ENVIRONMENTAL ENGINEERING SCIENCE REVIEW ARTICLE

Volume 27, Number 2, 2010

ª Mary Ann Liebert, Inc.
DOI: 10.1089=ees.2009.0100

Effective Biological Nitrogen Removal Treatment Processes

for Domestic Wastewaters with Low C=N Ratios: A Review
Sheng-Peng Sun,1,2,* Carles Pellicer i Nàcher,2 Brian Merkey,2 Qi Zhou,1
Si-Qing Xia,1 Dian-Hai Yang,1 Jian-Hui Sun,3 and Barth F. Smets2
1
State Key Laboratory of Pollution Control and Resources Reuse, College of Environmental Science and Engineering,
Tongji University, Shanghai, People’s Republic of China.
2
Department of Environmental Engineering, Technical University of Denmark, Kgs. Lyngby, Denmark.
3
Henan Key Laboratory for Environmental Pollution Control, College of Chemistry and Environmental Sciences,
Henan Normal University, Xinxiang, Henan, People’s Republic of China.

Received: March 22, 2009 Accepted in revised form: October 28, 2009

Abstract
Discharge of nitrogenous components to water bodies can cause eutrophication, deterioration of water quality,
toxicity to aquatic life, and pose a potential hazard to human and animal health. Biological nitrogen removal can
remove nitrogenous components via conversion to harmless nitrogen gas with high efficiency and relative low costs.
However, the removal of nitrogen from domestic wastewater with a low carbon=nitrogen (C=N) ratio can often be
limited in municipal wastewater plants (WWTPs) because organic carbon is a limiting factor for denitrification. The
present work reviews innovative bacterial nitrogen removal pathways such as shortcut nitrification=denitrification,
simultaneous nitrification=denitrification, and the nitritation Anammox process, which can remove nitrogen with
low or zero dosage of organic carbon sources. We conclude that advanced process control and some new biological
treatment processes including the modified anaerobic=anoxic=oxic (A2=O) process, the step-feed multistage an-
aerobic=oxic (A=O) process, and new reactors like the membrane bioreactors (MBRs) and the membrane-aerated
biofilm reactors (MABRs) can support the innovative biological nitrogen removal pathways. They can effectively
be used for nitrogen removal from low C=N domestic wastewater without external carbon addition. In addition,
conventional and alternative carbon sources for enhanced biological nitrogen removal were also reviewed. We
conclude that alternative carbon sources such as wine distillery effluent, the leachate of food waste, digested piggery
manure, hydrolyzed molasses, biologically hydrolyzed or mechanically disintegrated sludge offer the same or better
performance for nitrogen removal at reduced costs. Finally, we suggest that (1) these new processes and technol-
ogies are implemented at large scale for nitrogen removal from low C=N domestic wastewater, (2) further method
logic are explored to introduce the Anammox pathway into domestic wastewater treatment, and (3) alternative
carbon sources are explored and optimized for supporting the denitrification. With these efforts, cost-effective
nitrogen removal from low C=N ratio domestic wastewater can be obtained in the near future.

Key words: nitrogen removal; C=N ratio; nitrification; denitrification; Anammox; carbon source; domestic
wastewater; WWTP

Introduction Environmental protection has led to the implementation of

continuously increasing wastewater treatment plant (WWTP)

H uman-induced eutrophication occurs due to the

discharge of high nutrients loads. Eutrophication re-
duces water quality, alters the ecological structure and func-
tion of freshwater, and poses many potential hazards to
human and animal health (Ahn, 2006; Dodds et al., 2009).
*Corresponding author: State Key Laboratory of Pollution Con-
trol and Resources Reuse, College of Environmental Science and
Engineering, Tongji University, Shanghai 200092, People’s Republic
of China. Phone: þ86 21 6598 2692; Fax: þ86 21 6598 2689; E-mail:
sunsp@yahoo.cn
discharge standards for nutrients and has impelled efforts to
achieve better effluent quality especially with regard to ni-
trogen (Henze et al., 2008).
Biological nitrogen removal is preferred over physicochem-
ical processes because it is capable of removing fixed nitroge-
nous compounds to harmless dinitrogen gas (N2) in a more
effective and economical way (Ahn, 2006; Kim et al., 2008).
Nitrogenous matter in wastewater can be divided into two
categories: nonbiodegradable and biodegradable. The nonbio-
degradable nitrogenous fraction is associated with the nonbio-
degradable particulate chemicical oxygen demand (COD). The

111
112
biodegradable nitrogenous matter is subdivided into ammonia
(NH4þ ), soluble organic nitrogen, and particulate organic ni-
trogen. Particulate organic nitrogen is hydrolyzed to soluble
organic nitrogen. The soluble organic nitrogen is acted on by
heterotrophic bacteria and converted to NH4þ . The NH4þ
serves as the nitrogen supply for synthesis of heterotrophic
biomass and as the energy supply for growth of autotrophic
nitrifying bacteria (Henze et al., 2000). Presently, the most
commonly used approach for nitrogen removal in WWTPs
comprises a combination of aerobic autotrophic nitrification
of NH4þ to nitrite (NO2
) and nitrate (NO3
) and later anoxic
heterotrophic denitrification of the oxidized N-species to N2
(Pynaert et al., 2004). Biological denitrification is carried out
by heterotrophic bacteria (HB), which require a biodegrad-
able organic carbon source as an electron donor. Thus, the
denitrification potential of wastewater is primarily a function
of the available organic carbon (usually expressed as biode-
gradable chemical oxygen demand (bCOD)=nitrogen (N)
[carbon=nitrogen (C=N)] or biochemical oxygen demand
(BOD)=N ratio (Kujawa and Klapwijk, 1999). Without exter-
nal carbon sources, the specific denitrification rates (SDNR),
range from 0.2 to 0.6 mg-N=g-mixed liquor volatile sus-
pended solids (MLVSS)=h due to endogenous processes
(Kujawa and Klapwijk, 1999). Beccari et al. (1983) suggested
minimum C=N ratios of 13, and a similar value was inferred
with full-scale predenitrification experience (Komorowska-
Kaufman et al., 2006). Henze et al. (1994) calculated that a
minimum C=N ratio of 3.5–4.5 g-COD=g-N and proposed
that 6–11 g-COD=g-N could achieve good nitrogen removal
in single sludge systems with predenitrification. The C=N
ratio of domestic wastewaters is, however, often lower than
these prescribed values, and nitrogen removal is limited by
the lack of available organic carbon source (Ryu and Lee,
2009). It is desired for many municipal WWTPs to look for
practical and cost-effective alternative technologies for nitro-
gen removal.
Given the mentioned references above and the C=N ratio of
typical domestic wastewater (10.5–12.5) (Tchobanoglous et al.,
2004), we define a low C=N ratio domestic wastewater as a
wastewater with a C=N ratio lower than 8.0. To get satisfac-
tory nitrogen removal performance for wastewater with a
C=N ratio lower than the critical value, one way is to intro-
duce an innovative nitrogen removal pathway or treatment
processes that can support nitrogen removal with a low or
zero organic carbon demand; the other way is to add external
carbon for denitrification. Therefore, this article reviews in-
novative nitrogen removal pathways and available biological
treatment processes that are capable of nitrogen removal with
low or zero organic carbon demand. In addition, conventional
and alternative external carbon sources that can be used for
enhanced nitrogen removal are also discussed and summa-
rized. Hopefully, this review can provide guidance to mu-
nicipal WWTPs operators and planners to select cost-effective
nitrogen removal alternatives for the treatment of low C=N
ratio domestic wastewater.
Feasible N-Removal Pathway for Wastewater
with Low C/N Ratio
During the past 30 years several innovative nitrogen
removal pathways have been discovered such as shortcut
nitrification=denitrification, simultaneous nitrification=
SUN ET AL.
denitrification, and anaerobic ammonium oxidation (Ana-
mmox) (Ahn, 2006; Peng and Zhu, 2006; Xia et al., 2008b).
Shortcut nitrification=denitrification
Because NO2
is an intermediate compound in both nitri-
fication and denitrification, a pathway may be formed by
controlling the extent of NH4þ oxidation to NO2
(nitritation)
instead of to NO3
(nitratation) and then coupled by reduction
of the accumulated NO2
via denitrification (Ruiz et al., 2006;
Yang et al., 2007).
The stoichiometric equations of the conventional
nitrification=denitrification process and the shortcut
nitrification=denitrification process are shown below [Eqs.
(1)–(6)] (Henze et al., 2008).
Conventional nitrification=denitrification:
Nitrification: NH4þ þ 1:5 O2 ! NO2
þ H2 O þ 2H þ (1)
NO2
þ 0:5 O2 ! NO3
(2)
Denitrification: NO3
þ 4g COD þ H þ
(3)
! 0:5 N2 þ 1:5g biomass
Summary: NH4þ þ 2 O2 þ 4g COD ! 0:5 N2 þ H2 O
(4)
þ H þ þ 1:5g biomass
Shortcut nitrification=denitrification:
Nitrification: NH4þ þ 1:5 O2 ! NO2
þ H2 O þ 2H þ (1)
Denitrification: NO2
þ 2:4g COD þ H þ
(5)
! 0:5 N2 þ 0:9g biomass
Summary: NH4þ þ 1:5 O2 þ 2:4g COD ! 0:5 N2
(6)
þ H2 O þ H þ þ 0:9g biomass
From Equations (1)–(6), it can be seen that the shortcut ni-
trification followed by denitrification of NO2
can reduce
approximately 40% of carbon requirements, 25% of the oxy-
gen supply, and result in 40% less biomass produced com-
pared to oxidation to and denitrification of NO3
. The
reductions in the COD requirement by this pathway make
nitrogen removal from low C=N ratio domestic wastewater
feasible. On top of that, NO2
denitrification rates are 1.5–2
times faster than NO3
denitrification rates (Peng and
Zhu, 2006; Aslan and Dahab, 2008; Kornaros et al., 2008). In
full-scale WWTPs average maximum specific ammonia utili-
zation rates (nitrification) are from 0.03 to 0.18 g-NH4þ -N=g-
MLVSS  day (0.06–0.54 kg-NH4þ -N=m3=day), whereas the
observed specific denitrification rate (SDNR) are 0.04–
0.42 g-NO3
-N=g-MLVSS=day(0.08–1.36 kg-NO
3
3 -N=m =day)
for predenitrification with sufficient carbon source and
0.01–0.04 g-NO3
-N=g-MLVSS=day (0.02–0.12 kg-NO3
-N=
m3=day) for postdenitrification without an exogenous carbon
(Kristensen et al., 1992; Munch et al., 1996; Tchobanoglous
et al., 2004; Park et al., 2007).
NITROGEN REMOVAL FOR LOW C/N DOMESTIC WASTEWATER
Unfortunately, NO2
-N accumulation is difficult to attain
because nitrite oxidizing bacteria (NOB) generally have
higher substrate-utilization rates than ammonia oxidizing
bacteria (AOB) (Kim et al., 2008). However, it is technically
feasible to achieve shortcut nitrogen removal via NO2
by
specific inhibition or removal of NOBs while retaining AOBs
(Wu et al., 2007; Blackburne et al., 2008). These methods are
based on the fact that AOBs and NOBs have different physi-
ological characteristics and responses to environmental fac-
tors (Gali et al., 2006b). For instance:
1. Dissolved oxygen (DO) concentration. It has been
found that NOBs have lower affinity for oxygen than
AOBs, with DO half-saturation coefficients of 1.2–
1.5 mg=L and 0.2–0.4 mg=L, respectively. Therefore, the
growth of NOBs can be limited by lowering the DO
concentrations at values between 0.3 and 1.2 mg-O2=L
(Peng and Zhu, 2006; Chuang et al., 2007; Kim et al.,
2008; Aslan et al., 2009).
2. pH value. Free ammonia (FA) and free nitrous acid
(FNA) inhibit the growth of AOBs and NOBs at dif-
ferent concentration levels. The concentrations of FA
and FNA can be calculated according to Equations (7)
and (8) as function of the pH value (Anthonisen et al.,
1976),
17 TAN · 10pH
FA(mg=L) ¼ · (7)
14 [e6344=(273 þ T) þ 10pH ]
47 TNN
FNA(mg=L) ¼ · (8)
14 [e
2300=(273 þ T) · 10pH þ 1]
where TAN is the total ammonium as nitrogen
(TAN ¼ NH4þ - N þ NH3 - N, mg - N=L), TNN is the
total nitrite as nitrogen (TNN ¼ NO2
- N þ HNO2 - N,
mg - N=L), T is the temperature (8C). Anthonisen et al.
(1976) reported that from 0.1–1.0 mg-FA=L NOBs ac-
tivity is inhibited, whereas concentrations between 10
and 150 mg-FA=L are necessary to inhibit AOBs. In-
hibition of NOBs by FNA is reported at concentrations
between 0.011 and 2.8 mg-FNA=L (Anthonisen et al.,
1976; Vadivelu et al., 2006, 2007a, 2007b). Vadivelu et al.
(2006, 2007a, 2007b), however, also reported that
growth of the AOBs (Nitrosomonas) become inhibited at
approximately 0.10 mg-FNA=L and are completely
stopped at 0.40 mg-FNA=L. Although NOBs are in-
hibited at concentrations as low as 0.1 to 1 mg-FA=L, the
threshold inhibitory concentration of FA gradually
increases with time because NOBs can tolerate and
acclimate to the increasing levels of FA (up to 40 mg-
FA=L), resulting in an irreversible decline in NO2
ac-
cumulation efficiency (Wong-Chong and Loehr, 1978;
Turk and Mavinic, 1989; Villaverde et al., 2000).
3. Temperature. It is possible for AOBs to effectively
outcompete NOBs at temperatures above room tem-
perature (e.g., the specific growth rate of AOBs is ap-
proximately two times that of NOB at 358C) (Bougard
et al., 2006; Aslan and Dahab, 2008; Xue et al., 2009).
Therefore, NOBs can be washed out in a contriuous
stirred-tank reactor (CSTR) by controlling the sludge
retention time (SRT) at values between the critical SRT
113
for NOBs and AOBs. The SHARON (single reactor
high-activity ammonia removal over nitrite) process
relies on successfully washing out NOBs from a reactor
operated at high temperature between 30 and 408C and
an SRT of 1–1.5 days.
4. Inhibitors. Besides FA and FNA, some volatile fatty
acids such as formic, acetic, propionic and n-butyric
acid, and some heavy metals such as chromium (Cr),
nickel (Ni), copper (Cu), zinc (Zn), lead (Pb), and cad-
mium (Cd) preferentially inhibit NOBs (Peng and Zhu,
2006). Eilersen et al. (1994) showed that concentrations
of formic acid above 100 mg=L inhibited NOBs with
no adverse effect on AOBs. More recently, Blackburne
et al. (2008) used 40 mg=L of formic acid to successfully
initiate NO2
accumulation in a sequencding batch re-
actor (SBR) system. However, the use of inhibitors can
also bring an increase in the operation cost, the risk of
inhibiting other bacteria, and the possibility of them
appearing in the effluent as secondary pollutants.
Table 1 summarizes the nitrogen removal from domestic
wastewater based on shortcut nitrification=denitrification in
recent years. Total nitrogen (TN) removal rates between 0.08
and 0.238 kg-N=m3=day were obtained for the domestic
wastewater with a C=N ratio between 2.1 and 4.4 in a lab- and
pilot-scale SBR reactor or A=O process.
Simultaneous nitrification=denitrification
Simultaneous nitrification=denitrification was inferred from
the observation that TN losses of up to 30% are frequently
reported in aerated tanks of the activated sludge process (Hao
et al., 1997; Yoo et al., 1999; Zhao et al., 1999; Fuerhacker et al.,
2000; Ammary and Radaideh, 2005). One prevailing explana-
tion to this phenomenon is the simultaneous activity of auto-
trophic nitrification on the surface of the activated sludge flocs,
with denitrification occurring in the floc interior supported by
the existing DO gradients (Fig. 1) (Zhao et al., 1999; Holman
and Wareham, 2005). A second explanation is that the contin-
ued ability of heterotrophic denitrifiers to reduce nitrogen
under aerobic conditions (Holman and Wareham, 2005). A last
theory postulates that denitrifiers possess is greater physio-
logical variety than previously expected. In particular, it has
been suggested that some denitrifiers are also heterotrophic
nitrifiers and can directly convert NH4þ into gaseous nitrogen
end products (Zhao et al., 1999; Holman and Wareham, 2005).
It should also be pointed out that AOBs such as Nitrosomonas
europea and Nitrosomonas eutropha are able to denitrify in the
presence of small amounts of oxygen (Helmer and Kunst,
1998), and some NOBs strains can denitrify as well (Schmidt
et al., 2003). However, the contribution of these organisms
to denitrification is speculative.
COD concentration, DO concentration, and floc size
are significantly affecting the simultaneous nitrification=
denitrification performance in activated sludge systems
(Pochana and Keller, 1999). An optimal C=N ratio was re-
ported at 11.1, where the nitrification and the denitrification
reactions are in balance (Chiu et al., 2007). The optimal bulk
DO concentration was reported to be 0.5–1.5 mg=L (Bernat
and Wojnowska-Baryla, 2007) with an optimum floc size of
80–100 mm. Simultaneous nitrification=denitrification also oc-
curs in some other wastewater treatment processes besides
the activated sludge process, such as biofilm systems (Helmer
114
FIG. 1. Schematic diagram
of simultaneous nitrification=
denitrification in an activated
sludge floc.
and Kunst, 1998; Hibiya et al., 2003), slow sand filters (Nakhla
and Farooq, 2003), biological aerated filters (Lemoine et al.,
2006) and single packed bed batch reactors (Daniel et al.,
2009).
Compared to the conventional nitrification=denitrification
pathway, simultaneous nitrification=denitrification offers
several advantages (Andrade do Canto et al., 2008; Li et al.,
2008b): (1) it utilizes 22–40% less carbon source and reduces
sludge yield by 30%, (2) it exerts less alkalinity demand be-
cause alkalinity is consumed during nitrification but pro-
duced during denitrification, (3) it has smaller footprint
because it eliminates the need for either two separate tanks
operated in series or intermittent aeration operation in a sin-
gle tank, (4) it consumes less energy due to the reduction
in aeration requirement. Table 2 summarizes the different
processes for nitrogen removal from domestic wastewater
based on simultaneous nitrification=denitrification. TN re-
moval rates between 0.072 and 0.121 kg-N=m3=day have
been achieved for domestic wastewater with a C=N ratio of
2.7–4.9.
Anammox process
Anaerobic ammonium oxidation (Anammox) is an impor-
tant microbial pathway in the nitrogen cycle that could be
used to yield a new cost-effective technology to remove NH4þ
from wastewater. Anammox bacteria (AnAOB) have the un-
ique metabolic ability to oxidize NH4þ with NO2
as electron
acceptor to produce N2 and a small part of NO3
under anoxic
conditions without the requirement of an organic carbon
source (Date et al., 2008; Kuenen, 2008). AnAOBs belong to the
Planctomycetes. So far, two freshwater species, Candidatus
Brocadia anammoxidans and Candidatus Kuenenia stuttgartien-
sis, three marine species, Candidatus Scalindua sorokinii,
Candidatus Scalindua brodae, and Candidatus Scalindua wag-
neri, and a mixotrophic anammox bacterium Candidatus
Anammoxglobus propionicus have been described (Tsushima
et al., 2007). The stoichiometry of the Anammox reaction is
accepted to be as in Equation (9) (Strous et al., 1998).
SUN ET AL.
1 NH4þ þ 1:32 NO2
þ 0:066 HCO3
þ 0:13 H þ
! 1:02 N2 þ 0:26 NO3
þ 0:066 CH2 O0:5 N0:15
þ 2:03 H2 O (9)
Compared to the conventional biological nitrogen removal,
the Anammox process presents several advantages such as
100% savings on external organic carbon source, 63% less
oxygen demand, small surplus biomass production, and re-
duced carbon dioxide (CO2), nitrous oxide (N2O), and nitric
oxide (NO) emissions (Feng et al., 2007; Liu et al., 2008).
Anammox has attracted a lot of attention in the wastewater
engineering field in recent years as a key element in alterna-
tive nitrogen removal process. However, in order to take
advantage of the Anammox, partial nitritation is required. In
the oxidation of induced N-species (ammonia and organic-N),
the conversion of NO2
to NO3
should be avoided and the
amount of NH4þ converted to NO2
should be limited to ap-
proximately 60% (Yamamoto et al., 2008). Many novel bio-
logical nitrogen removal processes have been developed to
couple nitritation to Anammox. They can be grouped in two
categories: (1) two-reactor processes where a CSTR perform-
ing partial nitrification is followed by another reactor per-
forming the Anammox process (SHARON=Anammox) (Van
Dongen et al., 2001; Hwang et al., 2005), and (2) the one reactor
processes with simultaneously nitritation and Anammox oc-
curring, such as completely autotrophic nitrogen removal
over nitrite (CANON) (Sliekers et al., 2002; Third et al., 2005)
and oxygen-limited autotrophic nitrification denitrification
(OLAND) (Windey et al., 2005). Both processes have been
successfully used for the treatment of concentrated NH4þ
streams such as sewage sludge digester liquor, swine waste-
water digester liquor, livestock wastewater, seafood processing
wastewater, power plant wastewater, and other ammonium-
rich wastewaters at lab-, pilot-, and=or full scales (Lamsam
et al., 2008; Yamamoto et al., 2008). An outstanding case is the
first full-scale Anammox reactor with a volume of 70 m3 and
treatment ability of 750 kg-N=day at the sludge treatment plant
Sluisjesdijk (Rotterdam, The Netherlands) (Van der Star et al.,
 Table 1. Nitrogen Removal from the Low Carbon=Nitrogen Ratio Domestic Wastewater Based on Shortcut Nitrification and Denitrification

Wastewater characteristics Operating conditions Results and comments

TN
removal
NO2
= COD NH4þ- N TN rate
Process COD NH4þ- N TN HRT SRT DO Temperature NOx removal removal removal (kg-N=m3
Year and scale (mg=L) (mg=L) (mg=L) C=N (h) (days) pH (mg=L) (8C) (%) (%) (%) (%)  day) References

2003 SBR, Lab-scale 150TOC 50 50 3a 6 No 7.2–7.3 0–6.8 25 97% N=A 98% 98% 0.196 (Katsogiannis
control et al., 2003)
2007 SBR, Lab-scale 123.3–392.5 65.4–150 68.5–150 2.1a 6–12 14–18 7.35–7.9 0–7.0 28 80% N=A 80% 74% 0.222 (Wu et al., 2007)
2007 SBR, Lab-scale 160–310 58–108 59–110 2.8a 12 20 6.9–7.85 0.6a 23  1 95.6% 80% 95% 50% 0.080 (Peng et al., 2007)
2007 SBR, Pilot-scale 142.3a 60.05a 60.05* 2.4a 6 13 6.9–7.8 0–6.0 11.9–26.5 95% N=A 95% 95% 0.238 (Yang et al., 2007)
2007 A=O, Pilot-scale 163.2–389 65.4–105.7 68.5–110.2 2.9a 9 15 7.5 0.3–0.8 20–22 90% N=A 93% 63% 0.147 (Wang et al.,
2007)
2008 A=O, Lab-scale 160–500 50–100 60–110 4.4a 10–12 15–20 7–8 2.5–3.0 22  2 90% 88% 92% 87% 0.146 (Wang et al.,
2008b)
2009 A=O, Pilot-scale 232.6  67 76.9  15 78.2  15 3.0a 9–10 15 7.5 0.4–0.7 21  2 95% N=A 98% 70% 0.146 (Ma et al., 2009)
a
Mean value.
DO, dissolved oxygen; SRT, sludge retention time; HRT, hydraulic retention time; TN, total nitrogen; COD, chemical oxygen demand; NO
2, nitrite; NOx, nitrite plus nitrate; SBR, sequencing batch
reactor; A=O, anaeorbic=oxic process; C=N, carbon=nitrogen.

Table 2. Nitrogen Removal from the Low Carbon=Nitrogen Ratio Domestic Wastewater Based on Simultaneous Nitrification=Denitrification

Wastewater characteristics Operating conditions Results and comments

COD NH4þ- N TN TN removal

Process COD NH4þ- N TN HRT SRT DO Temperature removal removal removal rate (kg-N=
Year and scale (mg=L) (mg=L) (mg=L) C=N (h) (d) pH (mg=L) (8C) (%) (%) (%) m3  day) References

2006 Orbal oxidation 160–310 58–108 59–110 2.7a 16.5 N=A N=A 0.1–0.7 20–26 80% 90% 72% 0.085 (Gao et al., 2006)
ditch, Pilot-scale
2008 SHBR, Lab-scale 350a 65a 71a 4.9a 12 10 7.0–8.0 0.3–1.5 23–25 92% 93.5% 85% 0.121 (Wang et al., 2008a)
2008 BMBR, Pilot-scale 10–38TOC N=A 9–25 1.4a 4.7 29 N=A 0–6.5 10–15 85%TOC N=A 77% 0.102 (Kimura et al., 2008)
2008 SBBR, Lab-scale 100 50 N=A 3b 8 N=A 7.3–8.8 2.0 30  1 89% 97% 80% 0.072 (Andrade do Canto
et al., 2008)
2009 MBR, Pilot-scale 586a 125a 156a 3.8a 7.2 36 N=A 1.8–2.0 20–25 N=A 50% 37% 0.19 (Sarioglu et al., 2009)
a
Mean value.
b
The ratio was calculated by the addition of two different kinds wastewater (I: 100 mg=L COD and 50 mg=L NH4-N, II: 100 mg=L COD, VI=VII ¼ 2:1).
116
2007). Additional high TN removal rates of about 26 kg-
N=m3=day have been obtained in an up-flow fixed-bed Ana-
mmox biofilm column reactor (Tsushima et al., 2007).
Although Anammox can present a very cost-effective bio-
logical nitrogen removal pathway, there are no report on the
nitrogen removal from low C=N ratio domestic wastewater
by Anammox process. Some reasons for this maybe: (1) long
reactor start-up periods due to extremely slow growth rates of
Anammox bacteria (with doubling time of about 11 days)
(Strous et al., 1998). For the first full-scale Anammox reactor in
Rotterdam, no detectable Anammox activity was observed
during the first 800 days of operation, and stable conversion
did not take place until day 1,235 (Van der Star et al., 2007).
Recently, a new cultivation system for Anammox bacteria
was reported in which the organisms grow in suspension at
significantly higher enrichment, but the doubling time was
still approximately 8 days (Van der Star et al., 2008). (2) Their
growth is reversibly inhibited by oxygen concentrations
above 0.5% air saturation (about 0.042 mg-O2=L) and also ir-
reversibly inhibited by NO2
concentrations higher than
70 mg-NO2
-N=L (Schmidt et al., 2003; Kuenen, 2008). (3) Still
little is known about how the Anammox organisms interact
with denitrifiers, nitrifiers and dissimilatory nitrate reducers
when organic compounds are present (Kuenen, 2008). Because
AnAOBs have a much lower growth rate than denitrifiers, they
are easily outcompeted by denitrifiers, and Anammox reaction
cannot be sustained if organic carbon (COD=N > 2–5) is pres-
ent in the influent (Lackner et al., 2008). Therefore, more
experience is necessary before Anammox is successfully in-
corporated in the technologies for stable and reliable nitrogen
removal from low C=N domestic wastewater.
Figure 2 shows an indicative comparison between con-
ventional nitrification=denitrification and the rest of the de-
scribed microbial pathways. The reduction in organic carbon
demand in the denitrification step, the lower oxygen re-
quirements, and the reduced sludge production by these al-
ternate pathways are the main differential features of these
processes. Additionally, higher removal rate can also be ob-
tained by these pathways for low C=N ratio domestic
FIG. 2. Comparison the nitrogen removal by conventional
nitrification and denitrification, shortcut nitrification and
denitrification, simultaneous nitrification=denitrification, and
Anammox process.
SUN ET AL.
wastewater treatment compared to that by the conventional
nitrification=denitrification.
Alternative Novel Treatment Processes or Reactors
Common biological nitrogen removal processes occurs in
various treatment train configurations in WWTPs, including
the A=O process, the A2=O process, the University of Cape
Town (UCT) process, the Modified Bardenpho, and the Vir-
ginia Initiative Plant (VIP) process. All these processes rely on
a predenitrification zone where a portion of the nitrified
wastewater is recycled and mixed with the influent to serve as
an electron donor for denitrification. Disadvantages include
the need for high recirculation rates and the addition of ex-
ternal carbon substrate when the influent C=N ratio is not
high enough (Downing and Nerenberg, 2008b). To overcome
this situation, advanced process control methods, new bio-
logical treatment processes and reactors such as the modified
A2=O process, the multistage A=O step-feed process, mem-
brane bioreactors (MBRs) and membrane-aerated biofilm
reactors (MABRs) have been developed, many of them sup-
porting the biological removal pathways described in previ-
ous sections.
Advanced process control for achieving shortcut
nitrogen removal
Control of DO, pH, and temperature have been success-
fully used for shortcut nitrogen removal from high strength
ammonium containing wastewaters such as sanitary landfill
leachate, sewage sludge digester liquor, soybean wastewater,
and industrial wastewater (Wang et al., 2004; Ciudad et al.,
2005; Fux et al., 2006; Lai et al., 2004; Pedersen et al., 2003; Aslan
and Dahab, 2008; Spagni and Marsili-Libelli, 2009). However,
it is difficult and not economical to achieve shortcut nitrogen
removal for domestic wastewater treatment by maintaining
a relatively high pH and temperature for several thousand
or million tons=day of domestic wastewater in municipal
WWTPs.
The online monitoring of parameters such as pH, DO,
oxidation reduction potention (ORP), and N-species (NH4þ ,
NO2
and NO3
) can be helpful for achieving shortcut nitro-
gen removal in domestic wastewater treatment process.
Especially in batch mode, the end of nitrification and deni-
trification can be readily obtained from the typical bending
points of pH, DO, and ORP profiles (Yang et al., 2007).
Therefore, real-time control strategy regulating the operation
time of aerobic and anoxic phases can be successful to
achieving shortcut biological nitrogen removal (Fuerhacker
et al., 2000; Gao et al., 2003; Peng et al., 2004; Wu et al., 2007;
Tanwar et al., 2008). In addition, studies have indicated that
excessive energy and resource consumption could also be
avoided as well by using a real-time control strategy.
For instance, Yang et al. (2007) and Wang et al. (2008b) used
the real-time control strategy with pH (dpH=dt) and ORP
(dORP=dt) as the control parameter in a pilot-scale SBR
(working volume of 54 m3) and alternating activated sludge
system (AAS) for real domestic wastewater treatment. The
results showed that nitrogen removal via the nitrite pathway
could be successfully achieved with an average NO2
accu-
mulation rate above 95% at either normal or low temperature
(11.9–26.58C) in the SBR system and above 90% in the AAS
NITROGEN REMOVAL FOR LOW C/N DOMESTIC WASTEWATER 117

system. In addition, approximately 20% aeration energy was A2=O process, (1) the recycle sludge is directed to a separate
saved compared to the conventional steady-state control preanoxic basin where hydrolysis processes can release
systems. biodegradable organic carbon, which can be used in the de-
nitrification processes taking place downstream. (2) The in-
Modified A2=O process fluent wastewater goes directly to the anaerobic zone of the
reactor and gets mixed with the wastewater from the pre-
The A2=O process with separate anaerobic, anoxic, and
anoxic tank. Part of mixed wastewater in this reactor is re-
aerobic tanks is a suitable method for biological nitrogen re-
circulated to the postanoxic zone at a ratio of 0.4Qin to provide
moval (schema is shown in Fig. 3a). Denitrification of the
available organic carbon for denitrification. (3) In the first
NO3
recirculated from a downstream aerobic tank occurs in
aerobic zone, NH4þ is oxidized to NO2
and NO3
, and both
an anoxic tank where denitrifiers can utilize the organic
are fed continuously to the so-called postanoxic zone for de-
matter present in the influent, avoiding the need for an ad-
nitrification. (4) The treated wastewater passes through a final
ditional organic carbon source (Ouyang et al., 2000). However,
aerobic tank to minimize the amount of COD in the effluent,
the A2=O configuration normally requires a high mixed liquid
and enhances the settling ability of the sludge by minimiz-
return ratio (2–4Qin) from the aerobic zone to the anoxic zone
ing denitrification in the secondary settler. Pilot-scale re-
to bring more NO3
back for denitrification. High return ratios
sults showed more than 88% COD and 70% TN was removed
can result in DO concentration increase and COD dilution in
under a volumetric loading rate of 0.9 kg-COD=m3=day
the anoxic zone, which inevitably deteriorates the denitrifi-
and 0.12 kg-N=m3=day for the real domestic wastewater
cation efficiency, especially when the organic matter present
treatment.
in the influent wastewater is insufficient to deplete the DO
present in the recycled mixed liquor. In addition, high return
Step-feed multistage A=O process
ratios also lead to higher energy consumption and increased
operating costs. The step-feed multistage A=O process consists of two or
Yang et al. (2006) modified the A2=O process to avoid these more denitrification–nitrification units in series with waste-
disadvantages (schema is shown in Fig. 3b). In the modified water distributed at several reactor points so that an internal

FIG. 3. Schematic diagram of the anaerobic=anoxic=oxic (A2=O) process (a) and the modified A2=O process (b).
118
FIG. 4. Schematic diagram of the step-feed multistage anaerobic=oxic (A=O) process.
NO3
recirculation is unnecessary. A schema of step-feed
multistage A=O process is shown in Fig. 4. The biodegradable
organic material in the influent is utilized for denitrification,
and also simultaneous nitrification=denitrification may occur
in this process (Zhu et al., 2007b).
For the design of a step-feed multistage A=O process it is
important to optimize the influent flow distribution at each
stage, the relative split between anoxic and aerobic volumes,
and the final pass mixed liquor suspended solid (MLSS)
concentration (normally based on using an acceptable solids
loading for the secondary clarifier) (Tchobanoglous et al.,
2004). The volume or sludge loading in each stage can sig-
nificantly affect the nitrification rate. A small volume or
sludge loading might lead to a decreasing of the nitrification
rate, whereas a large load might cause overload and sludge
bulking (Zhu et al., 2007a, 2009).
Studies by Zhu et al. (2007a, 2009) indicated that a uniform l
can be used in this process for the treatment of wastewaters
with C=N ratio >9.0, by which it is simple for the process
control, optimization, and management. The optimal influent
flow distribution ratio (lOpt) can be obtained by trial method
and the influent flow rate in each stage can be calculated by
Equation (10).
n
1 n
2
QTotal ¼ q  kOpt þ q  kOpt þ K þ q  kOpt þ q (10)
where QTotal is the total influent flow rate (m3=day), q is the
influent flow rate distributed in the last stage (m3=day), and n
is the total number of the stage. However, the influent dis-
tribution ratio (l) between two adjacent stages should not be
uniform for treating domestic wastewaters with a low C=N
ratio. An optimal l with 2.0:2.1:2.5:3.4 (0.2Qin:0.21Qin:0.25-
Qin:0.34Qin) was proposed for the treatment of wastewater
with C=N ratios lower than 4.9 in a step-feed four-stage A=O
process. Under the given condition, 20% higher TN removal
efficiency could be achieved compared to the case with a
uniform l.
MBR
In an MBR, activated sludge treatment and membrane
separation are combined as a compact technology. MBRs have
SUN ET AL.
several advantages over conventional activated sludge sys-
tems including excellent removal of solids and organic matter,
ability to maintain high biomass condition, ability to handle a
wide range of SRTs at relatively short hydraulic retention
times (HRT), and a small footprint due to the elimination of
clarifier (Ersu et al., 2008; Kimura et al., 2008).
In submerged MBR, good nitrification performance can be
attained due to the high concentration of nitrifying biomass,
long SRTs, and the aerobic condition provided by the inten-
sive aeration used to mitigate membrane fouling. Almost
complete nitrification can be easily achieved in MBR, but
denitrification may limit TN removal, because a required
anoxic condition for denitrification cannot easily be created
with intensive aeration (Kimura et al., 2008; Li et al., 2008b). To
overcome these drawbacks, some modifications of MBR have
been proposed and developed:
1. A first approach involves installing an additional anoxic
reactor with MBR for denitrification, typically in pre-
denitrification configuration. Tan and Ng (2008) re-
ported almost 0.1 kg-TN=m3=day was removed from
raw domestic wastewater with a mean C=N ratio of
7.24 under mixed liquor recycle ratios of 3Qin using a
predenitrification MBR.
2. A second approach involves creating cyclic anoxic and
aerobic conditions in a single reactor MBR by inter-
mittent aeration (Choi et al., 2008).
3. A final method, the MBR’s chamber is modified to
create an anoxic or anaerobic zone within a single re-
actor along with internal mixed liquor recirculation.
Chae et al. (2006) proposed a vertical submerged membrane
bioreactor (VSMBR) (Fig. 5a). In a VSMBR, lower anoxic zone
is separated from the upper aerobic zone by a horizontal plate
with a hole in the center. The mixed liquor is recycled from the
aerobic to the anoxic zone, while raw wastewater also enters
the anoxic zone for supporting denitrification by using in-
fluent organic carbon. A pilot-scale study showed removal
rates of 0.093 kg-TN=m3=day when treating domestic waste-
water with a C=N ratio of 5.5 by the VSMBR (Chae and Shin,
2007). Similarly, Kimura et al. (2008) proposed a baffled
membrane bioreactor (BMBR) (Fig. 5b). With a baffle inserted
in the MBR’s chamber and an aeration bubbler placed inside
NITROGEN REMOVAL FOR LOW C/N DOMESTIC WASTEWATER
FIG. 5. Schematic diagram of the modified membrane bioreactor (MBR): vertical submergedmembrane bioreactor (VSMBR)
(a); baffled membrane bioreactor (BMBR) (b).
of it, aerobic and anoxic conditions are alternatively created in
the outer zone at constant intervals. A pilot-scale study
showed a 77% TN removal efficiency at a removal rate of
0.102 kg-TN=m3=day for the treatment of domestic waste-
water without external carbon addition.
Another possibility is to operate MBRs at very high MLSS
levels (25,000–30,000 mg=L), still allowing simultaneous nitri-
fication and denitrification due to the creation of an inner ag-
gregate anoxic zone to withstand heavy aeration (Sarioglu
et al., 2009). In addition, decay of the high concentration of
biomass can support endogenous denitrification. By control-
ling the bulk DO concentration at 1.5 mg-O2=L in a pilot MBR
operated without a separate anoxic volume, TN removal rates
of 0.19 kg-N=m3=day could be obtained for raw domestic
wastewater with a mean C=N ratio of 7.1 (Sarioglu et al., 2009).
Table 3 summarize the nitrogen removal results for the treat-
ment of low C=N ratio domestic wastewater by various MBRs
studies.
MABR
The MABR is another novel bioreactor technology that
combines biofilms and membrane technology. In an MABR,
gas-permeable hydrophobic membranes are used for bub-
bleless aeration and also as the support for biofilm growth.
Through control of the intramembrane oxygen supply pres-
sure and gas flow rate in MABR one can develop biofilms on
the membrane with oxic–anoxic–anaerobic zones (as shown
in Fig. 6), which support stratification of different functional
microbial populations in a single biofilm (Li et al., 2008a;
Syron and Casey, 2008a).
MABRs can support nitrification=denitrification biofilms
with higher activity compared to conventional biofilm reac-
tors because of the unique biofilm stratification and high ox-
ygen supply efficiency in MABRs. In a conventional biofilm
reactor all substrates follow the same route of transportation
and conversion processes within the biofilm. Because of their
higher growth rates and oxygen affinities, heterotrophic
bacteria generally outcompete autotrophs in the biofilm un-
less the organic carbon concentration is low. This organic
carbon fluxes to the inner anoxic area are usually insufficient
to support denitrification (Walter et al., 2005; Syron and
Casey, 2008a). In an MABR, however, oxygen and nutrients
119
are provided from two opposite sides of the biofilm. Oxygen
concentration is highest at the biofilm–membrane interface
and decreases toward the biofilm–liquid interface due to mi-
crobial activity. At the same time the nutrient concentration is
higher at the biofilm–liquid boundary, decreasing with depth
into the biofilm. With such concentration gradients, popula-
tions of nitrifiers and aerobic heterotrophs thrive adjacent to
the membrane while heterotrophic denitrifiers grow adjacent
to the liquid because of the higher carbon concentrations and
oxygen limitation there (Semmens et al., 2003; Satoh et al.,
2004; LaPara et al., 2006; Lemoine et al., 2006; Downing and
Nerenberg, 2008a; Syron and Casey, 2008b). Therefore, an
MABR can support complete nitrification=denitrification and
can offer a typically protected niche for nitrifiers against
heterotrophs, shock loads, and toxicants contained in the
wastewater influent (Syron and Casey, 2008a).
Hibiya et al. (2003) reported simultaneous nitrification=
denitrification in MABRs successful treating of domestic
wastewater. By controlling the intramembrane oxygen supply,
more than 80% TN and 90% TOC were removed under TN
and TOC loading rates of 0.16 kg-N=m3=day and 0.42 kg-
TOC=m3=day, respectively. FISH analysis showed that AOBs
were mainly located inside the biofilm while denitrifying
bacteria developed in the outer biofilm layers and in the sus-
pended sludge. Terada et al. (2003) reported 83% TN and 96%
TOC removal efficiencies in an MABR under TOC and TN
loading rates of 0.30 kg-TOC=m3=day and 0.27 kg-N=m3=day,
respectively. Matsumoto et al. (2007) modeled the effects of
C=N ratio, biofilm thickness, and surface loading of oxygen on
the efficiency of MABR performing simultaneous nitrification=
denitrification: by adjustment of the JO2 =JNH4þ flux ratio could
control simultaneous nitrification=denitrification and over
70% TN removal efficiency could be achieved at C=N ratios
ranging from 3.0 to 5.2 with biofilm thicknesses of 600–
1,200 mm. Hu et al. (2008) also reported that 86% COD, 94%
NH4þ and 84% TN removal in a carbon-membrane MABR
under COD and NH4þ loading rates of 0.698 kg-COD=m3=day
and 0.154 kg-NH4þ -N=m3=day, respectively.
In addition to traditional nitrification=denitrification,
completely autotrophic nitrogen removal can be achieved in
MABRs. Terada et al. (2007) developed a multipopulation
biofilm model to compare the conventional and MABR bio-
film reactor for autotrophic nitrogen removal: the MABR
 Table 3. Nitrogen Removal from the Low Cargon=Nitrogen Ratio Domestic Wastewater by Various Membrane Bioreactors

Results and comments

Operating conditions TN
Wastewater characteristics removal
COD NH4þ- N TN rate
COD NH4þ- N TN HRT SRT Temperature removal removal removal (kg-N=m3
Year Configuration and scale (mg=L) (mg=L) (mg=L) C=N (h) (d) pH (8C) (%) (%) (%)  day) References

1999 Additional anoxic tank 135BOD 20 34a 4.0a 13.4 72 6.7–6.9 19 99%BOD 99% 79% 0.048 (Ueda and Hata, 1999)
and sludge recirculation,
pilot-scale
2007 Vertical modified single 232  41 27  3 42  5 5.5a 8–10 60 7.3 13–25 88–97% 79–88% 63–74% 0.093 (Chae and Shin, 2007)
tank with continuous
aeration, pilot-scale
2007 Anaerobic, aerobic and 571–756 23–41 76–134 6.6a 12.2–13.7 20 N=A 18–23 96% 99% 76% 0.141 (Bracklow et al., 2007)
anoxic tanks with water
recirculation, lab-scale
2008 Additional hydrolysis 526  120 84.3  12.9 100 6.2a 48 50–200 N=A N=A 93% 98.7% 90% 0.045 (Abegglen et al., 2008)
chamber and sludge
recirculation, pilot-scale
2008 Additional anoxic tank 99–206 21–47 22–49 4.3a 4.2 N=A N=A N=A 93% 99% 90% 0.183 (Xia et al., 2008a)
and sludge recirculation,
pilot-scale
2008 Additional anoxic tank 365  74 N=A 51.7  5.0 7.2a 10 33.3 7.0–7.4 26–31 94% N=A 60–89% 0.110 (Tan and Ng, 2008)
and sludge recirculation,
pilot-scale
a
Mean value.
NITROGEN REMOVAL FOR LOW C/N DOMESTIC WASTEWATER 121

FIG. 6. Schematic diagram of the simultaneous nitrification=denitrification (a) and completely autotrophic nitrogen removal
(b) in membrane-aerated biofilm reactor (MABR) biofilm.

biofilm had a wider range for autotrophic TN removal than
that in conventional biofilm reactors and an optimal JO2 =JNH4þ
ratio of 1.73 g-O2=g-N was inferred. Furthermore, AnAOB
activity could be better maintained in the MABR biofilm
versus conventional biofilm under excess oxygen conditions
(JO2 =JNH4þ ratio >3.7). Gong et al. (2007) developed a labora-
tory-scale MABR where a gas-permeable carbon tube was
covered with nonwoven fabrics to support biofilm for single-
stage autotrophic nitrogen removal. Partial nitrification and
completely nitrogen removal were successfully achieved in
the single MABR, a max removal rate of 0.77 kg-TN=m3=day
was achieved with 88.5% TN removal efficiency after a 116-
day startup without need for any external organic carbon
source addition. Moreover, FISH analysis showed that AOBs
and AnAOBs coexisted in the MABR biofilm. The nitrifying
layer was located at the membrane–biofilm interface, whereas
the Anammox layer populated the anaerobic region closer
to the liquid–biofilm interface (Gong et al., 2008). Clearly,
MABRs are able to support single-stage completely autotro-
phic nitrogen removal, offering a promising perspective for
nitrogen removal from low C=N ratio wastewaters.
External Carbon Sources
Conventional carbon sources
Using the traditional nitrification=denitrification technol-
ogy, external carbon sources are often required to support
denitrification from nitrified low C=N domestic wastewater.

Table 4. Costs of Using Methanol, Ethanol, and Acetate for Postdenitrification

Cost C=N ratio Yield of biomass Average SDNR Cost

Carbon source ($=kg) (g COD=gNO3
- N) (g COD=g COD) (gNO3
- N=g VSS=day) ($=Kg - NO3
- N to N2 )

Methanol 0.33 5.19 0.45 0.24 1.14

Ethanol 0.84 6.08 0.53 0.71 2.43
Acetate 1.03 8.40 0.66 0.74 8.15
122 SUN ET AL.

Table 5. Chemical Characteristics of Different Alternative Carbon Sources

Pretreated Digested Nondigested Hydrolyzed Hydrolyzed Mechanically

olive oil mill piggery piggery Mecellulose Biodiesel primary secondary disintegrated
wastewater manure manure wasted liquor waste sludge sludge sludge
by lime (Obaja et al., (Obaja (Park et al., (Bodı́k (Gali et al., (Gali et al., (Kampas
Parameters (Tsonis, 1997) 2005) et al., 2005) 2005) et al., 2009) 2006a) 2006a) et al., 2007)

VFA (mg=L) 1,050 5,275 2,000  200 0 403–852

BOD5 (mg=L) 1,730 3,250
TCOD (mg=L) 68,200 3,085 7,450 58,846 1,600,000 60,000  1,000 1900  150
SCOD (mg=L) 65,800 54,494 3,000  200 55  5 4,440–5,810
NH4-N (mg=L) 1,600 720 180  10 35  5 50–60
NO2-N (mg=L) 30  3 <5
NO3-N (mg=L) 20  3 10  1
TN (mg=L) 720 19 2,000
PO4-P (mg=L) 42 147 120 100–326
TP (mg=L) 700
TS (g=L) 11.21 13.42 35  1 3.5  0.05
VS (g=L) 5.35 6.23 30  1 2.2  0.05
TSS (g=L) 2.58 3.10 0.162 35  1 2.2  0.05
VSS (g=L) 1.96 2.2 28  1 2  0.05

VFA, volatile fatty acids; BOD, biochemical oxygen demand; TP, total phosphorus; TCOD, total chemical oxygen demand; SCOD, soluable
chemical oxygen demand; VSS, volatile susplended solids; TSS, total suspended solids; VS, volatile solids.

During the past 30 years, commercially available chemicals
with methanol, ethanol, acetic acid, propionic acid, and glu-
cose have served to this end, with methanol typically pre-
ferred because of its lower cost (Table 4) (Mokhayeri et al.,
2009). Generally, only one type of carbon source is used, but
recent studies report that using mixed carbon sources might
be more effective than using a single one. For instance, Hag-
man et al. (2008) reported that a higher nitrate uptake rate was
observed by the addition of a mixture of acetate and methanol
than with each alone. Ahmed et al. (2008) also reported that
using methanol and acetate as a mixed carbon source was
more effective for nutrient removal and fouling problems
control in an MBR. This can be explained by the presence
of two different populations and each metabolizing only
one substrate, or a population of denitrifying bacteria that
utilized the two carbon sources simultaneously (Hagman
et al., 2008).
Nevertheless, the use of organic carbon sources increases
the operating cost significantly, and less-costly alternative
carbon sources for denitrification have been explored in some
WWTPs.
Alternative carbon sources
Various organic byproducts or waste rich in readily biode-
gradable organic matter and volatile fatty acids (VFA), such as
wine distillery effluent, the leachate of food waste, digested
piggery manure, mecellulose wasted liquor, biodiesel waste,
biologically hydrolyzed or mechanically disintegrated sludge,
etc., have been suggested and tested as alternative carbon
sources for denitrification (Table 5) (Kampas et al., 2007; Bodı́k
et al., 2009). A potential benefit of using organic carbon con-
tained in byproducts or waste materials as alternative carbon
sources is that they are produced in large amounts and can be
easily available at zero or even at negative cost.
The preparation of these carbon sources involves acid or
alkaline hydrolysis, thermal hydrolysis, thermochemical hy-
drolysis, or biological hydrolysis to yield a concentrated and
complex hydrolysate containing low molecular weight and
readily biodegradable matter. Quan et al. (2005) used hydro-
lyzed molasses as an external carbon source for nitrogen re-
moval from synthetic wastewater in an SBR with a TN
removal efficiency of 91.6%, which was slightly superior to
using methanol (85%). Obaja et al. (2005) used digested and
nondigested piggery manure as carbon source for biological
nitrogen and phosphorus removal from digested piggery
wastewater, same nutrient removal efficiencies (99.8% N and
97.8% P) was also obtained as when using acetate.
Gali et al. (2006a) examined hydrolyzed primary and sec-
ondary sludge as a carbon source for denitrification, but only
the hydrolyzed primary sludge supported stable denitrifica-
tion. Nitrogen removal efficiency was almost similar as when
using methanol as a carbon source; it could lead to a cost
saving of 0.2–0.3 euro for per kilogram-N removed. In addi-
tion, Kampas et al. (2009) reported that mechanically disin-
tegrated surplus activated sludge with energy inputs between
2,300 and 6,200 kJ=kg-TS, and a short disintegration time
(2–5 min) can also be used as an alternative carbon source
for denitrification.
However, it has to be pointed out that the addition of any
external organic carbon source to enhance nitrogen removal
may increase the load to the system, especially for using the
alternative carbon sources that may also introduce slowly
biodegradable COD, leading to an increase of total suspended
solids (TSS) in the process and compromise the effluent
quality. Therefore, the dosage of external organic carbon
source addition should be prior tested and not exceed the
requirement for the system.
Conclusions and Future Directions
Conclusions
The present work has reviewed innovative biological nitro-
gen removal pathways, shortcut nitrification=denitrification,
simultaneous nitrification=denitrification, and the Anammox
NITROGEN REMOVAL FOR LOW C/N DOMESTIC WASTEWATER
process that can remove nitrogen with low or zero dosage of
organic carbon sources.
It can be concluded that the advanced process control
based on the typical bending points of pH, DO and ORP
profiles is very helpful for achieving shortcut nitrogen re-
moval in an activated sludge system. In addition, the modi-
fied A2=O, step-feed mulistage A=O processes, MBR, and
MABR, can support the innovative biological nitrogen re-
moval pathways and they can be used to treat low C=N ratios
domestic wastewater without external carbon addition.
Furthermore, it has been shown that alternative organic
carbon sources can be potentially used for enhanced biologi-
cal nitrogen removal. These materials can offer the same or
better performance for enhanced nitrogen removal compared
to the common carbon source, allowing for a substantial re-
duction of the operating costs.
Overall, this review can provide guidance to municipal
WWTPs operators and planners to select cost-effective nitro-
gen removal alternatives for the treatment of low C=N ratio
domestic wastewater.
Future directions
Besides above, the presented results also show that further
studies are still required to obtain satisfying nitrogen removal
from low C=N ratio domestic wastewater in WWTPs. Some
research key points are proposed below: (1) go further and
apply these new processes and technologies mentioned in the
review at a large scale for the treatment of low C=N ratio
domestic wastewater; (2) explore the best alternatives to
introduce the Anammox pathway into domestic wastewater
treatment process because of its attractive metabolism; (3)
explore and optimize alternative carbon sources for enhanced
biological nitrogen removal. With these efforts, effective ni-
trogen removal from low C=N ratio domestic wastewater can
be achieved in the near future.
Acknowledgments
The authors thank the Shanghai Tongji Gao Tingyao En-
vironmental Science & Technology Development Foundation
(STGEF) and the China Scholarship Council (CSC) for sup-
porting this work. The authors also thank Lisbet Brusendorff
(DTU Environment, Denmark) for her contributions to the
graph design in the present work.
Author Disclosure Statement
No competing financial interests exist.
References
Abegglen, C., Ospelt, M., and Siegrist, H. (2008). Biological
nutrient removal in a small-scale MBR treating household
wastewater. Water Res. 42, 338.
Ahmed, Z., Lim, B.R., Cho, J., Song, K.G., Kim, K.P., and Ahn,
K.H. (2008). Biological nitrogen and phosphorus removal and
changes in microbial community structure in a membrane
bioreactor: Effect of different carbon sources. Water Res. 42, 198.
Ahn, Y.H. (2006). Sustainable nitrogen elimination biotechnolo-
gies: A review. Proc. Biochem. 41, 1709.
Ammary, B.Y., and Radaideh, J.A. (2005). Simultaneous nitrifi-
cation and denitrification in an oxidation ditch plant. Chem.
Biochem. Eng. Q. 19, 207.
123
Andrade do Canto, C.S, Rodrigues, J.A.D., Ratusznei, S.M.,
Zaiat, M., and Foresti, E. (2008). Feasibility of nitrification=
denitrification in a sequencing batch biofilm reactor with liq-
uid circulation applied to post-treatment. Bioresource Technol.
99, 644.
Anthonisen, A.C., Loehr, R.C., Prakasam, T.B.S., and Srinath,
E.G. (1976). Inhibition of nitrification by ammonia and
nitrous-acid. J. Water Pollut. Control Fed. 48, 835.
Aslan, S., and Dahab, M. (2008). Nitritation and denitritation of
ammonium-rich wastewater using fluidized-bed biofilm re-
actors. J. Hazard. Mater. 156, 56.
Aslan, S., Miller, L., and Dahab, M. (2009). Ammonium oxidation
via nitrite accumulation under limited oxygen concentration in
sequencing batch reactors. Bioresource Technol. 100, 659.
Beccari, M., Passino, R., Ramadori, R., and Tandoi, V. (1983).
Kinetics of dissimilatory nitrate and nitrite reduction in sus-
pended growth culture. J. Water Pollut. Control Fed. 55, 58.
Bernat, K., and Wojnowska-Baryla, I. (2007). Carbon source in
aerobic denitrification. Biochem. Eng. J. 36, 116.
Blackburne, R., Yuan, Z., and Keller, J. (2008). Demonstration of
nitrogen removal via nitrite in a sequencing batch reactor
treating domestic wastewater. Water Res. 42, 2166.
Bodı́k, I., Blšťáková, A., Sedláček, S., and Hutňan, M. (2009).
Biodiesel waste as source of organic carbon for municipal
WWTP denitrification. Bioresource Technol. 100, 2452.
Bougard, D., Bernet, N., Chèneby, D., and Delgenès, J.P. (2006).
Nitrification of a high-strength wastewater in an inverse tur-
bulent bed reactor: Effect of temperature on nitrite accumu-
lation. Proc. Biochem. 41, 106.
Bracklow, U., Drews, A., Vocks, M., and Kraume, M. (2007).
Comparison of nutrients degradation in small scale membrane
bioreactors fed with synthetic=domestic wastewater. J. Hazard.
Mater. 144, 620.
Chae, S.R., Kang, S.T., Watanabe, Y., and Shin, H.S. (2006).
Development of an innovative vertical submerged membrane
bioreactor (VSMBR) for simultaneous removal of organic
matter and nutrients. Water Res. 40, 2161.
Chae, S.R., and Shin, H.S. (2007). Characteristics of simultaneous
organic and nutrient removal in a pilot-scale vertical sub-
merged membrane bioreactor (VSMBR) treating municipal
wastewater at various temperatures. Proc. Biochem. 42, 193.
Chiu, Y.C., Lee, L.L., Chang, C.N., and Chao, A.C. (2007). Con-
trol of carbon and ammonium ratio for simultaneous nitrifi-
cation and denitrification in a sequencing batch bioreactor. Int.
Biodeteriorat. Biodegrad. 59, 1.
Choi, C., Lee, J., Lee, K., and Kim, M. (2008). The effects on
operation conditions of sludge retention time and carbon=
nitrogen ratio in an intermittently aerated membrane biore-
actor (IAMBR). Bioresource Technol. 99, 5397.
Chuang, H.P., Ohashi, A., Imachi, H., Tandukar, M., and Har-
ada, H. (2007). Effective partial nitrification to nitrite by down-
flow hanging sponge reactor under limited oxygen condition.
Water Res. 41, 295.
Ciudad, G., Rubilar, O., Muñoz, P., Ruiz, G., Chamy, R., Ver-
gara, C., and Jeison, D. (2005). Partial nitrification of high
ammonia concentration wastewater as a part of a shortcut
biological nitrogen removal process. Proc. Biochem. 40, 1715.
Daniel, L.M.C., Pozzi, E., Foresti, E., and Chinalia, F.A. (2009).
Removal of ammonium via simultaneous nitrification–
denitrification nitrite-shortcut in a single packed-bed batch
reactor. Bioresource Technol. 100, 1100.
Date, Y., Isaka, K., Sumino, T., Tsuneda, S., and Inamori, Y.
(2008). Microbial community of anammox bacteria immobilized
in polyethylene glycol gel carrier. Water Sci. Technol. 58, 1121.
124
Dodds, W.K., Bouska, W.W., Eitzmann, J.L., Pilger, T.J., Pitts,
K.L., Riley, A.J., Schloesser, J.T., and Thornbrugh, D.J. (2009).
Eutrophication of U.S. freshwaters: Analysis of potential eco-
nomic damages. Environ. Sci. Technol. 43, 12.
Downing, L.S., and Nerenberg, R. (2008a). Effect of bulk liquid
BOD concentration on activity and microbial community
structure of a nitrifying, membrane-aerated biofilm. Appl.
Microbiol. Biotechnol. 81, 153.
Downing, L.S., and Nerenberg, R. (2008b). Total nitrogen re-
moval in a hybrid, membrane-aerated activated sludge pro-
cess. Water Res. 42, 3697.
Eilersen, A.M., Henze, M., and Kløft, L. (1994). Effect of volatile
fatty acids and trimethylamine on nitrification in activated
sludge. Water Res. 28, 1329.
Ersu, B.C., Ong, S.K., Arslankaya, E., and Brown, P. (2008).
Comparison of recirculation configurations for biological
nutrient removal in a membrane bioreactor. Water Res. 42, 1651.
Feng, Y.J., Tseng, S.K., Hsia, T.H., Ho, C.M., and Chou, W.P.
(2007). Partial nitrification of ammonium-rich wastewater as
pretreatment for anaerobic ammonium oxidation (Anammox)
using membrane aeration Bioreactor. J. Biosci. Bioeng. 104, 182.
Fuerhacker, M., Bauer, H., Ellinger, R., Sree, U., Schmid, H.,
Zibuschka, F., and Puxbaum, H. (2000). Approach for a novel
control strategy for simultaneous nitrification=denitrification
in activated sludge reactors. Water Res. 34, 2499.
Fux, C., Velten, S., Carozzi, V., Solley, D., and Keller, J. (2006).
Efficient and stable nitritation and denitritation of ammonium-
rich sludge dewatering liquor using an SBR with continuous
loading. Water Res. 40, 2765.
Galiı́, A., Dosta, J., and Mata-Alvarez, J. (2006a). Use of hydro-
lyzed primary sludge as internal carbon source for denitrifi-
cation in a SBR treating reject water via nitrite. Indust. Eng.
Chem. Res. 45, 7661.
Galiı́, A., Dosta, J., Van Loosdrecht, M.C.M, and Mata-Alvarez, J.
(2006b). Biological nitrogen removal via nitrite of reject water
with a SBR and chemostat SHARON=denitrification process.
Indust. Eng. Chem. Res. 45, 7656.
Gao, D.W., Peng, Y.Z., Liang, H., and Wang, P. (2003). Using
oxidation-reduction potential (ORP) and pH value for process
control of shortcut nitrification–denitrification. J. Environ.
Sci.Health Part a Toxic=Hazard. Subst. Environ. Eng. 38, 2933.
Gao, S.Y., Peng, Y.Z., Wang, S.Y., and Yan, J. (2006). Novel
strategy of nitrogen removal from domestic wastewater using
pilot Orbal oxidation ditch. J. Environ. Sci. 18, 833.
Gong, Z., Liu, S., Yang, F., Bao, H., and Furukawa, K. (2008).
Characterization of functional microbial community in a
membrane-aerated biofilm reactor operated for com-
pletely autotrophic nitrogen removal. Bioresource Technol. 99,
2749.
Gong, Z., Yang, F., Liu, S., Bao, H., Hu, S., and Furukawa, K.
(2007). Feasibility of a membrane-aerated biofilm reactor to
achieve single-stage autotrophic nitrogen removal based on
Anammox. Chemosphere 69, 776.
Hagman, M., Nielsen, J.L., Nielsen, P.H., and Jansen, J.l.C.
(2008). Mixed carbon sources for nitrate reduction in activated
sludge-identification of bacteria and process activity studies.
Water Res. 42, 1539.
Hao, X., Doddema, H.J., and van Groenestijn, J.W. (1997).
Conditions and mechanisms affecting simultaneous nitrifica-
tion and denitrification in a Pasveer oxidation ditch. Bior-
esource Technol. 59, 207.
Helmer, C., and Kunst, S. (1998). Simultaneous nitrification=
denitrification in an aerobic biofilm system. Water Sci. Technol.
37, 183.
SUN ET AL.
Henze, M., Gujer, W., Mino, T., and Van Loosdrecht, M.C.M.
(2000). Activated Sludge Models ASM1, ASM2, ASM2D and
ASM3. London: IWA Publishing, p. 9.
Henze, M., Kristensen, G.H., and Strube, R. (1994). Rate-capacity
characterization of wastewater for nutrient removal processes.
Water Sci. Technol. 29, 101.
Henze, M., Van Loosdrecht, M.C.M., Ekama, G.A., and Brdja-
novic, D. (2008). Biological Wastewater Treatment: Principles,
Modelling and Design. London: IWA Publishing, p. 112, 139.
Hibiya, K., Terada, A., Tsuneda, S., and Hirata, A. (2003). Si-
multaneous nitrification and denitrification by controlling
vertical and horizontal microenvironment in a membrane-
aerated biofilm reactor. J. Biotechnol. 100, 23.
Holman, J.B., and Wareham, D.G. (2005). COD, ammonia and
dissolved oxygen time profiles in the simultaneous nitrification=
denitrification process. Biochem. Eng. J. 22, 125.
Hu, S., Yang, F., Sun, C., Zhang, J., and Wang, T. (2008). Simulta-
neous removal of COD and nitrogen using a novel carbon-
membrane aerated biofilm reactor. J. Environ. Sci. 20, 142.
Hwang, I.S., Min, K.S., Choi, E., and Yun, Z. (2005). Nitrogen
removal from piggery waste using the combined SHARON
and ANAMMOX process. Water Sci. Technol. 52, 487.
Kampas, P., Parsons, S.A., Pearce, P., Ledoux, S., Vale, P.,
Cartmell, E., and Soares, A. (2009). An internal carbon source
for improving biological nutrient removal. Bioresource Technol.
100, 149.
Kampas, P., Parsons, S.A., Pearce, P., Ledoux, S., Vale, P.,
Churchley, J., and Cartmell, E. (2007). Mechanical sludge
disintegration for the production of carbon source for bio-
logical nutrient removal. Water Res. 41, 1734.
Katsogiannis, A.N., Kornaros, M., and Lyberatos, G. (2003).
Enhanced nitrogen removal in SBRs bypassing nitrate gener-
ation accomplished by multiple aerobic=anoxic phase pairs.
Water Sci. Technol. 47, 53.
Kim, J.H., Guo, X., and Park, H.S. (2008). Comparison study of
the effects of temperature and free ammonia concentration on
nitrification and nitrite accumulation. Proc. Biochem. 43, 154.
Kimura, K., Nishisako, R., Miyoshi, T., Shimada, R., and Watanabe,
Y. (2008). Baffled membrane bioreactor (BMBR) for efficient nu-
trient removal from municipal wastewater. Water Res. 42, 625.
Komorowska-Kaufman, M., Majcherek, H., and Klaczynı́ski, E.
(2006). Factors affecting the biological nitrogen removal from
wastewater. Proc. Biochem. 41, 1015.
Kornaros, M., Marazioti, C., and Lyberatos, G. (2008). A pilot
scale study of a sequencing batch reactor treating municipal
wastewater operated via the UP-PND process. Water Sci.
Technol. 58, 435.
Kristensen, G.H., Jorgensen, P.E., and Henze, M. (1992). Char-
acterization of functional microorganism groups and substrate
in activated sludge and wastewater by AUR, NUR and OUR.
Water Sci. Technol. 25, 43.
Kuenen, J.G. (2008). Anammox bacteria: From discovery to ap-
plication. Nat. Rev. Microbiol. 6, 320.
Kujawa, K., and Klapwijk, B. (1999). A method to estimate de-
nitrification potential for predenitrification systems using
NUR batch test. Water Res. 33, 2291.
Lackner S., Terada A. and Smets B.F. (2008). Heterotrophic
activity compromises autotrophic nitrogen removal in
membrane-aerated biofilms: Results of a modeling study.
Water Res. 42, 1102.
Lai, E., Senkpiel, S., Solley, D., and Keller, J. (2004). Nitro-
gen removal of high strength wastewater via nitritation=
denitritation using a sequencing batch reactor. Water Sci.
Technol. 50, 27.
NITROGEN REMOVAL FOR LOW C/N DOMESTIC WASTEWATER
Lamsam, A., Laohaprapanon, S., and Annachhatre, A.P. (2008).
Combined activated sludge with partial nitrification (AS=PN)
and anammox processes for treatment of seafood processing
wastewater. J. Environ. Sci. Health Part a Toxic=Hazard. Subst.
Environ. Eng. 43, 1198.
LaPara, T.M., Cole, A.C., Shanahan, J.W., and Semmens, M.J.
(2006). The effects of organic carbon, ammoniacal-nitrogen,
and oxygen partial pressure on the stratification of membrane-
aerated biofilms. J. Indust. Microbiol. Biotechnol. 33, 315.
Lemoine, C., Payraudeau, M., and Meinhold, J. (2006). Aeration
control for simultaneous nitrification–denitrification in a bio-
logical aerated filter using internal model approach. Water Sci.
Technol. 54, 129.
Li, T.G., Liu, J.X., Bai, R.B., and Wong, F.S., (2008a). Membrane-
aerated biofilm reactor for the treatment of acetonitrile
wastewater. Environ. Sci. Technol. 42, 2099.
Li, Y.Z., He, Y.L., Ohandja, D.G., Ji, J., Li, J.F., and Zhou, T.
(2008b). Simultaneous nitrification-denitrification achieved by
an innovative internal-loop airlift MBR: Comparative study.
Bioresource Technol. 99, 5867.
Liu, S., Yang, F., Xue, Y., Gong, Z., Chen, H., Wang, T., and Su,
Z. (2008). Evaluation of oxygen adaptation and identification
of functional bacteria composition for anammox consortium in
non-woven biological rotating contactor. Bioresource Technol.
99, 8273.
Ma, Y., Peng, Y., Wang, S., Yuan, Z., and Wang, X. (2009).
Achieving nitrogen removal via nitrite in a pilot-scale con-
tinuous pre-denitrification plant. Water Res. 43, 563.
Matsumoto, S., Terada, A., and Tsuneda, S. (2007). Modeling of
membrane-aerated biofilm: Effects of C=N ratio, biofilm thick-
ness and surface loading of oxygen on feasibility of simulta-
neous nitrification and denitrification. Biochem. Eng. J. 37, 98.
Mokhayeri, Y., Riffat, R., Murthy, S., Bailey, W., Takacs, I., and
Bott, C. (2009). Balancing yield, kinetics and cost for three
external carbon sources used for suspended growth post-
denitrification. The 2nd IWA Specialized Conference on Nutrient
Management in Wastewater Treatment Processes, 6–9 September,
2009, Krakow, Poland.
Munch, E.V., Lant, P., and Keller, J. (1996). Simultaneous nitri-
fication and denitrification in bench-scale sequencing batch
reactors. Water Res. 30, 277.
Nakhla, G., and Farooq, S. (2003). Simultaneous nitrification-
denitrification in slow sand filters. J. Hazard. Mater. 96, 291.
Obaja, D., Mace, S., and Mata-Alvarez, J. (2005). Biological nu-
trient removal by a sequencing batch reactor (SBR) using an
internal organic carbon source in digested piggery wastewa-
ter. Bioresource Technol. 96, 7.
Ouyang, C.F., Chiou, R.J., and Lin, C.T. (2000). The character-
istics of nitrogen removal by the biofilter system. Water Sci.
Technol. 42, 137.
Park, C.H., Chung, C.W., Leek, Y.J., and Han, G.B. (2005). Fea-
sibility study of nitrogen removal with the mecellulose wasted
liquor as an external carbon source in the two-stage denitri-
fication process. Environ. Technol. 26, 591.
Park, S., Bae, W., Chung, J., and Baek, S. (2007). Empirical model
of the pH dependence of the maximum specific nitrification
rate. Proc. Biochem. 42, 1671.
Pedersen, P.D., Jensen, K., Lyngsie, P., and Johansen, N.H. (2003).
Nitrogen removal in industrial wastewater by nitration and
denitration—3 years of experience. Water Sci. Technol. 47, 181.
Peng, Y.Z., Chen, Y., Peng, C.Y., Liu, M., Wang, S.Y., Song, X.Q.,
and Cui, Y.W. (2004). Nitrite accumulation by aeration con-
trolled in sequencing batch reactors treating domestic waste-
water. Water Sci. Technol. 50, 35.
125
Peng, Y.Z., Gao, S., Wang, S., and Bai, L. (2007). Partial nitrifi-
cation from domestic wastewater by aeration control at am-
bient temperature. Chin. J. Chem. Eng. 15, 115.
Peng, Y.Z., and Zhu, G.B. (2006). Biological nitrogen removal
with nitrification and denitrification via nitrite pathway. Appl.
Microbiol. Biotechnol. 73, 15.
Pochana, K., and Keller, J. (1999). Study of factors affecting
simultaneous nitrification and denitrification (SND). Water
Sci. Technol. 39, 61.
Pynaert, K., Smets, B.F., Beheydt, D., and Verstraete, W. (2004).
Start-up of autotrophic nitrogen removal reactors via se-
quential biocatalyst addition. Environ. Sci. Technol. 38, 1228.
Quan, Z.X., Jin, Y.S., Yin, C.R., Lee, J.J., and Lee, S.T. (2005).
Hydrolyzed molasses as an external carbon source in biolog-
ical nitrogen removal. Bioresource Technol. 96, 1690.
Ruiz, G., Jeison, D., Rubilar, O., Ciudad, G., and Chamy, R. (2006).
Nitrification–denitrification via nitrite accumulation for nitro-
gen removal from wastewaters. Bioresource Technol. 97, 330.
Ryu, H.D., and Lee, S.I. (2009). Comparison of 4-stage biological
aerated filter (BAF) with MLE process in nitrogen removal from
low carbon-to-nitrogen wastewater. Environ. Eng. Sci. 26, 163.
Sarioglu, M., Insel, G., Artan, N., Orhon, D. (2009). Model
evaluation of simultaneous nitrification and denitrification
in a membrane bioreactor operated without an anoxic reactor.
J. Membr. Sci. 337, 17.
Satoh, H., Ono, H., Rulin, B., Kamo, J., Okabe, S., and Fukushi,
K.I. (2004). Macroscale and microscale analyses of nitrification
and denitrification in biofilms attached on membrane aerated
biofilm reactors. Water Res. 38, 1633.
Schmidt, I., Sliekers, O., Schmid, M., Bock, E., Fuerst, J., Kuenen,
J.G., Jetten, M.S.M., and Strous, M. (2003). New concepts of
microbial treatment processes for the nitrogen removal in
wastewater. FEMS Microbiol. Rev. 27, 481.
Semmens, M.J., Dahm, K., Shanahan, J., and Christianson, A.
(2003). COD and nitrogen removal by biofilms growing on gas
permeable membranes. Water Res. 37, 4343.
Sliekers, A.O., Derwort, N., Gomez, J.L.C., Strous, M., Kuenen,
J.G., and Jetten, M.S.M. (2002). Completely autotrophic nitrogen
removal over nitrite in one single reactor. Water Res. 36, 2475.
Spagni, A., and Marsili-Libelli, S. (2009). Nitrogen removal via
nitrite in a sequencing batch reactor treating sanitary landfill
leachate. Bioresource Technol. 100, 609.
Strous, M., Heijnen, J.J., Kuenen, J.G., and Jetten, M.S.M. (1998).
The sequencing batch reactor as a powerful tool for the study
of slowly growing anaerobic ammonium-oxidizing microor-
ganisms. Appl. Microbiol. Biotechnol. 50, 589.
Syron, E., and Casey, E. (2008a). Membrane-aerated biofilms for
high rate biotreatment: Performance appraisal, engineering
principles, scale-up, and development requirements. Environ.
Sci. Technol. 42, 1833.
Syron, E., and Casey, E. (2008b). Model-based comparative
performance analysis of membrane aerated biofilm reactor
configurations. Biotechnol. Bioeng. 99, 1361.
Tan, T.W., and Ng, H.Y. (2008). Influence of mixed liquor
recycle ratio and dissolved oxygen on performance of pre-
denitrification submerged membrane bioreactors. Water Res.
42, 1122.
Tanwar, P., Nandy, T., Ukey, P., and Manekar, P. (2008). Cor-
relating on-line monitoring parameters, pH, DO and ORP
with nutrient removal in an intermittent cyclic process biore-
actor system. Bioresource Technol. 99, 7630.
Tchobanoglous, G., Burton, F.L., and Stensel, H.D., Eds. (2004).
Metcalf & Eddy: Wastewater Engineering: Treatment and Reuse.
New York: McGraw-Hill.
126
Terada, A., Hibiya, K., Nagai, J., Tsuneda, S., and Hirata, A.
(2003). Nitrogen removal characteristics and biofilm anal-
ysis of a membrane-aerated biofilm reactor applicable to
high-strength nitrogenous wastewater treatment. J. Biosci.
Bioeng. 95, 170.
Terada, A., Lackner, S., Tsuneda, S., and Smets, B.F. (2007).
Redox-stratification controlled biofilm (ReSCoBi) for com-
pletely autotrophic nitrogen removal: The effect of co-versus
counter-diffusion on reactor performance. Biotechnol. Bioeng.
97, 40.
Third, K.A., Paxman, J., Schmid, M., Strous, M., Jetten, M.S.M.,
and Cord-Ruwisch, R. (2005). Treatment of nitrogen-rich
wastewater using partial nitrification and Anammox in the
CANON process. Water Sci. Technol. 52, 47.
Tsonis, S.P. (1997). Olive oil mill wastewater as carbon source in
post anoxic denitrification. Water Sci. Technol. 36, 53.
Tsushima, I., Ogasawara, Y., Kindaichi, T., Satoh, H., and Okabe,
S. (2007). Development of high-rate anaerobic ammonium-
oxidizing (anammox) biofilm reactors. Water Res. 41, 1623.
Turk, O., and Mavinic, D.S. (1989). Maintaining nitrite build-up
in a system acclimated to free ammonia. Water Res. 23, 1383.
Ueda, T., and Hata, K. (1999). Domestic wastewater treatment
by a submerged membrane bioreactor with gravitational fil-
tration. Water Res. 33, 2888.
Vadivelu, V.M., Keller, J., and Yuan, Z. (2007a). Free ammonia
and free nitrous acid inhibition on the anabolic and catabolic
processes of Nitrosomonas and Nitrobacter. Water Sci. Technol.
56, 89.
Vadivelu, V.M., Keller, J., and Yuan, Z. (2007b). Effect of free
ammonia on the respiration and growth processes of an en-
riched Nitrobacter culture. Water Res. 41, 826.
Vadivelu, V.M., Yuan, Z., Fux, C., and Keller, J. (2006). The in-
hibitory effects of free nitrous acid on the energy generation
and growth processes of an enriched Nitrobacter culture.
Environ. Sci. Technol. 40, 4442.
Van der Star, W.R.L., Abma, W.R., Blommers, D., Mulder, J.W.,
Tokutomi, T., Strous, M., Picioreanu, C., and Van Loosdrecht,
M.C.M. (2007). Startup of reactors for anoxic ammonium ox-
idation: Experiences from the first full-scale anammox reactor
in Rotterdam. Water Res. 41, 4149.
Van der Star, W.R.L., Miclea, A.I., Van Dongen, U.G.J.M.,
Muyzer, G., Picioreanu, C., and Van Loosdrecht, M.C.M.
(2008). The membrane bioreactor: A novel tool to grow ana-
mmox bacteria as free cells. Biotechnol. Bioeng. 101, 286.
Van Dongen, U., Jetten, M.S.M., and Van Loosdrecht, M.C.M.
(2001). The SHARON
-Anammox
process for treatment of
ammonium rich wastewater. Water Sci. Technol. 44, 153.
Villaverde, S., Fdz-Polanco, F., and Garciı́a, P.A. (2000). Nitri-
fying biofilm acclimation to free ammonia in submerged bio-
filters: Start-up influence. Water Res. 34, 602.
Walter, B., Haase, C., and Räbiger, N. (2005). Combined
nitrification=denitrification in a membrane reactor. Water Res.
39, 2781.
Wang, J., Peng, Y., Wang, S., and Gao, Y. (2008a). Nitrogen re-
moval by simultaneous nitrification and denitrification via
nitrite in a sequence hybrid biological reactor. Chin. J. Chem.
Eng. 16, 778.
Wang, S., Zhang, S., Peng, C., and Akio, T., (2008b). Intercross
real-time control strategy in alternating activated sludge
SUN ET AL.
process for short-cut biological nitrogen removal treating
domestic wastewater. J. Environ. Sci. 20, 957.
Wang, S.Y., Gao, D.W., Peng, Y.Z., Wang, P., and Yang, Q.
(2004). Nitrification-denitrification via nitrite for nitrogen re-
moval from high nitrogen soybean wastewater with on-line
fuzzy control. Water Sci. Technol. 49, 121.
Wang, X.L., Ma, Y., Peng, Y.Z., and Wang, S.Y. (2007). Short-
cut nitrification of domestic wastewater in a pilot-scale A=O
nitrogen removal plant. Bioproc. Biosyst. Eng. 30, 91.
Windey, K., Bo, I.D., and Verstraete, W. (2005). Oxygen-limited
autotrophic nitrification–denitrification (OLAND) in a rotat-
ing biological contactor treating high-salinity wastewater.
Water Res. 39, 4512.
Wong-Chong, G.M., and Loehr, R.C. (1978). Kinetics of micro-
bial nitrification: Nitrite–nitrogen oxidation. Water Res. 12, 605.
Wu, C., Chen, Z., Liu, X., and Peng, Y. (2007). Nitrification–
denitrification via nitrite in SBR using real-time control strategy
when treating domestic wastewater. Biochem. Eng. J. 36, 87.
Xia, S., Guo, J., and Wang, R. (2008a). Performance of a pilot-
scale submerged membrane bioreactor (MBR) in treating
bathing wastewater. Bioresource Technol. 99, 6834.
Xia, S., Li, J., and Wang, R. (2008b). Nitrogen removal perfor-
mance and microbial community structure dynamics response
to carbon nitrogen ratio in a compact suspended carrier bio-
film reactor. Ecol. Eng. 32, 256.
Xue, Y., Yang F., Liu S., and Fu Z. (2009). The influence of
controlling factors on the start-up and operation for partial
nitrification in membrane bioreactor. Bioresource Technol. 100,
1055.
Yamamoto, T., Takaki, K., Koyama, T., and Furukawa, K. (2008).
Long-term stability of partial nitritation of swine wastewater
digester liquor and its subsequent treatment by Anammox.
Bioresource Technol. 99, 6419.
Yang, D.H, Song, Y.H., Tan, Q.G., and Yang, Q.X. (2006). Re-
search on phosphorus and nitrogen removal by modified
A2=O process with low-carbon and low-energy consumption.
China Water Wasterwater 22, 18.
Yang, Q., Peng, Y., Liu, X., Zeng, W., Mino, T., and Satoh, H.
(2007). Nitrogen removal via nitrite from municipal waste-
water at low temperatures using real-time control to optimize
nitrifying communities. Environ. Sci. Technol. 41, 8159.
Yoo, H., Ahn, K.H., Lee, H.J., Lee, K.H., Kwak, Y.J., and Song,
K.G. (1999). Nitrogen removal from synthetic wastewater by
simultaneous nitrification and denitrification (SND) via nitrite
in an intermittently-aerated reactor. Water Res. 33, 145.
Zhao, H.W., Mavinic, D.S., Oldham, W.K., and Koch, F.A.
(1999). Controlling factors for simultaneous nitrification and
denitrification in a two-stage intermittent aeration process
treating domestic sewage. Water Res. 33, 961.
Zhu, G., Peng, Y., Wang, S., Wu, S., and Ma, B. (2007a). Effect
of influent flow rate distribution on the performance of step-
feed biological nitrogen removal process. Chem. Eng. J. 131, 319.
Zhu, G., Peng, Y., Wu, S., Wang, S., and Xu, S. (2007b).
Simultaneous nitrification and denitrification in step feeding
biological nitrogen removal process. J. Environ. Sci. 19, 1043.
Zhu, G., Peng, Y., Zhai, L., Wang, Y., and Wang, S. (2009).
Performance and optimization of biological nitrogen removal
process enhanced by anoxic=oxic step feeding. Biochem. Eng. J.
43, 280.
This article has been cited by:

1. Lan Wang, Ping Zheng, Tingting Chen, Jianwei Chen, Yajuan Xing, Qixing Ji, Meng Zhang, Jiqiang Zhang. 2012.
Performance of autotrophic nitrogen removal in the granular sludge bed reactor. Bioresource Technology 123, 78-85.
[CrossRef]
2. Samik Bagchi, Rima Biswas, Tapas Nandy. 2012. Autotrophic Ammonia Removal Processes: Ecology to Technology.
Critical Reviews in Environmental Science and Technology 42:13, 1353-1418. [CrossRef]
3. Li-Juan Feng, Jian Xu, Xiang-Yang Xu, Liang Zhu, Jing Xu, Wei Ding, Jing Luan. 2012. Enhanced biological nitrogen
removal via dissolved oxygen partitioning and step feeding in a simulated river bioreactor for contaminated source water
remediation. International Biodeterioration & Biodegradation 71, 72-79. [CrossRef]
4. Farah Naz Ahmed, Christopher Q. Lan. 2012. Treatment of landfill leachate using membrane bioreactors: A review.
Desalination . [CrossRef]
5. Xiangyang Xu, Gang Liu, Liang Zhu. 2011. Enhanced denitrifying phosphorous removal in a novel anaerobic/aerobic/anoxic
(AOA) process with the diversion of internal carbon source. Bioresource Technology . [CrossRef]
6. Libing Chu, Jianlong Wang. 2011. Nitrogen removal using biodegradable polymers as carbon source and biofilm carriers in
a moving bed biofilm reactor. Chemical Engineering Journal 170:1, 220-225. [CrossRef]
7. Qingmei Yan, Xuxiang Zhang, Tong Zhang, Herbert H. P. Fang. 2011. Seasonal microbial community shift in a saline sewage
treatment plant. Frontiers of Environmental Science & Engineering in China 5:1, 40-47. [CrossRef]
8. Akihiko Terada, Sheng Zhou, Masaaki Hosomi. 2011. Presence and detection of anaerobic ammonium-oxidizing (anammox)
bacteria and appraisal of anammox process for high-strength nitrogenous wastewater treatment: a review. Clean Technologies
and Environmental Policy . [CrossRef]
9. Da-wen Gao, Rui An, Yu Tao, Jin Li, Xin-xin Li, Nan-qi Ren. 2011. Simultaneous methane production and wastewater reuse
by a membrane-based process: Evaluation with raw domestic wastewater. Journal of Hazardous Materials 186:1, 383-389.
[CrossRef]
10. M. Ruscalleda Beylier, M.D. Balaguer, J. Colprim, C. Pellicer-Nàcher, B.-J. Ni, B.F. Smets, S.-P. Sun, R.-C. WangBiological
Nitrogen Removal from Domestic Wastewater 329-340. [CrossRef]