Simultaneous Removal of Aniline, Nitrogen and Phosphorus

8/16/2019 Simultaneous Removal of Aniline, Nitrogen and Phosphorus

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Simultaneous removal of aniline, nitrogen and phosphorus in

aniline-containing wastewater treatment by using sequencing

batch reactor

Yu Jiang, Hongyu Wang, Yu Shang, Kai Yang ⇑

School of Civil Engineering, Wuhan University, Wuhan 430072, China

h i g h l i g h t s

A sequencing batch reactor was used to treat aniline-containing wastewater.

Simultaneous removal of aniline, nitrogen and phosphorus was achieved.

Good removal performance was maintained during variation of operating conditions.

The accumulation of NH4+-N and TN during aniline biodegradation was alleviated.

a r t i c l e i n f o

Article history:

Received 6 December 2015

Received in revised form 1 February 2016

Accepted 4 February 2016

Available online 9 February 2016

Keywords:Aniline

Denitrifying phosphorus removal

Anaerobic/aerobic/anoxic condition

Sequencing batch reactor

Bioremediation

a b s t r a c t

The high removal efficiencies of traditional biological aniline-degrading systems always lead to accumu-

lation of ammonium. In this study, simultaneous removal of aniline, nitrogen and phosphorus in a single

sequencing batch reactor was achieved by using anaerobic/aerobic/anoxic (A/O/A) operational process.

The removal efficiencies of COD, NH4+-N, TN, TP were over 95.80%, 83.03%, 87.13%, 90.95%, respectively

in most cases with 250 mg L 1 of initial aniline at 6 h cycle when DO was 5.5 ± 0.5 mg L 1. Aniline was

able to be completely degraded when initial concentrations were less than 750 mg L 1. When DO

increased, the removal rate of NH4+-N and TP slightly increased along with the moderate decrease of removal efficiencies of TN. The variation of HRT had obvious influence on removal performance of pollu-

tants. The system showed high removal efficiencies of aniline, COD and nutrients during the variation of

operating conditions, which might contribute to disposal of aniline-rich industrial wastewater.

2016 Elsevier Ltd. All rights reserved.

1. Introduction

Aniline is hazardous to living beings and there has been more

concern about its disposal along with extensive applications

(Orge et al., 2015). Industrial wastewater, agricultural runoff and

urban sewage often contain aniline and various other pollutants.

Some of these compounds are toxic, carcinogenic, mutagenic and

teratogenic. They are able to remain in water and soil for a long

period of time, which has harmful impacts on environmental qual-

ity (Zhu et al., 2012). Therefore, it is important to treat wastewater

containing these compounds before discharging into water bodies.

China as well as USA has rated aniline as a persistent organic pol-

lutant of which the release amount is strictly limited ( Liu et al.,

2015).

The dispose of aniline and most of its derivatives can be

achieved by several physicochemical methods such as photode-

composition, electrolysis, ozone oxidation, resin adsorption and

electro-Fenton, which have limits because of relatively high eco-

nomic cost, energy consumption, and difficulty to remove pollu-

tants completely (Qi et al., 2002; Anotai et al., 2006). Biological

methods utilize microorganisms to thoroughly degrade aniline,

which are suitable for large-scale wastewater treatment (Wang

et al., 2007). Several biological technologies have been developed

to remove aniline in recent years such as aniline-degrading bacte-

ria, activated sludge reactors, moving bed biofilm reactors, which

showed relatively high removal rates (Campos et al., 2002; Li

et al., 2010; Dvoř ák et al., 2014). Most of these processes focus

on realizing good removal performance of aniline and COD. How-

ever, the fact that accumulation of ammonia nitrogen (NH4+-N)

and high concentration of total nitrogen (TN) coming from a large

amount of nitrogen in aniline is always ignored in previous studies.

Moreover, to our knowledge the phosphorus removal in these

treatment systems has not meant mentioned.

Nitrogen and phosphorus are common pollutants in sewerage

and industrial wastewater. Due to the fact that they are major

http://dx.doi.org/10.1016/j.biortech.2016.02.014

0960-8524/ 2016 Elsevier Ltd. All rights reserved.

⇑ Corresponding author. Tel.: +86 027 61218623; fax: +86 027 68775328.

E-mail address: [email protected] (K. Yang).

Bioresource Technology 207 (2016) 422–429

Contents lists available at ScienceDirect

Bioresource Technology

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causes of some environmental problems such as eutrophication

and several diseases which affect human beings, their release has

attached much attention. There have been increasing wastewater

treatment plants over the world which are committed to removal

of nitrogen and phosphorus (Li et al., 2014). Traditional nitrogen

biodegradation consists of two main processes, namely nitrifica-

tion and denitrification. Most of denitrifying bacteria are hetero-

trophic, which have a competitive relationship with phosphate-accumulating organisms (PAOs), leading to more difficulties in

simultaneous removal of nitrogen and phosphorus (Meinhold

et al., 1999). Unlike conventional phosphorus removal by release

under anaerobic conditions and uptake in aerobic environments,

denitrifying phosphate-accumulating organisms (DNPAOs) which

belong to PAOs, have capability of uptaking phosphorus by nitrate

and/or nitrite as electron acceptors in anoxic situation and utilizing

O2 under aerobic condition, resulting in denitrification and phos-

phorus removal at the same time (Zhang et al., 2011; Sun et al.,

2015). Additionally, the process has less carbon source consump-

tion as well as aeration costs and lower cell yield production,

which makes DNPAOs have advantages over general biological

methods to reduce nitrogen and phosphorus in wastewater

(Wang et al., 2015a).

In the present study, DNPAOs was enriched in SBR reactor oper-

ated by anaerobic/aerobic/anoxic (A/O/A) mode and one of the two

SBR reactors was fed with synthetic wastewater containing aniline.

The effect of organic loading rate, aeration as well as HRT on ani-

line, COD and nutrients removal was evaluated and the variations

of pollutants concentrations in two reactors within one cycle was

investigated. To our knowledge, this is the first report of simulta-

neous removal of aniline, nitrogen and phosphorus in the single

sequencing batch reactor. The work was conducted to remove both

aniline and nitrogen coming from aniline biodegradation. In the

meantime, the phosphorus was taken up in A/O/A operation mode.

It is reasonably expected that the removal of both aniline and

nutrients with high efficiency by the system might be an attractive

alternative to aniline-containing wastewater treatment.

2. Methods

2.1. Sludge and media

Seed sludge inoculated into the SBRs was obtained from the

secondary sedimentation tank of Shahu municipal wastewater

treatment plant in Wuhan, Hubei Province, China. The initial COD

loading rate fed in R1 and R2 was 0.9 g COD L 1 d1. The composi-

tion of initial synthetic wastewater for R1 was prepared with

analytical-grade chemicals as follow (per liter): aniline 125 mg,

NH4Cl 50.2 mg, NaNO3 79.7 mg, KH2PO4 21.9 mg, K2HPO43H2O

36.8 mg, CH3COONa 192.3 mg and 1 ml of trace element solution.

As for R2, the initial influent contained (per liter): CH3

COONa

576.9 mg, NH4Cl 86.0 mg, NaNO3 136.6 mg, KH2PO4 21.9 mg,

K2HPO43H2O 36.8 mg and 1 ml of trace element solution. The trace

element solution contained the following ingredients (per liter):

FeCl36H2O 5.0 g, H3BO3 0.10 g, CuSO45H2O 0.10 g, KI 0.20 g,

MnCl24H2O 0.50 g, Na2MoO42H2O 0.20 g, ZnSO47H2O 0.30 g,

CoCl26H2O 0.15 g, EDTA-2Na 10.0 g.

2.2. Reactor operation

Investigations were conducted in two parallel SBRs made of

plexiglass. As shown in Fig. 1, the reactor consisted of two cylindri-

cal columns with working volume of 9 L and the diameter/height

(D/H) was 6.67. Influent synthetic wastewater was fed into the sys-

tem through a pipe located at the top of the reactor by using a peri-staltic pump and aeration was carried out through a porous stone

diffuser at the bottom of the reactor. The sequential operation was

automatically controlled by timers in the initial cycle of 6 h and the

temperature of the reactors were maintained at 30 ± 2 C by using

water bath between the inner and outer columns. The initial oper-

ation was as follows: 5 min of feeding, 120 min of anaerobic agita-

tion phase, 120 min of aerobic phase, 90 min of anoxic agitation

phase, 20 min of settling time and 5 min of effluent withdrawal

phase. For effluent stage, 4.5 L of the supernatant was discharged

from the reactor by a diaphragm pump with 50% of volume

exchange ratio. Thus, the hydraulic retention time (HRT) was12 h and the dissolved oxygen (DO) in the anaerobic phase, aerobic

phase and anoxic phase were controlled at about 0.05, 5.5,

0.1 mg L 1, respectively. After the system was stable, the organic

loading rate, DO in aerobic phase as well as HRT were varied in

order to investigate the pollutants removal abilities of the two

reactors with the variations in operation parameters. The operating

conditions were shown in Table 1. The changes of process param-

eters of the two reactors are at the same time. Total nitrogen (TN)

in influent was adjusted accordingly to remain the same ratio of

carbon and nitrogen source throughout the whole operation. The

samples were taken from reactors at 1 or 1.5 h intervals and resid-

ual concentration of aniline, COD, NH4+-N, TN and TP were tested.

During the whole experiments, the concentrations of mixed liquor

suspended solids (MLSS) in the two reactors were adjusted to4200 ± 300 mg L 1.

Fig. 1. Schematic diagram of the reactors.

Table 1

Operating condition of each stage in two SBRs.

Stages Cycles Days COD loading rate

(g COD L 1 d1)

DO in aerobic

phase (mg L 1)

HRT

(h)

Stages 1 1–40 1–10 0.9 5.5 ± 0.5 12

Stages 2 41–80 11–20 1.5 5.5 ± 0.5 12

Stages 3 81–120 21–30 2.7 5.5 ± 0.5 12

Stages 4 121–160 31–40 3.9 5.5 ± 0.5 12

Stages 5 161–200 41–50 1.5 4.0 ± 0.5 12

Stages 6 201–240 51–60 1.5 5.5 ± 0.5 12

Stages 7 241–280 61–70 1.5 7.0 ± 0.5 12

Stages 8 281–340 71–80 1.5 5.5 ± 0.5 8

Stages 9 341–380 81–90 1.5 5.5 ± 0.5 12

Stages 10 381–410 91–100 1.5 5.5 ± 0.5 16

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2.3. Analytical methods

In all experiments, analysis of pollutants concentrations in

influent and effluent of the two reactors was carried out after the

samples were withdrawn and filtered. The concentrations of ani-

line, COD, NH4+-N, NO2

-N, NO3-N, TN and TP were measured

according to the standard methods (APHA, 2012). The tests were

done in triplicate. The settling properties of the sludge werereflected by sludge volume index (SVI) which was calculated as

the volume occupied by biomass after 30 min of settling ( Zhang

et al., 2015). Dissolved oxygen (DO) and temperature in the reactor

were measured by YSI550A DO meter. The batch tests for the pro-

portion of DNPAOs were conducted according to Zhang et al., 2015.

The pH values of samples were monitored by pH meter (PC-320).

3. Results and discussion

3.1. Aniline and COD biodegradation with different organic loading

rates

After 2 months of operation, the sludge in reactors had good

settling properties. SVI was less than 60 mL g1 and a large amountof DNPAOs were enriched. The removal performances of aniline as

well as COD by both R1 and R2 during the stable operation with

different organic loading rates were shown in Fig. 2. As for aniline

which could be seen in Fig. 2(a), over 99.8% could be degraded by

activated sludge in R1 even when the initial concentration of ani-

line was 500 mg L 1. It was also found from further tests that

750 mg L 1 of aniline was able to be completely degraded within

6 h cycles of operation (data not shown). Several pure isolates

and microbial consortia have been investigated for their capability

of aniline degradation as a promising alternate for treating

wastewater containing aromatic compounds by some other previ-

ous studies (Wang et al., 2007; Campo et al., 2011). The result in

this work was consistent with previous reports regarding to aniline

aerobic biodegradation in which pretty high concentration of ini-tial aniline could be thoroughly removed by microorganisms

(Xiao et al., 2009; Li et al., 2010). As shown in Fig. 2(b), both R1

and R2 had high removal efficiencies of COD when organic loading

rates ranged from 0.9 to 2.7 g COD L 1 d1. The removal rates of

COD in R1 and R2 were up to 94.22% and 93.53%, respectively.

The average residual COD concentrations with around 450, 750

and 1350 mg L 1 of COD in synthetic influent was 16.46, 26.10,

53.09 mg L 1, respectively for R1 and 12.54, 31.81, 54.43 mg L 1,

respectively for R2. Although the removal efficiencies of COD

remained relatively high during 30 days of operation, an obvious

increase of COD concentrations in effluent could be found when

initial COD concentration was 1350 mg L 1 in both of two reactors.

Rezaei et al. (2012) mentioned the phenomenon that the metabo-

lism of microorganisms in the system was slightly inhibited with

the increase of aniline concentration in inflow due to the toxicity

of the aromatic compound after gradual replacement of glucoseby aniline in aerobic granular bioreactor. However, the residual

concentrations of COD in effluents of R1 and R2 had no obvious dif-

ference at each COD level. The comparison of the COD removal per-

formance in this work between R1 and R2 showed that aniline

seemed to have no negative effect on COD removal. The results

indicated that the biomass in R1 developed the adaptability to

the toxic environment due to the gradual increase of aniline con-

centration in influent. This played an important role in acclimation

of microorganisms. The fast biodegradation by aniline-degrading

microorganisms contributed to the reduction of hazardous sub-

strates which might suppress the cell growth of other living beings

in the reactor. Since aniline concentration fluctuates wildly and

varies in different types of wastewater, the adaptation of the sys-

tem to a large range of aniline concentration might be advanta-

geous to practical wastewater treatment with pretreatment unit.

3.2. Simultaneous removal of nitrogen and phosphorus in the presence

of aniline

Nitrogen removal ability was also presented in two SBRs. As

seen in Fig. 3, the average removal efficiencies of NH4+-N in R2 with

22.5 and 37.5 mg L 1 of initial concentrations were 94.20% and

94.81%, respectively. The residual concentration in effluent was

less than 3 mg L 1. Compared to R2, R1 showed relatively lower

removal efficiency of NH4+-N and the average removal rates in these

two stages were 83.13% and 85.36%, respectively. The high removal

rate of NH4+-N owed to nitrification in aerobic phase of operation,

during which the NH4+-N was transferred to NO2

-N and NO3

-N(Chen et al., 2009; González et al., 2014). However, the residual

concentrations of NH4+-N significantly increased in both R1 and

R2 between day 21 and day 30. During this period, the residual

concentrations of NH4+-N in R1 and R2 were beyond 8.83 and

7.31 mg L 1, respectively. The higher residual concentrations of

NH4+-N in R1 than that in R2 might be due to higher concentration

of aniline in inflow. NH4+-N is one of the major by-products in

biological treatment of aniline-rich wastewater since it has been

Fig. 2. Aniline degradation (a) and COD consumption (b) at different initial aniline concentrations.

424 Y. Jiang et al. / Bioresource Technology 207 (2016) 422–429


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studied that the first step of aniline biodegradation causes releaseof nitrogen in the form of NH4

+-N (Lyons et al., 1984; O’Neill et al.,

2000). The NH4+-N production along with aniline degradation led to

more NH4+-N in the system. These NH4

+-N could not be completely

utilized by microorganisms within a limited time, resulting in high

concentration of NH4+-N in effluent. With regard to TN removal in

R1, the residual TN concentrations were below 10 mg L 1 after

6 h of biodegradation when initial aniline concentration was 125

or 250 mg L 1. However, with aniline concentration in inflow

increasing to 500 mg L 1, there was a notable decrease in TN

removal efficiency and average value was only 54.68% in this stage.

This might be due to the limited carbon source for denitrifying

microorganisms and DNPAOs to remove a large amount of nitrogen

in anoxic phase since aniline which provided the most of COD was

completely degraded by aniline-utilizing bacteria with activemetabolism in former aerobic phase. The phenomenon that the

lack of carbon substrate would reduce the removal efficiencies of

nutrients in this study could also be found in actual biological

treatment plants (Podedworna and Zubrowska-Sudol, 2012). This

might also explain the result that the residual concentrations of

TN in R2 were generally lower than that in R1 in the same stage.

Average removal efficiencies of R2 in the three stages were

92.04%, 91.94% and 87.81%, respectively. There were no toxic com-

pounds in R2 and COD removal in aerobic phase was not as good as

that in R1 by virtue of the fact that most of COD in R2 was provided

by sodium acetate instead of aniline. The good removal perfor-

mance of NH4+-N and TN by aerobic nitrification and anoxic denitri-

fication contributed to alleviating nitrogen accumulation in

wastewater in the presence of aniline. González et al. (2014)reported p-chloroaniline biodegradation in continuous biofilm sys-

tem which consisted of three main units. p-Chloroaniline removal

was achieved in the first continuous biofilm reactor and ammo-

nium released from p-chloroaniline was reduced through nitrifica-

tion in the subsequent aerobic up-flow fixed-bed reactor and

denitrification in the third anoxic reactor. However, the aniline

biodegradation, nitrification and denitrification were realized in

the single reactor in this work. Compared to the system described

by González et al. (2014), the process in this study was much sim-

pler and easier for operation, which might be promising in treating

wastewater containing aromatic compounds.

The result also showed excellent phosphorus reduction ability,

which could be seen from Fig. 4. In system fed with or without ani-

line, phosphorus removal efficiencies were generally over 88%,even in R1 when initial aniline concentration reached 500 mg L 1

or beyond. The average residual concentrations of phosphorus afterone cycle of operation in three stages were 0.64, 0.69, 0.98 mg L 1,

respectively in R1 and 0.53, 0.68, 0.78 mg L 1, respectively in R2. It

was obtained from the comparison of phosphorus removal perfor-

mances in the two reactors that the toxicity of aniline might not be

much harmful to PAOs and DNPAOs considering the fact that the

uptake of phosphorus seemed not to be significantly suppressed

by the increase of aniline. There were diverse microorganisms

working together such as genus Acinetobacter , Pseudomonas, Flex-

ibacter , Hyphomicrobium, Rhodobacter in aniline-degrading system.

These results revealed that aniline-degrading microorganisms,

nitrifying cells as well as DNPAOs were able to realize mutual

cooperation in aniline-containing wastewater treatment.

3.3. Effect of DO and HRT on pollutants removal performance

Effects of different DO concentrations in aerobic phase and HRT

on pollutants removal were investigated. As shown in Fig. 5, the

average removal efficiency of aniline within 30 days of operation

in R1 was 99.99% and high degradation rates remained when DO

concentrations ranged from 3.5 to 7.5 mg L 1, indicating the strong

activity of aniline-degrading microorganisms in R1. Both R1 and R2

Fig. 3. Nitrogen removal performance with different COD loading rates.

Fig. 4. Variation of phosphorus concentrations in influent and effluent of the twobioreactors and removal efficiencies over 30-day operational period.


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had high removal efficiencies of COD during the variation of DO.

The average residual concentrations of COD were 30.69, 26.59,

28.37 mg L 1, respectively in R1 when DO in the system was

adjusted to 4.0 ± 0.5, 5.5 ± 0.5, 7.0 ± 0.5 mg L 1. The removal effi-

ciencies of COD within one cycle were up to 94.35% in R2, even

at relatively low concentration of DO. The good removal perfor-

mance of both aniline and COD showed high efficiency of organic

substances biodegradation in fluctuation of DO, indicating that

the degradation of organic compounds in the system might be

adaptive to different working conditions in actual processes. It

Fig. 5. Variation of pollutants concentrations in effluent during changes of DO in aerobic phase.



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was found from Fig. 5(c) that NH4+-N removal efficiency had no sig-

nificant decrease with the drop of DO concentrations and average

removal efficiency in DO of 4.0 ± 0.5, 5.5 ± 0.5, 7.0 ± 0.5 mg L 1

was 89.90%, 90.67%, 91.52%, respectively in R1 and 90.83%,

92.14%, 92.84%, respectively in R2. Almost no NO2-N was detected

in effluent in both R1 and R2 during the variation of DO (data not

shown), revealing that NOB, AOB as well as denitrification bacteria

in the system were efficient and able to endure the toxicity of ani-

line. The reduction of TN in the system relied on denitrification

processes in anaerobic and anoxic conditions, which were hardly

influenced by aeration rate in aerobic phase. However, high DO

concentrations might take more time to decrease in anoxic phase,

which led to a slight increase of residual TN concentration. The

average removal efficiency of TN decreased from 87.42% to

75.39% in R1 and 92.38% to 84.23% in R2 with the DO concentration

increased from 4.0 ± 0.5 to 7.0 ± 0.5 mg L 1. The phenomenon that

nitrogen removal was enhanced at low DO levels was also men-

tioned in reports of Sayi-Ucar et al. (2015), in which TN was thor-

oughly removed under low DO conditions. Phosphorus

assimilation was also examined when DO concentration altered.

The residual concentration of phosphorus had no obvious fluctua-

tion with DO concentration of 5.5 ± 0.5 or 7.0 ± 0.5 mg L 1 in the

two systems. Both R1 and R2 showed good removal performance

in these two stages and phosphorus concentrations in effluent

were generally below 0.8 mg L 1. However, when DO concentra-

tions decreased to 4.0 ± 0.5 mg L 1, the average residual concentra-tion of phosphorus increased to 2.13 and 1.79 mg L 1, respectively

in R1 and R2. This indicated that in the systems aerobic uptake of

phosphorus was one of major ways to achieve phosphorus removal

in wastewater. The remarkable decrease of DO concentration

would inhibit this process, which caused increase of residual phos-

phorus concentration (Wang et al., 2015b). The test of pollutants

removal performance at different HRT was carried out by operating

the two reactors over three runs for 30 days. As seen in Table 2,

aniline was able to be completely degraded when HRT was 8 h,

12 h or 16 h. There was no notable distinction in COD removal

among these three conditions. However, the NH4+-N removal effi-

ciency increased with the delay of reaction time, especially that

in the aerobic phase. As with TN and TP, there was a noticeable

drop of removal efficiency when the cycle was reduced to 4 h whilethere was an increase when the HRT was adjusted from 12 h to

16 h, which revealed that the removal of nutrients improved along

with the extension of HRT (Liang et al., 2015). Considering the

residual concentrations of pollutants as well as energy saving,

12 h might be optimal HRT for both of the two systems.

3.4. Residual concentration profiles of pollutants during one cycle of

operation

In order to investigate the specific process of nutrients removal

in R1 and R2, typical profiles of pollutants compounds in reactors

during SBR cycle are presented in Fig. 6. As shown in Fig. 6(a), ani-

line concentration in R1 decreased moderately in anaerobic phasesince aniline was able to be utilized as the sole source of carbon,

nitrogen and energy for denitrifying bacteria in the presence of

NO3-N (Hu et al., 2014). It was also noted that there was a slight

drop in NO3-N concentration during this period (data not shown),

suggesting that a fraction of aniline was degraded through denitri-

fication. However, reduction of aniline mainly depended on aerobicbiodegradation. Aniline was able to be completely removed before

anoxic phase started. Over 95.51% of initial aniline was degraded in

aerobic phase. Aerobic biodegradation of aniline basically relied on

two steps, namely transformation of aniline into catechol via

aniline dioxygenase firstly and conversion into cis,cis-muconate

through the ortho-cleavage pathway by the catechol

1,2-dioxygenase or the meta-cleavage pathway by the catechol

2,3-dioxygenase afterward (Hyung-Yeel et al., 2000). NH4+-N was

liberated from aniline in first step of aniline biodegradation

(O’Neill et al., 2000; Dvoř ák et al., 2014). However, most of

NH4+-N coming from influent as well as aniline was removed in

aerobic phase in the present study, which successfully solved the

problem of excess NH4+-N in effluent. Both R1 and R2 showed high

removal rate of NH4+

-N in this stage and generally residual concen-trations at the end of the aerobic phase were less than 3 mg L 1. As

Fig. 6. Nutrients removal at the end of each phase in a typical operation cycle.

Table 2

Removal efficiencies of pollutants during variation of HRT.

R1 R2

8 h 12 h 16 h 8 h 12 h 16 h

COD 96.76 96.28 97.03 96.92 97.15 97.44

Aniline 100.00 100.00 100.00

NH₄+-N 78.96 92.27 97.21 85.47 90.18 96.56

TN 81.99 84.87 90.92 86.65 92.73 92.12

TP 89.95 93.81 93.45 76.47 92.96 93.04


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for COD concentrations in R1 and R2, after a slight decrease in

anaerobic phase there was a significant drop in the aerobic phase.

More than 65% of COD was removed in aerobic conditions and

residual concentrations were always below 40 mg L 1. The

consumption of COD in anaerobic conditions might be the result

of heterotrophic denitrification, which led to the noticeable

decrease of TN concentrations in the two systems (Bassin et al.,

2012). In anoxic phase, phosphorus uptake and denitrification tookplace simultaneously. It could be found from Fig. 6(a) and (b) that

TP reduction mainly depended on two ways in these two systems.

One was the uptake of phosphorus in aerobic conditions, which

was also described by Zhang et al. (2014), who used A/O/A

sequencing batch reactor to cultivate aerobic granular sludge. After

the release of phosphorus in anaerobic conditions by traditional

PAOs and DNPAOs, a large amount of phosphorus was accumulated

in the aerobic phase (Merzouki et al., 2005; Sun et al., 2015).

Another way was achieved by DNPAOs in anoxic conditions. After

energy conservation and polyhydroxyalkanoates (PHA) synthesis

in anaerobic condition in the presence of organic matters, PAOs

use the stored PHA to remove phosphorus during the aerobic or

anoxic period. This was quite different with the conventional SBR

process since this kind of microorganisms were able to perform

phosphorus uptake and denitrification simultaneously with nitrate

or nitrite instead of oxygen as electron acceptors, which could be

proved by the decrease of phosphorus during anoxic phase

(Wang et al., 2009). The decrease of residual TN during this period

might be due to metabolism of DNPAOs, ordinary denitrifying

bacteria as well as the process of anaerobic ammonium oxidation.

The phenomenon that the removal of NO2-N and NO3

-N with

relatively low concentration of COD in anoxic conditions could also

be found in the report of simultaneous denitrifying polyphosphate-

accumulating system described by Liu and Li, 2015. The good

removal performance of organic compounds, nitrogen and phos-

phorus in the system illustrated the cooperative relationship

among functional microorganisms. According to the results of the

present study, no additional devices were required for the

subsequent removal of nitrogen and phosphorus. The removal of both aniline and nutrients with high efficiency by a single reactor

could save the occupation area of the system and simplify

operation as well as management, which might contribute to the

practical application of the process.

4. Conclusions

In this work, simultaneous removal of aniline, nitrogen and

phosphorus in a single sequencing batch reactor was achieved.

Aniline in inflow was able to be completely degraded within every

cycle of operation during the whole test when initial concentra-

tions were less than 750 mg L 1. Under optimal conditions (Aniline

250 mg L 1, DO 5.5 ± 0.5 mg L 1, HRT 12 h), the removal efficien-

cies of COD, NH4+-N, TN, TP were generally over 95.80%, 83.03%,87.13%, 90.95%, respectively. The system in the present study

might have significance for nutrient removal from wastewater in

the presence of aniline.

Acknowledgements

This work was financially supported by the National Natural

Science Foundation of China (NSFC) (NO. 51378400), the National

Science and Technology Pillar Program (2014BAL04B04), the Natu-

ral Science Foundation of Hubei Province, China (NO. 2013CFB289).

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Simultaneous Removal of Aniline, Nitrogen and Phosphorus

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