Front. Environ. Sci. Engin.

China 2007, 1(1): 49–52

DOI 10.1007/s11783-007-0009-1
RESEARCH ARTICLE

Simultaneous nitrification and denitrification in activated

sludge system under low oxygen concentration
ZHANG Peng1, ZHOU Qi ( )2
1 College of Environmental and Material Engineering, Yantai University, Yantai 264025, China
2 State key Laboratory of Pollution Control and Resources Reuse, Tongji University, Shanghai 200092, China

© Higher Education Press and Springer-Verlag 2007

Abstract Simultaneous nitrification and denitrification found that some principal factors predominantly influence
(SND), which is more economical compared with the tradi- SND. These are carbon supply, oxygen concentrations, and
tional method for nitrogen removal, is studied in this paper. In floc size [9,11]. Pochana et al. [9,10] proposed that the addi-
order to find the suitable conditions of this process, a mixed tion of readily biodegradable COD source resulted in a sig-
flow activated sludge system under low oxygen concentration nificant increase of SND activity. The increased dissolved
is investigated, and some key control parameters are exam- oxygen concentrations in the reactor bulk liquid negatively
ined for nitrogen removal from synthetic wastewater. The affected SND and SND activities, which were increased in
results show that SND is accessible when oxygen concentra- reactors with larger activated sludge floc size.
tion is 0.3–0.8 mg/L. The nitrogen removal rate can be Currently in China, most studies on the mechanisms of
obtained up to 66.7% with solids retention time (SRT) SND are focused on nitrification and denitrification as sepa-
of 45 d, C/N value of 10, and F/M ratio of 0.1 g COD/ rate independent phases, spatially conducted in different reac-
(g MLSS · d). Theoretical analysis indicates that SND is a tors like traditional A/A/O, and A/O process or temporally
physical phenomenon and governed by oxygen diffusion in conducted in a sequencing batch reactor or oxidation ditch
flocs. under aerobic and anaerobic conditions. However, authentic
mechanisms of SND have not been proposed. Therefore, in
Keywords low oxygen concentration, simultaneous this study, a complete mixing activated sludge system was
nitrification and denitrification (SND), aerobic nitrification, used to further explore the SND process to understand its
microenvironment theory mechanisms sufficiently.

1 Introduction 2 Materials and methods

2.1 Experimental set-up and reactor operation

There are a number of dominant mechanisms for simultane-
ous nitrification and denitrification (SND) such as the micro-
Figure 1 shows the experimental set-up of this study. As
environment theory [1], heterotrophic nitrification [2,3],
shown in the figure, the aeration tank is aerated with diffused
aerobic denitrification [4,5], and short-cut nitrification-
air by pore aeration, and gas flow meter is used to control the
denitrification [6,7]. Based on the micro-environment theory,
airflow to maintain the dissolved oxygen (DO) concentration
SND occurs in a process when oxygen diffusion is limited
within 0.3–0.8 mg/L. The maximum working volume of the
within the micro-flocs, thus the anoxic micro-zone inside the
aeration tank is 8.5 L. The top of the reactor is open to air.
flocs for heterotrophic denitrification is created. Rittmann
Reactors were kept in a room with constant temperature of
et al. [8] and Pochana et al. [9,10] discovered the phenome-
23–28°C. Bicarbonate buffer was added to supplement
non of SND in an oxidation ditch and in a sequencing batch the exhausted alkalinity and maintain pH of 7.5–8.0 in the
reactor (SBR), respectively. They suggested that SND would
reactors.
occur according to the micro-environment theory. It was
2.2 Wastewater characteristics
Translated from Environmental Pollution and Control, 2004, 26(1):
11–13 [译自: 环境污染与防治]
The synthetic wastewater used in the experiment consisted of
E-mail: [email protected] glucose, ammonium chloride, and monopotassium phosphate
50
were selected as the variable factors, and three levels (states)
had been picked for each factor. The orthogonal design table
L4(34) is shown in Table 1.

Table 1 Design of orthogonal method

Item Factors

SRT/d(1) C/N(2) F/M(3)

1 60 10 0.30
2 60 5 0.10
3 60 2 0.05
4 45 10 0.10
5 45 5 0.05
6 45 2 0.30
7 30 10 0.05
8 30 5 0.30
9 30 2 0.10
Fig. 1 Schematic diagram of the experimental equipment

as the carbon, nitrogen, and phosphorous in the wastewater,

respectively. Bicarbonate, calcium chloride, ferrous sulphate,
3 Results and discussion
magnesium sulphate, and manganese sulphate as trace
The results of orthogonal method are shown in Table 2.
elements were added.
3.1 The nitrification process
2.3 Sludge acclimation
Based on the traditional theory, successful nitrification
The activated sludge was obtained from the secondary sedi-
requires autotrophic nitrifying bacteria under strict aerobic
mentation tank of the Quyang Waste Water Treatment Plant
in Shanghai, China. After aerating for one day without extra conditions. Dissolved oxygen (DO) concentration is usually
nutrients, a certain amount of activated sludge was added into above 0.5 mg/L in the mixing activated sludge system [12].
the aeration tank to maintain the initial mixed liquor suspend- However, as shown in Table 2, high ammonia nitrogen
ed solids (MLSS) concentration at about 3.5 g/L. After oper- removal efficiency went up to 93% under experimental con-
ating for 12 d, the sludge’s settling characteristics was good ditions of 2, 3, 4, 5, and 7 where DO concentration was main-
when the sludge volume index (SVI) in the aeration tank was tained at 0.3–0.8 mg/L. Nitrification also occurred to a certain
88. At the time, COD removal rate was 89%, and ammonia extent in other experimental conditions.
nitrogen removal rate was 82%, indicating the ending of the Many studies have observed the phenomenon of heterotro-
sludge acclimation period. phic nitrification. In this study, when F/M ratio was 0.30 g
COD/(g MLSS · d), the removal rate of ammonia nitrogen
2.4 Operational conditions was 55.4%, 7.3%, and 48.1% under experimental conditions
of 1, 6, and 8, respectively, which were lower in all experi-
Orthogonal design method was used in this study. Organic mental conditions. When F/M ratio was as low as 0.05 g
load (F/M ratio), C/N ratio, and solids retention time (SRT) COD/(g MLSS · d), ammonia nitrogen removal rate was

Table 2 Results of orthogonal method

Item HRT/h COD NH4+-N TN NO2−-N NO3−-N

Influent Effluent Removal Influent Effluent Removal Influent Effluent Removal Effluent Effluent
/(mg · L−1) /(mg · L−1) rate/% /(mg · L−1) /(mg · L−1) rate/% /(mg · L−1) /(mg · L−1) rate/% /(mg · L−1) /(mg · L−1)

1 8 301.3 40.2 86.7 27.0 12.4 55.4 31.5 14.1 55.1 1.15 0.55
2 12 311.7 30(22) 89.5(93) 62.4 1.0 98.5 66.8 32.1 43.0 7.0 24.1
3 16 120.1 20.7 82.8 62.4 0.0 100.0 62.3 60.2 8.3 0.1 60.1
4 24 317.4 28.9 90.3 26.9 0.0 100.0 29.8 10.1 66.7 0.7 9.4
5 42 319.5 15.5 96.4 60.5 4.1 93.2 63.0 24.8 60.0 0.3 20.4
6 3.2 118.2 31.6 73.3 68.9 57.0 17.3 69.4 58.7 14.7 1.5 0.2
7 12 334.6 37.3 88.9 25.7 0.0 100.0 31.5 14.0 54.4 1.2 12.8
8 8 320.7 62.9 80.4 56.6 29.4 48.1 60.1 32.5 45.4 0.1 3.0
9 9.6 126.7 46.7(33.5) 63.1(73.6) 60.9 21.4 64.9 62.7 38.7 37.8 11.6 5.7

Note: The data showed within parenthesis were COD and COD removal rate after deducted by NO2−-N. The deduction was calculated according to 1 mg/L
NO2−-N consuming 1.14 mg oxygen
 51
above 93%, and the effluent ammonia nitrogen was lower Another experiment was conducted to further confirm that
than 5 mg/L under experimental conditions of 2, 5, and 7 the nitrification that occurred in this study was an aerobic
indicating that the heterotrophic nitrification did not occur in reaction. Experimental conditions were the same as the
this experiment. According to the traditional theory, nitrifiers experimental condition 2 except that DO concentrations were
are autotrophic aerobes, and their growth may be inhibited 0.0–0.1 mg/L. The results (see Table 3) show that the ammo-
in conditions with high F/M ratio. Nitrogen-containing nia nitrogen removal rate was reduced from 98.5% to 74.7%
substrates will be utilized by the dominant heterotrophic with decreasing DO concentration. To maintain normal activ-
microbes for synthetic metabolism while a large number of ity of nitrifying bacteria, DO concentration should be no less
DO is consumed after the synthetic metabolisms by the dom- than 0.3 mg/L.
inant heterotrophic microbes using the nitrogen, suppressing
the nitrification process. However, autotrophic nitrification 3.2 The denitrification process
was observed under low oxygen concentrations in this study.
Three assumptions are provided to explain this. The total nitrogen (TN) removal rate, as shown in Table 2,
Firstly, the smaller biomass floc was observed through was mainly between 40%–50% and the maximum was 66.7%,
microscopic analysis, compared to those in general complete confirming the occurrence of simultaneous nitrification and
mixing systems, which facilitate the transfer of DO at low denitrification. Under the experimental conditions of 3, 6, and
concentration through the small size flocs easily, and enlarge- 8, the TN removal rate was lower because of the uncompleted
ment of the aerobic micro-zone inside the flocs to supply nitrification process. Under other experimental conditions,
enough oxygen for the nitrifiers. the concentration of nitrate in the effluent was higher, ranging
Secondly, SRT in this study was long (30 d, 45 d, and 60 d, from 10 to 60 mg/L. The DO concentration should not be
respectively) enough to reduce the degradation rate of carbon a limiting factor for denitrification. As shown in Table 3, the
containing organics by heterotrophic aerobe. Therefore, a denitrification performance was not improved with the
certain amount of DO inside the flocs could be maintained, decrease of DO concentration. The available carbon may be
and consumed by the nitrification process. Additionally, the potential reason. Insufficient carbon can inhibit the het-
stronger competition of nitrifiers under the condition of erotrophic denitrifiers, which need organic carbon as electron
longer SRT contributes to higher nitrogen removal rate. donor under low oxygen concentration. In this study, organic
Thirdly, under the five experimental conditions, the load was 0.05–0.1 g COD/(g MLSS · d) under some condi-
organic loadings were very low, only 0.05–0.10 g CODcr/ tions, and most of the organic carbon was utilized by other
(g MLSS · d). Figure 2 schematically presents the ratio of heterotrophic aerobes, therefore limiting the denitrification
aerobic zone/anoxic zone in activated sludge flocs under the process.
influence of F/M ratio and DO concentration [13]. As shown An experiment was performed to confirm the influence of
in Fig. 2, the flocs could maintain aerobic condition under organic carbon on the denitrification process. Taking the
low F/M ratio and low DO concentration. Although the DO experimental condition 4 as an example, the effluent from the
concentration was 0.3–0.4 mg/L, below 1.0 mg/L, the small secondary settling tank with the addition of glucose was
flocs are attributable to oxygen penetration. rotated for 12 h in a rotary bed, and after that, concentrations
of nitrite and nitrate were analyzed. Results (see Table 4)
showed that nitriate was removed completely with the addi-
tion of carbon source, and the denitrification process was per-
formed well, indicating that the process could be influenced
by available carbon.

3.3 Nitrate and nitrite in the effluent

Fig. 2 Effects of F/M and DO on aerobic/anoxic zone ratio in Experimental results (see Table 2) showed that the effluent
schematic activated sludge floc nitrite concentration was higher in the experimental
Table 3 Effect of DO on the nitrification process
DO/(mg·L−1) COD Ammonia nitrogen Total nitrogen Influent Influent
NO2−-N NO3−-N
Influent Effluent Removal Influent Effluent Removal Influent Effluent Removal /(mg · L−1) /(mg · L−1)
/(mg · L−1) /(mg · L−1) rate % /(mg · L−1) /(mg · L−1) rate % /(mg · L−1) /(mg · L−1) rate %

0.5 311.7 30 89.5 62.4 1.0 98.5 66.8 32.1 43 7.0 24.1
(22) (93)
0. 298.1 43.2 85.5 66.5 16.8 74.7 70.3 41.7 40.8 4.04 20.81
(38.6) (87.1)

Note: The data showed within parenthesis were COD and COD removal rate after deducted by NO2−-N. The deduction was calculated according to 1 mg/L
NO2−-N consuming 1.14 mg oxygen
52
Table 4 Effect of available carbon on nitrogen removal rate
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