Simultaneous Nitrification, Denitrification, and Phosphorus Removal in Single-Tank Low-

Dissolved-Oxygen Systems Under Cyclic Aeration

Author(s): Lu-Kwang Ju, Lin Huang and Hiren Trivedi
Source: Water Environment Research, Vol. 79, No. 8 (August 2007), pp. 912-920
Published by: Water Environment Federation
Stable URL: http://www.jstor.org/stable/23804181
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 Simultaneous Nitrification, Denitrification,
and Phosphorus Removal in Single-Tank
Low-Dissolved-Oxygen Systems
Under Cyclic Aeration
Lu-Kwang Jur, Lin Huang1, Hiren Trivedi2
ABSTRACT: Simultaneous nitrification and denitrification (SND or
SNdN) may occur at low dissolved oxygen concentrations. In this study,
bench-scale (approximately 6 L) bioreactors treating a continuous feed of
synthetic wastewater were used to evaluate the effects of solids retention time
and low dissolved oxygen concentration, under cyclic aeration, on the
removal of organics, nitrogen, and phosphorus. The cyclic aeration was car
ried out with repeated cycles of 1 hour at a higher dissolved oxygen concen
tration (HDO) and 30 minutes at a lower (or zero) dissolved oxygen
concentration (LDO). Compared with aeration at constant dissolved oxygen
concentrations, the cyclic aeration, when operated with proper combinations
of HDO and LDO, produced better-settling sludge and more complete
nitrogen and phosphorus removal. For nitrogen removal, the advantage
resulted from the more readily available nitrate and nitrite (generated by
nitrification during the HDO period) for denitrification (during the LDO
period). For phosphorus removal, the advantage of cyclic aeration came from
the development of a higher population of polyphosphate-accumulating
organisms, as indicated by the higher phosphorus contents in the sludge
solids of the cyclically aerated systems. Nitrite shunt was also observed to
occur in the LDO systems. Higher ratios of nitrite to nitrate were found in the
systems of lower HDO (and, to less dependency, higher LDO), suggesting
that the nitrite shunt took place mainly because of the disrupted nitrification at
lower HDO. The study results indicated that the HDO used should be kept
reasonably high (approximately 0.8 mg/L) or the HDO period prolonged, to
promote adequate nitrification, and the LDO kept low (<0.2 mg/L), to
achieve more complete denitrification and higher phosphorus removal. The
above findings in the laboratory systems find strong support from the results
obtained in full-scale plant implementation. Two plant case studies using the
cyclic low-dissolved-oxygen aeration for creating and maintaining SND are
also presented. Water Environ. Res., 79, 912 (2007).
KEYWORDS: simultaneous nitrification and denitrification, low dis
solved oxygen concentration, solids retention time, cyclic aeration, nitrite
shunt, enhanced biological phosphorus removal,
doi: 10.2175/106143007X175942
Introduction
Regulations on the nitrogen and phosphorus contents in
wastewater discharge are increasingly more stringent for controlling
the rate of eutrophication in the aquatic environment. Biological
reduction of the nitrogen contents in wastewater relies primarily on
1 Department of Chemical and Biomolecular Engineering, The University of
Akron, Akron, Ohio.
2 Enviroquip, Inc., Austin, Texas.
* Department of Chemical and Biomolecular Engineering, The University of
Akron, Akron, OH 44325-3906; e-mail: Ju@uakron.edu.
912
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two mechanisms—aerobic nitrification and anoxic denitrification
(Gee and Kim, 2004; Grady et al., 1999; Schmidt et al., 2003).
Generally, the two processes are carried out in physically separated
oxic and anoxic treatment zones (Barnard, 1975; Ju et al., 1995;
Ludzack and Ettinger, 1962). They can also be achieved in the same
reactor, but temporally separated with alternating oxic and anoxic
periods, by turning the aeration on and off in repeated cycles
(Alientan and Irvine, 1980; Randall et al., 1992; Sedlak, 1991;
Silverstein and Schroeder, 1983).
Recent studies have revealed that nitrification and denitrification
can occur simultaneously in the same reactor (Trivedi and Heinen,
2000), arising from the following mechanisms (Kaempfer et al.,
2000; Satoh et al., 2003; Stensel, 2001):
(1) Presence of microscopic anoxic/oxic zones within sludge floes.
Aeration promotes the dissolution of oxygen into the water. The
dissolved oxygen subsequently diffuses into the floes and,
along the diffusion path, is consumed by the organisms. The
diffusion and consumption cause a gradient of dissolved
oxygen concentration (DO) in the floes and, at suitably low
dissolved oxygen concentrations, create floes that nitrify in the
oxic outer layer and denitrify in the anoxic inner core.
(2) Presence of macroscopic anoxic/oxic zones within bioreactors.
Such zones are commonly formed as a result of nonhomoge
neous mixing and aeration, particularly in bioreactors with
surface aerators, where the dissolved oxygen concentration is
high near the aerators and low or zero away from the aerators.
(3) Presence of novel microorganisms. For example, Robertson
et al. (1988) reported that Thiosphaera pantotropha could
simultaneously nitrify and denitrify under aerobic conditions. In
addition, several bacteria were found to perform aerobic
denitrification (Davies et al., 1989). More recently, Chen et al.
(2003) confirmed, with continuous culture of Pseudomonas
aeruginosa, that aerobic denitrification functioned as an
electron-accepting mechanism supplementary to aerobic respi
ration. Some nitrifiers, such as Nitrosomonas europea and
N. eutropha, could also denitrify at low dissolved oxygen
concentrations (Zart et al., 1996).
Single-tank simultaneous nitrification and denitrification (SND)
processes have potential advantages in eliminating the need for
separate tanks and for recycling the mixed liquor from oxic
nitrifying zones to the typically upstream anoxic zones for
denitrification. In addition to the simpler process design, SND has
Water Environment Research, Volume 79, Number 8
 Ju et al.

Synthetic
Synthetic wastewater
wastewater _ shortcut biochemical pathway with partial nitrification of ammonia
(NH3) to nitrite (N02~), followed by the partial denitrification of
Pu N02~ to N2, bypassing the formation and consumption of nitrate
, m
1N NaOHmV Tdrfi I, ii ™
^ i(Yoo et al., 1998).
pumpsplim„, Slidins
Air
pumps
r-J■
INNaoT.^ Pumps ; Sliding
1NHC1 , tOO—711
I /baffle
: y Methodology
Experimental Setup and Procedure. Two modified Ecken
c±r] rrin felder's reactors (approximately 6 L) were used simultaneously in

I
Timers
pH controller j * § | this study. The reactor setup is shown schematically in Figure 1. A
pH controller

sliding baffle was used to divide the bioreactor into a mixing zone
pHprobe
pH probe » | and a settling/clarifying zone, with a volume ratio of 5:1. A pump
was used to slowly return the settled sludge from the settling zone to
□ the mixing zone, to avoid the sludge loss resulting from flotation
DO probe
caused by the denitrification-generated bubbles in the accumulated
Air diffuser ^ blanket of sludge. Depending on the studied SRT (5, 10, or 20
Air diffuser

days), a calculated volume of mixed liquor was removed once to

Figure 1—Schematic of experimental setup.
thrice daily, after pulling up the dividing baffle to allow thorough
mixing of the reactor contents.
been estimated to require smaller total tank sizes (Kaempfer et al., Mixing in the mixing zone was achieved by mechanical agita
2000; Stensel, 2001). The SND also helps to maintain a relatively tion and diffused-air aeration. A marine propeller operated at a con
neutral pH in the bioreactor, without the addition of an external acid stant speed (approximately 300 r/min) provided the mechanical
or base. The alkalinity consumed by nitrification is partially agitation. The aeration, through a diffuser placed at the bottom of
recovered by denitrification (Grady et al., 1999). the mixing zone, was supplied by two air pumps, which were
While offering the above potential benefits, SND faces several separately controlled by two timers to allow the study of various
challenges in design, control, and operation. The single-stage, aeration schemes. In this study, the aeration was applied in three
continuously stirred tank reactor configuration and the low different modes. One experimental run (run 1, Tables 1 to 3) was
dissolved oxygen environments required for SND processes are carried out under constant aeration, to maintain constant dissolved
conventionally considered more susceptible to sludge bulking, oxygen levels. Some experiments (runs 2 to 4) were aerated in
primarily because of the excessive growth of filamentous bacteria repeated on/off cycles, with an air pump being turned on for 1 hour
(Grady et al., 1999; Jenkins et al., 2003; Martins et al., 2004). A and then off for 30 minutes. Aeration to the third group of exper
study conducted in this laboratory concluded that the cyclic iments (runs 5 to 7) was also done in repeated l-hour/30-minute
aeration, with repeated cycles of 1 hour at a higher dissolved cycles. In this group, however, both air pumps were turned on for 1
oxygen concentration (HDO approximately 0.8 mg/L), followed by hour to obtain a certain dissolved oxygen concentration. One of the
30 minutes at a lower dissolved oxygen concentration (LDO < pumps was then turned off for 30 minutes, to maintain a lower
approximately 0.2 mg/L), was appreciably better than constant dissolved oxygen concentration in the mixing zone. Different dis
aeration in avoiding bulking (Huang and Ju, 2007). Shortening the solved oxygen levels were studied for each aeration mode, as sum
system solids retention time (SRT) was also found to improve marized in Table 1.
sludge settling significantly. The bioreactors were operated at room temperature (22 ± 1°C).
In addition to the potential bulking problem, the performance of By controlling the wastewater feeding rate, the hydraulic retention
SND relies on achieving a dynamic balance between nitrification time was maintained at 24 hours for all of the experiments. The
and denitrification. The objective of this study was to evaluate the influent feed was continuous during the experiment. A synthetic
treatment performance (i.e., the removal of organic matter and wastewater was used in this study, with the following composition
nutrients [nitrogen and phosphorus]) of the cyclically aerated low- modified from that used by Kiso et al. (2000): 500 mg/L skim milk,
dissolved-oxygen systems maintained at different SRT and HDO 80 mg/L NH4C1, 20 mg/L K2HP04, 580 mg/L NaHC03, 100 mg/L
and LDO. The potential occurrence of "nitrite shunt" and the MgS04 • 6H20, and 100 mg/L CaCl2 • 2H20. The wastewater had
development of sludge with elevated biophosphorus contents in approximately 480 mg/L chemical oxygen demand (COD), 50 mg/L
such systems were also examined. Nitrite shunt referred to the total Kjeldahl nitrogen (TKN), and 8 mg/L total phosphorus. Fresh

Table 1—Experimental results obtained under different operating conditions, SVI, and solids concentrations.*

Run HDO (mg/L) LDO (mg/L) SRT (days) DO (mg/L) SVI (ml/g) Total solids (mg/L) Volatile solids (mg/L)

1 0.8 0.8 5 0.8 60 1394.4 ± 55 721 ± 53

2 2 0 20 1.33 140 1660 ± 130 862 ± 160
3 0.8 0 5 0.53 35 1580 ± 100 720 ± 93
4 0.8 0 10 0.53 75 1300 ± 73 644 ± 89
5 0.4 0.2 10 0.33 140 1490 ± 340 542 ± 36
6 0.6 0.2 10 0.47 65 1510 ± 410 620 ± 160
7 0.8 0.2 10 0.6 45 1637 ± 38 537 ± 31

Note: HDO = Higher dissolved oxygen used during the 1-hour period of cyclic aeration, LDO = Lower dissolved oxygen used during the
0.5-hour period of cyclic aeration.

August 2007

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Ju et al.

Table 2—Measured data on mixed-liquor and effluent properties.*

Run COD
CODNH3-N
NH3-NN02"-N
N02--NN03"-N
N03"-NTPmu
TPmuTPe
TPeTKNml
TKNml TKNe

(mg/L)
1 17.9
17.9± 6.1 ± 6.1 7.4
7.4± 1.0±1.0 3.6
3.6 ± ±
0.4 0.4 2.4
2.4 ±±
0.90.9 18.5
18.5 ±±1.21.2 8 8
±± 0.3
0.3 102.1
102.1 9.4
2 32.4
32.4 0.4 ± ±
± 1.4±1.4 0.4 0.2 0.2 0.2
0.2 ± ±0.1 0.1 3.8
3.8 ±±0.9
0.9 38.8 ±3.7
38.8 ± 3.7 7.3
7.3 ±±0.40.4 81.4
81.4 3.8
3 44.8
44.8± 8.5± 8.5 9.1
9.1 ± ±
2.5 2.5 0.1
0.1 ±±0.03
0.03 0.1
0.1 ±± 0.01 23.8
0.01 23.8 ± ±
2.5 2.5 2.9
2.9 ± ±
0.3 0.3 87.9
87.9 17.8
4 22.0
22.0± 7.8± 7.8 0.5
0.5 ± 0.04 0.8
± 0.04 0.8 ± 0.2 1.4
± 0.2 ± 0.2 26.8
1.4 ± 0.2 26.8 ± ±
2.5 2.5 3.4
3.4 ± ±
0.20.2 57.6
57.6 4.6
5 22.8
22.8 ± 3.3± 3.3 10.9
10.9± 2.1 ± 2.1 6.4
6.4 ± 0.4 1.6
± 0.4 ± 0.8 25.8
1.6 ± 0.8 25.8 ± ±
1.3 1.3 7.0
7.0 ± ±
0.50.5 58.7
58.7 11.1
6 25.5
25.5± 3.5± 3.5 0.7
0.7 ± 0.2
± 0.2 7.5 1.0 1.0 5.4
7.5 ± ± Oi 26.4 ±
l+ ±o 000.8 26.4 ±0.20.2 6.7
6.7 ±±0.90.9 43.1
43.1 3.6
7 24.7
24.7 ± 3.3± 3.3 0.9
0.9 ± 0.04 4.0
± 0.04 4.0 ± 0.2 5.4
± 0.2 ± 0.2 28.7
5.4 ± 0.2 28.7 ± ±
5.1 5.1 4.4
4.4 ±±
0.50.5 57.1
57.1 3.5

' Subscripts "ML" and "E" refer to "mixed liquor" and "effluent", respectively. The laborious TKN measurements were conducted with only
one set of mixed liquor and effluent samples in many of the experimental runs. Therefore, standard deviations were not reported.

sludge samples were taken from a secondary clarifier of the nearby nitrate-nitrogen [N03~-N], nitrite-nitrogen [N02--N], and ortho
Water Pollution Control Division Station at Akron, Ohio. The phosphate [P¡]). All tests were conducted in accordance with
sludge was diluted with tap water to make an initial concentration Standard Methods (APHA et al., 1995), as follows: TSS by method
of total solids (TS) of 1000 to 1500 mg/L. After seeding, the 2540 B, VSS by method 2540 E, COD by the closed reflux
bioreactor was maintained for a period of 3 SRTs. The mixed-liquor titrimetric method (5220 D), NH3-N by using an ammonia selective
samples were then taken every 1 to 2 days to monitor the electrode (method 4500-NH3 D), combined N03~-N and N02"-N
concentrations of TS and volatile solids (VS). After TS and VS by the titanous chloride reduction method (4500-N03~ G), N02~-N
stabilized (with random fluctuations within ± 1 standard deviation), by the colorimetric method (4500-N02~ B), P, by method 4500-P
more samples (3 to 6 per day) were taken for the additional analyses E, total phosphorus first digested by the Persulfate Digestion
of ammonium (NH4+) and nitrate (N03~) concentrations. The Method then followed by method 4500-P E, and TKN by method
system was considered to have reached (pseudo)steady state when 4500-Norg C.
the NH4+ and N03 concentrations also only fluctuated randomly
within ±1 standard deviation. Afterwards, the bioreactor was
maintained for an additional period of 1 to 2 SRTs, during which, Results
more frequent samples were taken and analyzed to evaluate the Overall Treatment Performance. Sludge bulking occurred in
process characteristics and performance. some experimental runs, particularly those under constant aeration
The pH in the mixing zone tended to increase because of the and/or with a longer SRT (20 days). Only the results from the
protein-rich synthetic wastewater used in this study. The pH was nonbulking runs (sludge volume index [SVI] <150 mL/g) are
therefore maintained at 7.2 ± 0.3 using an assembly of pH probe, summarized in Tables 1 to 3—Table 1 for the operating conditions,
meter/controller, and pump for automatic addition of 1 N hydro- SVI, and solids concentrations; Table 2 for the experimentally
chloric acid (or, occasionally, 1 N sodium hydroxide). Dissolved measured data on mixed-liquor and effluent properties; and Table 3
oxygen concentration was measured continuously using a YSI5739 for the calculated/derived results. As shown in Table 3, the COD
dissolved oxygen probe with a meter (model 58, YSI Inc., Yellow removal (91 to 96%) was good and did not differ much under
Springs, Ohio). various operating conditions. Nitrogen removal, however, ranged
Analytical Methods. The sample analyses included those for from 62 to 86%. Phosphorus removal varied even more, from
the sludge properties (TS, VS, TKN, and total phosphorus contents) approximately 3 to 65%. Among all the operating conditions
and those for water quality (COD, ammonia-nitrogen, [NH3-N], examined, run 4 had the highest nitrogen removal and the second
highest phosphoms removal. It produced good overall treatment
results for the synthetic wastewater used in this study.

Table 3—Calculated
Table 3—Calculatedtreatment
treatmentperformance
performanceand
and sludge
sludge „ Variations of Nitrogen Species and Phosphate Concentrations
properties
properties.* * During Aeration Cycles. To monitor the variation of water prop
erties during the alternating HDO and LDO aeration cycle, a 15-mL
COD Nitrogen
COD Nitrogen Phosphorus
Phosphorus mixed-liquor sample was taken every 10 minutes, totaling 18
Run removal removal
removal removal
removal TPS TKNS
removal samples over 2 full aeration cycles. The supernatants collected by
TPS
—— centrifugation were frozen and later analyzed for concentrations of
nitrogen species (ammonium, nitrite, and nitrate) and inorganic
1 96.3
96.3 69.2
69.2 3.3
3.3 1.5
1.5 12.9
phosphate. The above sampling was repeated on multiple days to
2 93.2
93.2 84.4
84.4 11.7
11.7 3.5
3.5 9.0
90.7
confirm the reproducible profiles obtained. The profiles obtained for
3 90.7 64.0
64.0 64.9
64.9 2.7
2.7 9.7
95.4 nitrogen species are described first.
4 95.4 86.4
86.4 58.9
58.9 3.7
3.7 8.2
5 95.3
95.3 61.8
61.8 15.3
15.3 3.5
3.5 8.8 As an example, the profiles from run 4 are shown in Figure 2a.
6 94.7
94.7 67.0
67.0 19.0
19.0 3.2
3.2 Opposite trends were observed for NH3-N and NOx~-N (i.e.,
8.6
7 94.9
94.9 74.2 74.2 46.8 46.8 4.34.3 10.0 combined N03~-N and N02~-N) concentrations. During the
LDO period, the NH3-N concentration increased and NOx~-N
* TPS: P content in solids, calculated as (TPML - TPE)/VS; and concentration decreased, because the low dissolved oxygen
TKNs: N content in solids, calculated as (TKNML — TKNE)/VS. concentration (0 mg/L, in this case) prevented nitrifiers from

914 Water Environment Research, Volume 79, Number 8

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 Ju et al.

LDO ; LDO •

0.4 0.6 0.8 1

0.6 0.8 1 1.2

Time-Averaged DO(mg/L)
Time-Averaged DO (mg/L)

OI Constant DO O LDO = 0 mg/L • LDO = 0.2 mg/L

Figure 3—Effects of operating conditions (SRT and time

averaged dissolved oxygen concentration) on nitrogen
removal, with
removal, with the
thebubble
bubblesize
sizeindicating
indicating the
the percent
percent
nitrogen removal attained.

Discussion
The implications associated with the observed variations in
nitrogen and phosphorus removal are discussed in more details in
separate sections below.
Balance of Nitrification and Denitrification. Successful
nitrogen removal in low-dissolved-oxygen SND depends on the
90 90 180
balance between nitrification and denitrification occurring in the
Time
Time (min)
(min) process. A three-dimensional bubble plot is shown in Figure 3 for
the effects of dissolved oxygen concentration (time averaged for
-NH3 —N03- -A-N02-
-A-N02- -X-NOx
-X-NOx
runs with cyclic aeration) and SRT on nitrogen removal, where the
Figure2—Variations
Figure 2—Variationsof of
(a) (a) nitrogen
nitrogen species
species and
and (b) (b) size represents the magnitude of nitrogen-removal percent
bubble
phosphate-phosphorus
phosphate-phosphorus concentrations
concentrations in centrifuged
in centrifuged a8e- Within the investigated ranges, the nitrogen removal tended to
supernatantsofofsamples
supernatants samples taken
taken in run
in run 4 during
4 during two two
increase with increasing dissolved oxygen concentration and SRT.
HDO-LDO
HDO-LDOcycles
cyclesofofaeration (samples
aeration were
(samples taken
were at Because
taken at high dissolved oxygen concentrations favor nitrification
aa 10-minute
10-minute interval).
interval). and low dissolved oxygen concentrations favor denitrification, the
observed trend indicated that the overall nitrogen removal in the
low-dissolved-oxygen SND process is more susceptible to the limi
converting the influent NH4+ to NOx", while promoting de- tation in nitrification. This susceptibility to nitrification limitation
nitrification. During the HDO period, the higher dissolved oxygen is a'so consistent with the observed trend of higher nitrogen re
concentration enabled nitrification and inhibited denitrification. The moval at longer SRTs. The chemoautotrophic nitrifiers are typi
influent NH4+ was converted to NOx~, causing NH3-N concentra- cally slow growers that require a longer SRT to develop properly
tion to decrease and the NOx~-N concentration to increase. in the bioreactors (Grady et al., 1999), particularly under the low
Similar profiles were seen for other runs (data not shown), dissolved-oxygen environments of SND processes,
although the range of variation differed depending on the operating However, the time-averaged dissolved oxygen concentration is
conditions, particularly SRT (which affected nitrification, because not a perfect parameter to describe the effects of cyclic aeration. For
nitrifiers grew much slower at low dissolved oxygen concentrations example, comparing runs 4 and 7 of the same SRT (10 days), the
and required longer SRT to establish in the sludge, as described former had a slightly lower time-averaged dissolved oxygen
more in the Discussion section) and the difference between HDO concentration (0.53 mg/L) than the latter (0.6 mg/L), but achieved
and LDO (i.e., ADO = HDO — LDO). an appreciably higher nitrogen removal (86 versus 74%). As shown
The variation of the phosphate-phosphorus concentration over in Table 2, the less ideal nitrogen removal in ran 7 resulted from the
two aeration cycles is shown in Figure 2b. The phosphate incomplete denitrification, giving much higher N02--N and N03~-N
concentration increased during the LDO period, and then decreased concentrations in the effluent (4.0 and 5.4 mg/L, respectively). More
during the HDO period. The phenomenon was similar to that effective denitrification was obtained in the cyclically aerated
observed in the common biophosphorus removal processes, with systems with larger differences between the higher dissolved
sequential anaerobic and aerobic stages, where the phenomenon oxygen concentration (HDO) used during the 1-hour aeration
was attributed to the activity of phosphorus-accumulating organ- period and the lower dissolved oxygen concentration (LDO) in the
isms (PAOs) (Grady et al., 1999). As shown in Table 3, the solids subsequent 30-minute period. This can be explained as follows,
from the systems under cyclic aeration had higher phosphorus The effectiveness of denitrification in SND is affected by how the
contents (2.7 to 4.3%) than the system under constant aeration did nitrification-generated NOx~ (N02 and N03 ~ ) becomes available
(run 1, 1.5%). This finding strongly supported the feasibility of to the microorganisms in the anoxic zone of floes. This availability
enriching PAOs in the low-dissolved-oxygen SND process with may occur in two mechanisms—(1) diffusion of NOx from the
cyclic aeration. outer aerobic zone of floes to the inner anoxic zone and (2) change

August 2007 915

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Ju et al.

25
bubble size corresponds to the magnitude of the N02--N/N03--N
(A) ratio. There was no clear effect of SRT on nitrite shunt, although the
20
observation was inconclusive because of limited data (only 3 SRTs
studied). As for the effects of DO, according to the results from runs
£ 15 5 to 7 with an SRT of 10 days, LDO of 0.2 mg/L, and different
HDO (0.4, 0.6, and 0.8 mg/L), the ratio clearly decreased with
ûi 10
</)
increasing DO (or HDO). The trend, however, did not apply to the
experiments conducted under different aeration modes; for example,

O—© for the two runs with an SRT of 5 days (runs 1 and 3), the ratio was
larger under constant aeration, despite the larger DO used (Figure
1 1 1 1 1 1 4a). Therefore, DO is not an ideal parameter for predicting the
1

0.2
0.2 0.40.4
0 6 0.8
0.6 1 1.2
0.8 1 4
1 1.6
1.2occurrence
1.4 1.6 of nitrite shunt. Instead, the HDO and LDO used should
Time-Averaged
Time-Averaged DO (mg/L)
DO (mg/L) be considered, as suggested by the more consistent trends in Figure
4b. The ratio appeared to increase with decreasing HDO and
increasing LDO, and the dependency on HDO was more pro
(B) nounced. The observations could be explained as follows.
0.8
Nitrite accumulation could result from disrupted nitrification or,
if nitrification was complete, unbalanced denitrification. The
accumulation resulting from disrupted nitrification would occur
when ammonia oxidation (to nitrite) was faster than nitrite oxidation
O (to nitrate). This might result from the higher sensitivity of nitrite
Q

0.2 oxidizing species to low dissolved oxygen concentrations than the

ammonia-oxidizing species (Ju and Nallagatla, 2003; Yu et al.,

0.5
O
o 1 1.5
1.5
2000). On the other hand, the nitrite accumulation resulting from
unbalanced denitrification would appear when nitrite reduction
(eventually to N2) was slower than nitrate reduction (to nitrite). The
HDO
HDO (mg/L)
(mg/L) unbalanced denitrification might be a result of the higher
susceptibility of nitrite reductases to oxygen inhibition/repression
|O
O Constant
Constant DO
DO O
O LDO
LDO =
=00 mg/L
mg/L 9• LDO
LDO =- 0.2
0.2 mg/L
mg/L |
than nitrate reductases (Chayabutra and Ju, 2000; Drury et al.,
4—Effects of
Figure 4—Effects of operating
operatingconditions
conditionson
onthe
theextent
extentofof1991). Accordingly, nitrite accumulation resulting from disrupted
nitrite shunt indicated by the ratio of N02~-N:N03_-N.
N02~-N:N03~-N. nitrification would be more significant at a lower HDO, while that
resulting from unbalanced denitrification would be more significant

of the aerobic zone to anoxic zone as the dissolved oxygen at a h,gher LDa These expectations were reflected in the trends
concentration drops from HDO to LDO under cyclic aeration. The observed in FiSure 4b' and the more Pronounced dependency on
, ^ , rc . , . , t .. HDO suggested that the nitrite shunt took place mainly because of
latter mechanism is more effective, because it does not require the y j
i j-rr • rxTA - r™ - _ , , ..i the disrupted nitrification at low-dissolved-oxygen conditions,
slow diffusion of NOv . The latter mechanism can be promoted by «. . . o. . ^ ^ . L- ,, ,
_ , r • Biophosphorus Sludge Establishment. It is generally agreed
larger differences between HDO and LDO, because larger fractions A . ,. , , . „ , nA^uij
& ° that, in biological nutrient removal processes, PAOs break down
of floe volume are involved in the dynam.c swing between aerobic intracel|u|ar polyphosphates in the ^obic stage, to obtain the
and anoxic conditions. energy for taking up readily biodegradable organic substrates and
Nitrite Shunt. Nitrite shunt is known to require lower COD to
storing them as polyhydroxyalkanoates (PHAs). The phosphate
drive the nitrate-bypassed denitrification. The COD/NOx~-N ratio
. rA , produced from the polyphosphate hydrolysis is released to the
required for complete denitrification of nitrate was 5.0 to 6.0 (w/w) „ , , „ . T .1
n r water, causing an increase in phosphate concentration. In the
(Tarn et al., 1992), while complete denitrification in a sequencing , , , . «a^a a- *u * a nu a
v » /■» r n & subsequent aerobic stage, PAOs oxidize the stored PHAs to
batch reactor with nitrite shunt occurring was achieved at a much generate energy for growth> g)ycogen synthesiSi and phosphate
lower ratio of 2.9 (Yu et al., 2000). Therefore, the low-dissolved- uptake Thjs causes remQval of phosphate from the water> while
oxygen SND process may be particularly beneficial for wastewater replenishing the intracellular polyphosphate pool depleted during
with low carbon-to-nitrogen ratios. More importantly, the nitrite the previous anaerobic stage.
shunt pathway was also reported to require less aeration energy jn tde low-dissolved-oxygen systems maintained under cyclic
(Yoo et al., 1998). ^ aeration in this study, a portion of the population/volume inside
As shown in Figure 2a for run 4, N02 -N constituted a rather each floe also experienced the anaerobic and aerobic cycles. The
high fraction (approximately 36%) of the total NOx -N. This observed profile of phosphate concentration, shown in Figure 2b,
observation was consistent with the nitrite shunt reported in low- suggested the establishment of PAOs under such conditions. The
dissolved-oxygen systems (Gee and Kim, 2004; Yoo et al., 1998; effects of operating conditions on phosphorus removal in the
Yu et al., 2000; Zeng et al., 2003). The extent of nitrite shunt systems evaluated in this study are shown in Figure 5. Several
occurrence was expected to correlate with the ratio of N02 -N/ observations can be made. First, the run with constant aeration
N03~-N, which would be larger at a higher extent of nitrite shunt (dissolved oxygen concentration = 0.8 mg/L) clearly gave the
(because of the bypassed involvement of N03 ). Accordingly, the lowest phosphorus removal, confirming the beneficial effects of
N02 -N/N03~-N ratio was plotted against SRT and time-averaged cyclic aeration in enhanced biophosphorus removal. Second, for the
dissolved oxygen concentration (DO) (Figure 4a) and against HDO three runs of SRT = 10 days and LDO = 0.2 mg/L, phosphorus
and LDO (Figure 4b) in three-dimensional bubble plots, where the removal increased with increasing DO (or HDO). Thus, larger

916 Water Environment Research, Volume 79, Number 8

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 Ju et al.

U) 8 ■

0.4 0.6 0.8

0.6 0.8 1
1 1.2

Time-Averaged DO (mg/L)

I Constant DO O LDO = 0 mg/L • LDO = 0.2 mg/L

£>£>.e>,o.ç>£>£>£>,ç>ç>.e>.e>.e>ç>

Figure 5—Effects of operating conditions (SRT and time ^ V V <iy <a V <tf <tf <$•

averaged
averaged dissolved oxygen concentration)
dissolved on phospho
oxygen concentration) on phospho- Date
Date
rus removal, with the bubble size indicating the percent
phosphorus
phosphorus removal
removal attained. Figure 6—Results Figure
attained. of effluent
6—ResultsTIN in theTIN
of effluent plant atplant
in the BigatBear,
Big Bear,
California, following the implementation of a low
dissolved-oxygen SND process.
differences between HDO and LDO in the cyclic aeration had
potentially positive effects on phosphorus removal. Third, phos
phorus removal was low (approximately 12%) in run 2 (Tables 1 Big Bear, California. Big Bear Area Regional Wastewater
to 3), with SRT = 20 days, LDO = 0 mg/L, and HDO = 2 mg/L. Agency in California operates an oxidation ditch plant with a design
Two factors might have effected the low phosphorus removal. flow of 12 000 m3/d (3.2 mgd). Of the three oxidation ditches at the
One was the long SRT (20 days) involved, although the effect of facility, only two are normally operated in parallel. Each ditch has
SRT was inconclusive, because run 2 was the only nonbulking a volume of 6000 m3 (1.6 mil. gal) and uses brush aerators (total
system attained at this long SRT. The other factor was the high 134 000 W [180 hp] installed aeration capacity in each ditch) for
HDO (2 mg/L) involved. Development of PAOs requires the aeration and mixing. The plant was designed as a conventional
existence of cyclic aerobic and anaerobic states. In run 2, active nitrification plant based on ammonia removal requirements in the
nitrification during the 1-hour aeration at HDO = 2 mg/L would past. As the effluent requirement changed to 10.0 mg/L total
yield appreciable amounts of nitrate and/or nitrite, which sustained inorganic nitrogen (TIN = ammonia-nitrogen + nitrite-nitrogen +
longer periods of anoxic denitrification and, thus, allowed shorter nitrate-nitrogen), the plant opted for the cyclic on/off low-dissolved
anaerobic periods and smaller anaerobic cores in the floes during the oxygen aeration to introduce simultaneous denitrification within the
0.5-hour LDO operation. Such conditions would be less favorable ditches. Initially, during a 3-month trial period in the summer of
for enhanced biophosphorus removal (but achieving effective 2000, the brush aerators were controlled in on/off mode, with
nitrogen removal). dissolved oxygen concentrations in the range 0.2 to 0.8 mg/L during
Plant Case Studies. The above findings in laboratory low- the "on" period. The modified operation generated effluent TIN
dissolved-oxygen SND systems with alternating aeration between values below 2.0 mg/L, while the ammonia concentrations were
HDO and LDO phases are largely consistent with the phenomena maintained below 0.5 mg/L. This clearly indicated that effective
observed in full-scale wastewater treatment plants that incorporated nitrification was maintained, while simultaneous denitrification was
alternating low-dissolved-oxygen aeration in their operation. Some introduced using the low-dissolved-oxygen SND technology. Over
case studies are presented in the following sections. 30% of aeration energy savings were achieved.

Table 4—Effluent results at Big Bear, California, plant before and after installation of low-dissolved oxygen SND SymBio
system (Enviroquip, Inc.).*

Flow Biochemical oxygen TSS Ammonia-nitrogen Nitrate-nitrogen

Year Location (m3/d [mgd]) demand (BOD) (mg/L) (mg/L) (mg/L) (mg/L) TIN (mg/L)

Before installation

1999 Influent 8000 (2.12) 193 219 N/A N/A N/A

Effluent 8000 (2.12) 5 14 1.2 8.5 9.7

After installation

2001 to 2002 Influent 10 300 (2.72) 275 371 N/A N/A N/A
Effluent (2.72)
10 300 (2.72) 6 6 0.6 2.3 2.8
2002 to 2003 Influent 7900 (2.09) 286 287 N/A N/A N/A
Effluent 7900 (2.09) 8 7 1.4 1.5 3.3

* N/A = not available.

August 2007

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Ju et al.

10.0 i provided in Figure 6. Table 4 provides the average plant effluent

results during this period. In addition, the plant showed enhanced
biophosphorus removal, as observed in the laboratory systems and
explained earlier. Typical influent total phosphorus levels were
7 mg/L, and the effluent total phosphorus averaged approximately
K /\ 1 mg/L.
Rochelle, Illinois. This 18 400-m3/d (4.87-mgd) plant, operated
1 Avg. TKN 1.5 mg/L / by Rochelle Municipal Utilities, uses a parallel operation between
\ I -4 A four plug-flow reactors, each using a two-pass system. Only two
reactors are typically used at a given time. Fine-bubble diffusers
coupled with centrifugal, multistage blowers are used for aeration.
The plant treats a combination of industrial (food processing) and
¿5.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0.0,0.0.0.0
domestic wastewater. Currently, it is required to perform only
^ ri-
^ £J J& <&
<,-{• /A-<£- /v<o"
¿A- <©" 'vCo'V/v/V^V
<d &cA<£-
rA rAJrA<$
rS- c^
rK •^ e
nitrification and not denitrification. The city, however, decided
Date
to install the SymBio system in 2001 at this facility, to maximize
results in
Figure 7—Effluent TKN results in the
the plant
plant at
at Rochelle,
Rochelle, the energy savings (and to use the NAD(P)H measurement for
Illinois, following
Illinois, followingthe
the implementation
implementation of aof a low-dissolved
low-dissolved- monitoring the organic loading fluctuations from a food-processing
oxygen
oxygen SND process.
SND process. industry). Further, a denitrification requirement is expected in the
future. Because of the industrial contribution, the influent TKN
loading is relatively high, in the range 50 to 60 mg/L, and,
In this case, the above aeration control was based on the online correspondingly, the effluent nitrates were high before the intro
monitored fluorescence profile of intracellular NAD(P)H (commer- duction of the low-dissolved-oxygen SND process. Simultaneous
cially known as the SymBio technology, Enviroquip, Inc., Austin, denitrification (potentially more than 70%) in the plug-flow reactors
Texas). The NAD(P)H (i.e., NADH + NADPH) are the reduced was achieved after ,he installation. The effluent TKN results are
forms of the coenzymes nicotinamide adenine d,nucleotides. The shown m F¡gure ? Thc overa„ plam effluent results for 2001 t0
NAD(P)FI fluorescence intensity changes significantly when the 2003 are iven in Table 5
heterotrophic microorganisms in the sludge experience "local" " TheTlanftefore using SND, had settling issues during summer
environmental changes that cause them to switch their respiratory .. .„ . ., , .„ „ „
° months, as a result of denitrification in the clanfiers. Because ol
mechanisms among aerobic respiration, anoxic denitrification, and
fluctuating loadings, the overaeration caused the pin-floc phenom
anaerobic fermentation. More detailed descriptions are available
enon, at times. This also had negative effect on solids separation.
elsewhere for the effects of aeration on the NAD(P)FI profile in
One additional benefit with the low-dissolved-oxygen SND
wastewater treatment processes (Arunachalam et al., 2005; Ju et al.,
, . . , , ,. , , . operation for this facility has been the improvement in sludge
1995) and, particularly, in low-dissolved-oxygen SND systems ' 3 v &
(Huang and Ju 2007) settling. As indicated in Table 5, the SVI, based on 30-minute
Subsequently, in March 2001, variable frequency drives were settling tests, was maintained at approximately 130. No filamentous
installed on the brush aerators for better aeration control. The bulking was observed. A significant increase in phosphorus removal
NAD(P)H fluorescence was monitored in the biomass, and the resulting from the low-dissolved-oxygen SND operation was also
speed of the aerators was regulated to match the oxygen demand observed in this plant. Phosphorous removal efficiency improved
and vary the dissolved oxygen concentration between HDO (<0.8 by 43%, with the influent total phosphorus levels in the range 55 to
mg/L) and LDO (>0.2 mg/L). Effluent nitrogen concentration 60 mg/L, as a result of the food-processing-industry waste contri
results from this automatic operation during 2001 to 2003 are bution (data not shown).

Table 5—Effluent results at Rochelle, Illinois, plant after installation of low-dissolved-oxygen SND SymBio system
(Enviroquip, Inc.).

Flow BOD TSS Ammonia-nitrogen Nitrate-nitrogen

Year Location (m3/d [mgd]) (mg/L) (mg/L) (mg/L) (mg/L) TKN

2001 to 2002 Influent 11 500 (3.04) 252 148 N/A N/A 73

Effluent 11 500 (3.04) 1.6 1.0 0.3 17.9 N/A
2002 to 2003 Influent 7870 (2.08) 285 166 N/A N/A 42
Effluent 7870 (2.08) 3.2 2.8 0.6 9.0 3

Mixed-liquor
Year suspended solids (mg/L) SVI Sludge age (days)

2184 131 15
2001 to 2003 (average)

* N/A = not available.

918 Water Environment Research, Volume 79, Number I

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Conclusions
For the laboratory low-dissolved-oxygen SND studies, the
systems with a 5-day SRT (runs 1 and 3) exhibited poor nitri
fication, presumably as a result of the slow growth of nitrifiers at
low dissolved oxygen concentrations. Compared with constant low
dissolved-oxygen aeration, cyclic aeration, when operated with
proper combinations of HDO and LDO, achieved more effective
nitrogen and phosphorus removal. For nitrogen removal, the
advantage of cyclic aeration presumably resulted from the more
readily available nitrate and nitrite (generated by nitrification during
the HDO period) for denitrification (during the LDO period). Under
constant low-dissolved-oxygen aeration, denitrification would
rely on the slow diffusion of nitrate and nitrite from the outer
nitrifying zone of the floes into the inner denitrifying zone. None
theless, nitrogen removal in the systems investigated was more
susceptible to nitrification limitation than denitrification limita
tion. Therefore, the HDO used should be kept reasonably high
(0.8 mg/L or higher) or the HDO period prolonged, to promote
adequate nitrification. Nitrite shunt was also observed in low
dissolved-oxygen SND systems, with higher ratios of N02~/N03~
in systems of lower HDO (and, to less dependency, higher LDO).
The results suggested that the nitrite shunt took place mainly
because of the disrupted nitrification at low dissolved oxygen
concentrations. For phosphorus removal, the advantage of cyclic
aeration came from the development of higher polyphosphate
accumulating populations, as indicated by the higher phosphorus
contents in the sludge solids established in the systems under cyclic
aeration. The above findings in the laboratory systems were largely
consistent with the observations in full-scale wastewater treatment
plants, as indicated in the two case studies. The feasibility of
simultaneous nitrification, denitrification, and enhanced phosphorus
removal in single-tank, low-dissolved-oxygen, cyclically aerated
systems was clearly demonstrated in both laboratory-scale systems
and full-scale plants.
Credits
This research was supported by Enviroquip, Inc. (Austin, Texas).
The authors are grateful for the technical assistance of Shuyan Qiu,
Nicholas J. Hamilton, Andrew S. Lash, Kristen M. Dudak, and
Elizabeth A. Amaddio.
Submitted for publication April 6, 2006; revised manuscript
submitted November 13, 2006; accepted for publication November
28,2006.
The deadline to submit Discussions of this paper is November 15,
2007.
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