Biochemical Engineering Journal 43 (2009) 191–196

Contents lists available at ScienceDirect

Biochemical Engineering Journal

journal homepage: www.elsevier.com/locate/bej

Simultaneous nitrification and denitrification coupled with phosphorus removal

in an modified anoxic/oxic-membrane bioreactor (A/O-MBR)
Zhimin Fu, Fenglin Yang ∗ , Yingyu An, Yuan Xue
Key Laboratory of Industrial Ecology and Environmental Engineering, MOE, School of Environmental and Biological Science and Technology,
Dalian University of Technology, Linggong Road 2, Dalian, Liaoning 116024, PR China

a r t i c l e i n f o a b s t r a c t

Article history: In this study, simultaneous carbon, nitrogen and phosphorus (PO4 3− -P) removals using an anoxic/oxic-
Received 25 January 2008 membrane bioreactor (A/O-MBR) system without any circulation was investigated. The total nitrogen (TN)
Received in revised form 24 August 2008 removal efficiency was above 84.6%, and the average removal efficiencies of COD and PO4 3− -P were 94.6
Accepted 29 September 2008
and 90.0%, respectively. Ammonium (NH4 + -N) removal without nitrite (NO2 − -N) and nitrate (NO3 − -N)
accumulation in the reactor suggested that simultaneous nitrification and denitrification (SND) occurred.
Keywords:
The phosphorus-accumulating organisms (PAOs) accumulated in the reactor. AnOx/Oxic test indicated
Biological nitrogen removal
that intracellular organic carbon might be utilized as denitrification electron donor in nitrogen removal
Biological assimilation
Phosphorus-accumulating organisms
process.
Poly-␤-hydroxybutyrate © 2008 Elsevier B.V. All rights reserved.
Membrane bioreactor

1. Introduction of an anoxic zone to enable denitrification was investigated to

Nitrogen removal is achieved by aerobic nitrification and
anoxic denitrification using autotrophic nitrifying and het-
erotrophic denitrifying bacteria. Thus two separate reactors are
required to provide different environments for the two kinds
of bacteria in nitrogen removal process. Conventional biological
nitrification–denitrification process combined anoxic and aerobic
units with NO3 − -N recycle, because the circulation of liquid that
contains NO3 − -N and NO2 − -N was necessary. Researchers found
that when recycle ratio increased, denitrification efficiency would
be strengthened [1].
Presently, the membrane bioreactor (MBR) process has attracted
considerable interest due to various advantages compared to a
conventional activated sludge process, including high concentra-
tions of biomass; the separation of solids retention time (SRT) and
hydraulic retention time (HRT); good effluent quality, which orig-
inate from the use of a membrane for solid–liquid separation [2].
Earlier studies had already shown that treatment performances of
MBR were better than conventional treatment [3].
Biological nitrogen removal has been achieved by various
configurations of MBR systems [4–7] and pre-denitrification is
always the preferred configuration. A pilot MBR plant comprised
∗ Corresponding author. Tel.: +86 411 8470 6172; fax: +86 411 8470 6171.
E-mail address: fuzhimin2004@yahoo.com.cn (F. Yang).
treat municipal wastewater. The results showed that nitrification
was complete and up to 82% of the total nitrogen (TN) could be
denitrified when a 300% recycle ratio was used [8]. Tan and Ng [9]
had illustrated that the internal mixed liquor recycle flow from
the oxic to the anoxic zone played an important role during the TN
removal process in a pre-denitrification system. Calculations from
the nitrification–denitrification model showed that denitrification
was limited by mixed liquor recycle ratio under low aeration rate.
In recent years, more studies have shown that nitrification
and denitrification could occur concurrently in one reactor under
aerobic conditions with low dissolved oxygen (DO), through the
so-called simultaneous nitrification and denitrification (SND) pro-
cess [10]. SND relied on concurrent aerobic NH4 + -N oxidation
and anaerobic denitrification under identical operating conditions.
Additionally, it required a slowly degradable organic substrate to
provide reducing power for denitrification.
Wastewater containing organic substances affected nitrification
process, for the sake of growth competition between autotrophic
and heterotrophic microorganism populations. Reporters [11]
showed that, during the feast period (i.e., external organic carbon
present), heterotrophic respiration activity was high and nitrifi-
cation was prevented due to the inability of nitrifying bacteria to
compete with heterotrophs for oxygen. Nitrification took place only
after external organic carbon exhaustion, when the oxygen compe-
tition was low. In anoxic/oxic process, organic matter was mainly
consumed in anoxic stage, which apparently resulted in lower com-

1369-703X/$ – see front matter © 2008 Elsevier B.V. All rights reserved.
doi:10.1016/j.bej.2008.09.021
192 Z. Fu et al. / Biochemical Engineering Journal 43 (2009) 191–196
petition between nitrifiers and heterotrophs in the next aerobic
stage. Previous studies [11–16] had shown that storage polymers
(e.g., poly-␤-hydroxybutyrate, PHB) were degraded much slower
than soluble substrate and could be used as the electron donor for
denitrification when no external substrate was available. If carbon
source could be stored as polymers (e.g., PHB) under anoxic condi-
tion and then be utilized as electron donor for denitrification under
oxic condition, the circulation of liquid from aerobic to anoxic zone
would be unnecessary.
Until now, most of successful SND recorded were obtained in
sequencing operation process [15], but few studies were achieved
in continuous-flow processes. Furthermore, although there were
numerous reports on carrying out nitrogen removal in differ-
ent kinds of reactor, such as circulating fluidized bed bioreactor
(CFBB) [17], horizontal-flow biofilm reactor (HFBR) [18] and air-
lift bioreactor [19], little information was currently available on
the behaviors of carbon and nitrogen removal in A/O-MBR with-
out NO3 − -N liquid recycle. Therefore, this study attempted to
develop an A/O-MBR without NO3 − -N recycle for simultaneous
carbon, nitrogen and PO4 3− -P removals for treating high strength
wastewater.
2. Materials and methods
2.1. Experimental set-up
In phases I and II, the laboratory scale A/O-MBR (Fig. 1a) con-
sisted of three Plexiglas units with each volume of 16 L, an anoxic
zone with a stirrer, an oxic zone and a membrane department
with a submerged hollow fiber membrane module (polypropylene,
Fig. 1. Sketch map of A/O-MBR. (a) A/O-MBR consisted of anoxic/oxic/membrane
units. (b) A/O-MBR consisted of anoxic/oxic units. (1) Feed tank; (2) balance tank;
(3) stirrer; (4) A/O-MBR; (5) suction pump; (6) membrane model; (7) air flow meter;
(8) air pump.
hydrophilic, pore size: 0.1 ␮m, Hangzhou Kaihong Membrane Tech-
nology Co. Ltd., China). In phase III, the clapboard between aerobic
and membrane departments in the A/O-MBR dropped acciden-
tally. Thus the valid volumes of anoxic zone and aerobic zone were
artificially adjusted to 6 and 12 L, respectively. The schematic dia-
gram of reconstructive A/O-MBR was shown in Fig. 1b. The influent
was continuously fed into the anoxic department and then flow to
the MBR. The permeation was continuously extracted by a suction
pump. Hydraulic retention time (HRT) was kept as 1.5 d. The con-
stant fluxes were 3.8 L/m2 h in phases I, II and 2.5 L/m2 h in phase
III.
2.2. Reactor operating conditions
The seed sludge was taken from a native sewage wastewater
treatment plant (Chun-liu, Dalian) and the initial sludge concen-
tration was about 3800 mg/L. The pH was maintained at 7.5–8.5 in
the reactor. All experiments were performed at room temperature
(25 ◦ C). The DO concentration in anoxic zone and aerobic zone were
<0.3 and 0.4–2.0 mg/L, respectively. There was no sludge discharged
except sampling to maintain large amounts of biomass.
The continuous operation periods were divided into three
phases for different influent NH4 + -N concentration, denoted as I, II
and III, respectively. A synthetic wastewater comprising of sucrose,
NH4 Cl and KH2 PO4 was used in this study. Alkalinity was sup-
plied according to 7.07 g CaCO3 /g NH4 + -N. One liter of nutrient
solution contained: 0.8 g FeSO4 ·7H2 O; 0.02 g CuSO4 ·5H2 O; 0.4 g
MnCl2 ·4H2 O; 0.1 g Na2 MoO4 ·2H2 O; 0.08 g ZnSO4 ·7H2 O and 0.002 g
CoCl2 ·6H2 O. Table 1 shows the general components and operation
conditions for phases I, II and III.
Membrane washing was performed when the transmembrane
pressure (TMP) reached 40 kPa. During phase I, the membrane suc-
tion pressure was found to increase slowly, therefore, it was not
cleaned chemically until the phase stopped. In phases II and III, the
membrane was cleaned chemically every 20–22 d. Furthermore, it
was noticeable that in phases II and III the membrane was daily
artificially cleaned by wiping off sludge cake.
2.3. Analytical methods
COD, NH4 + -N, NO2 − -N, NO3 − -N, PO4 3− -P and mixed liquid
volatile suspend solid (MLVSS) were measured according to stan-
dard methods [20]. DO and pH were measured by DO (Modle 55/12,
YSI, USA) and pH (PB-10 Sartorius, Germany) meters, respectively
(Aqualytic). The value for TN was based on the sum of NH4 + -N,
NO2 − -N and NO3 − -N rather than an independent test.
2.4. Batch tests
Mixed liquor was regularly taken from the reactor to analyze the
character and elucidate the mechanism of nitrogen removal. The
activated sludge sample after centrifugation at 3000 × g for 10 min
was washed three times and then diluted to about 3000 mg/L with
deionized water. Certain sucrose, NH4 Cl and KH2 PO4 were added
to the mixed liquor, resulting in concentrations of 1000 mg COD/L,
100 mg NH4 + -N/L and 10 mg PO4 3− -P/L. During batch test, the solu-
tion pH was maintained at 7.5–8.5 using NaHCO3 . Liquid samples
were intermittently removed to analyze COD, NH4 + -N, NO2 − -N,
NO3 − -N and PO4 3− -P. Tests were performed as follows.
AnOx/Oxic test. At the start of the experiment, 500 mL of mixed
liquor were added to 1000 mL Erlenmeyer flasks (Pyrex). It was then
agitated on a thermostatic shaker at 200 rpm. Two hours later, air
was sparked to start the aerobic stage, which lasted for 4 h. DO was
controlled at 2.5 mg/L.
 Z. Fu et al. / Biochemical Engineering Journal 43 (2009) 191–196
Table 1
Components and operation conditions of different phases.
Phase COD (mg/L) NH4 Cl (mg N/L)
I 1985.1 ± 250.3 112.2 ± 7.6
II 2014.7 ± 132.7 169.7 ± 18.3
III 1990.6 ± 332.4 212.3 ± 21.3
2.5. Fluorescence in situ hybridization (FISH)
Sample fixation and FISH was performed as previously described
in Amann et al. [21] with Cy3-labelled EUB338 probes (for most bac-
teria) and either FITC-labeled PAOMIX probes (comprising equal
amounts of probes PAO462, PAO651 and PAO846) [22]. FISH prepa-
rations were visualized with a leica TCS ST2 (Germany) confocal
laser scanning microscope (CLSM).
3. Results
3.1. COD, NH4 + -N, PO4 3− -P removal
Fig. 2a presents the performance of the organic carbon removal
through the experiment. A stable and high average COD removal
efficiency of 94.6% was maintained; even the COD concentration
fluctuated between 1250 and 3000 mg/L. The average membrane
effluent COD was below 100 mg/L through all the operation, which
suggested the system with a capability of counteracting influent
fluctuation after membrane filtration to offer a high effluent quality.
The average COD removal efficiencies were 95.4, 94.6 and 95.8%, at
phases I, II and III, respectively. It was noticeable that the membrane
separation contributed 1.8–12.6% to COD removal, which indicated
the overall COD removal was mainly due to biological degradation
rather than membrane separation.
Fig. 2b displays the performance of NH4 + -N concentration in
the influent, effluent and removal efficiency under various influ-
ent concentrations. The average influent NH4 + -N concentration
increased gradually to investigate its impact on the nitrogen
removal performance of the A/O-MBR. A high average NH4 + -N
removal efficiency of 85.6% was maintained; even the max influent
NH4 + -N concentration increased to 275 mg/L.
During phases I and II, the average influent NH4 + -N concen-
trations were 112.2 ± 7.6 and 169.7 ± 18.3 mg/L, respectively. The
average NH4 + -N removal efficiencies were 94.8 and 94.7%, and the
residual concentrations stabilized at 2.3–10.9 and 8.5–14.6 mg/L,
respectively. During phase III, the influent average NH4 + -N concen-
tration was increased to 212.3 ± 21.3 mg/L. The NH4 + -N removal
efficiency decreased to around 75.2% and then increased to
87.8%, the effluent NH4 + -N concentration was finally decreased to
25.2 mg/L. Although the influent NH4 + -N fluctuated greatly, the
membrane effluent NH4 + -N was always below 75 mg/L.
As shown in Fig. 2b, the most of NH4 + -N was removed in the A/O-
MBR and no obvious NOX − -N accumulation was observed. Namely,
the TN loss occurred in the aerobic zone suggesting that SND took
place. The average NOX − -N concentration was stabilized at low
level (4.6 ± 0.6 mg/L) in aerobic zone all through the operation. TN
removal efficiencies were 91.8, 91.9 and 84.6%, respectively.
Table 2
Summary of nitrogen removal during the operation time.
Phase NH4 + -Ninf (mg/L) NH4 + -Neff (mg/L) TNeff (mg/L)
I 112.2 ± 7.6 5.9 ± 5.9 9.1 ± 6.1
II 169.7 ± 18.3 7.2 ± 1.7 9.2 ± 8.5
III 212.3 ± 21.3 29.9 ± 9.5 37.9 ± 11.1
TNeff : total nitrogen concentration in effluent; NLR: nitrogen loading rate; ANRLR: NH4 + -N removal loading rate; TNRLR: TN removal loading rate.
193
KH2 PO4 (mg P/L) Days (d) HRT (d) SRT (d)
20.0 ± 5.0 0–40 1.5 ∞
20.0 ± 5.0 40–82 1.5 ∞
20.0 ± 5.0 82–160 1.5 ∞
Fig. 2c presents the PO4 3− -P concentration in influent, anoxic,
oxic, effluent and PO4 3− -P removal efficiency during the whole
operation period. The average effluent concentration was 1.8 mg/L
and PO4 3− -P removal efficiency was 90.4%. Under anoxic con-
ditions, PO4 3− -P release was observed periodically as shown in
Fig. 2c. The average PO4 3− -P concentration in anoxic department
was larger than that in the influent while the aerobic PO4 3− -P
concentration was extremely low (<1.3 mg/L) suggesting that the
PO4 3− -P uptake efficiency was excellent (>95.8%). Consequently, it
was speculated that PAOs might be accumulated in the A/O-MBR
system. As shown in Fig. 3, PAOs were gathered in the A/O-MBR sys-
tem, though the results demonstrated that the abundance of PAOs
identified by the probes was not very large. It should be pointed
out that the effluent PO4 3− -P concentration was slightly higher
than that in aerobic zone. The reason might be that the cake layer
formed on the fouled membrane module could facilitate PO4 3− -
P release, as it was [23] reported that PAOs could release internal
poly-phosphate (poly-P) under anaerobic conditions.
3.2. Batch tests results
A simulated cycle study (AnOx/Oxic experiment) was car-
ried out in order to explore the mechanism of TN removal.
Fig. 4 shows the typical profiles obtained in cycle study. During
anoxic phase (0–2 h), organic carbon (account as COD) was con-
sumed by heterotrophic organism and the rate was accounted to
421.3 mg/L h. The NH4 + -N concentration increased slightly from
94.5 to 105.2 mg/L. The PO4 3− -P released rapidly from 12.2 to
26.4 mg/L, the released rate was accounted to 27.8 mg/L h, in the
first 0.5 h. Then PO4 3− -P release reaction slowed down. Finally, the
PO4 3− -P concentration was increased to 31.7 mg/L.
In the aerobic phase, NH4 + -N was consumed accompanied by
NOX − -N forming, while PO4 3− -P was taken up from the medium.
The organic carbon decreased from 123.4 to 62.3 mg/L after aera-
tion for 1 h (2–3 h) and then remained steady for about 3 h (3–6 h).
Totally, 64.3 mg/L NH4 + -N was removed while 13.4 mg/L NO2 − -N
and 5.7 mg/L NO3 − -N were accumulated, respectively, during aera-
tion period. A rapid PO4 3− -P uptake occurred in the first 1 h (2–3 h),
the PO4 3− -P concentration decreased from 31.7 to 9.4 mg/L. After
1 h, the PO4 3− -P uptake rate turned down, 8.6 mg P/L was taken up
in the following 3 h.
4. Discussion
During the operation, the influent NH4 + -N concentration
increased gradually, therefore the nitrogen loading rate (NLR)
increased accordingly (Table 2). However, the sludge loading rate
did not increase, due to the increase of sludge concentration
TN removal efficiency (%) NLR (mg N/L d) ANRLR (mg N/L d) TNRLR (mg N/L d)
91.8 74.8 ± 5.1 70.9 ± 6.1 68.8 ± 6.1
91.9 113.1 ± 12.2 107.0 ± 10.9 103.9 ± 11.5
84.6 141.5 ± 13.3 120.1 ± 22.4 119.2 ± 22.1
194 Z. Fu et al. / Biochemical Engineering Journal 43 (2009) 191–196
Fig. 2. Nutrients removal changes during the operation. (a) COD concentration in the
influent, anoxic, oxic, effluent and COD removal efficiency in different phase. (b) The
nitrogen element concentration in the influent, effluent and removal efficiency in
different phase. (c) The PO4 3− -P concentration in the influent, anoxic, oxic, effluent
and PO4 3− -P removal efficiency in different phase.
Fig. 3. Confocal laser scanning micrographs of FISH. PAO appear yellow (hybridized
with FITC-labelled PAOMIX) and other bacteria appear red (hybridized with Cy3-
labelled EUB338). Bar = 200 ␮m. (For interpretation of the references to color in this
figure legend, the reader is referred to the web version of the article.)
(accounted as MLVSS). The average MLVSS in different phases were
shown in Fig. 5. In oxic zones, the concentrations were 8.0, 13.0
and 13.2 g/L, at phases I, II and III, respectively. While in the anoxic
zones, they were 6.4, 9.7 and 11.1 g/L. The MLVSS of oxic depart-
ment was always higher than that in the anoxic. The HRT was kept
similar at approx. 1.5 d, whereas organic sludge loading rate (OSLR,
273.9–122.2 mg COD/g MLVSS d) and nitrogen sludge loading rate
(NSLR, 8.2–11.1 mg N/g MLVSS d) differed somewhat between the
three phases of the A/O-MBR as shown in Fig. 5. The sludge removal
loading rates were found to have varied considerably in line with
the influent and the differences between them were slight which
indicated that the removal efficiency was high.
Nitrogen could be removed only by assimilation into biomass
or by nitrification–denitrification. Stephenson et al. [24] reported
that the sludge yield was often very low in MBRs which was
operated in a long SRT of more than 20 days. Thus the con-
tribution by assimilation to the TN removal would be low and
nitrification–denitrification would therefore play a more important
role in the removal of nitrogen from wastewater. Tan and Ng [9]
had observed that cell assimilation was estimated to remove about
15–20% of the influent TN concentration in a pre-denitrification
MBR, which demonstrated that TN removal efficiency operating at
long SRT was not due to cell assimilation, but rather contributed
more significantly by the nitrification–denitrification process.
In the A/O-MBR reactor, the NOX − -N recycle liquid was removed.
Furthermore, NH4 + -N was oxidized while no NOX − -N accumulated
in the oxic zone during the operation (Fig. 2b). Thus the nitrogen
removal was through SND process occurred in the oxic zone. The DO
concentration was heterogeneity in the A/O-MBR; therefore, there
existed anoxic area in the sludge under aerobic condition which
could promote the denitrification process.
The relationship between organic matter consumed and nitro-
gen reduced could be determined by denitrification process
stoichiometry and organic matter composition. It was reported
that the stoichiometric requirement for denitrification was
4.2 g COD/g N [25]. Therefore, organic matters consumed by den-
itrification process were 435, 660 and 750 mg COD/L, respectively.
However, the differences of COD concentrations between anoxic
and oxic zones were not tremendous (Fig. 2a); the average val-
ues were 132.6, 64.3 and 35.3 mg/L, respectively. Obviously, the
 Z. Fu et al. / Biochemical Engineering Journal 43 (2009) 191–196 195

Fig. 4. COD, N and PO4 3− -P concentration changes in A/O batch test.

consumed COD was lower than the denitrification required. It
was reasonable to speculate that endogenous carbon source was
utilized for denitrification process. Bernat et al. [16] had shown
that storage polymers (e.g., poly-␤-hydroxybutyrate, PHB) could
be used as the electron donor for denitrification when no external
substrate was available.
Previous studies had shown that biomass subjected to alter-
nating anoxic and aerobic conditions (activated sludge was cycled
between anaerobic and aerobic phases) would promote the accu-
mulation of PAOs [23,26,27]. In this study, sludge recycle line
was omitted suggesting that external anoxic and aerobic condi-
tions alternated for biomass was absence. However, results showed
that PAOs existed in the A/O-MBR system. Thus it was reason-
able to consider that there was an inner sludge recycles between
anoxic and oxic zone which induced PAOs’ accumulation. The liq-
uid recycle flow was mainly dependent on the extent of activated
sludge mixing which was droved by differential concentrations
between the anoxic and oxic zones. Because of the possibility of
inner recycle in the experiment, traditional denitrification might
have been responsible for the TN removal. However, since the
NOX − -N concentration was stabilized at low level (4.6 ± 0.6 mg/L)
in aerobic zone all through the operation, the NOX − -N contained
in recycling liquid was negligible comparing to TN removal. The
contribution of traditional denitrification occurred in anoxic zone
to TN removal could be ignored. Hence, SND occurred in aero-
bic zone played important role in the removal of nitrogen from
wastewater.
In AnOx/Oxic experiment, SND occurred and the efficiency
accounted according to Eq (1) was 69.9%. Totally, 45.2 mg N/L was
removed during aerobic operation period (4 h) (Fig. 4). Theoret-
ically, 189.6 mg COD/L was required, since organic stoichiometric
requirement for denitrification was 4.2 g COD/g N. However, only
61.1 mg COD/L was removed during the reaction (Fig. 5) which was
obviously lower than 189.6 mg COD/L. Hence, it was speculated that

Fig. 5. The changes of MLVSS and sludge loading rates.

196 Z. Fu et al. / Biochemical Engineering Journal 43 (2009) 191–196
part of carbon source stored as polymers (e.g., PHB) under anoxic
conditions was utilized as denitrification electron donor in the next
aerobic stage. The elimination of nitrogenous compounds by SND
was likely transformed to N2 O and N2 gases.
 
NOX − -Nremained
SND (%) = 1− × 100 (1)
NH4 + -Noxidized
where NOX − -Nremained was the NOX − -N concentration in effluent
(mg/L), NH4 + -Noxidized was the NH4 + -N concentration difference
between influent and effluent (mg/L).
Zeng et al. [28] operated a lab-scale SBR in alternating
anaerobic–aerobic mode with a low DO concentration (0.5 mg/L)
during the aerobic period, and demonstrated to accomplish nitrifi-
cation, denitrification, and PO4 3− -P removal. In our AnOx/Oxic test,
the COD, NH4 + -N and PO4 3− -P cycle profile seemed coincident with
their investigation.
5. Conclusions
This paper investigated carbon, nitrogen and phosphorus
removal in the A/O-MBR system without liquid circulation. Results
showed that the removal efficiencies of carbon and PO4 3− -P were
94.6 and 90.0%, respectively, at all different operating phases. The
average NH4 + -N removal efficiencies were 94.8, 94.7 and 85.8%,
respectively, when the average influent NH4 + -N concentrations
were 112.2 (phase I), 169.7 (phase II) and 212.3 mg/L (phase III).
Results showed that the NH4 + -N removed without NOX − -N accu-
mulation in the reactor suggesting that SND occurred. The TN
removal efficiencies were 91.85, 91.87 and 84.60%, respectively.
AnOx/Oxic test indicated that organic carbon stored as polymers
was utilized as denitrification electron donor.
References
[1] J.A. Baeza, D. Gabriel, J. Lafuente, Effect of internal recycle on the nitrogen
removal efficiency of an anaerobic/anoxic/oxic (A2 /O) wastewater treatment
plant (WWTP), Process. Biochem. 39 (2004) 1615–1624.
[2] C. Visvanathan, R. Ben Aim, K. Parameshwaran, Membrane separation biore-
actors for wastewater treatment, Crit. Rev. Environ. Sci. Technol. 30 (1) (2000)
1–48.
[3] B. Marrot, A. Barrios-Martinez, P. Moulin, N. Roche, Industrial wastewater treat-
ment in a membrane bioreactor: a review, Environ. Prog. 23 (1) (2004) 59–69.
[4] Y. Wang, X. Huang, Q.P. Yuan, Nitrogen and carbon removals from food process-
ing wastewater by an anoxic/aerobic membrane bioreactor, Process. Biochem.
40 (2005) 1733–1739.
[5] J.H. Shin, S.M. Lee, J.Y. Jung, Y.C. Chung, S.H. Noh, Enhanced COD and nitro-
gen removals for the treatment of swine wastewater by combining submerged
membrane bioreactor (MBR) and anaerobic upflow bed filter (AUBF) reactor,
Process. Biochem. 40 (2005) 3769–3776.
[6] B. Farizoglu, B. Keskinler, E. Yildiz, A. Nuhoglu, Simultaneous removal of C, N, P
from cheese whey by jet loop membrane bioreactor (JLMBR), J. Hazard. Mater.
146 (2007) 399–407.
[7] Y.T. Ahn, S.T. Kang, S.R. Chae, C.Y. Lee, B.U. Bae, H.S. Shin, Simultaneous high-
strength organic and nitrogen removal with combined anaerobic upflow bed
filter and aerobic membrane bioreactor, Desalination 202 (2007) 114–121.
[8] S. Rosenberger, U. Krüger, R. Witzig, W. Manz, U. Szewzyk, M. Kraume, Perfor-
mance of a bioreactor with submerged membranes for aerobic treatment of
municipal waste water, Water Res. 36 (2) (2002) 413–420.
[9] T.W. Tan, H.Y. Ng, Influence of mixed liquor recycle ratio and dissolved oxy-
gen on performance of pre-denitrification submerged membrane bioreactors,
Water Res. 42 (4–5) (2008) 1122–1132.
[10] E.V. Münch, P. Lant, J. Keller, Simultaneous nitrification and denitrification in
bench-scale sequencing batch reactors, Water Res. 30 (2) (1996) 277–284.
[11] K.A. Third, N. Burnett, R. Cord-Ruwisch, Simultaneous nitrification and denitri-
fication using stored substrate (PHB) as the electron donor in a SBR, Biotechnol.
Bioeng. 83 (6) (2003) 706–720.
[12] W.L. Jones, E.D. Schroeder, P.A. Wilderer, Denitrification in a batch wastewater
treatment system using sequestered organic substances, Res. J. Water Pollut.
Control Fed. 62 (3) (1990) 259–267.
[13] W.L. Jones, P.A. Wilderer, E.D. Schroeder, Operation of a three-stage SBR system
for nitrogen removal from wastewater, Res. J. Water. Pollut. Control. Fed. 62 (3)
(1990) 268–274.
[14] J.J. Beun, F. Paletta, M.C.M. van Loosdrecht, N removal in a granular sludge
sequencing batch airlift reactor, Biotechnol. Bioeng. 75 (1) (2001) 82–92.
[15] K.A. Third, B. Gibbs, M. Newland, R. Cord-Ruwisch, Long-term aeration man-
agement for improved N-removal via SND in a sequencing batch reactor, Water
Res. 39 (15) (2005) 3523–3530.
[16] K. Bernat, I. Wojnowska-Baryła, A. Dobrzyńska, Denitrification with endoge-
nous carbon source at low C/N and its effect on P(3HB) accumulation, Bioresour.
Technol. 99 (7) (2008) 2410–2418.
[17] P. Ajay, Z. Jesse, N. George, Simultaneous carbon, nitrogen and phosphorous
removal from municipal wastewater in a circulating fluidized bed bioreactor,
Chemosphere 65 (7) (2006) 1103–1112.
[18] H. Guo, J. Zhou, J. Su, Z. Zhang, Integration of nitrification and denitrification in
airlift bioreactor, Biochem. Eng. J. 23 (1) (2005) 57–62.
[19] L.W. Xiao, M. Rodgers, J. Mulqueen, Organic carbon and nitrogen removal from
a strong wastewater using a denitrifying suspended growth reactor and a
horizontal-flow biofilm reactor, Bioresour. Technol. 98 (4) (2007) 739–744.
[20] APHA, Standard Methods for the Examination of Water and Wastewater, Amer-
ican Public Health Association, 1995.
[21] R.I. Amann, B.J. Binder, R.J. Olson, S.W. Chisholm, R. Devereux, D.A. Stahl, Com-
bination of 16S rRNA-targeted oligonucleotide probes with flow cytometry for
analyzing mixed microbial populations, Appl. Environ. Microbiol. 56 (6) (1990)
1919–1925.
[22] G.R. Crocetti, P. Hugenholtz, P.L. Bond, A. Schuler, J. Keller, D. Jenkins, L.L. Black-
all, Identification of polyphosphate accumulating organisms and design of
16S rRNA-directed probes for their detection and quantitation, Appl. Environ.
Microbiol. 66 (3) (2000) 1175–1182.
[23] T. Mino, M.C.M. Van Loosdrecht, J.J. Heijnen, Microbiology and biochemistry of
the enhanced biological phosphate removal process, Water Res. 32 (11) (1998)
3193–3207.
[24] T. Stephenson, S. Judd, B. Jefferson, K. Brindle, Membrane Bioreactors for
Wastewater Treatment, IWA Publishing, London, 2000.
[25] V. Mateju, S. Cizinska, J. Krejci, T. Janoch, Biological water denitrification—a
review, Enzyme Microbiol. Technol. 14 (3) (1992) 170–183.
[26] V. Arun, T. Mino, T. Matsuo, Biological mechanism of acetate uptake mediated
by carbohydrate consumption in excess phosphorus removal system, Water
Res. 22 (5) (1988) 565–570.
[27] C.D.M. Filipe, G.T. Daigger, Development of a revised metabolic model for the
growth of phosphorus-accumulating organisms, Water Environ. Res. 70 (1)
(1998) 67–79.
[28] R.J. Zeng, R. Lemaire, Z.G. Yuan, J. Keller, Simultaneous nitrification, denitri-
fication, and phosphorus removal in a lab-scale sequencing batch reactor,
Biotechnol. Bioeng. 84 (2) (2003) 170–178.