ARTICLE IN PRESS

International Biodeterioration & Biodegradation 59 (2007) 1–7

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Review

Control of carbon and ammonium ratio for simultaneous nitrification

and denitrification in a sequencing batch bioreactor
Ying-Chih Chiua,, Li-Ling Leeb,1, Cheng-Nan Changb, Allen C. Chaoc
a
Department of Environmental Engineering, National Ilan University, I-Lan City, Taiwan 26041, ROC
b
Graduate Institute of Environmental Science and Engineering, Tunghai University, Box 5-819, Taichung City, Taiwan 40704, ROC
c
Department of Civil Construction and Environmental Engineering, North Carolina State University, Raleigh, NC 27695, USA
Received 11 April 2006; received in revised form 13 July 2006; accepted 1 August 2006
Available online 2 October 2006

Abstract

This study shows how the carbon and nitrogen (C/N) ratio controls the simultaneous occurrence of nitrification and denitrification in a
sequencing batch reactor (SBR). Data demonstrated that a low C/N ratio resulted in a rapid carbon deficit, causing an unbalanced
simultaneous nitrification–denitrification (SND) process in SBR. When the initial COD/NH+ 4 -N ratio was adjusted to 11.1, the SND-

based SBR achieved complete removal of NH4-N and COD without leaving any NO2-N in the effluent. The nitrogen removal efficiency
decreases gradually with increasing ammonium-loading rate to the SND–SBR system. Altogether, data showed that appropriate controls
of carbon and nitrogen input are required to achieve an efficient SND–SBR. An established SND technology can save operation time
and energy, and might replace the traditional two-stage biological nitrification and denitrification process.
r 2006 Elsevier Ltd. All rights reserved.

Keywords: Carbon source; Heterotrophic nitrification; Oxidation–reduction potential (ORP); Simultaneous nitrification and denitrification (SND)

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
2. Materials and methods. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2.1. Composition of the synthetic wastewater. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2.2. Operational conditions of the SND-based SBR system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2.3. Analytical methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
3. Results and discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
3.1. Effect of insufficient carbon source . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
3.2. Influence of ammonium-loading rate. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
4. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7

1. Introduction

Corresponding author. Tel.: +886 3 9357954; fax: +886 3 9367642.
E-mail address: ycchiu@niu.edu.tw (Y.-C. Chiu).
1
Present address: Energy Strategy Research Lab., Industrial Technology
Research Institute, Chutung, Hsinchu, Taiwan 310, ROC.
The traditional biological process used in wastewater
treatment to achieve nitrogen removal involves separate
aerobic and anaerobic phases that are generally carried out
in separate bioreactors or by different aeration intervals

0964-8305/$ - see front matter r 2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.ibiod.2006.08.001
 ARTICLE IN PRESS
2 Y.-C. Chiu et al. / International Biodeterioration & Biodegradation 59 (2007) 1–7
(Metcalf and Eddy, 1991). However, Ferguson (1994)
proposed that heterotrophic microorganisms may be
involved in nitrification and simultaneous aerobic deni-
trification. Some heterotrophic nitrifiers have been re-
ported to denitrify nitrite (NO 2 ) and nitrate (NO3 )

aerobically (Robertson et al., 1988; Zart and Eberhard,
1998). Other studies show that nitrification and denitrifica-
tion occur concurrently in a single reactor under aerobic
conditions (von Münch et al., 1996; Zeng et al., 2003),
which is often referred to as the simultaneous nitrification
and denitrification (SND) process. The SND process
represents a significant advantage over the conventional
separated nitrification and denitrification processes.
SND occurs naturally inside microbial biofilms and
flocks due to the oxygen gradient that establishes across the
biomass (von Münch et al., 1996). Nitrifiers are active in
areas of high dissolved oxygen (DO) concentration,
whereas denitrifiers are active in areas of very low DO
concentration. The uneven distribution of DO inside the
biomass allows simultaneous proliferation of nitrifying and
denitrifying bacteria.
On the other hand, some heterotrophic bacteria, e.g.,
Alcaligenes faecalis (van Niel et al., 1992) and Thiosphaera
pantotropha (Robertson and Kuenen, 1988), are capable of
performing SND by using organic substrates aerobically as
sources of carbon and energy to convert ammonium
(NH+ 4 ) aerobically into nitrogen gas (Stouthamer et al.,
1997). Pathways of heterotrophic SND metabolism are
different from those of autotrophic nitrifiers and hetero-
trophic denitrifiers. Ammonium is hydroxylated to hydro-
xylamine (NH2OH) by ammonium monooxygenase under
aerobic conditions. Subsequently, hydroxylamine is oxi-
dized to nitrite by hydroxylamine oxidase. Finally, nitrite is
directly transformed into N2. The SND process is also
termed ‘‘aerobic deammonification’’ (Bock et al., 1995; van
Loosdrecht and Jetten, 1998).
Hellinga et al. (1998) designed an SND-based reactor in
which aerobic heterotrophic bacteria removed nitrite
produced from ammonia by Nitrosomonas in the same
reactor. Robertson and Kuenen (1988) studied the
nitrification and denitrification ability of T. pantotropha,
and observed that this facultative anaerobic and auto-
trophic sulfur bacteria can perform simultaneous hetero-
trophic nitrification and aerobic denitrification by using O2
and NO 3 simultaneously. Their observation implies that T.
pantotropha is capable of carrying out SND. Pochana and
Keller (1999) investigated the efficiency of nitrogen
removal from wastewaters by an SND-based sequencing
batch reactor (SBR). They observed that higher DO
concentrations enhanced nitrification rates. Simulta-
neously, high DO concentrations inhibited the denitrifica-
tion process, causing an accumulation of nitrite and nitrate
in the reactor. On the other hand, at lower DO concentra-
tions the nitrification process was slowed down and the
denitrification process was enhanced. Therefore, the DO
level is a factor critical to the SND process; it must be
maintained at an appropriate level in the SND reactor in
order to reach a balanced equilibrium between the
nitrification and denitrification processes.
Under either oxygen-limiting or oxygen-free conditions,
Nitrosomonas europaea transformed NH+ 4 into NO 2,
which was simultaneously transformed to nitrous oxide
(N2O) and N2 (Shrestha et al., 2002). von Münch et al.
(1996) observed that an efficient SND process occurred at a
DO concentration lower than 0.5 mg L1. The importance
of maintaining a low DO concentration was confirmed by
Pochana and Keller (1999), who showed that NH+ 
4 , NO2 ,

NO3 , and soluble COD were removed in 4 h under DO
conditions between 0.3 and 0.8 mg L1. This suggests that
some heterotrophic nitrifiers have the ability to denitrify
under low oxygen conditions to affect an SND reaction.
When Paracoccus denitrificans was grown under conditions
favoring the coupled heterotrophic nitrification–denitrifi-
cation reaction, a small increase in the maximum microbial
growth rate and a drastic reduction in cell yield were
observed compared to cultures that were not growing
under SND conditions (Stouthamer et al., 1997). Similar
results related to other pure cultures have also been
reported by van Niel et al. (1992) and Robertson et al.
(1988).
The available literature shows that under suitable
conditions, the SND process will occur in the wastewater
system. Wastewater treatment based on an SND–SBR
process would represent a simpler biological nitrogen
removal system than the conventional two-step nitrifica-
tion and denitrification process. However, the conditions
that may result in the occurrence of an efficient SND-based
SBR process are not yet well established. In this investiga-
tion, an SND-based SBR process was operated to resolve
some of the conditions under which significant SND
activities can occur.
2. Materials and methods
2.1. Composition of the synthetic wastewater
A synthetic wastewater was used for investigating the SND at different
influent COD/NH+ 4 -N (C/N) ratios and ammonium-loading rates with the
formula listed in Table 1. The optimal COD/nitrate ratio to support
conventional biological denitrification was reported to be 6.0–8.0 by Chiu
and Chung (2003). Zhao et al. (1999) suggested that heterotrophic
nitrifiers, which can coexist with autotrophic nitrifiers under a wide range
of conditions, play a significant role in the oxidation of ammonia at a
BOD/N ratio of 6.9 or greater. Therefore, three C/N ratios of a synthetic
wastewater (6.3, 11.1, and 19.7) were tested in this study by adjusting the
sodium acetate dosage. In order to evaluate the influence of NH+ 4 -N
loading on the nitrification efficiency of the SND–SBR system, quantities
of ammonium chloride (NH4Cl) and sodium acetate were adjusted at
different levels in three different operational phases during the experi-
mental period. Concentrations of relevant constituents contained in the
synthetic wastewater are summarized in Table 1. The high NH+ 4 -N content
was used to simulate the inorganic nitrogen variation in domestic effluents.
2.2. Operational conditions of the SND-based SBR system
A schematic diagram of the experimental SBR system comprised of two
reactors is shown in Fig. 1. Each reactor was a rectangular acrylic tank,
 ARTICLE IN PRESS
Y.-C. Chiu et al. / International Biodeterioration & Biodegradation 59 (2007) 1–7 3

Table 1
Operational conditions of an SBR system set to obtain an SND process

Operation conditions Phase I Phase II Phase III

Reactor A B A B A B

Day 60–160 60–80 81–114 115–160 161–241 242–312

SRT (day) 20 20 20 20
HRT (day) 1 1 1 1
MLVSS (mg L1) 41007200 41007200 41007200 41007200
DO (mg L1) 0.640.68 0.640.68 0.640.68 0.640.68
F/M (g-COD g-VSS1 day1) 0.34 0.18 0.34 0.59 0.34 0.14 0.34 0.49
Ammonium loading rate (g-NH+
4 -N/g-VSS/day) 0.029 0.029 0.029 0.012 0.029 0.046
COD(mg L1) 1400720 750 1320 2350 1400717 570 1400719 2000
NH+ 1
4 -N (mg L ) 119.271.5 119.271.5 119.271.7 50.4 119.271.8 187.0
1
NOX-N (mg L ) ND ND ND ND
TKN (mg L1) 15472.1 15472.1 — —
Org-N (mg L1) 34.871.6 34.871.6 — —
T–N (mg L1) 153.871.6 153.871.6 — —
PO 1
3 -P (mg L ) 9.170.4 9.170.4 9.170.3 9.170.5
Alkalinity (mg-CaCO3 L1) 460710 460710 460710 460710

Sample size ¼ 7.

60 cm (D)  40 cm (W)  40 cm (L) with a maximum working volume of
30 L. Both bioreactors were agitated (120 rpm) and equipped with an
aerator, a peristaltic pump, and a water level sensor, as well as probes to
follow variations of the oxidation/reduction potential (ORP), the pH, and
the DO concentration.
Sludge collected from the aerobic pond of a swine farm wastewater
treatment system (Hamburger Changhua, Taiwan) was used as inoculum.
The sludge was elutriated with clean water three times to remove fine
suspended solids that may interfere with the analyses. It was then added to
each of two reactors at a final concentration of approximately 1.1 g L1 of
synthetic wastewater. Both reactors were operated as SBR. Fifteen liters of
the reactor content were replaced twice daily by the same volume of fresh
synthetic wastewater. The total cycling time of the SBR operation was
12 h, which was comprised of four stages: 60 min anoxic conditions,
540 min oxic conditions, 90 min of settling, and 30 min drawing. The two
reactors were operated in parallel with a hydraulic retention time of 24 h
and a sludge retention time of 20 days in a temperature-controlled room at
2572 1C.
The reactors became stable after 60 days and they were operated for an
additional 250 days. Reactor B was successively set at three different
ammonium-loading rates representing each of phase I, II and III which
were, respectively, 0.012 g-NH+ 1
day1 during day 60–160;
4 -N g-VSS
0.029 g-NH+ 4 -N g-VSS
1
day1 during day 161–241; and 0.046 g-NH+ 4 -
N g-VSS1 day1 during day 242–312 (Table 1). Reactor A, a control, was
set at an ammonium-loading rate of 0.029 g-NH+ 4 -N g-VSS
1
day1
throughout the 250 days of operation. Other relevant loadings and
operational conditions are also listed in Table 1.
2.3. Analytical methods
The electrodes for measuring the pH (Mettler-Toledo pH), the DO
(WTW TriOxmatic 690), and the ORP (Mettler-Toledo ORP) were
connected to a computer, and the on-line data were processed with the
LabVIEW (National Instruments, USA) software. Dissolved organic
carbon (DOC) was measured using a TOC analyzer (Model TOC-5000,
Shimadzu Co., Kyoto, Japan). All samples for DOC analysis were filtered
through 0.45-mm glass fiber filters before analysis. Ammonium nitrogen
(NH+4 -N) was determined by titration. Nitrate and nitrite concentrations
were measured with an ion chromatograph (Model L-3700, Hitachi Ion
Chromatograph, Japan). The procedures in Standard Methods (APHA,
1998) were followed for determining the mixed liquor suspended solids
(Method 2540D), the mixed liquor volatile suspended solid (MLVSS)
(Method 2540E), and the COD (Method 5220C). N2O and N2 were
quantified by gas chromatography using a China Chromatograph 8900
(Taipei, Taiwan) equipped with a Supelco 2.5-m column (80/100 mesh
Porapak Q, 250 1C) and a thermal conductivity detector. High-purity
helium (99.999%) was the carrier gas (flow rate, 30 mL min1). The
injector, oven, and detector temperatures were controlled at 70, 45, and
70 1C, respectively.
3. Results and discussion
3.1. Effect of insufficient carbon source
Fig. 2 shows the variations of biological nitrogen
transformation in a reactor subject to three different initial
C/N ratios (6.3, 11.1, and 19.7) during phase I where the
ammonium-loading rate was maintained at 0.029 g-NH+ 4 -
N g-VSS1 day1. Under an initial C/N ratio of 6.3, the
ammonium nitrogen removal was not significant (Fig. 2a);
41% of initial NH4-N were removed after 240 min and only
11% of the remaining NH+ 4 were removed after
240–720 min. Hydroxylamine (Fig. 2b) and nitrite were
produced, but nitrate was not (Fig. 2c). This implies that
the C/N ratio was inadequate for establishing an efficient
SND process. The SND process requires sufficient low-
degradable carbon substrate to provide the reducing power
required for the denitrification reaction that occurs after
the nitrification reaction. Therefore, due to the rapid
carbon depletion in the early oxic stage (Fig. 2d), at low
initial C/N ratio, after 240 min of incubation, NH+ 4
concentration was high in the effluent at 36 mg-NH+ 4 -
N L1 and in addition, 5 mg-NO 2 -N L
1
were generated.
+
In the 240–720-min-period, NH4 -N concentration de-
creased by only 7 mg L1 (36–29 mg L1) because there was
not enough carbon remaining to support the heterotrophic
biomass. Both NO 
2 -N and NO3 -N increased during this
 ARTICLE IN PRESS
4 Y.-C. Chiu et al. / International Biodeterioration & Biodegradation 59 (2007) 1–7

Anoxic Oxic Settling and Draw

80
5
60

NH4+--N (mg/L)
4

40 C/N=6.3
LabVIEW
20 C/N=19.7 C/N=11.1

0
DO pH ORP (a) 0 120 240 360 480 600 720
DO pH ORP 0.8

0.6 C/N=19.7

NH2OH (mg/L)
1 1
0.4

0.2 C/N=6.3
C/N=11.1

0.0
(b) 0 120 240 360 480 600 720
12 3

2 2
9

NO3- -N (mg/L)
C/N=6.3 (NO2- -N)
NO2-N (mg/L)

6
C/N=6.3 (NO3- -N)
-

1
3 C/N=11.1(NO-2-N)
C/N=19.7
3 0 0
3 (c) 0 120 240 360 480 600 720
1,200

900 C/N=19.7
COD (mg/L)

Reactor A Reactor B
600
Fig. 1. Schematic diagram of the SND–SBR systems: (1) Stirrer, (2) pH, C/N=11.1
ORP, and DO meters, (3) Air compressor, (4) AD/DA interface card, and
300
(5) Computer. Reactor A is a control set with an ammonium-loading rate
of 0.029 g-NH+ 4 -N g-VSS
1
day1. Reactor B was operated with ammo- C/N=6.3
nium-loading rates of 0.012, 0.029, and 0.046 g-NH+
4 -N g-VSS
1
day1 for 0
different phases. 0 120 240 360 480 600 720
(d) Time (min)

Fig. 2. Profiles of NH4-N, NH2OH, and NO2-N concentrations and COD

period, which indicates that nitrification by autotrophic during one 12-h cycle of an SBR when the ammonium-loading rate was
maintained at 0.029 g-NH+ 4 -N g-VSS
1
day1 and the initial C/N ratio
bacteria was the major process occurring at low C/N ratio
was adjusted at 6.3, 11.1, or 19.7.
(Hooper et al., 1997).
When the initial C/N ratio was maintained at higher
levels (11.1 and 19.7), at the end of the 12-h period, all
concentrations of NH+ 
4 -N, NH2OH, NO2 -N, and NO3 -N

The equation proposed by Katie et al. (2003) was used to
in effluent were lower than the analytical method detection calculate the efficiency of the SND process (EfficiencySND)
limits; intermediate products were not observed to accu-  
NO x remained
mulate either (Figs. 2b and c). This corroborates previous EfficiencySND ¼ 1  100, (1)
NHþ 4 oxidized
reports showing that an efficient SND process occurs when
the nitrification and denitrification rates are in a balanced where NOx remained is the NOx

remained after reaction,
þ +
equilibrium producing very low amounts of nitrate and NH4 oxidized the NH4 oxidized after reaction.
nitrite in the system (von Münch et al., 1996; Zeng et al., Table 2 summarizes the calculated nitrification and
2003). Moreover, the effluent COD reached values below denitrification rates and the SND efficiency. von Münch
detectable limits when the system was operated with an et al. (1996) obtained efficient SND conditions under
initial C/N ratio of 11.1. nitrification and denitrification rates that were lower than
 ARTICLE IN PRESS
Y.-C. Chiu et al. / International Biodeterioration & Biodegradation 59 (2007) 1–7 5

Table 2
SND efficiency under different initial C/N ratios and comparison with reported values

C/N ratio Nitrification rate (mg- Denitrification rate (mg- Efficiency of SND (%) References
NH+ 4 -N L
1 1
h ) NOx-N L1 h1)

6.3 3.45 9.62 1.02 This study

11.1 9.44 9.23 98.7 This study
19.7 11.33 11.01 97.1 This study
14.5 15 12 80 Pochana and Keller
(1999)
11.2 3 3 100 von Münch et al. (1996)
— 3.3 — 78 Katie et al. (2003)

ours. Pochana and Keller (1999) estimated an 80% SND Anoxic Oxic Settling and Draw
120
efficiency at an initial C/N ratio of 14.5. These results Ammonium Loading Rate=0.012
Ammonium Loading Rate=0.029
demonstrate that under carbon deficit conditions, the rates 90 Ammonium Loading Rate=0.046

NH4+-N (mg/L)
of the nitrification and denitrification reactions are not in a
balanced equilibrium, resulting in an inefficient SND 60
process. The highest nitrification rate (11.33 mg-NH+ 4 -
N L1 h1) was found at an initial C/N ratio of 19.7 when 30
the carbon source was in excess (Fig. 2d). In practice, 11.1
(a) 0
was the optimal value for the initial C/N ratio, allowing the 0 120 240 360 480 600 720
system to reach equilibrium between the nitrification and 8 8
AmmoniumLoading Rate=0.012 (NO2--N)
AmmoniumLoading Rate=0.029 (NO2--N)
the denitrification reactions and resulting in optimal AmmoniumLoading Rate=0.046 (NO2--N)

NH2OH (mg/L)
6 6
NO2--N (mg/L)

removal of both nitrogen and organic carbon. AmmoniumLoading Rate=0.012 (NH2OH)

AmmoniumLoading Rate=0.029 (NH2OH)
AmmoniumLoading Rate=0.046 (NH2OH)
4 4
3.2. Influence of ammonium-loading rate
2 2
Fig. 3 depicts the NH+ 4 -N removal in the SBR system
and the resulting effluent quality with respect to NH2OH, (b) 0 0
0 120 240 360 480 600 720
NO 
2 -N, and NO3 N concentrations. Fig. 4 displays the 3
corresponding real-time monitoring of ORP, pH, and DO.
At the highest influent NH+ 4 -N concentration (0.046 g-
N03--N (mg/L)

2 Ammonium Loading Rate=0.012

NH+ 4 -N g-VSS
1
day1) with a COD loading rate main-
tained at 0.49 g-COD g-VSS1 day1, the DO was rapidly
1
depleted and its level remained close to zero (Fig. 4c) from
60 to 600 min. This agrees with the finding of Ng et al.
(c)
(2000) that during the oxic stage, low DO concentrations in 0
SBR system were obtained at high COD and NH+ 4 -N 1,000
0 120 240 360 480 600 720
loadings. However, when the DO level was low, the
conversion of NO 2 -N from NH2OH was inhibited, 750
COD (mg/L)

resulting in NH2OH accumulation (Fig. 3b). This phenom- Ammonium Loading Rate=0.046

enon confirms that high ammonium-loading rates create 500

oxygen limitation conditions that have a negative effect on Ammonium Loading
250
the SND process. Since the carbon source was not Rate=0.029
Ammonium Loading
completely utilized in the SND process, the residual 0 Rate=0.012

COD at the end of the oxic stage was around 120 mg L1 0 120 240 360 480 600 720
(Fig. 3d). (d) Time (min)
At an ammonium-loading rate of 0.029 g-NH+ 4 -N
Fig. 3. Profiles of NH4-N, NO2-N, and NO3-N concentrations and COD
g-VSS1 day1, NH+ 4 -N (Fig. 3a) was almost completely during one 12-h cycle of an SBR when the ammonium-loading rate was
oxidized, leaving no nitrite or nitrate. At the lower ammo- adjusted at either 0.012, 0.029, or 0.046 g-NH+
4 -N g-VSS
1
day1.
nium-loading rate of 0.012 g-NH+ 4 -N g-VSS
1
day1,
 
NO2 -N and NO3 -N were detected (3.08 and 1.04 mg L1,
respectively) after 360 min (Fig. 3b and c). Under the latter COD was exhausted and the DO increased to values above
conditions, the heterotrophic nitrifiers may have domi- 2.21 mg L1 (Fig. 4c), which inhibited the SND process.
nated and utilized the available organic carbon and DO. Hence, NO 
2 -N and NO3 -N were observed to increase
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6 Y.-C. Chiu et al. / International Biodeterioration & Biodegradation 59 (2007) 1–7

Anoxic Oxic Settling and Draw Table 3

Comparison of Ks values of an SBR system ran at different ammonium
Ammonium Loading Rate=0.012 loading rates and F/Ma ratios
300 Ammonium Loading Rate=0.029
Ammonium Loading Rate=0.046
Operation condition
a
200 F/M Ammonium MLVSS Ks (day1) R2
ORP (mV)

loading (mg L1)

rateb

100 0.136 0.012 4200 0.190 0.95

0.326 0.029 4300 1.112 0.96
0.489 0.046 4100 0.382 0.96
0 a
F/M: g-COD g-VSS1 day1.
b
(a) Ammonium loading rate: g-NH+
4 -N g-VSS
1
day1.
0 120 240 360 480 600 720

not shown). The fact that COD was completely exhausted

7.6
in the effluent at the ammonium-loading rates of both
0.012 and 0.029 g-NH+ 4 -N g-VSS
1
day1 demonstrated
that the carbon source was entirely utilized in the SND
7.2 system.
pH

The Grau model can be used to obtain the first-order

COD removal rate constant (Ramalho, 1997) as described
6.8
by Eq. (2):
S0  Se Se
q¼ ¼ Ks , (2)
X V th S0
(b)
6.4 where q is the specific rate of COD removal (g-COD g-
0 120 240 360 480 600 720 VSS1 day1), S0 the influent COD concentration
8
(mg L1), Se the effluent COD concentration (mg L1),
Xv the biomass in reactor (mg-MLVSS L1), th the reaction
time (day), Ks the Grau model first-order COD removal
6
rate constant (day1).
The multiple correlation coefficients (R2) of Eq. (2) were
DO (mg/L)

4
0.95, 0.96, and 0.96, respectively, under the three different
ammonium-loading rates of 0.012, 0.029, and 0.046 g-
NH+ 4 -N g-VSS
1
day1 and F/M ratios of 0.136, 0.326,
2
and 0.489 g-COD g-VSS1 day1, respectively (Table 3).
The COD removal rate constant (Ks) was 0.190, 1.112, and
0.382 day1 and the highest Ks of 1.112 day1 was obtained
0
for F/M ¼ 0.326 day1 , showing that a suitable ammo-
0 120 240 360 480 600 720
nium-loading rate and F/M ratio are necessary for
achieving effective removal of both nitrogen and COD.
(c) Time (min)

Fig. 4. Profiles of ORP, pH, and DO during one 12-h cycle of an SBR
when the ammonium-loading rate was adjusted at either 0.012, 0.029, or
0.046 g-NH+4 -N g-VSS
1
day1.
gradually between 150 and 300 min (Fig. 3b and c). von
Münch et al. (1996) proposed that a balanced SND system
should not generate any nitrite or nitrate. Fig. 3b shows
that NO2 -N decreased slightly from 3.08 to 1.18 mg L
1

while Fig. 3c shows an increase of NO3 -N from 1.04 to
2.28 mg L1 during the 300–600-min period. The results
show that part of NO 
2 -N was converted to NO3 -N by
autotrophic nitrification during this period, which is also
evidenced by the observed consumption of alkalinity (data
4. Conclusions
In this study, we showed that both the ammonium and
the organic loading rates affect the occurrence of the SND
process in the SBR system. The SBR system was operated
as an efficient SND-based SBR system when the COD/
NH+ 4 -N ratio was controlled at 11.1, resulting in nearly
complete removal of both organic matter and NH+ 4 -N with
no accumulation of intermediate byproducts (NO 2 -N).
The nitrogen removal efficiency decreased gradually with
increasing ammonium-loading rates at constant COD
loading. Therefore, appropriate control of the carbon
source concentration can stimulate the SND in a tradi-
tional SRB to optimize biological nutrient removal.
 ARTICLE IN PRESS
Y.-C. Chiu et al. / International Biodeterioration & Biodegradation 59 (2007) 1–7
Acknowledgments
We thank the National Science Council of the Republic
of China for providing financial assistance (NSC 93-2211-
E-029-004) to complete this work.
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