Journal of Water and Environment Technology, Vol. 9, No.

3, 2011

Significance of Reactor Type for High-Rate Nitrification

Using Aerobic Granular Sludge

Yoshiaki HASEBE*, Masahiro EGUCHI*, Hiroaki MEGURO*, Satoshi TSUNEDA**

*R&D Center, Organo Corporation, Kanagawa 252-0332, Japan

**Department of Life Science and Medical Bio-Science, Waseda University, Tokyo 162-8480,
Japan

ABSTRACT
Nitrifying granules were applied to high-speed nitrification of wastewater discharged from an
electronic industrial factory. The behavior of the formation of nitrifying granules and the start-up
performance of nitrification process were examined with three types of reactors: a column with a
small diameter, a column with a large diameter and a rectangular-type reactor. As a result, the
performance of the three types of reactors did not give significant differences in terms of the
formation of nitrifying granules and the start-up trend of the reactor. A nitrification rate of about
2.0 kgN/m3/day was achieved in all reactors.

Keywords: aerobic upflow fluidized bed (AUFB), high-rate nitrification, inorganic wastewater,
nitrifying granule.

INTRODUCTION
Biological nitrogen removal by means of nitrification and denitrification is a key
technology in advanced wastewater treatment process. Many of the nitrogen removal
processes have been done using conventional activated sludge process. However, it is
difficult to retain a large amount of nitrifying bacteria within the reactor, and it is
difficult to increase the volumetric ammonia loading rate. Consequently, the nitrification
process is the rate-determining step in the entire nitrogen removal process. A fluidized
bed reactor with a carrier (ex. sponge or gel materials) is used to speed-up the
nitrification process. On the other hand, new techniques of aerobic granulation have
been developed for the treatment of organic pollutants in wastewater using a column-
type sequencing batch reactor (SBR) (Morgenroth et al., 1997; Beun et al., 1999;
Etterer and Wilderer, 2001; Tay et al., 2001) or using an aerobic upflow sludge blanket
(AUSB) with front aeration (Mishima and Nakamura, 1991). Furthermore, the
techniques for producing granules of nitrifying bacteria using a conical-shaped AUSB
with front aeration (de Beer et al., 1993; Schramm et al., 1998, 1999) or using an SBR
(Tay et al., 2002) have been proposed. Although the mechanism of granulation is still a
subject of discussion, it has been thought that key factors of granulation are settling time
(selection of sludge by settling velocity), shear force, volumetric loading rate and feast-
famine regime (Beun et al., 1999; Tay et al., 2001; McSwain et al., 2004; Liu and Tay,
2006).

Although many of the formation of aerobic granules have been studied by using SBR,
we successfully produced nitrifying granules with a diameter of 200 µm within 100
days using an aerobic upflow fluidized bed (AUFB) reactor in continuous flow
condition (Tsuneda et al., 2003). Since nitrifying bacteria could be retained with a high
density in the AUFB reactor due to the form of granules, inorganic wastewater
containing 500g/m3 NH4+-N was effectively and stably treated with an ammonia

Address correspondence to Yoshiaki Hasebe, R&D Center, Organo Corporation,

Email: hasebe-y@organo.co.jp
Received May 10, 2010, Accepted May 30, 2011.
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removal rate of 1.6 kgN/m3/day and ammonia removal efficiency of more than 90%
throughout 500 days (Tsuneda et al., 2003). Although a column-type reactor was used in
this research, the internal flow condition possibly receives a big influence for the scale-
up of the reactor.

Meanwhile, in the electronic industry, especially in semiconductor manufacturing

factories, different kinds of ammonia compounds such as ammonium persulfate and
ammonium fluoride are used to wash silicon wafers, producing wastewater that contains
ammonium and fluoride ion. Biological treatment system is mainly used for the removal
of nitrogen from electronic industrial wastewater, and the fluoride ion was removed
from the wastewater before nitrogen removal process so that the existing fluoride ion
may have a harmful effect on the biological treatment. To precipitate and remove
fluoride, calcium ion is added to the wastewater. As a result, the influent water of
biological nitrogen removal process contains calcium ion besides ammonium ion.

In the present study, we attempted to treat such wastewater with nitrifying granulation
processes. As the feasibility of scale-up was of major interest, we compared the start-up
and treatment performances of three types of reactors with different shapes.

MATERIALS AND METHODS

Wastewater composition
The composition of the synthetic wastewater used throughout the study contained 0.5
kg/m3 of NH4+-N (as NH4Cl), 0.005 kg/m3 of Fe (as FeSO4·7H2O), 0.010 kg/m3 of PO43-
-P and 0.5 kgCa/m3 of Ca (as CaCl2·2H2O) in tap water. Table 1 shows the chemical
composition of synthetic wastewater.

Reactor configuration and operating conditions

Nitrification using column-type reactor with a small diameter (Run 1)
An AUFB reactor that has a gas-solid separator (GSS) at the top was used (Fig. 1). The
reactor had a diameter of 0.05 m and a height of 3.2 m (effective volume: 6 L).
Activated sludge collected from a food industry was inoculated to the AUFB reactor
with an initial mixed liquor suspended solids (MLSS) concentration of 5 kg/m3.

Table 1 - Chemical composition of synthetic wastewater

Component Concentration [g/m3]
NH 4Cl 3200
FeSO4 ·7H2O 25
K2HPO4 56
CaCl2 ·2H 2O 1800
KCl 10
MgSO 4·7H 2O 3.5
AlCl 3·6H 2O 0.32
NaCl 0.32
CaCl 0.32
CuSO 4·5H 2O 0.32
ZnSO4·H 2O 0.32
MnSO4·H2O 0.32
FeCl3 0.28

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Moreover, Fe2O3 used as aggregation accelerant was added to the reactor with an initial
concentration of 0.25 kg/m3. From the bottom of the reactor, synthetic wastewater and
air were continuously fed. The initial nitrogen volumetric load and air flow rate were set
to 0.2 kgN/m3-reactor/day and 1.0 L/min, respectively. The feeding flow rate was
increased when stable and mostly complete nitrification was achieved and the
settleability of the biomass was good enough. The pH sensor was set at the middle of
the column and the pH was controlled at 7.1 - 7.3 by the addition of NaOH.

Nitrification using a column-type reactor with a large diameter (Run 2)

An AUFB reactor with a solid-liquid separator, diameter of 0.15 m and height of 3.2 m
(effective volume: 57 L) was used. During the start-up of the reactor, activated sludge
and Fe2O3 were inoculated to the AUFB reactor in the same manner as Run 1. To have a
shear stress equal to that of Run 1, the air flow rate was set to 9.0 L/min. Other
experimental conditions followed those of Run 1.

Nitrification using a rectangular-type reactor (Run 3)

The schematic diagram of the reactor is shown in Fig. 1. When the column-type reactor
is used, the influence of internal flow by the scale-up is very large. On the other hand,
the rectangular-type reactor was mainly used for wastewater treatment. The scale-up of
the reactor was thought to be so easy that the rectangular-type reactor was used in Run 3.
The size of the reactor was 0.3 m in length, 0.6 m in width, 1.2 m in height (effective
volume: 180 L). A solid-liquid separator was set up in the top of reactor. Synthetic
wastewater was continuously fed from the top of the reactor and air was fed from the
bottom of the reactor. The initial flow rate was set to 15 L/min and increased to 30
L/min in a stepwise manner. Other experimental conditions followed those of Run 1.

Chemical analysis
All samples obtained from the reactors except those for the measurement of MLSS and
suspended solids (SS) were filtered with a filter paper (5C, ADVANTEC, Japan) and
used in water quality measurement. Concentrations of NH4+-N, MLSS and SS were

pH
controller

Gas-Solid separator

pH
Influent controller
Effluent
Gas-Solid separator

Effluent
pH sensor
Air
Air pH sensor
Diffuser Air flow meter

Influent Airflow meter

Diffuser

AUFB-type reactor rectangular-type reactor

(Effective volume:6L in Run 1 (Effective volume:180L)
57L in Run 2)

Fig. 1 - Schematic illustration of the AUFB reactor used in Runs 1 and 2, and the
rectangular-type reactor used in Run 3

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measured in accordance with the standard method (APHA, 1995). Both NO2--N and
NO3--N concentrations were measured using an ion chromatograph (DX-120, Dionex,
Japan). Dissolved oxygen (DO) concentration and pH were measured using a portable
DO meter (LDO, HACH, USA) and a pH meter (Twin pH, HORIBA, Japan),
respectively.

RESULTS AND DISCUSSION

Performance of column-type reactor with a small diameter
The time courses of water quality data (the influent and effluent concentrations of NH4+-
N) and the ammonia removal rate of Run 1 using the AUFB with a small diameter are
shown in Fig. 2. To exclude the influence of water temperature, the ammonia removal
rate in Fig. 2 was converted to the ammonia removal rate at 20°C by using the following
equation.

K20°C = KT / θ (T - 20) (1)

K20°C: ammonia removal rate at 20°C
KT: ammonia removal rate at T°C
θ: temperature correction factor (= 1.057)

When the effluent concentration reached a steady state, the flow rate was increased in a
stepwise manner to elucidate the maximum ammonia removal rate. The ammonia
removal rate was 0.2 kgN/m3/day during the start-up of the reactor, and then, it reached
1.3 kgN/m3/day after 60 days. The ammonia removal rate reached 2.0 kgN/m3/day (flow
rate was about 0.015 L/min) or more after 110 days although the increase of the
ammonia removal rate stagnated for about 30 days from day 60. The stagnation was due
to the competition between the increase of sludge settling velocity (it leads to the
increase in MLSS) and the decrease of HRT (it leads to the decrease in MLSS).
Moreover, the concentration of NH4+-N in the effluent was always less than 0.01
kgN/m3, and most of NH4+-N was converted to NO3--N, while NO2--N concentration in

4 700
NH4+-N concentration [mgN/L]

3.5 600
NH4+-N removal rate

3
500
[kgN/m3/day]

2.5
400
2
300
1.5
1 200
ammonia removal rate
influent NH4-N
NH4-N 100
0.5
effluent NH4-N
NH4-N
0 0
0 20 40 60 80 100 120 140 160
Time [day]

Fig. 2 - Time courses of ammonia removal rate and NH4+-N concentrations of influent
and effluent (Run 1). Open and closed diamonds in the figure indicate NH4+-
N concentration of influent and effluent, respectively. Closed circles indicate
NH4+-N removal rate at 20°C

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the effluent was always less than 0.005 kgN/m3. These effluent water qualities were
almost equal in all runs.

Comparison between column-type reactors with small and large diameters

(Dependence property of aspect ratio)
While the height of the reactor used in Run 1 was similar to those of full scale plants, its
diameter was significantly less. In order to demonstrate the influence of the scale-up of
the AUFB reactor, the reactor used in Run 2 had three times bigger diameter than that
used in Run 1. The time courses of the ammonia removal rate of Run 1 and Run 2 are
shown in Fig. 3. Although high ammonia removal rate was temporarily obtained from
day 20 to 30 because of the rapid increase in ammonium load in Run 2, the overflow of
sludge was superior, resulting in stagnation of the increase of the ammonia removal rate
around day 30 - 60. However, after the stagnation period, the ammonia removal rate has
increased again. It was thought that the influence of the scale-up was little in terms of
ammonia removal rate because the trend was similar to Run 1. On the other hand, the
distribution of MLSS concentration and pH in the reactor as shown in Figs 4 and 5,
respectively, differed greatly between Run 1 and Run 2. Because the mixing in the
column from top to bottom was little, the MLSS concentration was high at the lower
part of the column in Run 1. In contrast, the difference of the MLSS concentration at the
top and bottom of the column was small in Run 2, suggesting that the bulk in the
column was close to a completely mixed condition. The pH as well as the MLSS
concentration difference between the top and bottom of the column was observed in
Run 1. In this study, the pH was measured and controlled in the middle of the column,
and NaOH was added at the bottom of the column. As a result, the pH was high in the
lower part and low in the upper part of the column in Run 1. In contrast, the pH
difference as well as the difference of the MLSS concentration between the upper and
lower parts of the column was not observed in Run 2. The distributions of the NH4+-N
concentration were measured but they were less than 0.005 kgN/m3 in all sampling
points for both Run 1 and Run 2. Therefore, the influence of the differences of column
diameters was thought to be small in terms of the nitrification rate even though the
internal flow condition was significantly different.

2
NH4+-N removal rate [kgN/m3/day]

1.8
1.6
1.4
1.2
1
0.8
0.6
0.4
0.2
0
0 20 40 60 80 100
Time [day]

Fig. 3 - Time courses of converted ammonia removal rates (20°C) of Run 1 and Run
2. Circles and triangles indicate Run 1 and Run 2, respectively

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300 300

250 250

200 200

Height [cm]
Hight [cm]

150 150

100 100

50 50

0 0
0 5000 10000 15000 6.5 7 7.5 8

MLSS [mg/L] ｐH [-]

Fig. 4 - Distribution of MLSS in the Fig. 5 - Distribution of pH in the

reactor of Run 1 and Run 2 at reactor of Run 1 and
day 100. Circles and triangles Run 2 at day 100.
in the graph indicate MLSS Circles and triangles in
concentrations of Run 1 and the graph indicate pH of
Run 2, respectively. The Run 1 and Run 2,
height in the graph was respectively measured
measured from the bottom of from the bottom of
column column

Comparison between column-type reactor (AUFB) and rectangular-type reactor

(Dependence property of state of flow)
In order to demonstrate the influence of the shape of the reactor, the nitrification
performance using the rectangular-type reactor was examined (Run 3). The scale-up of
the rectangular-type reactor is thought to be easier than that of the column-type reactor.
This reactor has a solid-liquid separator (SLS) at the top to separate sludge from the
effluent, and gives almost complete mixing by aeration in the main part of the reactor.
The time courses of the ammonia removal rate of Run 3 are shown in Fig. 6. In Run 3,
since the increase in ammonia load was too fast, the stagnation period of the ammonia
removal rate was observed in the same manner as Run 2. The ammonia removal rate
increased with the improvement of the settling velocity of sludge and reached 1.9
kgN/m3/day (flow rate was about 0.58 L/min) after the stagnation period. The formation
of nitrifying granules was observed in Run 3, and the ammonia removal rate was almost
equal to Run 1 and Run 2. The flow condition in the column changes to a complete
mixture as the diameter changes to large. As a result, it was considered that the shape of
the reactor and the flow condition are not a big factor for the formation of nitrifying
granules. If it is possible that the rectangular-type reactor is used, there is a possibility
that the initial cost of the system falls greatly compared with the column-type reactor.
So, the successful formation of granules by using rectangular-type reactors in
continuous-flow conditions is a very important result.

The photographs of nitrifying granules formed in Runs 1, 2 and 3 are shown in Fig. 7.
In all the reactors, granulation was achieved. It was suggested that the feast-famine
condition is not a significant factor in forming nitrifying granules though the condition
was thought as an important factor in forming aerobic granules mainly composed of
heterotrophic bacteria.

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NH 4+-N removal rate [kgN/m3/day]

1.8
1.6
1.4
1.2
1
0.8
0.6
0.4
0.2
0
0 20 40 60 80 100
Time [day]

Fig. 6 - Time courses of converted ammonia removal rates (20°C) of Run 1 and Run
3. Circles and diamonds indicate Run 1 and Run 3, respectively

(a) (b) (c) (d)

Fig. 7 - Photographs of nitrifying granules in the reactor viewed by optical microscopy

(a) Seed sludge; (b) Run 1 (day 100); (c) Run 2 (day 100); (d) Run 3 (day 100)
Bar = 400 µm

CONCLUSIONS
In this study, the influence of the reactor scale and the reactor type on the formation of
nitrifying granules and start-up trend of ammonia removal rate were investigated in the
nitrification of synthetic wastewater from semiconductor manufacturing factories by
using nitrifying granules. The experimental results are summarized as follows:

(1) Nitrifying granules were formed in the AUFB reactor under continuous-flow
condition and the ammonia removal rate reached about 2.0 kgN/m3/day.
(2) The influence of the ratio of the column diameter and height was small during the
start-up of the nitrification process and the formation of nitrifying granules
although the flow conditions greatly influenced the distribution of MLSS and pH in
the AUFB reactor. The internal flow condition of the large column (in Run 2) was
closed to a complete mixing flow. Therefore, the scale-up of the column-type
reactor seems easy.
(3) The nitrifying granules were formed in the rectangular-type reactor under
continuous-flow condition, and the large difference in terms of start-up trend of the
ammonia removal rate between AUFB and rectangular-type reactor was not
confirmed. This outcome is very important in the sense that the use of cheap and
conventional rectangular-type tanks could be possible.

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