Chemical Engineering Journal 345 (2018) 22–30

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Chemical Engineering Journal

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

Intensified nitrogen removal in intermittently-aerated vertical flow T

constructed wetlands with agricultural biomass: Effect of influent C/N ratios
⁎
Lixia Jiaa, Ruigang Wanga, Likui Fenga, Xu Zhoua, Jialong Lva, Haiming Wua,b,
a
College of Resources and Environment, Northwest A&F University, Yangling, Shaanxi 712100, PR China
b
Key Laboratory of Plant Nutrition and the Agri-environment in Northwest China, Ministry of Agriculture, PR China

H I GH L IG H T S G R A P H I C A L A B S T R A C T

• Addition of agricultural biomass could

intensify nitrogen removal sig-
nificantly.
• TN +
High removal of NH -N (99%) and
4
(96%) was obtained in VFCWs
with wheat straw.
• TN
Best removals of COD, NH -N and 4
were detected at an influent C/N
+

ratio of 0.5.
• Dosage of biomass addition and in-
fluent C/N should be further opti-
mized.

A R T I C LE I N FO A B S T R A C T

Keywords: The poor nitrogen removal due to the low denitrification caused by lack of carbon sources in vertical flow
Agricultural biomass constructed wetlands (VFCWs) especially for treating low carbon sewage. In this study, agricultural biomass
Constructed wetland materials including wheat straw, apricot pit and walnut shell had been tested as substrates in intermittently-
Carbon source aerated VFCWs for enhancing the nitrogen removal, and the effect of C/N ratios on the nitrogen removal was
C/N ratio
also studied. The results showed that intermittently-aerated VFCWs by adding agricultural biomass could in-
Nitrogen removal
tensify the denitrification process significantly, and the system with wheat straw performed much better than the
systems with walnut shell and apricot pit. Four microcosms with different agricultural biomass showed different
trends with the rising C/N ratio of influent. Moreover, the best removals (COD 97%, NH4+-N 99% and TN 96%)
have been obtained at an influent C/N ratio of 0.5, which indicated that the dosage of biomass addition and
influent C/N should be further optimized for achieving the sustainable nitrogen removal in VFCWs for low
carbon wastewater treatments.

1. Introduction
Water pollution problem is getting serious increasingly due to the
advancement of society. The access of clean water has become a
pressing issue influencing human health and surrounding environment
directly [1,2]. In recent years, more and more attention has been paid
to constructed wetlands (CWs) wastewater treatments because of their
lower cost and less energy input [3,4]. The CW is recognized as a
comprehensive treatment system for removing various pollutants (such
as organic and nutrients) by various processes such as sedimentation,
filtration, precipitation, volatilization, adsorption, plant uptake, and
microbial degradation [3,5]. Typically, subsurface flow constructed
wetlands (SFCWs) can be classified into two types: vertical flow con-
structed wetlands (VFCWs) and horizontal flow constructed wetlands

⁎
Corresponding author at: College of Resources and Environment, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
E-mail address: haimingwu14@nwsuaf.edu.cn (H. Wu).

https://doi.org/10.1016/j.cej.2018.03.087
Received 27 January 2018; Received in revised form 15 March 2018; Accepted 16 March 2018
Available online 17 March 2018
1385-8947/ © 2018 Elsevier B.V. All rights reserved.
L. Jia et al. Chemical Engineering Journal 345 (2018) 22–30

A B C D

15 cm
(1~2 mm)
Simulated
sewage
Wheat Apricot Walnut 15 cm
straw pit shell (10~20 mm)

25 cm
(20~30 mm)

Flowmeter Flowmeter
10 cm
(30~40 mm)
Air pump

Fig. 1. The schematic diagram of experimental laboratory-scale constructed wetland system (A: CW with wheat straw; B: CW with apricot pit; C: CW with walnut
shell; D: CW without addition).

Table 1
Characteristics of the influent during the experimental period (Mean ± SD, n = 10).
Parameters COD/N = 0.5 COD/N = 1 COD/N = 3 COD/N = 5

−1
COD (mg L ) 24.18 ± 1.57 45.40 ± 1.95 123.95 ± 2.47 222.36 ± 7.60
NH4+-N (mg L−1) 42.49 ± 0.88 42.57 ± 0.66 41.11 ± 1.09 40.67 ± 1.39
NO3−-N (mg L−1) 1.43 ± 0.25 1.44 ± 0.05 1.32 ± 0.14 1.49 ± 0.08
TN (mg L−1) 44.22 ± 0.27 44.24 ± 0.66 41.79 ± 0.37 42.45 ± 0.47
TP (mg L−1) 5.49 ± 0.22 5.36 ± 0.25 5.39 ± 0.39 5.63 ± 0.27
COD/N 0.57 ± 0.05 1.07 ± 0.04 3.02 ± 0.07 5.47 ± 0.10

(HFCWs) in terms of flow direction [6]. However, nitrogen removal in
common SFCWs often exhibits substantial variations that could not
meet the stringent effluent discharge standards when treating some low
carbon sewages.
Among various nitrogen removal processes in CWs, microbial ni-
trification and subsequent denitrification are known as the key routes
for ultimate nitrogen removal [6]. Thus, it would be difficult to achieve
high nitrogen removal efficiency if either nitrification or denitrification
process was obstructed. The nitrification process in CWs can be influ-
enced by multifarious factors, of which available oxygen is always a key
one. The additional dissolve oxygen (DO) is necessary for eliminating
nitrogen. Some researchers have showed that intermittent aeration can
create an alternative anaerobic and aerobic environment to enhance
nitrogen removal. Moreover, this method outperformed conventional
continuous aeration. Ong et al. [7] found that the supplementary
aeration only at the upper beds of CWs enhanced the biodegradation of
organic matters and the removal of ammonia nitrogen (NH4+-N). Dong
et al. [8] found that higher total nitrogen (TN) removal (57%) could be
obtained under condition of intermittent aeration. Fan et al. [9] also
proved that intermittent aeration could be an effective method and
optimal mode for TN removal by providing an alternative environment
respectively for nitrification and denitrification processes. Uggetti et al.
[10] have demonstrated the high potential of the intermittent aeration
to increase wastewater treatment efficiency of CWs providing an ex-
treme benefit in terms of the energy consumption. Zhang et al. [11]
investigated the intermittent aeration operation mode in a mature
landfill leachate and proved this method can effectively improve ni-
trogen removal. The inadequate carbon source or labile organic matters
could be also a major influencing factor, which can lead to an in-
efficient denitrification in CWs [12]. The inherent carbon source
coming from wastewater, substance and the root excretion of plants is
always deficient, so the external carbon addition (such as liquid carbon
source and solid carbon source) is necessary to enhance TN removal
[13,14]. Many studies reported that liquid carbon sources such as
methanol, fructose and glucose played a great role in CWs treating low
C/N wastewaters [15–17]. However, those carbon sources are generally
high cost and readily biodegraded through aerobic decomposition and
the CW system is difficult to be controlled [18,19] Synthetic polymers
and natural materials are two typical solid carbon sources in which the
former is expensive generally [17]. Natural materials such as agri-
cultural biomass (maize cobs, wheat straw and cotton), crab-shell chitin
and plant litter have proven to be suitable carbon media to improve
nitrogen removal course completely [12,19–21]. Wen et al. [12] dis-
cerned that nitrate removal was greatly affected by the composition of
the plant biomass as well as the quantity and quality of the available
organic matters. Shen et al. [19] found that Cornstarch/poly-
caprolactone (SPCL) blends as external carbon sources could improve
the nitrogen elimination. However, there are few studies on evaluating
comparatively the potential role of different kinds of agricultural bio-
mass materials as carbon source to enhance nitrogen removal in CWs
for treating low carbon sewage.
Besides above two crucial factors, the fluctuating influent C/N af-
fected significantly integral treatment performance likewise [22,23].
Zhao et al. [24] found that when influent C/N ratios changed from 2.5:1
to 5:1, the TN removal efficiency could reach 62% in conventional
VFCWs. Fan et al. [25] studied the effect of C/N on the treatment
performance of VFCWs in the presence of intermittent aeration and
obtained high TN removal (90%) under the influent C/N of 10:1. From
the above analysis, there are more studies on intermittent aeration
method applied in CWs, and the researches about agricultural materials

23
 Table 2
Characteristics of effluents and respective removal efficiencies (Mean ± SD, n = 10).
L. Jia et al.

Parameters System1 RE (%) RR (mg m−2 d−1)

COD NH4+-N TN TP COD NH4+-N TN TP

COD/N = 0.5 A 37.62 ± 11.05c 98.78 ± 0.25a 96.89 ± 1.28a 64.18 ± 3.82a 410.37 ± 68.18c 2219.14 ± 40.32a 2261.03 ± 44.57a 186.79 ± 13.01a
B 55.81 ± 13.12b 98.30 ± 0.50a 19.42 ± 0.13c 35.58 ± 5.42b 648.16 ± 70.89b 2207.46 ± 40.61a 481.72 ± 57.11c 103.83 ± 15.52b
C 68.39 ± 10.42ab 98.83 ± 0.58a 62.88 ± 0.30b 49.05 ± 5.09b 748.41 ± 135.65ab 2218.39 ± 42.23a 1459.45 ± 57.97b 142.79 ± 14.66b
D 76.07 ± 1.17a 98.82 ± 0.07a 3.98 ± 2.12d 34.63 ± 7.58b 911.66 ± 71.94a 2220.10 ± 38.20a 110.39 ± 53.39d 101.03 ± 20.17b

COD/N = 1.0 A 67.55 ± 2.41b 99.83 ± 0.07a 59.15 ± 3.19a 72.85 ± 5.54a 1594.84 ± 91.30b 2230.62 ± 26.05a 1314.07 ± 68.36c 211.59 ± 25.19a
B 78.76 ± 8.36b 99.51 ± 0.12a 24.88 ± 3.15c 44.44 ± 2.41b 1679.55 ± 109.17b 2222.83 ± 22.69a 471.00 ± 69.97c 128.62 ± 11.14b
C 81.21 ± 7.77b 99.46 ± 0.20a 47.76 ± 1.13b 52.90 ± 2.33b 1768.63 ± 115.42b 2226.56 ± 30.30a 1019.76 ± 75.84b 153.05 ± 11.98ab
D 87.28 ± 6.01a 99.70 ± 0.25a 15.41 ± 0.66d 42.02 ± 3.95b 1974.41 ± 75.43 a 2228.87 ± 27.07a 333.08 ± 37.65d 122.10 ± 15.88b

COD/N = 3.0 A 85.46 ± 1.35d 99.46 ± 0.12a 69.27 ± 0.22a 65.87 ± 3.56a 5583.11 ± 117.15d 2191.03 ± 43.37a 1532.07 ± 34.35a 188.88 ± 19.75a
B 88.97 ± 0.83c 99.29 ± 0.13a 28.10 ± 1.78c 61.20 ± 2.31c 5820.41 ± 118.07c 2187.20 ± 43.22a 701.34 ± 31.49c 175.45 ± 16.62c
C 92.14 ± 0.39b 99.19 ± 0.26a 58.84 ± 0.42b 64.96 ± 3.83b 6034.34 ± 69.90b 2184.62 ± 41.64a 1311.35 ± 47.39b 186.27 ± 19.83b
D 97.65 ± 0.84a 99.63 ± 0.27a 20.48 ± 4.95d 58.59 ± 2.05d 6403.17 ± 57.58a 2194.51 ± 42.95a 478.19 ± 50.43d 167.82 ± 14.42d

COD/N = 5.0 A 92.71 ± 0.79a 98.87 ± 0.87a 63.02 ± 1.64a 59.21 ± 0.88b 10880.33 ± 237.95a 2135.32 ± 60.47a 1391.88 ± 39.24a 177.14 ± 9.40b
B 95.33 ± 0.77ab 98.49 ± 0.75a 46.27 ± 3.04b 66.70 ± 0.93ab 11215.25 ± 261.52ab 2126.15 ± 57.25a 1019.22 ± 33.74b 199.31 ± 6.94ab
C 94.96 ± 0.45ab 99.01 ± 0.68a 46.27 ± 3.04a 70.67 ± 1.37a 11183.51 ± 256.24ab 2137.73 ± 58.08a 1398.84 ± 65.76a 211.44 ± 11.93a
D 96.61 ± 1.01b 98.93 ± 0.30a 32.64 ± 2.78c 66.76 ± 1.87ab 11413.23 ± 256.71b 2135.87 ± 51.85a 710.60 ± 70.67c 199.70 ± 11.42ab

Different letters indicate significant differences between the treatments (P < 0.05).
RE: Removal Efficiency; RR: Removal Rate.
1
A: CW with wheat straw; B: CW with apricot pit; C: CW with walnut shell; D: CW without addition.

24
+ -1 -1
NH4-N Con. (mg L ) COD Con. (mg L )
0
10
20
30
40
50
150
200
250

20
30
40
50

0.0
0.5
1.0
1.5
0

0
6

6
COD/N=0.5
COD/N=0.5

COD/N=0.5

2. Material and methods

2.1. Agricultural biomass materials

Influent
Influent
COD/N=1.0

COD/N=1.0

Effluent B
Effluent D
Effluent B
Effluent A

Effluent D
Effluent C

Class (A)

Time (d)
Time (d)
Class (A)
COD/N=3.0

COD/N=3.0

Effluent A
Effluent C

apricot pit; C: CW with walnut shell; D: CW without addition).

apricot pit; C: CW with walnut shell; D: CW without addition).

Discharge standard(GB18918-2002)

provide the operating parameters for the practical application.

COD/N=5.0
COD/N=5.0

12 18 24 30 36 42 48 54 60 66 72 78 84
12 18 24 30 36 42 48 54 60 66 72 78 84
Discharge standard(GB18918-2002)

different biomass materials and intermittent aeration is still scarce.

0
5

straw, apricot pit and walnut shell as substrates in VFCWs to enhance

Fig. 3. The influent and effluent concentrations of NH4+-N in intermittently-

carbon substrates to drive denitrification. The agricultural biomass

spread obtainability and lower cost to serve as prospective natural
Fig. 2. The influent and effluent concentrations of COD in intermittently-aer-

Three kinds of agricultural biomass materials including wheat

sibility and efficiency by using agricultural biomass such as wheat

effect of the C/N ratios on the nitrogen removal was also investigated to
The main objectives of the present study were to explore the fea-
as carbon source improving nitrogen removal can also be found.
aerated VFCWs under different C/N ratios (A: CW with wheat straw; B: CW with
ated VFCWs under different C/N ratios (A: CW with wheat straw; B: CW with

the nitrogen removal in treating low carbon sewage. Moreover, the

enhancing removal performance in VFCWs with the application of
Nonetheless, the research about the effect of influent C/N ratios on the

straw, apricot pit and walnut shell were selected based on their wide-
Chemical Engineering Journal 345 (2018) 22–30
L. Jia et al. Chemical Engineering Journal 345 (2018) 22–30

COD/N=0.5 COD/N=1.0 COD/N=3.0 COD/N=5.0 was installed at the bottom of the wetlands and was connected to an air
50 compressor for oxygen supply with a flow meter for setting the daily
aeration volume. Each VFCWs system was planted with Iris pseudacorus
45
at a density of 4–6 rhizomes. The reasons of choosing this macrophyte
40 specie are based on the following points: (1) large plants to provide
more biomass; (2) strong adaptability to environment; (3) high removal
35
efficiency of pollutant; (4) high resistance to pests and diseases. Before
TN Con. (mg L )

Class (A) planting, the plants were cultivated by improved Hoagland Nutrient
-1

30
Discharge standard(GB18918-2002) Solution for about two weeks [26]. When the plants were growing well,
25
they were transplanted into each system. All systems were then flooded
20 with simulated wastewater for 30 d, and acclimated for inoculating
microorganisms by adding the supernatant from the secondary sedi-
15
Influent mentation tank in Yang ling sewage treatment plant.
10 Effluent A
Effluent B
5 2.3. Experimental procedure
Effluent C
Effluent D
0 To investigate the nitrogen removal performance under different C/
0 6 12 18 24 30 36 42 48 54 60 66 72 78 84
N ratios, the influent with the different COD/N (chemical oxygen de-
Time (d)
mand/TN concentration) was manipulated by changing the amount of
Fig. 4. The influent and effluent concentrations of TN in intermittently-aerated sucrose to create four COD/N ratios (0.5:1, 1:1, 3:1, 5:1). The synthetic
VFCWs under different C/N ratios (A: CW with wheat straw; B: CW with apricot wastewater used in this experiment was mainly composed of dissolving
pit; C: CW with walnut shell; D: CW without addition). sucrose, (NH4)2SO4, KH2PO4, MgSO4, CaCl2 and FeSO4. The char-
acteristics of the influents in each parallel experiment are shown in
materials were collected from the local farms near Yang ling city, Table 1. The hydraulic retention time (HRT) was 72 h based on the
China. After washing with deionized water and air-drying for a week to previous study [27]. All systems were intermittently aerated at an air-
a constant weight, the collected materials were then cut into 1–2 cm flow rate of 1.0 L min−1 for 4 h (hours 0–1, 6–7, 12–13 and 18–19) each
pieces. Finally, the prepared biomass litter was stored in a dryer until day [28]. Sequencing fills-and-draw batch mode was applied in this
used. study, and each system was manually drained at about 9:00 am after an
operating cycle (72 h), and then re-filled with the synthetic sewage. The
temperature during the whole experiment was 25 ± 1 °C.
2.2. Design of experimental CW systems

The experiment was carried out in Northwest A&F University. A
schematic diagram of system is shown in Fig. 1. Four parallel micro-
cosm VFCWs were constructed and filled with different substrates (A:
aerated with wheat straw; B: aerated with apricot pit; C: aerated with
walnut shell; D: aerated without addition). The wetland bed system was
divided into two layers, and the lower is 50 cm thick gravel layer with
the following upper layer of 15 cm thick sand. The size of gravel par-
ticle was gradually decreasing from the bottom to the top in order to
improve the utilization rate of aeration. From the bottom to the top, the
first section was 10 cm thick gravel (Φ = 5–7 cm). The following was
25 cm medium gravel section (Φ = 2–4 cm). The next part was 15 cm
small gravel (Φ = 1–2 cm) with addition of prepared biomass materials,
and each biomass material should be mixed the gravel at 1:1 propor-
tion. The particle size of sand in the top was less than 1 cm. To facilitate
the in situ measurement of DO inside the wetland system, a PVC per-
forated tube (66 cm high and 3 cm in diameter) was inserted in the
center of the each system. A high temperature-sintered bubble stone
2.4. Water sampling and analysis
The water samples of influent and effluent in each system were
collected at 9:00 am after 72 h to analyze the concentration of chemical
oxygen demand (COD; HACH DR 2008TM Spectrophotometer, USA),
ammonia nitrogen (NH4+-N), nitrate (NO3−-N), nitrite (NO2−-N), total
nitrogen (TN) and total phosphorus (TP). The dissolved oxygen (DO)
was measured by a DO meter (HQ 30d 53LEDTM HACH USA).
Particularly, in order to investigate the transformation of pollutants in
the typical cycle, water samples were also collected at different time
intervals (1, 2, 4, 6, 10, 14, 24, 28, 32, 38, 48, 54, 60 and 72 h). All
analysis was according to standard methods [29].
2.5. Statistical analysis
The statistical analysis was carried out using SPSS version 16.0
software (SPSS Inc., Chicago, USA). All results were expressed as the

Table 3
Comparative comparison of treatment performance in CWs by using various external carbon sources.
System1 External carbon source Wastewater type Removal Efficiency (%) References

+
COD NH4 -N TN

VF CW Agricultural biomass Synthetic wastewater 97.7 99.7 96.9 This study

SSF CW cattail litter nitrate-dominated water – – 29.9–80.3 [12]
(NO3−-N)
VF CW Cornstarch/polycaprolactone (SPCL) blends nitrate-contaminated wastewater – – 98.2 [19]
(NO3−-N)
VF CW Biochar Synthetic wastewater 94.9 99.1 52.7 [28]
SSF CW sludge-ceramsite Synthetic wastewater 97.2 98.9 85.8 [33]
CW immobilized pellets Synthetic wastewater 79.6–82.9 67.1–78.3 49.1–59.9 [34]
VF CW Plant-based carbon source Artificial sewage 91.5 92.8 [35]
SSF CW Plants litter secondary effluent 11.1 – 75.2 [36]
HSSF CW Plant fermentation broth The secondary effluent 71.6 72.2 96.7 [37]

1
SSF CW-Subsurface Flow CW; VF CW-Vertical Flow CW; HSSF CW-Horizontal Subsurface Flow CW.

25
L. Jia et al. Chemical Engineering Journal 345 (2018) 22–30

45 30 45 30
A + B
40 NH4-N 40
-
NO3-N 25 25
35 - 35
NO2-N
30 TN 20 30 20
Nitrogen Con. (mg L )

Nitrogen Con. (mg L )

COD Con. (mg L )

COD Con. (mg L )

-1

-1
-1

-1
+
NH4-N
COD -
25 25 NO3-N
15 NO2-N
-
15
20 20 TN
COD
15 10 15 10

10 10
5 5
5 5

0 0 0 0
0 6 12 18 24 30 36 42 48 54 60 66 72 0 6 12 18 24 30 36 42 48 54 60 66 72
Time (h) Time (h)

45 30 45 30
C NH4-N
+ D
40 40
-
NO3-N 25 25
35 NO2-N
- 35

30 TN 20 COD Con. (mg L ) 30 20

Nitrogen Con. (mg L )

Nitrogen Con. (mg L )

+
-1
-1

-1
NH4-N

COD Con. (mg L )

COD

-1
-
25 25 NO3-N
15 NO2-N
- 15
20 20
TN
10 COD 10
15 15

10 10
5 5
5 5

0 0 0 0
0 6 12 18 24 30 36 42 48 54 60 66 72 0 6 12 18 24 30 36 42 48 54 60 66 72
Time (h) Time (h)

Fig. 5. Dynamic transformations of organics and nitrogen in intermittently-aerated CW systems at C/N rate = 0.5:1 during the typical operating period (A: CW with
wheat straw; B: CW with apricot pit; C: CW with walnut shell; D: CW without addition).

mean values. All of the variables were tested to ensure that they were was more efficient in organic matters degradation under low C/N
normally distributed. Significance levels are reported as non-significant condition. The average removal efficiency of system without any ex-
(p > 0.05), significant (p < 0.05). ternal matters increased from 76% to 96% (911.66 to
11413.23 mg m−2 d−1), which was higher than that of other three
systems with addition of biomass materials. A possible explanation is
3. Results and discussion
that organic matters released from the biomass itself was partially used
for enhancing the removal of nitrogen. On the other hand, these agri-
3.1. Overall treatment performance
cultural biomass might have some components such as lignin making
the COD concentration increase which is difficult for microorganisms to
3.1.1. Impact of influent C/N ratio on organics removal
degrade [31,32].
The removal performance of COD, NH4+-N, TN and their dynamic
changes in VFCWs during the experiment are shown in Table 2 and
Fig. 2. Values of COD, NH4+-N, TN and TP differed significantly among 3.1.2. Impact of influent C/N ratio on NH4+-N removal
four types of wetlands under different influent C/N ratios. As can be Nitrification is indispensable and the primary step when NH4+-N
seen from the Fig. 2, the effluent COD concentration was almost stable, was transformed into NO3−-N for the TN elimination. An efficient ni-
and did not show an increasing or decreasing trend in each system with trification often requires high DO concentration [3,6]. Previous studies
the increase of C/N ratio. However, an obvious discrepancy could be have shown that much more nitrifying bacteria can be detected in in-
found among four system adding wheat straw was highest than that of termittently aerated CWs [25]. As shown in Table 2, the NH4+-N re-
other systems but far lower than the ClassⅠ(A) of Wastewater Discharge moval efficiencies of four systems could reach above 98% under various
Standard (GB18918-2002) in China (50 mg L−1). This phenomenon C/N ratios. Furthermore, the effluent NH4+-N concentrations in all
could be explained by the reason that wheat straw can release more systems were below 1 mg L−1 which was far lower than the ClassⅠ(A) of
available carbon for promoting denitrification theoretically, followed Wastewater Discharge Standard (GB18918-2002) in China (5 mg L−1).
by walnut shell, and apricot pit release the least. It was observed that Obviously from Fig. 3, the change of influent C/N ratios as well as the
average COD removal rates were increased with the rise of C/N ratios addition of external biomass carbon sources had little effect on the
from 0.5 to 5.0. Fig. S3 shows that there were strong positive correla- removal of NH4+-N. As a whole, there was no significant difference
tions between the COD/N ratio and organics removal rate both in CWs (P > 0.05) for effluent NH4+-N concentrations among CWs with dif-
with agricultural biomass and in CW without addition. This tendency is ferent external carbon source under various influent C/N ratios. It can
in agreement with the result of a previous study [30]. But it also should also be indicated that gradually increase of carbon resulted from higher
be noted that the conventional wetlands with none biomass materials influent C/N did not deplete all available oxygen by intermittent

26
L. Jia et al. Chemical Engineering Journal 345 (2018) 22–30

45 50 45 50
A +
NH 4-N B
40 -
45 40 45
NO 3-N
35 - 40 35 40
NO 2-N
TN 35 + 35
30 30 NH 4-N

Nitrogen Con. (mg L )

Ntrogen Con. (mg L )

-1
-1

CODCOD

COD Con. (mg L )

COD Con. (mg L )

-1

-1
-
30 NO 3-N 30
25 25 -
25 NO 3-N 25
20 20 TN
20 20
CODCOD
15 15
15 15
10 10
10 10
5 5 5 5

0 0 0 0
0 6 12 18 24 30 36 42 48 54 60 66 72 0 6 12 18 24 30 36 42 48 54 60 66 72
Time (h) Time (h)

45 50 45 50
+
C NH 4-N D
40 -
45 40 45
NO 3-N
-
NO 2-N 40 35 40
35
TN 35 35

Nitrogen Con. (mg L )

-1
+
30 30
Nitrogen Con. (mg L )

NH 4-N
-1

CODCOD COD Con. (mg L )

COD Con. (mg L )

-1

-1
30 - 30
25 25 NO 3-N
-
25 NO 2-N 25
20 20 TN
20 20
COD
15 15
15 15
10 10
10 10
5 5 5 5

0 0 0 0
0 6 12 18 24 30 36 42 48 54 60 66 72 0 6 12 18 24 30 36 42 48 54 60 66 72
Time (h) Time (h)

Fig. 6. Dynamic transformations of organics and nitrogen in CW systems at C/N rate = 1:1 during the typical operating period (A: CW with wheat straw; B: CW with
apricot pit; C: CW with walnut shell; D: CW without addition).

aeration, thus aeration can provide sufficient oxygen to enhance ni- vigorous denitrification. The following was C system and B system. This
trification. Therefore, the oxygen concentration is crucial to the NH4+- phenomenon might be attributed to the more available carbon source
N removal in aerated VFCWs with agricultural biomass. released from biomass materials, particularly wheat straw [31]. In solid
carbon sources supporting systems, denitrification rate is strongly de-
pendent on the biodegradability of solid carbon sources. A higher de-
3.1.3. Impact of influent C/N ratio on TN and TP removal nitrification rate will be received when the solid carbon sources have
To achieve TN removal completely, the nitrate accumulated from better biodegradability [19]. The TN removal efficiency in this study
nitrification should be converted into N2 or N2O by denitrification [3]. rested with the nitrate accumulated in the denitrification process.
Fig. 4 and Table 2 also show the average effluent TN concentration and Therefore, the wheat straw might provide more carbon source that can
the removal performance in each system throughout the whole ex- easily degraded by microorganism.
periment. It is worthwhile mentioned that only the effluent TN con- However, each system has a different variation with the increase of
centration of system A with wheat straw complied with the ClassⅠ(A) of influent C/N ratio. Meanwhile, there was significant difference
Wastewater Discharge Standard (GB18918-2002) in China (15 mg L−1). (P < 0.05) for effluent TN concentrations among CWs with different
The removal efficiencies were 96.89%, 59.15%, 69.27%, 63.02% from external carbon source under various influent C/N ratios. The effluent
the influent C/N rate = 0.5 to 5. This demonstrated a great improve- TN concentration of the CW with wheat straw showed a trend of first
ment for the TN removal in VFCWs with agricultural biomass compared increasing but decreasing when the C/N rate = 3.0, which was similar
to the results of study by Zhou et al. [28] in which the TN removal with the trend in the CW adding walnut shell. Furthermore, relation-
efficiency were only 52.7% in biochar-amended VFCWs. From the table ships between the COD/N ratio and nitrogen removal rate in four mi-
3, we can find that our results showed a substantial improvement for crocosms were examined via simple linear regression analyses. It is
the TN removal comparing with other enhanced CWs in previous stu- clear that there were positive correlations between the COD/N ratio
dies [33–37,12,28]. Chen et al. [36] revealed that the relative abun- and nitrogen removal rate in CW B, C and D except CW A. (Fig. S3). This
dances of nitrifier, denitrifier and sulfate-reducing bacteria were posi- suggested high nitrogen removal rate may be related to the type of
tively correlated with the removal efficiencies of ammonia, nitrate and biomass-derived carbon. In addition, this result could be confirmed by
sulfate observed by adding plant biomass into CW. The TN removal other studies. Zhang et al. [31] observed that although significant
efficiency could be improved to 75.2% in Chen’s study [36]. In addi- amounts of carbon source could be released from wheat straw in initial
tion, Zhang et al. [37] adapted a self-supplying carbon source by fer- stage, the organic carbon concentration rapidly decreased to a lower
menting plant litter. The experiment proved that the addition of fer- value relatively in 15 days. To a certain extent, this can explain the
mentation broth greatly improved the nitrate removal rate. From our phenomenon obtained in the CW system A and C in this study. Different
research, it is noted that among the four systems, the A system with from the system A and C, the TN concentration of system B and D varied
wheat straw showed the most significant decrease in TN and the most

27
L. Jia et al. Chemical Engineering Journal 345 (2018) 22–30

45 125 50 +
135
A +
NH 4 -N B NH 4 -N
40 - 45 - 120
NO 3 -N NO 3 -N
-
35
-
NO 2 -N 100 40 NO 2 -N 105
TN 35 TN
30 90
Nitrogen Con.(mg L )

Nitrogen Con.(mg L )
COD 1)
-1

-1
COD 1)

COD Con.(mg L )

COD Con. (mg L )

-1

-1
75 30
25 75
25
20 60
50 20
15 45
15
10 30
25 10
5 5 15

0 0 0 0
0 6 12 18 24 30 36 42 48 54 60 66 72 0 6 12 18 24 30 36 42 48 54 60 66 72
Time (h) Time (h)

45 135 45 135
C +
NH 4 -N
D
40 120 40 120
-
NO 3 -N
35 -
NO 2 -N 105 35 105

30 TN 30

Nitrogen Con. (mg L )

90 90
Nitrogen Con.(mg L )

-1
-1

COD Con. (mg L )

COD Con. (mg L )

-1
COD 1)

-1
+
NH 4 -N
25 75 25 75
-
NO 3 -N
20 60 20 NO 2 -N
- 60

15 15 TN
45 45
COD 1)
10 30 10 30

5 15 5 15

0 0 0 0
0 6 12 18 24 30 36 42 48 54 60 66 72 0 6 12 18 24 30 36 42 48 54 60 66 72
Time (h) Time (h)

Fig. 7. Dynamic transformations of organics and nitrogen in CW systems at C/N rate 3:1 during the typical operating period (A: CW with wheat straw; B: CW with
apricot pit; C: CW with walnut shell; D: CW without addition).

regularly which decreased with the increasing of influent C/N ratio. concentration in the CW with none addition was lower than that of
This was might because apricot pit can provide less organic carbon but other CWs with biomass materials. Overall, the increasing of influent C/
continuously like the biodegradable plastic (BP) researched in Zhang N from 0.5 to 5.0 improves the efficiencies of organic matters de-
et al. [31]. gradation.
The average TP removal efficiencies in four CW systems can be
shown in Table 3 and Fig. S3. The CWs with agricultural biomass ex-
3.2.2. Dynamic of nitrogen transformation
hibited a better TP removal performance than the CW with none ad-
The nitrogen transformation in CWs mainly includes volatilization,
dition. It is also noted that TP removal rate increased in CW systems B,
ammonification, nitrification, denitrification, plant uptake and matrix
C and D except CW system A as the influent C/N ratio increased like the
adsorption [22]. Denitrification as a key process to remove nitrogen
variation of TN.
relied largely on the amount of nitrate, organic carbon and the anae-
robic environment [6]. The role of enhancing denitrification by various
3.2. Organics and nitrogen transformations in the typical operating period biomass materials added in the wetland systems was mainly considered
in this study. The time-profiles of the NH4+-N, NO3−-N and NO2−-N
3.2.1. Variation of DO and COD concentrations concentration in four kinds of VFCWs at different influent C/N ratios
The variations of DO and COD concentrations in four systems under during typical period are presented in Figs. 5 to 8. It was observed that
different influent C/N ratios during the typical experiment cycle are NH4+-N concentration decreased sharply in the first day of the cycle
photographically shown from Figs. 5 to 8 and Fig. S1. The DO con- and then reduced slightly. Finally, it was stable in the end of the period.
centration in all systems at various influent C/N ratios exhibited similar The average effluent concentrations of NH4+-N in each system at var-
changing tendency that is strict alternative anaerobic and aerobic en- ious influent C/N ratios were all close to 0.5 mg L−1, which could be
vironment at the initial stage of cycle. However, the values of DO attributed to sufficient DO provided by intermittent aeration. This
concentration decreased slower gradually when stopping aeration. As a suggested that increasing sucrose and the different external carbon did
result, the DO concentration of CWs could reach above 2 mg L−1 at the not affect significantly the removal performance of NH4+-N.
end of the period, which can form an aerobic environment. Previous After the conversion of NH4+-N to NO3−-N and NO2−-N via ni-
studies described that the effect of aeration always led to higher DO trification, the denitrification must transform the NO3−-N and NO2−-N
concentration and caused low efficiencies in denitrification [23,38]. into N2 to finish N removal completely [3]. It can be seen that large
Therefore, the denitrification was restrained because anaerobic condi- amount of NO3−-N was accumulated immediately and increased until
tion was not available, and thus inhabited the activity of denitrifying at the end of cycle in the system D without external matters under
bacteria in the latter period of the cycle. As presented in results during different C/N ratios. Compared with none-biomass-aerated VFCWs, the
the typical cycle, COD concentration decreased rapidly after feeding effluent NO3−-N concentrations were lower in the systems adding
wastewater, but approximately 10 h later, the COD concentration sta- agricultural biomass under different influent C/N ratios. This suggested
bilized at a low level. Zhou et al. [28] also obtained a similar conclusion that these biomass materials could improve the denitrification process
in the previous study. However, in the terminal stage, the COD at different levels. On the other hand, the value of TN was almost equal

28
L. Jia et al. Chemical Engineering Journal 345 (2018) 22–30

50 250 50 250
A +
NH 4 -N B +
NH 4 -N
45 -
225 45 - 225
NO 3 -N NO 3 -N
40 NO 2 -N
- 200 40 -
NO 3 -N 200

35 TN 175 35 TN 175

Nitrogen Con. (mg L )

Nitrogen Con. (mg L )

-1
-1

COD F COD 1)

COD Con. (mg L )

COD Con. (mg L )

-1

-1
30 150 30 150

25 125 25 125

20 100 20 100

15 75 15 75

10 50 10 50

5 25 5 25

0 0 0 0
0 6 12 18 24 30 36 42 48 54 60 66 72 0 6 12 18 24 30 36 42 48 54 60 66 72
Time (h) Time (h)
50 250 50 250
+
C NH 4 -N
+
D NH 4 -N
45 225 45 - 225
NO 3 -N
- NO 3 -N
-
40 - 200 40 NO 2 -N 200
NO 2 -N
35 TN 35 TN
175 175
Nitrogen Con. (mg L )

Nitrogen Con. (mg L )

COD Con. (mg L )
-1

-1
COD F

-1

COD Con. (mg L )

COD

-1
30 150 30 150

25 125 25 125

20 100 20 100

15 75 15 75

10 50 10 50

5 25 5 25

0 0 0 0
0 6 12 18 24 30 36 42 48 54 60 66 72 0 6 12 18 24 30 36 42 48 54 60 66 72
Time (h) Time (h)

Fig. 8. Dynamic transformations of organics and nitrogen in CW systems at C/N rate = 5:1 during the typical operating period (A: CW with wheat straw; B: CW with
apricot pit; C: CW with walnut shell; D: CW without addition).

to NO3−-N indicating that denitrification is the limiting step for effec- biomass as a potential substrate in VFCWs could be a feasible strategy
tive TN removal due to inadequate carbon source in the initial stage for improving denitrification in low C/N wastewater treatments.
when the alternative anaerobic and aerobic environment was guaran-
teed. However, for other three CWs with various biomass materials, Acknowledgements
NO3−-N increased slightly in the early and accumulated rapidly in the
later of cycle. Moreover, there was no accumulation of NO2−-N We gratefully acknowledge financial support by the Water
throughout the whole typical period under different C/N ratios. The Resources Science and Technology Program of Shaanxi Province
above phenomenon can be explained from the following reason. The (2017slkj-6), the National Science Foundation of China (NSFC, Project
wetland system formed an alternative anaerobic and aerobic environ- No. 51508466), and the Fundamental Research Funds for the Central
ment because of intermittent aeration, so nitrate accumulated slowly Universities (No. 2452016067).
and less in initial stage [25]. But with the consumption of NH4+-N and
organic matter, DO concentration in wetlands gradually increased that Appendix A. Supplementary data
can be observed from the Fig. S2. Thus at the terminal stage, the de-
nitrification was obstructed due to the aerobic environment and NO3−- Supplementary data associated with this article can be found, in the
N accumulated quickly. online version, at http://dx.doi.org/10.1016/j.cej.2018.03.087.
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