities act as a passage for the transportation of substrate, oxygen, and nutrients into the inner
cores of the granules and ensure the stability within the granules composition). Different methods to
measure the granule density include the pycnometer method, the Percoll density gradient method, the
dextran blue method, and the settling velocity method [13]. Settling velocity method was used in this
study. The granule’s settling velocity is approximately thirty times faster than the actual activated sludge
(actual activated sludge 0.6 m/h and AGS 18.04 m/h). The granules minimum settling velocity is 30 m/h
and can reach up to 70 m/h depending on the compactness and density of the granules [14]. The SVI30
of granules (average 60 mL/g is lower than actual activated sludge with SVI30 value of 150 mL/g). The
SVI of AGS is lower than 100 mL/g while activated sludge flocs, 150 mL/g and above [15]. Figure 8
and Figure 9 shows SVI30 for 1st and 2nd aeration setting time.

3.3. Removal Efficiency of Granules

During the 32 days development parameter such as BOD, COD, Ammoniacal Nitrogen (AMN), NO3-,
TSS, TP, MLSS and MLVSS were monitored daily and weekly. The performance of the reactor from
1st to 2nd aeration setting time improved satisfactory. Average percentage BOD removal was about 33%
to 76.7%. COD removal 42% to 89.7%. These results indicate the removal performance of BOD and
COD improved at different operation setting time, due to the transformation of the flocculent sludge
into granular sludge took place in the bioreactor system. These results are comparable to the previous
study by Ab Halim et al. (2018) [16] who also conducted experiment by using actual domestic
wastewater for the biological activity of granules. AMN removal 95.5% to 74.2% slightly decreased (it
may due to the influent which has high AMN content) but still complied with Malaysia Environmental
Quality Act (design effluent values discharge to river/stream for Standard A < 5 mg/L). The removal
of NO3 was decreased 22.6% to -95.9%. However, the value of 6.5 mg/L of NO3- still complied with
Malaysia Environmental Quality Act (design value required was 10 mg/L). TSS and TP the removal
performance percentage were 40% to 91.9% and -74% to 96.3%, respectively (refer Figure 10, Figure
11 and Table 3).

Figure 4. Microscope images of granules at 32 days.

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The 9th AUN/SEED-Net Regional Conference on Natural Disaster (RCND 2021) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1091 (2022) 012059 doi:10.1088/1755-1315/1091/1/012059

Figure 5. Photo of microbes captured within the granules (Cocci-

shaped) using scanning electron microscope.

Figure 6. Photo of EPS captured within the granules using scanning

electron microscope.

Figure 7. Photo of cavities captured within the granules using scanning

electron microscope.

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The 9th AUN/SEED-Net Regional Conference on Natural Disaster (RCND 2021) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1091 (2022) 012059 doi:10.1088/1755-1315/1091/1/012059

SVI Profile for SBRA, Feed and Aerate Pumps Operated Simultaneously

Figure 8. SVI30 value until day 16 with 1st aeration setting time.

SVI Profile for SBRB, Feed and Aerate Pumps Operated Separately

Figure 9. SVI30 value until day 32 with 2nd aeration setting time.

Removal Performance for SBRA, Feed and Aerate Pumps Operated Simultaneously

Figure 10. SBR removal performance for 1st aeration setting time.

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The 9th AUN/SEED-Net Regional Conference on Natural Disaster (RCND 2021) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1091 (2022) 012059 doi:10.1088/1755-1315/1091/1/012059

Removal Performance for SBRB, Feed and Aerate Pumps Operated Separately

Figure 11. SBR removal performance for 2nd aeration setting time.

Table 3. Removal Performance for SBRA and SBRB

No Parameters SBRA SBRB

1. BOD 33% 76.7%
2. COD 42% 89.7%
3. AMN 95.5% 74.2%
4. NO3 22.6% -95.9%
5. TSS 40% 91.9%
6. TP -74% 96.3%

4. Conclusion
The developed aerobic granular sludge (AGS) in SBRA and SBRB were harvested using 0.3mm,
0.6mm and 1.0mm sieve after 32 days and settling velocity for both reactors were analysed. From the
finding, it is suggested that it would be better for feed and aerate pumps to be operated separately.

Acknowledgements
This research was collaborated between MJIIT Universiti Teknologi Malaysia and Indah Water
Research Centre. Indah Water Konsortium (IWK) is Malaysia’s national sewerage company which has
been entrusted with the task of developing and maintaining sewerage system. The authors are very
grateful to IWK for facilitating samples collection and lab facilities.

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 The 9th AUN/SEED-Net Regional Conference on Natural Disaster (RCND 2021) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1091 (2022) 012059 doi:10.1088/1755-1315/1091/1/012059

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