REVIEW

Ghulam Abbas et al. [Link]., Vol. 36, No. 4, 2014 775

Ammonia Nitrogen Removal by Single-stage Process: A Review

1,2
Ghulam Abbas, 1 Ping Zheng*, 1 Lan Wang, 1 Wei Li, 2 Khurram Shahzad
1
Hongtao Zhang, 1 Muhammad Zaffar Hashmi, 1 Jiqiang Zhang and 1 Meng Zhang
1
Department of Environmental Engineering, Zhejiang University, Hangzhou 310058, China.
2
Department of Chemical Engineering, University of Gujrat, Gujrat, Pakistan.
pzheng@[Link]*

(Received on 27th March 2013, accepted in revised form 19th September 2013)

Summary: Single-stage nitrogen removal from ammonium-rich streams has been studied
extensively during the last decade in view of rising cost of traditional treatment methods and
stringent discharge standards. This system has been developed under different names (DEMON,
CANON, OLAND, SNAP and SNAD) to carry out simultaneous aerobic, anaerobic ammonium
oxidizing and denitrification reactions in the same reactor. The maximal influent ammonia
concentration is 431.7 ± 25.5 mg N/L, the minimal HRT is 4 hr, the maximal nitrogen loading rate is
5.44 kg N/m3day and the maximal nitrogen removal rate is 2.57 kg N/m3day for the process. The
single-stage process has distinct advantages in terms of saving configuration, aeration and carbon
sources. This paper reviews working principles, operational characteristics, efficiency of the single-
stage system, key control factors of the process and its applications for nitrogen removal.

Key words: Single-stage, CANON, DEMON, OLAND, SNAP, SNAD, Efficiency.

Introduction

Generally, ammonia is the main nitrogen stage system and seems competitive in terms of
compound present in most of the wastewaters. process simplicity and investment which may include
Ammonia removal has become a worldwide concern engineering, construction, and operating costs [3].
because: it causes eutrophication, gives rise to This paper discusses operational characteristics,
oxygen depletion and poisons aquatic life. Ammonia efficiency of the single-stage system, key control
removal by the conventional biological process, factors of the process and its applications for nitrogen
nitrification followed by denitrification, requires both removal.
extensive energy for aeration to carry out nitrification
to nitrate and an external carbon source for Principle of Single-Stage Process
denitrification. The conventional biological nitrogen
removal process is used for treating wastewaters with The working principle of single-stage
relatively low nitrogen concentrations (total nitrogen nitrogen removal process is based on microbiology
concentration less than 100 mg N/L). The and application technology of functional
requirement of added electron donors (methanol, microorganisms.
acetate etc) makes full-scale denitrification quite
expensive for wastewater with higher nitrogen Microbiology Principle
concentrations [1]. Furthermore, most existing
wastewater treatment facilities were not designed for The process relies on the harmonious and
nitrogen removal, and meeting the demands of the balanced interaction of Aerobic Ammonium
nitrification-denitrification steps in these facilities Oxidizing Bacteria ([Link]) and Anaerobic
can be difficult. Thus, many wastewater treatment Ammonium Oxidizing Bacteria ([Link]) that
plants do not meet the current discharge standard of perform two sequential reactions of partial
10 mg N/L [2]. nitrification and anoxic oxidation of ammonia
simultaneously in the same reactor under oxygen-
Whereas single-stage autotrophic nitrogen limited conditions, and result in an almost complete
removal has several advantages over conventional conversion of ammonium to nitrogen gas along with
nitrification-denitrification (1) a 63% reduction in small amounts of nitrate.
energy consumption for aeration (2) no organic donor
requirement for denitrification; (3) a 90% reduction AOB: AOB contain Genus I Nitrosomonas,
in sludge handling and transportation costs; and (4) Genus II Nitrosococcus, and Genus III Nitrosospira.
less production of N2O, a powerful green house gas However, Nitrosomonas europaea/eutropha,
[2]. In addition, single-stage process generally has Nitrosomonas oligotroph/uera, Nitrosomonas
higher volumetric nitrogen removal rate than two- communis, and Nitrosospira sp. are the well known
*
To whom all correspondence should be addressed.
Ghulam Abbas et al. [Link]., Vol. 36, No. 4, 2014 776

aerobic ammonia oxidizers reported in the lab-scale

studies of single-stage process. Classification of Single-Stage Process
[Link]: [Link] include Candidatus
Two different reactors are used in Sharon-
Brocadia, Candidatus Anammoxoglobus, Candidatus
Anammox combined system for partial nitrification
Jettenia, Candidatus Kuenenia, and Candidatus
and Anammox processes. Partial nitrification and
Scalindua. Candidatus Kuenenia stuttgartiensis,
Anammox processes can be accomplished in a single-
Brocardia anammoxidans, and Candidatus Brocadia-
stage system for successful elimination of ammonium
related anaerobic ammonium oxidizing bacteria have
nitrogen. The single-stage nitrogen removal process
been found as predominant functional microbes in
was observed by different research groups and
various studies of one reactor system [4-7].
following names have been given in different parts of
the world.
The cooperation of these two groups of
ammonium-oxidising bacteria is the microbial basis • Aerobic/Anoxic Deammonification (DEMON)
of the sustainable one reactor system. • Oxygen-Limited Autotrophic Nitrification-
Denitrifcation (OLAND)
Overall reaction for nitrogen removal in a • Completely Autotrophic Nitrogen removal over
single-stage process can be represented by following Nitrite (CANON)
equation [8]. • Single-stage Nitrogen removal using Anammox
and Partial nitritation (SNAP)
NH4++0.85O2 0.44N2+0.11NO3-+1.43H2O+ 0.14H+
• Simultaneous partial Nitrification, Anammox
Technology Principle and Denitrification (SNAD)
DEMON and OLAND processes were
Single-stage process can be accomplished in developed at Hanover University (Germany) and
the reactor based either on granular sludge or bio- Ghent University (Belgium) respectively [10-12].
film. The biomass in a reactor can be grown in the CANON and SNAP processes were developed at
form of granular sludge or bio-film. Granular sludge Delft University (Netherlands) and Kumamoto
is formed by self-aggregation in the granules and University (Japan) respectively [13-15]. Recently,
consists of dense, fast-settling granules with a large SNAD process [16] was developed at Dalian
surface area and bio-film is formed when the carrier University of Technology (China) which also
material is submerged in a liquid that contains incorporated denitrification process.
nutrients and microorganisms. Then, microorganisms
attach to the surface and start to grow. Functional Reactor Configuration
microbes work in different zones of the granular Several kinds of high biomass retention
sludge or bio-film but they interact harmoniously to reactors have been tested for the single-stage process
remove nitrogen from ammonium-rich wastewaters. on laboratory scale including sequencing batch
Both processes (partial nitrification and Anammox) reactor (SBR), air lift reactor, fixed bed, moving bed
occur in the same reactor as a result of dissolved bio-film reactor, rotating biological contactor (RBC),
oxygen (DO) concentration gradients within the and up-flow granular bed reactor. The efficient
granular sludge or bio-film due to diffusional retention of biomass in a SBR makes it possible to
limitations. The presence of oxygen concentration cultivate slowly growing bacteria. However, higher
gradients has been verified by microelectrode nitrogen removal rates were obtained in a RBC [17, 6]
measurements and 15N tracer techniques [3]. The DO and in an air lift reactor [8]. For full biomass
ranges for lab-scale single-stage systems and for pilot retention, membrane bioreactors can also be used but
or full-scale systems are 0.1 mg/L to 1.8 mg/L and bio-film or granular reactors are preferred for full-
0.3 mg/L to 3 mg/L, respectively [1]. Ammonium is scale applications because sludge granules or bio-
converted to nitrite in the outer oxic regions, while films are easily developed. Bio-film or granule
Anammox is active in the inner anoxic regions of the reactors function in the same fashion. In bio-film or
granule or bio-film. The concentration of substrates granule reactors the ammonium oxidizers are active
(ammonium and oxygen) determines the conversion in the outer layers of the bio-film or granule,
of ammonia to nitrite. However, ammonia to nitrite producing a suitable amount of nitrite for the
conversion with the maximal nitrogen loading rate Anammox organisms that are active in the inner
and the maximal nitrogen removal rate is 55.4% [9]. layers. This way the Anammox organisms are
protected from oxygen, which is consumed in the
outer layers [1].
Ghulam Abbas et al. [Link]., Vol. 36, No. 4, 2014 777

Functional Microorganisms converted almost completely into nitrogen gas and

At first, nitrifiers were assumed to perform small amounts of nitrate. Too much or too little
ammonium oxidation process in DEMON and ammonium affects the performance of single-stage
OLAND processes under micro-aerobic conditions system. If excess ammonium is oxidized to nitrite,
[10, 11]. But later studies [4, 6] confirmed by FISH Anammox would be baffled because of the shortage
analysis that both processes are Anammox-based. of ammonium. If too much ammonium is left,
This is also corroborated by the term Anammox would also be blocked because of the
OLAND/CANON used by Van Hulle et al. [18]. It shortage of nitrite.
has been clearly found that AOB and Anammox Accumulation: Single-stage process involves
bacteria co-exist and co-operate to remove production of nitrite due to partial oxidation of
ammonium nitrogen in CANON and SNAP processes ammonium in the first step and leftover ammonium
[19, 7]. Most of the ammonium-rich wastewaters combines with the produced NO2 to yield nitrogen
(such as old landfill leachates) have a certain gas in the denitrification step. As long as ammonium
concentration of chemical oxygen demand (COD). concentration outside the bio-film or granule is
Denitrification process was added to the single-stage higher than the oxygen or nitrite concentration,
process to remove nitrogenous and carbonaceous ammonium diffusion into the bio-film or granule will
compounds. It was observed in the SNAD process not limit the process rate. If the nitrite produced in
that denitrifying bacteria population worked in the outer layer is mainly consumed in the inner layer,
harmony with AOB and Anammox to remove oxygen is the main limiting factor controlling the
simultaneously ammonium and COD from the overall rate. The nitrite oxidizing bacteria (NOB)
wastewater under oxygen-limited conditions [16]. have a lower affinity [21] for oxygen than AOB and
Process Analysis NOB require a high DO concentration in comparison
to AOB. Therefore, AOB dominate the NOB at low
Inflow: Generally, synthetic wastewater DO concentration which causes accumulation of
prepared with tap water is employed to feed the NO2- and occurrence of partial nitrification and
single-stage reactor. The medium may contain denitrification via NO2-.
(NH4)2SO4, KH2PO4, KHCO3 CaCl2.2H2O, MgSO4,
FeSO4, NaCl, KCl and NH4Cl etc and trace element Outflow: Besides nitrogen gas, the effluent
solution. The trace element solution includes: EDTA, from the one reactor system may contain small
ZnSO4·7H2O, CoCl2·6H2O, MnCl2·4H2O, amounts of nitrate, nitrite and free ammonia. The
CuSO4·5H2O, NaMoO4·2H2O, NiCl2·6H2O, reactor is provided with drum to collect the effluent.
NaSeO4·10H2O, H3BO4, NaWO4·2H2O [20, 5, 19]. Process Efficiency
The reactor is provided with drum of suitable size to
Reaction Rate: As growth rates of AOB and
supply the influent continuously with the help of a
Anammox bacteria (10-12 days doubling time;
peristaltic pump.
specific growth rate µ=0.00648/d and low biomass
O2 Transfer: Aeration devices and reactor yield, 0.11-0.13 g VSS/g NH4+-N) are low [22] so
configuration determine the transfer of air to the bulk reactors based on Anammox should have long
liquid. AOB require oxygen to carry out nitrification sludge retention time to maintain high biomass
and synthesis of cellular material. Micro-aerobic concentrations, especially at the start-up stage. The
conditions in bio-films or granular systems are performance of single-stage systems can be enhanced
maintained by controlled supply of oxygen. Due to its by using carrier materials (glass beads, kaldnes rings)
low solubility in aqueous solution, continuous by developing bio-film or by self-aggregation in
transfer of oxygen is ensured that results in good granules. Generally saying, increase of temperature
mixing conditions in the reactor. Oxygen is increases both the rate of chemical and biological
transferred by air pump. processes. But increase of temperature to high levels
Conversion: Partial nitrification depends can be harmful for biological organisms. Most
upon the concentration of substrates, which are studies suggest a maximum temperature of 40°C
ammonium (electron donor) and oxygen (electron which enhances rate of Anammox and shows
acceptor), and Anammox reaction depends upon negative effect on activity beyond 45°C.
concentration of ammonia, nitrite and DO which acts Conversion Percentage
as inhibitor for Anammox bacteria. Under oxygen-
limited condition, only part of the ammonium is Several lab-scale studies have been
nitrified to intermediate nitrite during partial performed to evaluate the performance of single-
nitrification and the produced nitrite is utilized with stage process for nitrogen removal. Conversion
remaining ammonium by Anammox bacteria and percentages and nitrogen removal rates of lab-scale
single-stage systems are given in Table-1 [23-30].
Ghulam Abbas et al. [Link]., Vol. 36, No. 4, 2014 778

Table-1: Conversion percentages and nitrogen removal rates of lab-scale single-stage systems.
Reactor type Reactor name NRR kgN/m3day Conversion percentage (%) Reference
SBR CANON 0.06 50 [5]
RBC OLAND 1.05 89 [6]
Airlift CANON 1.5 42 [8]
Fixed Bed SNAP 0.86 52-80 [14]
RBC OLAND 1.80 88 [17]
RBC OLAND 0.42 42 [17]
Up-flow granular bed DEMON 0.77 89 [23]
SBR CANON 0.06 76 [24]
Moving bed bio-film CANON 0.77 89 [25]
SBR CANON 0.5 40-80 [26]
SBR CANON 0.49 80 [27]
SBR SNAD 0.197 - [28]
Non-woven
SNAD 94%(COD Removal Efficiency ) 79 [29]
RBC
SBR SNAD 87%(COD Removal Efficiency ) 96 [30]

ammonium surface load. They reported optimal bulk

Reaction Products: Sliekers et al. [5] found oxygen concentration for a CANON bio-film reactor
absence of undesirable by-products like N2O, NO as 1mg O2/L, although this optimum concentration
during CANON process, although production of depends on the bio-film thickness, density, boundary
these gases by AOB at low oxygen conditions was layer thickness, COD content of influent and
reported by different studies. The main product of temperature. At low DO, accumulation of nitrite was
single-stage process comprises of 90 % nitrogen gas observed [35, 36] and NOB were out-competed due
and 10% nitrate. As the nitrite also serves as an to a lower oxygen affinity than ammonia oxidizers.
electron donor for the formation of biomass from For fluctuating ammonium loading rates in
carbon dioxide, the formation of nitrate in the engineering operations, DO can be regulated through
reaction is stoichiometrically coupled to cell growth. online feed-back control [37]. Liao et al. [38]
The nitrate produced is a pollutant and causes reported optimal DO of 0.5-0.7 mg/L at an ammonia
environmental problems. Therefore, further oxidation surface loading of 120 g/m2 day. Oxygen supply
of nitrite to nitrate should be prevented or reduced to under different strategies has been found efficient and
a minimum during the reaction. However, in some economical for nitritation process [39-41]. Samik et
cases Anammox reaction can proceed with little al. [42] used a lab-scale fixed film bioreactor and
nitrate formation as long as the oxygen supply rate is termed feed alkalinity and DO concentrations as
maintained at or below the benchmark oxygenation critical controlling parameters for single-stage
rate [31]. This suggests the possibility of higher total process. They stated that maintaining an alkalinity to
nitrogen removal rates and higher level of safety in ammonia ratio of less than 8 and DO loading of less
terms of nitrate discharge from single-stage reactor. than 0.06 mg O/mg N day are necessary to inhibit
nitratation and enhance CANON process. Cema et al.
Key Control Factors [43] investigated the effect of DO concentrations on
the nitrogen removal rates in a pilot plant and
DO documented average nitrogen removal rates of
1.8±0.31 g N/m2day at oxygen concentrations around
Under micro-aerobic conditions, different 3 g O2/m3. However, lab-scale studies indicate
reactor configurations have been employed to obtain optimal DO values in the range of 0.5-1 mg/L for
higher AOB growth rate than the NOB in single- single-stage process [1]. Frequent monitoring by DO
stage system. AOB can not produce a sufficient meter and adjusting the air flow rate accordingly may
amount of NO2-N for Anammox bacteria under too be a useful strategy to maintain optimum DO
low DO concentrations. However, too high DO concentration in the reactor.
concentrations in the bulk phase may cause inhibition
effects on the Anammox bacteria [32]. Sliekers et al. Ammonia, Nitrite and pH
[8] and Szatkowska et al. [33] reported oxygen
The experimental results indicated that an
transfer as the limiting factor for a lab-scale air-lift
ammonia loading rate of 14 mg/[Link] provided a
and a pilot-scale moving bed reactor, respectively.
sufficient ammonia source in CANON [44]. The
This oxygen limitation can be attributed to the slow
interaction between aerobic AOB and [Link]
diffusion into the bio-film or granules. With a
affects single-stage process [44, 45].These two types
simulation study, Hao et al. [34] observed that
of bacteria live on different substrates, with aerobic
maximum nitrogen removal rate was achieved only
AOB requiring ammonia and oxygen and anaerobic
when DO concentration kept pace with the
AOB requiring ammonia and nitrite. Therefore, the
Ghulam Abbas et al. [Link]., Vol. 36, No. 4, 2014 779

process will be disrupted by the presence of NOB denitrifiers [55]. However, Kartal et al. [56] and
because, NOB use oxygen and nitrite substrates and Güven et al. [57] both observed that some organic
compete with aerobic AOB for oxygen and anaerobic carbon sources (acetate, propionate) do not have an
AOB for nitrite. The lowest ammonium concentration inhibition effect on the Anammox activity.
for a stable CANON system was found to be 0.1 kg Ruscalleda et al. [58] stated that co-existence of
N/m3day. If the influx of nitrogen is lower than the Anammox and denitrifiers can treat streams with high
critical NH4 + influx, the stoichiometry of reaction is quantities of slowly biodegradable organic carbon.
affected, and this causes a temporary decrease of Growth of heterotrophic denitrifying is limited by the
nitrogen removal from 92% to 57% [46]. low availability of easily biodegradable organic
carbon and denitrifiers are not able to dominate in
Higher concentrations of ammonium and
these systems and could not out-compete Anammox
nitrite were identified as inhibitors of the Anammox
organisms. Ni et al. [59] revealed that low organic
bacteria activity [47, 48]. Fux et al. [49] reported
matter concentration did not affect ammonia and
serious inhibition of Anammox activity at nitrite
nitrite removal significantly and improved total
concentration of 30-50 mg NO2-N/L during six days.
nitrogen removal via denitrifiers. However, high
However, Strous et al. [47] and Dapena-Mora et al.
organic matter could suppress Anammox activity,
[48] identified different ammonium and nitrite
resulting in a lower ammonia removal.
concentrations that inhibited the Anammox activity.
If nitrite is consumed at about the same rate as it is The threshold concentration in which
produced, then, inhibition effect is not significant. No denitrifiers out-compete Anammox bacteria differ
negative effect of nitrite was observed by Vazquez- from report to report. Güven et al. [57] stated that
Padin et al. [26] although during the first 100 days of Anammox bacteria are not longer able to compete
operation a mean nitrite concentration of 25 mg N/L with heterotrophic denitrifying bacteria at C/N ratio
was registered. Probably a concentration gradients above 1 while Chamchoi et al. [60] stated that an
inside the granules resulted in a low nitrite organic matter concentration above 300 mg COD/L
concentration at the location of the Anammox or COD to N ratio of over 2.0 inactivates Anammox
bacteria. Jaroszynski et al. [50] revealed that nitrite organisms. Milenuevo et al. [61] observed a complete
toxicity has been overestimated in some literature and inhibition of the Anammox process at COD
nitrite as high as 170-250 mg NO2-N/L did not cause concentrations up to 292 mg/L while Tang et al. [62]
deactivation of Anammox consortium despite 2 days stated that denitrifiers became dominant at high
of exposure time. influent COD:NO2-N ratio of 2.9:1. Anammox
activity is completely and irreversibly inhibited by
Lab-scale studies have shown 7-8 as
low concentrations of methanol (15 mg/L) and
optimum pH range for single-stage process [1]. pH
ethanol [57].
affects the substrate availability, quality of effluent
and causes inhibition [51]. Jubany et al. [52] and Status and Applications of Single-stage Process
Tora et al. [53] reported that free ammonia affects
NOB more than AOB populations and it can be used Status: Ample experimental knowledge on
to enrich AOB and wash-out NOB. Shan Li et al. [54] single-stage autotrophic nitrogen removal has been
studied the inhibition of [Link], AOB, and NOB documented by research groups around the world.
activities by pH-related free ammonia under different The maximal influent ammonia concentration for
conditions. single-stage process is 431.7 ± 25.5 mg N/L, the
maximal nitrogen loading rate is 5.44 kg N/m3day,
Organic Matter the maximal nitrogen removal rate is 2.57 kg
N/m3day and the minimal HRT is 4 hr [63, 9]. The
Most of the real wastewaters contain both lab-scale efforts resulted in a growing number of
organic carbon and nitrogen. Several studies reported pilot and full-scale applications.
that presence of organic matter has a negative impact
on Anammox growth [1]. Many genera of Applications: Single-stage nitrogen removal
denitrifying bacteria can use NO3− and NO2− to technology has been implemented in several pilot and
degrade organic matter. So, in the presence of full-scale applications with promising results.
organic carbon, Anammox organisms are no longer
able to compete for nitrite with heterotrophic Cema et al. [64] reported a nitrogen removal
denitrifiers. Thermodynamically, denitrification rate of 1.9 g/m2day in a pilot plant and 3 g N/m2day
reaction is more feasible than Anammox reaction and in the parallel batch tests, where the pilot plant’s
higher growth yield of denitrifiers also renders the conditions were simulated.
Anammox species less competitive with the
Ghulam Abbas et al. [Link]., Vol. 36, No. 4, 2014 780

Table-2: Performance of pilot and full-scale single-stage nitrogen removal systems.

Reactor type Vol.(m3) Influent type DO(mg O2/L) NRR(kgN/ m3d) Removal efficiency (%) Reference
500 Sludge liquor 0.3 0.6 84 [68]
SBR 400 Sludge liquor 0.3 0.4 90 [69]
4.1 Sludge liquor 0.5–1.0 0.65 90 [70]
600 Sludge liquor 2.0–3.0 1.3 75–80 [70]
Up-flow reactor
Full scale Landfill leachate 0.5 0.33 73 [71]
MBR
Full scale Landfill leachate 0.3 0.33 84 [72]
0.04 Sludge liquor <1.0 0.5 60–70 [73]
0.04 Sludge liquor 0.8–2.0 0.12–0.22 60–70 [74]
Moving bed 21 Sludge liquor 1.2–2.6 0.38 62 [75]
Full scale Sludge liquor 3 0.35 64 [76]
Full scale Sludge liquor _ 0.21 72 [76]
265 Landfill leachate 0.7–1.0 0.15–0.26 40–70 [77]
RBC 33 Landfill leachate 1.0–2.0 0.25–0.57 30–70 [78]
240 Landfill leachate 0.8–1.2 1.7 30–70 [79]

The plant in Strass treats the wastewater of retention options and protection from adverse
200,000 population equivalents, and is equipped with shocks.
a 500m3 SBR for deammonification of reject-water 4. Full-scale applications of single-stage system
originating from digested sludge dewatering [65]. have been realized at different locations but
Joss et al. [66] used digester liquid in a full-scale using only sludge digester effluents and landfill
single suspended growth SBR plant (1400m3) to leachates for ammonium nitrogen removal.
achieve ammonium oxidation rates of up to 500 g More scientific knowledge is needed to extend
N/m3day with a nitrogen conversion of over 90%. applicability of this technology to other areas
Bertino, [67] performed pilot plant study on CANON and promote its worldwide industrial
process for four months at 25°C in a 200 L applications.
continuous stirred tank reactor, filled with 40% of
Kaldnes media. He reported nitrogen removal rate of Acknowledgment
2.85 g N/m2day under ammonium surface loading of
3.45 g N/m2day. Removal efficiencies of 95%, 85% The study was supported by the National High-
and 83% were achieved for NH4+-N, inorganic Tech Research and Development (R&D) Program of
nitrogen, and total nitrogen, respectively. China (2009AA06Z311), the Natural Science
Performance of pilot and full scale single-stage Foundation (31070110), Zhejiang Provincial Natural
nitrogen removal systems is given in Table 2[68-79]. Science Foundation (Z5110094), and Higher
Education Commission (HEC) of Pakistan.
Conclusions and Future Prospects

1. Single-stage technology is more economical and References

environment friendly than conventional
technology and it offers advantages of less 1. W.H.S. Van Hulle, J.P.H Vandeweyer, B.D.
construction, operating and maintenance costs. Meesschaert, P.A. Vanrolleghema, P. Dejans, A.
Process simplicity, little aeration and no Dumoulin, Chemical Engineering Journal, 162,1
additional organic carbon source are the added (2010).
advantages which can save much energy and 2. T. Khin, A.P. Annachatre. Biotechnology
resources. Advances, 22, 519 (2004).
2. Single-stage technology has some good 3. S. Wyffels, K. Pynaert, P. Boeckx, W. Verstraete,
characteristics compared with two-stage O.V. Cleemput, Water Research, 37, 1252
technology such as higher nitrogen removal rate, (2003).
no need to control nitrite ammonia ratio, no risk 4. C. Helmer-Madhok, [Link], E. Filipov, T.
to overload the system with NO2-N and lower Gaul, A. Hippen, K.H. Rosenwinkel, C.F.
HRT. But as single-stage systems are Anammox Seyfried, M. Wagner, S. Kunst, Water Science
based, so, further research is required to shorten Technology, 46, 223 (2002).
the start-up period. 5. A.O. Sliekers, N. Derwort, J.L. Campos Gomez,
3. Ammonium oxidation rate in one reactor system [Link], J.G. Kuenen, M.S.M. Jetten. Water
depends greatly on the nature of nitrifying Research, 36, 2475 (2002).
culture and operational characteristics which 6. K. Pynaert, B.F. Smets, S. Wyffels, D. Beheydt,
necessitates more studies on oxygen transfer, S.D. Siciliano, W. Verstreate Applied
microbial communities, effective biomass Environmental Microbiology, 69, 3626 (2003).
Ghulam Abbas et al. [Link]., Vol. 36, No. 4, 2014 781

7. T. Liu, D. Li, H. Zeng, X. Li, T. Zeng, X. Chang, 27. Y.T. Lv, L. Wang, T. Sun, X.D. Wang, Y.Z.
Y. Cai, J. Zhang, Bioresource Technology, 118, Yang, Z.Y. Wang, Chemosphere, 79, 180 (2010).
399 (2012). 28. A. Daverey, S.H. Su, Y.T. Huang, J.G. Lin,
8. A.O. Sliekers, K.A. Third, W. Abma, J.G. Bioresource Technology, 113, 225 (2012).
Kuenen, M.S.M. Jetten, FEMS Microbiology 29. H. Chen, S. Liu, F. Yang, Y. Xue, T. Wang,
Letters, 218, 339 (2003). Bioresource Technology, 100, 1548 (2009).
9. L. Wang, P. Zheng, T. Chen, J. Chen, Y. Xing, 30. C.J. Lan, M. Kumar, C.C. Wang, J.G. Lin,
Q. Ji, M. Zhang, J. Zhang, Bioresource Bioresource Technology, 102, 5514 (2011).
Technology, 123, 78 (2012). 31. W. Kwak, P.L. McCarty, J. Bae, Y.T. Huang,
10. L. Kuai, W. Verstraete, Applied Environmental P.H. Lee, Bioresource Technology, 123,400
Microbiology, 64, 4500 (1998). (2012).
11. C. Helmer, C. Tromm, A. Hippen, K.H. 32. B. Kartal, W.R.L vander Star, M. C. Schmid, V.
Rosenwinkel, C.F. Seyfried, S. Kunst, Gwf, 140 Pas-Schoonen, K.C. Picioreau, W. Abma, R.O.H.
Nr. 9, 622 (1999). Camp, M.S.M. Jetten, V. Loosdrecht, M. Strous,
12. A. Hippen, K.H. Rosenwinkel, G. Baumgarten, Proceeding of the CLONIC Final Workshop,
C.F. Seyfried, Water Science Technology, 35 Barcelona, Spain, (2007).
111 (1997). 33. B. Szatkowska, PhD Thesis, Performance and
13. D. Paredes, P. Kuschk, T. S. A. Mbwette, F. control of biofilm systems with partial nitritation
Stange, R. A. Muller, H. Koser. Engineering Life and Anamox for supernatant treatment, Royal
Sciences, 7, 13(2007). Institute of Technology, Stockholm, Sweden,
14. K. Furukawa, H. Tokitoh, P.K. Lieu, T. Fujii (2007).
Proceeding of the Sino Japanese forum on 34. X. Hao, J.J. Heijnen, M.C.M. van Loosdrecht,
protection and restoration of water environment, Biotechnology Bioengineering, 77, 266 (2001).
Beijing (2004). 35. N. Bernat, N. Dangjong, J.P. Delgenes, R.
15. P.K. Lieu, R. Hatozaki, H. Homan, K. Furukawa, Moletta, Journal Environmental
Japanese Journal Water Treat Biology, 41,103 Engineering,127, 266 (2011).
(2005). 36. H.P. Chuanga, A. Ohashia, H. Imachib, M.
16. H. Chen, S. Liu, F. Yang, Y. Xue, T. Wang, Tandukara, H. Haradaa, Water Research, 41, 295
Bioresource Technology, 100, 1548 (2009). (2007).
17. K. Pynaert, B.F. Smets, D. Beheydt, W. 37. M. Nielsen, A. Bollmann, A.O. Sliekers, M.
Verstraete, Environmental Science Technology, Jetten, M. Schmid, M. Strous, I .Schmidt, L.H.
38, 1228 (2004). Larsen, L.P. Nielsen., N.P. Revsbech, FEMS
18. S. Van Hulle, PhD Thesis, Modelling, simulation Microbiology Ecology, 51, 247 (2005).
and optimization of autotrophic nitrogen 38. D.X. Liao X.M. Li, G.M. Zeng, Q. Yang, G. J.
removal processes, Ghent University, Belgium, Yang, China Environmental Science, 25, 222
(2005). (2005).
19. K. Furukawa, P.K. Lieu, H. Tokitoh, T. Fujii, 39. J. GUO, G. Yang, F. FANG, Y. QIN, Frontiers
Water Science Technology, 53, 83 (2006). Environmental Science Engineering in China,
20. K.A. Third, A.O. Sliekers, J.G. Kuenen, M.S.M. 439 (2008).
Jetten., Systematic Applied Microbiology, 24,588 40. C. Wantawin, J. Juateea, P.L. Noophan, J.
(2001). Munakata-Marr, Water Science Technology,
21. G. Zhu, Y. Peng, B. Li, J. Guao, Q. Yang, S. IWA Publishing (2008).
Wang, Reviews Environmental Contamination 41. M. Zubrowska-Sudol, J. Yang, J. Trela, E. Plaza,
Toxicology, 192,159 (2008). Water Science Technology, IWA Publishing
22. M. Strous, V. Gerven, Z.E. Ping, J.G. Kuenen, (2011).
M.S.M. Jetten, Water Research, 31,1955 (1997). 42. S. Bagchi, R. Biswas, T. Nandy, Journal
23. T. Gaul, E. Filipov, N. Schlosser, S. Kunst, C. Industrial Microbiology Biotechnology,
Helmer-Madhok, Water Science Technology,46 37,871(2010).
157 (2002). 43. G. Cema, E. Płaza, J. Trela, J. Surmacz-Górska,
24. Y. Ahn, H.C. Choi, Process Biochemistry, 41, Water Science Technology, IWA Publishing
1945 (2006). (2011).
25. Z. Gong, F. Yang, S. Liu, H. Bao, S. Hu, K. 44. K.A. Third, A.O. Sliekers, J.G. Kuenen, M.
Furukawa., Chemosphere, 69,776 (2007). Jetten, Systematic Applied Microbiology, 24,588
26. J.R. Vazquez-Padin, I. Figueroa, A. Mosquera- (2001).
Corral, J.L. Campos, R. Méndez, Water Science
Technology, 60, 1135 (2009).
Ghulam Abbas et al. [Link]., Vol. 36, No. 4, 2014 782

45. K.A. Third, J. Paxman, M. Schmid. M. Strous, 64. G. Cema, B. Szatkowska, J. Trela, Surmacz-
M. Jetten, R. Cord-Ruwisch, Microbiology Górska, Water Science Technology, 54, 209
Ecology, 49, 236 (2005). (2006).
46. T. Khin, A.P. Annachatre, Biotechnology 65. G. Innerebner, H. Insama, I.H. Franke-Whittle, B.
Advances, 22, 519 (2004). Wett, Systematic Applied Microbiology, 30, 408
47. M. Strous J.G. Kuenen, M. Jetten, Applied (2007).
Environmental Microbiology, 65, 3248 (1999). 66. A. Joss, D. Salzgeber, J. Eugster, R. Knig, K.
48. A. Dapena-Mora, I. Fernández, J.L. Campos, A. Rottermann, S. Burger, P. Fabijan, S. Leumann,
Mosquera-Corral, R. Mendez, M.S.M Jetten, J. Mohn, H. Siegrist, Environmental Science
Enzyme Microbial Technology, 40, 859 (2007). Technology, 43 (2009).
49. C. Fux, H. Siegrist, Water Science Technology, 67. A. Bertino, PhD Thesis, Study on one-stage
50, 19 (2004). Partial Nitritation-Anamamox process in
50. L.W. Jaroszynski, N. Cicek R. Sparling, J.A. Moving Bed Biofilm Reactors: a sustainable
Oleszkiewicz, Bioresource Technology, 102, nitrogen removal KTH, Sweden (2010).
7051 (2011). 68. B. Wett, Water Science Technology, 53, 121
51. A. Magri, L.I. Corominas, H. Lopez, E. Campos, (2006).
M. Balaguer, J. Colprim, Environmental 69. G. Nyhuis, V. Stadler, B. Wett, I. derersten, S.
Technology, 28, 255 (2010). Technik, Aachen, ATEMIS GmbH, Germany
52. I. Jubany, J. Lafuente, J.A. Baeza, J. Carrera, (2006).
Water Research, 43, 2761 (2009). 70. S.E. Vlaeminck, L.F.F. Cloetens, H. De
53. J.A. Tora, J. Lafuente, J.A. Baeza, J. Carrera, Clippeleir, M. Carballa, W. Verstraete, Water
Bioresource Technology, 101, 6051 (2010). Science Technology, 59, 609 (2009).
54. [Link], Y.P. Chen, C. Li, J.S. Guo, F. Fang, X. Gao, 71. V. Rekers, M. Denecke, U. Walter, Depotech,
Applied Biochemical Biotechnology, 167,694 Leoben (2008).
(2012). 72. M. Denecke, V. Rekers, U. Walter, Muell und
55. M. Kumar, J.G. Lin, Journal of Hazardous Abfall, 39, 4 (2007).
Materials, 178, 1 (2010). 73. P. Johansson, A. Nyberg, M. Beier, A. Hippen,
56. B. Kartal, [Link], L.A. van Niftrik, [Link] de C.F. Seyfried, K.H. Rosenwinkel, Royal Institute
Vossenberg, M.C. Schmid, R.I. Webb, S. of Technology, Stockholm, Report 3048 (1998).
Schouten, J.A. Fuerst, J.S. Damsté, M.S.M. 74. A. Hippen, C. Helmer, S. Kunst, K.H.
Jetten, M. Strous, Systematic Applied Rosenwinkel, C.F. Seyfried, Water Science
Microbiology, 30,39 (2007). Technology, 44, 39 (2001).
57. D. Güven, A. Dapena, B. Kartal, M.C. Schmid, 75. B. Szatkowska, G. Cema, E. Plaza, J. Trela, B.
B. Maas, K. van de Pas-Schoonen, S. Sozen, R. Hultman, Water Science Technology, 8, 19
Mendez, H.J.M. Op den Camp, M.S.M. Jetten, (2007).
M. Strous, I. Schmidt, Applied Environmental 76. M. Beier, Y. Schneider, Leibniz, Hannover,
Microbiology, 71, 1066 (2005). Germany (2008).
58. M. Ruscalleda, H. López, R. Ganigué, S. Ouig, 77. A. Hippen, K.H. Rosenwinkel, G. Baumgarten,
M. Balaguer, J. Colprim, Water Science C.F. Seyfried, Water Science & Technology, 35,
Technology, 58, 1749 (2008). 111 (1997).
59. S.Q. Ni, J.Y. Ni, D.L. Huc, S. Sung, Bioresource 78. H. Siegrist, S. Reithaar, G. Koch, P. Lais, Water
Technology, 110, 701 (2012). Science Technology, 38, 241 (1998).
60. N. Chamchoi, S. Nitisoravut, J.E. Schmidt, 79. M. Schmid, K. Walsh, R. Webb, W.I.C. Rijpstra,
Bioresource Technology, 99, 3331(2008). K. van de Pas-Schoonen, M.J. Verbruggen, T.
61. B. Molinuevo, M.C. Garcia, D. Karakashev, I. Hill, B. Moffett, J. Fuerst, S. Schouten, J.S.
Angelidaki, Bioresource Technology, 100, 2171 Sinninghe Damsté, J. Harris, P. Shaw, M. Jetten,
(2009). M. Strous, Systematic Applied Microbiology, 26,
62. C. Tang, P. Zheng, C. Wang, Q. Mahmood, 529 (2003).
Bioresource Technology, 101, 1762 (2010).
63. S. Bagchi, R. Biswas, T. Nandy, Journal
Industrial Microbiology Biotechnology, 37,871
(2010).