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Article history:
Received 19 November 2007
Received in revised form 12 July 2008
Accepted 14 July 2008
Keywords:
Airlift reactor
Granular sludge
Nitrication
Anaerobic ammonium oxidation
a b s t r a c t
Since nitrication is the rate-limiting step in the biological nitrogen removal from wastewater, many studies have been conducted on the immobilization of nitrifying bacteria. A laboratory-scale investigation was
conducted to examine the effectiveness of a continuous-ow airlift reactor (ALR) on the granulation of
nitrifying sludge and the nitrication efciency of the reactor after granulation. The results showed that
the granular sludge began to appear on day 30 and matured in 75 days. The mature granules had an average
diameter of 1.54 mm, settling velocity higher than 82.4 m h1 and specic gravity of 1.07. The granules cultured in the present study had aerobic ammonia oxidation activity of 13.3 mg NH4 + -N (g VSS)1 day1 and
anaerobic ammonium oxidation (ANAMMOX) activity of 3.22 mg NH4 + -N (g VSS)1 day1 , which demonstrated that the nitrifying granules possessed the potential to be used as seed sludge for ANAMMOX and
CANON (completely autotrophic nitrogen-removal over nitrite) reactors. After granulation, the ALR exhibited an excellent nitrication performance. It had strong tolerance to inuent NH4 + -N of 1100 mg L1 .
When operated at inuent NH4 + -N concentration of 546 mg L1 , the reactor could remove over 94.4%
of ammonium even at a nitrogen loading rate (NLR) of 2.37 f m3 day1 with a short hydraulic retention time (HRT) of 5.4 h. With the inuent NH4 + -N concentration of 547 mg L1 , HRT 6.9 h and NLR of
1.90 kg m3 day1 , superior efuent quality could be achieved robustly, with an efuent NH4 + -N of less
than 5 mg L1 , satisfying the national primary discharging standard of China (GB 8978-1996).
2008 Elsevier B.V. All rights reserved.
1. Introduction
Nitrogenous compounds like ammonium are prevalent in many
wastewaters and need to be removed to prevent oxygen depletion and eutrophication of surface waters. Biological nitrogen
removal from wastewater using nitricationdenitrication is a
well-known and cost-effective treatment process [1,2]. Because of
extremely low growth rate, it is generally accepted that retaining a
large number of nitrifying bacteria within the reactor is difcult to
achieve, thereby making the nitrication a rate-limiting step in the
entire nitrogen removal process [2,3].
Many researches have been carried out to develop the physical
or ecological methods of immobilizing nitrifying bacteria including
cell-entrapping and cell-attaching techniques [46]. However, the
immobilized cells created by gel entrapping are easy to be suffered
from mass transfer resistance [6]. Previous researches also demonstrated that construction of a nitrifying biolm on the surface of
carrier materials takes a long time, particularly when the wastewater contains few organic compounds [7]. Moreover, the matrices
and carriers used for cell immobilization inevitably occupy sig-
Corresponding author. Tel.: +86 571 86971709; fax: +86 571 86971709.
E-mail address: pzheng@zju.edu.cn (P. Zheng).
1383-5866/$ see front matter 2008 Elsevier B.V. All rights reserved.
doi:10.1016/j.seppur.2008.07.012
671
2.1. ALR
Fig. 1. Schematic illustration of the ALR used in the experiment: (1) inuent tank, (2)
peristaltic pump, (3) efuent tank, (4) riser, (5) downcomer, (6) settler, (7) overow
weir, (8) air pump.
2.2. Inoculum
Aerobic sludge from a local municipal wastewater treatment
plant was used as inoculum for the present study and the initial
volatile suspended solid (VSS) concentration was 10 g L1 .
2.3. Synthetic wastewater
The composition of synthetic wastewater is listed in Table 1. For
alkalinity and carbon source supplement, the theoretical NaHCO3
requirement for nitrication (7.1 g as CaCO3 (g NH4 + -N)1 ) was
added to the wastewater.
2.4. Analytical procedures
Ammonium, nitrite, nitrate, VSS and pH were determined using
the standard methods [16]. DO was measured by JPB-607 dissolved
oxygen meter. The average diameter of granules was measured
using an optical microscope (YS2-H, Nikon, China) as a circleTable 1
Composition of synthetic ammonium-containing wastewater
Compound
Concentration
KH2 PO4
MgSO4 7H2 O
CaCl2
NaHCO3
(NH4 )2 SO4
Trace elements Ia
Trace elements IIb
0.027 g L1
0.300 g L1
0.136 g L1
0.8413.3 g L1
0.335.23 g L1
1.25 mL L1
1.25 mL L1
672
Fig. 2. Optical microscope photograph of nitrifying sludge: (A) seed sludge (10 10), (B) oc-like sludge (10 10), (C) granular sludge (10 10), (D) granular sludge (10 20).
Diameter (mm)
<0.5
0.51.0
1.00.5
>1.5
30
45
60
75
22
2
0
3
49
24
27
16
23
45
35
34
6
29
38
47
Table 3
Physical characteristics of nitrifying granular sludge
Time(day)
30
45
60
75
0.82
55.5
1.01
1.30
75.6
1.08
1.40
74.9
1.05
1.54
82.4
1.07
effectively retained when used in a continuous stirring tank reactor under perfectly mixed-ow conditions at a short HRT of 5.4 h.
The excellent setting capability of granules in the present study
was assumed to be due to the seed sludge characteristics or/and
the operational conditions used which were still unclear and are
subjected to further clarication.
The granules possessed excellent ammonia oxidation activity.
The batch test results shown in Fig. 4 indicated that, under given
experimental conditions, the time taken for complete oxidation of
168 mg N L1 ammonium by nitrifying granules was 32 h, in contrast to 200 h for seed sludge. The aerobic ammonia oxidation
activity of nitrifying granules (13.3 mg NH4 + -N (g VSS)1 day1 )
was 5.8 times higher than that of seed sludge (1.96 mg NH4 + N (g VSS)1 day1 ).
The ANAMMOX activity was detected in nitrifying granules on
day 120, which indicated the presence of ANAMMOX bacteria. The
granules conducted ANAMMOX reaction at a rate of 3.22 mg NH4 + N (g VSS)1 day1 and the observed ratio of ammonia oxidation rate
to nitrite reduction rate was 0.96, near the stoichiometry of ANAMMOX. The ANAMMOX process is a biological reaction in which
ammonia is oxidized to nitrogen gas using nitrite as the electron
acceptor under anoxic conditions [28].
The ANAMMOX reaction appears to occur only under strict
anaerobic conditions. As little as 1.1 mM O2 was sufcient to
inhibit ANAMMOX activity completely in a bioreactor [29]. The
inhibition was reversible, and in experiments with intermittent aeration, the rate of ANAMMOX before and after aeration was identical.
The presence of ANAMMOX bacteria in granules detected in the
present study may be due to the layered structure of granules [10],
where the nitrifying bacteria in surface layer almost utilize all of the
oxygen and the resistance for oxygen mass transfer is high enough,
keeping a strict anaerobic environment for ANAMMOX organisms.
The ANAMMOX process should be combined with partial
nitrication stage, in which around 50% of ammonia could be
converted to nitrite. The process offers advantages for the treatment of efuents decient in organic matter, compared with the
673
due to the higher growth rate of aerobic sludge, the seed nitrifying
granular sludge can be harvested in large quantities subsequently.
The aerobic and anaerobic ammonium oxidizing bacteria could
coexist in granular sludge under investigation, even at a high dissolved oxygen level above 5.0 mg L1 . When the reactor is operated
under oxygen-limiting conditions, the ANAMMOX organisms are
able to grow and partial nitrication and ANAMMOX are engineered
in a single reactor. The process has been named CANON, an acronym
for completely autotrophic nitrogen-removal over nitrite [31]. In
this setup, aerobic ammonia-oxidizing bacteria consumed all the
oxygen, thus maintaining a very low oxygen concentration in the
granules and produced nitrite as the electron acceptor for ANAMMOX. In CANON process, the entire nitrogen removal can be carried
out in a single reactor with very low aeration, greatly reducing space
and energy requirements. Its feasibility, effectiveness and stability have been established by some studies, achieving a nitrogen
removal rate of 1.5 kg m3 day1 in a gaslift reactor [31], which is
more than sufcient to start application trials. The results in present
study suggest that CANON reactor could be started up directly by
seeding the nitrifying granules.
3.2. Performance of ALR
3.2.1. Performance during start-up
As shown in Fig. 5, the reactor was started at 24 h HRT and inuent NH4 + -N concentration of 70 mg L1 , corresponding to a nitrogen
nitricationdenitrication process. This process allows the reduction in the oxygen requirements and carbon dioxide emission to the
atmosphere. However, the practical application of the ANAMMOX
process is still limited by its long start-up period due to the very
low growth rate (0.072 day1 measured at 32 C) of bacterial communities involved and biomass yield generated per gram ammonia
nitrogen consumed (0.088 g g1 ) by these microorganisms [30]. For
these reasons, the inoculum with high ANAMMOX activity is desirable, especially in situations where the industrial scale ANAMMOX
systems are operated. The present study suggested that the nitrifying granules were potential seed sludge of ANAMMOX reactor, and
674
Table 4
Performance of airlift reactor with different working HRTs
HRT (h)
22.8
19.5
15.9
13.1
11.8
9.61
9.01
7.38
7.02
6.34
6.25
6.02
5.41
NH4 + -N (mg L1 )
Inuent
Efuent
528
566
570
535
560
512
534
520
531
519
553
571
594
2.52
4.06
0.56
0
6.86
0.56
12.3
28.6
29.0
0.42
14.4
31.5
21.1
99.5
99.2
99.9
100
98.8
99.9
97.7
94.4
94.6
99.9
97.4
94.5
96.5
0.556
0.697
0.864
0.979
1.14
1.28
1.42
1.69
1.82
2.00
2.10
2.27
2.37
0.553
0.691
0.863
0.979
1.13
1.28
1.39
1.60
1.72
1.99
2.04
2.15
2.29
0
0
44.7
1.68
51.2
0
0
276
251
73.6
140
232
111
377
440
427
486
472
491
337
142
194
383
295
179
353
Table 5
Performance of airlift reactor at inuent NH4 + -N of 547 mg L1 and HRT of 6.9 h
Time (day)
1st
2nd
3rd
4th
5th
6th
7th
8th
9th
10th
NH4 + -N (mg L1 )
Inuent
Efuent
524
567
546
542
560
567
528
558
508
570
2.38
4.76
4.48
4.76
3.92
2.80
0.14
0
0
0
Removal
efciency (%)
Nitrogen
loss (%)
99.6
99.2
99.2
99.1
99.3
99.5
100
100
100
100
1.94
1.89
2.03
1.96
1.85
1.90
1.79
1.91
1.83
1.93
1.89
1.83
1.97
1.90
1.80
1.85
1.75
1.87
1.79
1.89
450
404
368
389
343
187
155
86.4
69.9
98.1
68.7
54.2
115
100
179
328
319
365
434
452
0.6
18.4
10.9
8.9
6.1
8.8
10.4
19.2
0.9
3.5
675