Front. Environ. Sci. Eng.

2018, 12(6): 5
https://doi.org/10.1007/s11783-018-1047-6

RESEARCH ARTICLE

Nitrogen removal performance of municipal reverse osmosis

concentrate with low C/N ratio by membrane-aerated biofilm
reactor

Xiao Quan1,2,3,4, Kai Huang1,2,3,4, Mei Li1,2,3,4, Meichao Lan1,2,3,4, Baoan Li (✉)1,2,3,4
1 Chemical Engineering Research Center, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China
2 State Key Laboratory of Chemical Engineering, Tianjin University, Tianjin 300072, China
3 Tianjin Key Laboratory of Membrane Science and Desalination Technology, Tianjin University, Tianjin 300072, China
4 Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin 300072, China

HIGHLIGHTS GRAPHIC ABSTRACT

• MABR exhibits excellent TN removal perfor-
mance for treating ROC with low C/N ratio.
• Operating conditions should be properly con-
trolled to achieve optimal TN removal.
• Denitrifying bacteria and NOB are proved
notably inhibited by high salinity stress.
• The TN removal rate remains over 70% when the
NaCl addition amount is below 20 g/L.

ARTICLE INFO
Article history:
Received 15 December 2017 ABSTRACT
Revised 7 March 2018
Accepted 20 April 2018 A membrane-aerated biofilm reactor was employed to investigate the nitrogen removal of one typical
Available online 30 May 2018 municipal reverse osmosis(RO) concentrate with a high total nitrogen (TN) concentration and a low C/
N ratio. The effects of operational conditions, including the aeration pressure, the hydraulic retention
time and the C/N ratio, on the systematic performance were evaluated. The nitrogen removal
Keywords: mechanism was evaluated by monitoring the effluent concentrations of nitrogen contents.
Membrane-aerated biofilm reactor (MABR) Furthermore, the microbial tolerance with elevated salinity was identified. The results indicated that
the optimal TN removal efficiency of 79.2% was achieved of the aeration pressure of 0.02 MPa,
Salinity hydraulic retention time of 24 h, and the C/N ratio of 5.8, respectively. It is essential to supplement the
Total nitrogen carbon source for the targeted RO concentrate to promote the denitrification process. The inhibitory
Reverse osmosis concentrate effect of salinity on denitrifying bacteria and nitrite oxidizing bacteria was significant, revealing the
limited TN removal capacity of the conditions in this work. The TN removal efficiency remained more
than 70% with the addition of salt (NaCl) amount below 20 g/L. This work preliminarily demonstrated
the MABR feasibility for the nitrogen removal of municipal RO concentrate with low C/N ratio and
provided technical guidance for further scale-up application.

© Higher Education Press and Springer-Verlag GmbH Germany, part of Springer Nature 2018

1 Introduction for seawater desalination, wastewater treatment and

With high purification efficiency and simple operation,
reverse osmosis (RO) technology has been widely applied
✉ Corresponding author
E-mail: libaoan@tju.edu.cn
reclamation in recent years (Malaeb and Ayoub, 2011).
Due to the high recovery of the RO process, RO
concentrates contain great amounts of salts and refractory
organics (Pérez-González et al., 2012). For RO applied in
municipal wastewater reclamation, organics such as
natural organic matter, personal care products, pharma-
ceuticals, synthetic dyes, pesticides and other organics
2 Front. Environ. Sci. Eng. 2018, 12(6): 5
accumulate within the municipal RO concentrate, further
damaging the bio-ecological environment (Long and
Schäfer, 2006; Benner et al., 2008).
Effective organics removal performance could be
achieved by utilizing physicochemical techniques in the
treatment of RO concentrates, mainly including advanced
oxidation processes (AOPs) and adsorption (Dialynas et
al., 2008; Zhou et al., 2011b). However, high energy
consumption and demand for chemical agents have
restrained applicability for large-scale and long-term
operation (Muruganandham et al., 2014; Boczkaj and
Fernandes, 2017) in addition to the possible secondary
pollution of the environment. With a powdered activated
carbon dose of 0.6 g/L, the chemical oxygen demand
(COD) removal rate was 70% when refinery RO
concentrate was treated by powdered activated carbon
adsorption (Zhao et al., 2012). Electrochemical oxidation
on three different electrodes was employed to treat a high
saline RO concentrate. COD could be totally removed on a
boron-doped diamond anode when the energy consump-
tion (0.158 kWh/gCOD) was the highest. For the Ti/IrO2–
RuO2 anode, the energy consumption (0.048 kWh/gCOD)
was the least with only partial COD removal (Zhou et al.,
2011a). These findings reflect the enormous challenges of
achieving higher efficiency with lower energy consump-
tion in the electrochemical oxidation process.
Combinations of AOPs and biological methods were
attempted to enhance the nitrogen substances and organic
removal of RO concentrates. However, the extremely high
salt level and refractory organics of RO concentrates, as
well as oxidant residuals, would invalidate the conven-
tional microbial systems of wastewater treatment with
limited nitrogen removal efficiency (Vendramel et al.,
2013; Pradhan et al., 2015). Some biological processes that
show promising performance for organics removal in RO
concentrate have been reported. In addition, for effective
total nitrogen (TN) removal by biological methods, a
sufficient carbon source is essential for the denitrification
process during the treatment of RO concentrate with low
C/N ratio. Although the operation was relatively compli-
cated, a sequencing batch reactor process achieved 25.7%
COD removal and 87.3% TN removal when it was
employed to treat RO concentrate from a wastewater
reclamation plant with methanol supplied as the supple-
mentary carbon source (Kim et al., 2016).
Characterized by the biofilm immobilized on the surface
of gas-permeable membrane, the membrane-aerated bio-
film reactor (MABR) is a novel technology for wastewater
treatment (Brindle et al., 1998). The counter-diffusional
mass transfer of substrates within the biofilm promotes the
microbial niche, allowing the simultaneous removal of
carbon and nitrogen substances in a single reactor. High
theoretical oxygen utilization efficiency could be achieved
with the bubbleless aeration (Shanahan and Semmens,
2004; Syron and Casey, 2008). During the past decades,
MABR has been employed in the treatment of multiple
types of wastewater, indicating its significant applicability
and feasibility when combined with other methods to
enhance the water quality (Sun et al., 2015). An MABR-
O3-BAC combined system was applied for oil-field
wastewater treatment (Li et al., 2015). The concentrations
of COD, oil content, TN and ammonia nitrogen (NH4+-N)
in the effluent were 45 mg/L, 2.8 mg/L, 6.8 mg/L and 3.6
mg/L, respectively, meeting the discharge standards.
Membrane bioreactors which achieved degrees of
success in terms of organic removal for high saline sewage
have been reported. Over 99% NH4+-N was removed
when a membrane bioreactor was employed to treat
wastewater with high salinity up to 4%, but the removal
performance was further limited when the salinity
increased to 7% (Wang et al., 2016). A pilot-scale
membrane bioreactor achieved 42% TN removal for the
treatment of synthetic shipboard slops with supplementary
sodium chloride (NaCl) amount of 20 g/L (Mannina et al.,
2016). In comparison, a recent study indicated that an
MABR could exhibit notable TN removal efficiency of
70.6% for synthetic sewage (CODCr 220 mg/L, NH4+-N
32 mg/L) when salinity was up to 3% (Tian et al., 2017).
However, few investigations were available on the
application of MABRs for the treatment of practical RO
concentrates, and it is crucial to evaluate systematic
performance under different operational parameters.
In this work, an MABR was utilized in the treatment of a
typical municipal RO concentrate with high TN concen-
tration and poor biodegradability. The effects of opera-
tional parameters, including the hydraulic retention time
(HRT), the aeration pressure and C/N ratio by external
carbon source addition, on the systematic nitrogen removal
performance were evaluated after the membrane-aerated
biofilm acclimation for RO concentrate. Subsequently, the
nitrogen removal mechanism was illustrated. Microbial
tolerance of the salinity was determined by stepwise salt
(NaCl) supplementation to the targeted RO concentrate.
The results of the work could be conducive to providing
valuable technique guidance for further scale-up MABR
application for wastewater with similar water quality.
2 Materials and methods
2.1 The MABR setup
In this study, the designed MABR system was composed
of the following four parts: MABR reactor, air supply
device, circulation device and water storage equipment
(Fig. 1). The effective volume of the main reactor was 6 L,
where 80 hollow-fiber membranes (effective length: 130
cm; outer diameter: 750 mm; and wall thickness: 175 mm)
specific for MABR (Hydroking Sci. & Tech, Ltd. (Tianjin,
China)) were arranged in an orderly fashion. The
commercialized hollow-fiber gas-permeable membrane
has been demonstrated to be appropriate for long-term
 Xiao Quan et al. Nitrogen removal of ROC with MABR
Fig. 1 Experimental set-up diagram of the MABR system
use in both research or engineering application. During the
investigation, the aeration pressure was manipulated by
adjusting the pressure maintaining valve of the air
compressor, the gas flow indicator and the atmospheric
valve.
2.2 Biofilm acclimation and MABR start-up
Activated sludge with great settleability and abundant
nitrifying microbes from the secondary sedimentation tank
of a wastewater treatment plant (Tianjin, China) was
seeded as the biofilm inoculum. Then the synthetic nutrient
solution was formulated in accordance with the composi-
tion of domestic sewage during the inoculation and
acclimation phase with influent concentrations of CODcr
and NH4+-N simultaneously- adjusted according to the
experimental requirements. In addition, small amounts of
substances containing KH2PO4 (30 mg/L), NaCl (45 mg/
L), NaHCO3 (50 mg/L), Na2CO3 (110 mg/L), K2SO4 (45
mg/L), MgSO4$7H2O (55 mg/L), FeSO4 (0.2 mg/L),
ZnSO4 (0.2mg/L), Cu(NO3)2 (0.2 mg/L) and MnSO4 (0.2
mg/L) were added to the sewage to promote the biofilm
formation. The main operational parameters for biofilm
inoculation and acclimation included the HRT of 12 h and
the aeration pressure of 0.03 MPa.
In this work, the RO concentrate was sampled from the
RO concentrate pool of a municipal water reclamation
plant (Redsun Technology Development Co., Ltd., Tianjin,
China), where an ultrafiltration unit was employed as the
pretreatment process for an RO unit. The mean character-
istics of the RO concentrate are detailed in Table 1.
Subsequently, the RO concentrate was gradually fed to the
MABR to replace the synthetic domestic sewage under an
aeration pressure of 0.03 MPa and an HRT of 24 h.
To illustrate the effects of operational parameters on
nitrogen removal and optimize the process, the aeration
pressure (0.01, 0.02, 0.03, 0.04 and 0.05 MPa, respec-
tively), HRT (18, 24 and 30 h, respectively) and C/N ratio
Table 1 Characteristics of the RO concentrate
Parameter CODcr TN NH4+-N NO3–-N
Value 80–90 74–76 0.054 7.2
Unit mg/L mg/L mg/L mg/L
3
(1.1, 3.8, 5.8 and 10.8) were regulated and controlled in
continuous mode. The C/N ratio in this study was the value
of CODcr divided by TN. Then under the optimal operating
conditions, the effluent concentrations of TN, NH4+-N,
NO3–-N, nitrite nitrogen (NO2–-N), COD and dissolved
oxygen (DO) were monitored at intervals of two hours for
24 h in batch mode. Furthermore, the salinity of the RO
concentrate was elevated by adding NaCl (10, 20, 30 and
40 g/L, respectively) in continuous mode to identify the
systematic salinity-shock resistance in terms of the activity
changes of nitrogen removal-related microbes.
2.3 Analysis of the water quality
The concentrations of TN, NH4+-N, NO3–-N, NO2–-N and
CODcr were measured through the spectrophotometry
using a Multiparameter Bench Photometer for Laboratories
(DR3900, Hach Instruments, Inc., USA). DO was
measured by a multiparameter water quality analyzer
using the DO probe (HQ30d, Hach Instruments, Inc.,
USA). Biochemical oxygen demand (BOD5) was mon-
itored by a BOD analyzer (BODTrak™, Hach Instruments,
Inc., USA). Electrical conductivity and TDS were
measured by a conductivity meter (DDSJ-308A, Inesa
Instrument Inc., China). Each measurement was conducted
in triplicate.
3 Results and discussion
3.1 Startup of MABR and microbial acclimation
After the inoculation of activated sludge, the MABR
accomplished biofilm acclimation by using synthetic
domestic sewage as the influent in continuous mode. The
reactor was operated with an aeration pressure of 0.03MPa
and an HRT of 12h. During the acclimation process, the
influent concentrations of CODcr and NH4+-N were
simultaneously elevated to enhance the MABR treatment
efficiency and improve the steady-state formation of a
multi-population biofilm.
As seen in Fig. 2, the concentrations of CODcr and
NH4+-N in the effluent remained low despite the increased
influent organic loading. The removal efficiencies of
CODcr and NH4+-N increased to 92% and 98.5% with
the increase of CODcr and NH4+-N concentrations to 400
mg/L and 60 mg/L, respectively, suggesting the improve-
ment of the treatment efficiency and the formation of a
steady biofilm in the MABR for further application of the
MABR to the real RO concentrate.
NO2–-N BOD Conductivity TDS TP pH
0.190 20–22 10.16 5.5 23.5 7.9
mg/L mg/L mS/cm g/L mg/L –
4 Front. Environ. Sci. Eng. 2018, 12(6): 5
Fig. 2 Variation of effluent concentrations of CODcr and
NH4+-N during the acclimation period
3.2 Biofilm acclimation with RO concentrate
The targeted RO concentrate was characterized by a low
C/N ratio of 1.1. Considering the adverse impact of a low
C/N ratio on the denitrification process, glucose was added
into the RO concentrate as supplementary carbon source
(Yang et al., 2012). The C/N ratio was elevated to 5.8,
which was relatively conducive to achieving the successful
biofilm acclimation to RO concentrate. Using RO
concentrate as influent, the MABR was operated in
continuous mode while the aeration pressure and HRT
were controlled at 0.03 MPa and 24 h.
As indicated in Fig. 3, the TN removal efficiency in the
first four days was below 30%, suggesting the short-term
reduction of nitrogen removal performance under condi-
tions of a drastic change in water quality containing
salinity. Then, the removal rate gradually increased to
75.6% during the next days. The result shows that MABR
could achieve effective nitrogen removal for RO concen-
trate with a short adaptation period. In addition, the COD
removal efficiency was slightly influenced by salinity
compared to the TN efficiency. Overall, the MABR
exhibited an outstanding tolerance of salinity shocks and
significant adaptability to the RO concentrate.
3.3 Systematic performance under different operational
parameters
The effects of major process conditions (HRT, the aeration
pressure, and C/N ratio) were evaluated in order to attain
the optimal operation parameters for better TN removal
performance.
3.3.1 Effects of HRT on nitrogen removal
Under an aeration pressure of 0.03 MPa and a C/N ratio of
Fig. 3 (a) Variations of the effluent COD concentration and
removal efficiency during the RO concentrate acclimation period.
(b) Variations of the effluent TN concentration and removal
efficiency during the RO concentrate acclimation period
5.8, the nitrogen removal performance was investigated
with different HRTs (18, 24 and 30 h), which were
regulated by controlling the influent flow rate.
The decreased HRT corresponded to an increase in the
influent organic load for the MABR, which could promote
the aerobic growth of heterotrophic bacteria in the biofilm
and result in higher oxygen consumption. As illustrated in
Fig. 4(c), the effluent concentration of NH4+-N showed a
decreasing tendency from 1.78 mg/L to 0.127 mg/L with
HRT increasing from 18 h to 30 h. This was possibly
because that nitrifying microbes, including ammonia-
oxidizing microbes and nitrite-oxidizing bacteria, out-
performed aerobic heterotrophic bacteria for oxygen
utilization under a reduced organic load (Rodríguez et
al., 2012). This indicates that the nitrification process is
greatly enhanced under higher HRTs. In contrast, the
denitrification process was substantially inhibited by the
elevated HRT. As indicated in Fig. 4(b), the NO3–-N
concentration at HRT of 30 h was approximately 7.2 mg/L
which was higher than 3.3 mg/L at 24 h and 1.9 mg/L at 18
 Xiao Quan et al. Nitrogen removal of ROC with MABR
Fig. 4 Effects of HRT on the performance of MABR: (a) TN
concentration in effluent under different HRTs; (b) NO3–-N
concentration in effluent under different HRTs; (c) NH4+-N
concentration in effluent under different HRTs
h, which suggested that the inhibition of denitrification was
possibly caused by the permeation of dissolved oxygen
5
(Tian et al., 2015). The result also demonstrated that the
denitrifying bacteria in the MABR was predominantly
facultative anaerobes. NO2–-N concentrations in the
effluent maintained a certain low level (below 0.7 mg/L)
without marked change under the aeration pressures which
have been investigated. The TN removal rate reached
76.5% at an HRT of 24 h greater than the 73.1% at an HRT
of 18 h and 68.1% at an HRT of 30 h (Fig. 4(a)). A
moderate HRT is required for the effective TN removal and
an excessively high HRT would inhibit the TN removal
owing to the limited carbon source for denitrification.
Consequently, the optimal HRT for RO concentrate
treatment to conduct the further investigation on TN
removal performance was 24 h.
3.3.2 Effects of aeration pressure on nitrogen removal
Under an HRT of 24 h and a C/N ratio of 5.8, the aeration
pressure effect on the TN removal was evaluated.
From Fig. 5(a), the TN removal rate achieved a
maximum of 79.2% at an aeration pressure of 0.02 MPa
and reduced from 79.2% to 67.8% with the aeration
pressure ranging from 0.02 to 0.05 MPa. The reduction of
the TN removal rate is highly relevant to the accumulation
of NO3–-N. The effluent NO3–-N concentration increased
from 1.9 to 9.9 mg/L with the aeration pressure increasing
from 0.02 to 0.05 MPa (Fig. 5(b)), revealing that
incomplete denitrification was affected by the dissolved
oxygen permeation throughout the biofilm, i.e., the
increased DO concentration in the bulk liquid. The DO
concentration was greater than 1 mg/L when pressure was
up to 0.04 MPa, furthermore, the DO concentration at 0.05
MPa was close to 2 mg/L (Fig. 5(d)), which caused an
almost aerobic environment in the outer layer of the
biofilm leading to the intensive inhibition of denitrification
(LaPara et al., 2006).
As shown in Fig. 5(c), the effluent NH4+-N showed an
obvious accumulation at the aeration pressure of 0.01 MPa
(2.3 mg/L) compared to that at 0.05 MPa (0.08 mg/L),
which indicated inefficient nitrification. This is probably
because nitrifying bacteria is inferior to heterotrophic
bacteria for oxygen competition at low oxygen concentra-
tions (DO below 0.3 mg/L, Fig. 5(d)) (Sliekers et al., 2005;
Hwang et al., 2009). Owing to the accumulation of NH4+-
N, the TN removal efficiency at 0.01 MPa (74.8%) was
lower than those at 0.02 MPa (79.2%) and 0.03 MPa
(76.5%). The NO2–-N concentration in the effluent was
relatively stable (below 0.150 mg/L) when aeration
pressure varied from 0.01 to 0.05MPa. Moderate aeration
pressure enables the reactor to form a unique stratification
of nitrifying bacteria and denitrifying bacteria within the
biofilm to achieve simultaneous nitrification and denitri-
fication in a single reactor (Cole et al., 2004; Downing and
Nerenberg, 2008). In summary, 0.02 MPa was the optimal
aeration pressure of the reactor for TN removal under the
HRT of 24 h and C/N ratio of 5.8.
6 Front. Environ. Sci. Eng. 2018, 12(6): 5

Fig. 5 Effects of aeration pressure on the performance of the MABR: (a) TN concentration in effluent at different aeration pressures;
(b) NO3–-N concentration in effluent at different aeration pressures; (c) NH4+-N concentration in effluent at different aeration pressures;
(d) DO concentration in the bulk liquid at different aeration pressures

3.3.3 Effects of C/N ratio on nitrogen removal
The research concerning the influence of C/N ratio on TN
removal performance was conducted with the HRT of 24 h
and aeration pressure of 0.02 MPa. The C/N ratios were
controlled at 1.1 (RO concentrate without carbon supple-
ment), 3.8, 5.8 and 10.8 by regulating the supplemented
amount of glucose.
As seen from Fig. 6 (e), the COD concentration in
effluent maintained a relatively stable level, except for the
temporary increase in the first 2 days with a C/N ratio of
10.8. It was noteworthy that the NO3–-N concentration in
effluent was significantly affected by the C/N ratio. The
NO3–-N concentration in effluent presented an extreme
increase of 15.5 mg/L when the C/N ratio decreased from
10.8 to 1.1 (Fig. 6 (b)). It was closely associated with the
deficient quantity of electron donor for denitrification,
which resulted from the insufficiency of the carbon source
(Kumar et al., 2012). The changing tendency of NO2–-N
concentration in effluent was similar to that of NO3–-N (Fig.
6(d)). Furthermore, the insufficiency of the carbon source
was prejudicial to the effective degradation of organic
nitrogen by heterotrophic bacteria, which impaired the
removal of TN. The removal efficiency of TN was 43.4% at
the C/N ratio of 1.1, with a marked reduction compared to
the value (79.2%) at the C/N ratio of 5.8 (Fig. 6 (a)).
Since heterotrophic bacteria outperforms the ammonia-
oxidizing microbes for oxygen utilization under high C/N
ratio, nitrification is inhibited as an aerobic reaction, which
leads to the accumulation of NH4+-N (Nogueira et al.,
2002). For instance, the NH4+-N concentration increased
from 0.07 to 4.35 mg/L when the C/N ratio varied from 1.1
to 10.8 (Fig. 6 (c)). As a result, the TN concentration in
effluent at C/N ratio of 10.8 increased significantly
 Xiao Quan et al. Nitrogen removal of ROC with MABR 7

compared to that at a C/N ratio of 5.8, with a 5% decrease addition leads to unnecessary economic costs in practical
in the TN removal efficiency (Fig. 6(a)). In addition, a application. To summarize, the optimal C/N ratio for
meaningless increase in supplementary carbon source MABR nitrogen removal was 5.8.

Fig. 6 Effects of C/N ratio on the performance of MABR: (a) TN concentration in effluent at different C/N ratios; (b) NO3–-N
concentration in effluent at different C/N ratios; (c) NH4+-N concentration in effluent at different C/N ratios; (d) NO2–-N concentration in
the bulk liquid at different C/N ratios; (e) COD concentration at different C/N ratios
8 Front. Environ. Sci. Eng. 2018, 12(6): 5

3.4 Tracking analysis of nitrogen removal in the MABR
To illustrate the mechanism of nitrogen removal in the
MABR system, the MABR was operated in batch mode
with the aeration pressure and C/N ratio being controlled at
0.02 MPa and 5.8. The effluent concentrations of TN,
NH4+-N, NO3–-N and NO2–-N were monitored at intervals
of two hours during the entire 24 h experiment.
From Fig. 7(a), the TN concentration decreased
gradually throughout the operation, and the TN removal
efficiency stabilized at 75.5%. Initially, aerobic hetero-
trophic bacteria and ammonia oxidizing bacteria (AOB)
exhibited high activity with adequate carbon and nitrogen
sources and consumed a large amount of oxygen. This was
conducive to the formation of an anaerobic environment in
the outer layer of the biofilm, as proven by the DO
concentration in the bulk liquid (less than 0.5 mg/L) in the
first 12 h (Fig. 7(b)). Appropriate oxygen gradients could
promote the formation of a unique stratified biofilm,
enhancing simultaneous nitrification and denitrification.
The nitrification efficiency was lower than the ammo-
nification efficiency in the early stage, as indicated by the
brief NH4+-N accumulation in the first 8 h. Then aerobic
heterotrophic bacteria outcompeted AOB for dissolved
oxygen with sufficient carbon sources, further inhibiting
the nitrification in the early stage. Compared to the initial
value, the NH4+-N concentration showed an increase of
3.6 mg/L at the 2nd hour (Fig. 7(b)). The DO concentration
gradually increased in the last 12 h (Fig. 7(b)), caused by
the decreasing oxygen consumption given the deficiency
of carbon and nitrogen sources. The DO value was
ultimately 1.93mg/L, indicating that the entire biofilm was
an aerobic environment. Hence, the reactor achieved
sufficient nitrification in the last 12 h. The NH4+-N
concentration gradually decreased to 0.4 mg/L gradually
during the first 12 h and then remained relatively stable in
the next 12 h (Fig. 7(b)), indicating that most of the organic
nitrogen had already been transformed into NH4+-N, and
the production amount of NH4+-N was relatively low in
the later stage.

Fig. 7 (a) Changes of TN concentration in the effluent and removal efficiency at different times. (b) The effluent concentrations of NO3–-N,
NO2–-N and NH4+-N at different times. (c) Changes of COD concentration in the effluent and removal efficiency at different times. (d) Changes
of DO concentration in MABR at different times
 Xiao Quan et al. Nitrogen removal of ROC with MABR 9

NO3–-N and NO2–-N accumulated gradually in the initial high salinity level of 40 gNaCl/L. As the NaCl addition
stage of the experiment and reached the maximum at 6 h amount ranged from 0 to 20 g/L, denitrifying bacteria were
and 8 h, respectively (Fig. 7(b)). The maximal concentra- not notably inhibited, as demonstrated by the insignificant
tions of NO3–-N and NO2–-N were 25.6 and 21.4 mg/L
(Fig. 7(b)), respectively, further revealing the inhibition of
denitrification by salinity stress. Hence, the denitrification
efficiency was far lower than the ammonification and
nitrification in the MABR system. The NO2–-N accumula-
tion resulted from the inhibition of nitrite-oxidizing
bacteria (NOB), illustrating that NOB were sensitive to
the salinity elevation. Previous research has discovered the
inhibitory effects of high salinity on NOB, which exerts a
positive influence on shortcut nitrification and denitrifica-
tion (Liu et al., 2008; She et al., 2016). A study observed
that a NaCl concentration of 0.1 mol/L caused 40%
inhibition of the maximum ammonia oxidizing activity
compared to no salt addition (Mosquera-Corral et al.,
2005). In this MABR, the rate of ammonia oxidation
decreased significantly with the inhibition of NOB activity,
which led to the accumulation of NO2–-N in the process
(Fig. 7(b)). Nevertheless, the NOB activity was incomple-
tely inhibited, and the reactor could not maintain the
process at the maximum accumulation of NO2–-N due to
the continuous oxygen supply. Hence, the complete
inhibition of NOB and the promotion of shortcut
nitrification and denitrification should be further studied
in this reactor. As illustrated in Fig. 7(b), the denitrification
process still continued at a relatively low rate, as
demonstrated by the decreased values of NO3–-N and
NO2–-N when the DO concentration increased up to 1 mg/
L after 18 h. This result suggests that aerobic denitrification
occurred in this reactor. In the end, the effluent concentra-
tions of NH4+-N, NO3–-N and NO2–-N were 0.44, 4.2 and
0.471 mg/L, respectively (Fig. 7(b)).

3.5 Effects of elevated salinity on nitrogen removal

performance

The paper studied the effect of elevated salinity on MABR

nitrogen removal performance and the salinity tolerance of
microbial communities by supplementing NaCl with the
gradient of 10 g/L under an HRT of 24 h, and an aeration
pressure of 0.02 MPa and a C/N ratio of 5.8.
As illustrated in Fig. 8, the impact of salinity elevation
on the removal of COD and NH4+-N was insignificant.
After acclimation with salinity elevation, the concentra-
tions of COD and NH4+-N in effluent remained below 70
mg/L and 0.5 mg/L, respectively. This proved that
heterotrophic carbon-oxidizing bacteria and AOB pos-
sessed a strong tolerance for the high salinity tested in this
study (NaCl addition amount up to 40 g/L) (Deorsola et al.,
2013). In terms of denitrifying bacteria, the significant Fig. 8 (a) The concentrations of NO3–-N, NO2–-N, NH4+-N in
increase of NO3–-N at 40 gNaCl/L (15.6 mg/L) compared effluent at different salinity. (b) The TN concentration in effluent at
to that (1.9 mg/L) at ROC without NaCl addition illustrated different salinity. (c) The COD concentration in effluent at different
that denitrifying bacteria was intensively inhibited by the salinity
10 Front. Environ. Sci. Eng. 2018, 12(6): 5
accumulation of NO3–-N in the effluent. The NO2–-N
concentration was ultimately stable in the effluent (below
0.5 mg/L, Fig. 8(a)) ultimately, while it showed a notable
increase (greater than the change of NH4+-N) in the initial
stages of each acclimation period with salinity elevation,
indicating that NOB was more sensitive to the salinity
variation than AOB in this reactor (Mannina, et al., 2016).
The MABR maintained a TN removal efficiency level of
over 70% when the NaCl addition amount ranged from 0 to
20 g/L, verifying the significant effectiveness and stability
of the MABR under relatively high salinity conditions.
Nevertheless, the further addition of salts induced a
marked decline in TN removal performance. The TN
removal efficiency reduced to 57.2% and 47.5% at 30
gNaCl/L and 40 gNaCl/L(Fig. 8(b)). In addition, the
increase of the TN concentration (17.5 mg/L) in the
effluent was much greater than the increase of NO3–-N
(12.3 mg/L) in the effluent at 40 gNaCl/L compared to
those with the RO concentrate without NaCl supplement,
implying the inhibition of ammonification caused by high
salinity.
Overall, the MABR exhibited a notable tolerance and
superior treatment capacity for TN removal under high
salinity, up to a NaCl addition amount of 20 g/L. At higher
salts addition amounts, the TN treatment capacity of the
MABR was obviously reduced.
4 Conclusions
In this work, the applicability of an MABR for the nitrogen
removal of a typical municipal RO concentrate with high
TN content and a low C/N ratio was investigated, the
nitrogen removal mechanism was specified, and the
inhibitory effects of salinity were illustrated. The optimal
process parameters included the HRT of 24h, the aeration
pressure of 0.02MPa and the C/N ratio of 5.8, under which
the carbon source was supplemented to enhance the TN
removal (79.2%). The MABR system exhibited a remark-
able tolerance to salinity and potential treatment capacity
for RO concentrate with high salinity stress. The TN
removal efficiency maintained high (greater than 70%)
when the NaCl addition amount was below 20 g/L, but
decreased to 57.2% and 47.5% when the NaCl addition
amounts increased to 30 g/L and 40 g/L, respectively. The
MABR is a promising technique for removing nitrogen
contents from municipal RO concentrates under certain
high salinity levels. In addition, NOB inhibition by salinity
stress provided the feasibility of achieving shortcut
nitrification-denitrification in the future research.
Acknowledgements This study was financially supported by the National
Natural Science Foundation of China (Grant No. 51478304) and the
Technical Demonstration Project of Ministry of Water Resources (No. SF-
201605).
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