Wat. Res. Vol. 33, No. 1, pp.

145±154, 1999
# 1998 Elsevier Science Ltd. All rights reserved
Printed in Great Britain
PII: S0043-1354(98)00159-6 0043-1354/98 $ - see front matter

NITROGEN REMOVAL FROM SYNTHETIC WASTEWATER

BY SIMULTANEOUS NITRIFICATION AND
DENITRIFICATION (SND) VIA NITRITE IN AN
INTERMITTENTLY-AERATED REACTOR
HYUNGSEOK YOO1*, KYU-HONG AHN1*
M
, HYUNG-JIB LEE1,
KWANG-HWAN LEE , YOUN-JUNG KWAK and KYUNG-GUEN SONG1
2 2

1
Environment Research Center, Korea Institute of Science and Technology (KIST), P.O. Box 131
Cheongryang, Seoul, South Korea and 2Department of Environmental Engineering, Seoul National
Polytechnic University, Gongneung-Dong, Nowon-Ku, Seoul, South Korea

(First received August 1997; accepted in revised form March 1998)

AbstractÐAn intermittently aerated and decanted single-reactor process was proposed and some key
control parameters investigated for nitrogen removal from wastewater by simultaneous nitri®cation and
denitri®cation (SND) via nitrite. Two types of synthetic wastewater with acetate as the main carbon
source, and chemical oxygen demand (COD):nitrogen (N) ratios of approximately 5:1 and 10:1 were
used. For both types of wastewater the average COD removal eciency reached above 95%, and under
optimal conditions nitrogen removal eciency also reached above 90%. This process consisted of
72 min aeration, 48 min settling and 24 min e‚uent decanting. In¯uent wastewater was fed from the
bottom of the reactor, and did not require a separate mixing phase. In this process, nitritation (1st step
of nitri®cation) was induced but nitratation (2nd step of nitri®cation) was e€ectively suppressed and
denitri®cation was carried out using nitrite. Most important parameters for e€ective SND via nitrite in
the proposed process were: (1) the increase rate and the minimum and maximum dissolved oxygen
(DO) concentration during the aeration period, (2) length of one cycle, and of aeration per cycle, taking
advantage of the lag-time of nitrite-oxidizers behind ammonia-oxidizers when adjusting from anoxic/an-
aerobic to aerobic condition, and (3) close contact between the mixed liquor suspended solids (MLSS)
in the reactor and the in¯uent wastewater under anoxic/anaerobic condition. During the aeration
period, DO concentration dropped initially reaching the minimum value after several minutes, and then
rose in 2nd-order fashion. The optimal maximum DO concentration (at the end of the aeration period)
for nitrogen removal was determined to be around 2.0±2.5 mg/l in this proposed process. # 1998 Else-
vier Science Ltd. All rights reserved

Key wordsÐnitrogen removal, simultaneous nitri®cation and denitri®cation (SND), nitrate accumu-
lation, nitritation, nitratation, intermittently decanted extended aeration (IDEA)

INTRODUCTION up to 40% could result (Turk and Mavinic, 1986,

Simultaneous nitri®cation and denitri®cation (SND)
which has advantages over the separated nitri®ca-
tion and denitri®cation processes, means that nitri®-
cation and denitri®cation occur concurrently in the
same reaction vessel under identical operating con-
dition. In continuously operated plants, SND o€ers
the potential to save cost for a second (anoxic)
tank, or at least reduce its size, if it can be ensured
that a considerable amount of denitri®cation takes
place together with nitri®cation in the aerated tank.
If SND is accompanied by the inhibition of the
second step of nitri®cation (oxidation of nitrite to
nitrate), theoretically a saving of organic energy of
*Author to whom all correspondence should be addressed.
Hyung Yoo, Dept of Civil and Environmental
Engineering, Stanford University, Stanford, CA 94305-
4220, USA. [Tel: +1(650)723-0315; Fax: +1(650)725-
3162; E-mail: hyungseok@yahoo.com].
145
1987; Abeling and Seyfried, 1992). This is of par-
ticular interest when biologically removing nitrogen
from wastewater with low COD:nitrogen ratio.
Research by Sutherson and Ganczarczyk (1986)
and Turk and Mavinic (1989) has shown the inter-
est of mastering nitritation and achieving the direct
coupling of nitritation and denitri®cation by per-
forming a nitrate shunt, which was achieved by
reducing the activity of Nitrobacter and giving
Nitrosomonas species growth advantages.
Nitri®cation and denitri®cation via nitrite has been
investigated in detail (Abeling and Seyfried, 1992;
Yang and Alleman, 1992; Akunna et al., 1993;
Turk and Mavinic, 1986; Prakasam and Loehr,
1972; Murray et al., 1975; Laudelout et al., 1976;
Sauter and Alleman, 1980; Blaszczyk et al., 1981).
Investigation of Turk and Mavinic (1986, 1987) of
the shortened nitrogen removal pathway via nitrite
revealed: (1) 40% reduction of COD demand
146 Hyungseok Yoo et al.
during denitri®cation; (2) 63% higher rate of deni-
tri®cation; (3) 300% lower biomass yield during an-
aerobic growth; and (4) no apparent nitrite toxicity
e€ects for the microorganisms in the reactor.
Abeling and Seyfried (1992) discovered in their
half-technical experiences, that saving cost was also
possible with the reduced oxygen demand. For
nitritation only 75% of the oxygen was required
compared to that of the complete oxidation to
nitrate.
SND is also e€ective in maintaining neutral pH
level in the reactor, without the addition of external
acid/base source. This is important since a narrow
optimal range between pH 7.5±8.6 is known to exist
for the nitrifying bacteria. During nitri®cation alka-
linity is consumed, but alkalinity is produced during
denitri®cation. The optimal pH lies between 7 and
8 for denitri®cation with di€erent optimums for
di€erent bacterial populations.
However, high nitrite build-up may, in many
cases, be harmful to the operation of the waste-
water treatment plant, and therefore its concen-
tration in the e‚uent must be kept low due to its
toxicity to some species of ®sh; therefore, SND via
nitrite must accompany far-reaching denitri®cation.
This investigation proposes a simple, single-reac-
tor nitrogen-removal process utilizing SND via
nitrite, without a separate mixing period, evaluates
its performance, and suggests some key control par-
ameters for e€ective operation. The design of the
proposed process is based on the following litera-
ture review; key control parameters and their opti-
mal values were determined in the laboratory.
FACTORS FOR NITRITE ACCUMULATION DURING
NITRIFICATION
Some of the best works investigating nitrite
build-up during nitri®cation were performed by
Anthonisen (1976), Sutherson and Ganczarczyk
(1986), Jayamohan and Ohgaki (1988) and Hanaki
and Wantawin (1990), which focused on a number
of factors, such as the free ammonia (FA) concen-
tration, the free hydroxylamine (FH) concentration,
the pH, the temperature, and the dissolved oxygen
(DO) concentration, on the transient build-up of
the nitrite ion.
High free ammonia (NH3) concentration
According to the research by Anthonisen (1976),
the non-ionized forms of the ammonium and nitrite
have Ð as ammonia (NH3) and nitrous acid
(HNO2) Ð an inhibition e€ect on both the
Nitrosomonas and the Nitrobacter. The Nitrobacter
react more sensitively so that concentration of NH3
in relatively low range is sucient for inhibition.
Abeling and Seyfried (1992) reported that concen-
tration of 1±5 mg NH3/l inhibited nitratation but
not the nitritation. In the research of Turk and
Mavinic (1986, 1987) nitrite build-up was achieved
with intermittent contact to high FA levels (5 mg
NH3±N/l) in the ®rst cell of a four-cell system.
However, nitrite build-up could not be sustained
inde®nitely, due to the acclimation of the
Nitrobacter to FA. Wong-Chong and Loehr (1978)
observed that the Nitrobacter acclimated to FA
could tolerate concentrations as high as 40 mg
NH3±N/l, while unacclimated ones were inhibited
at concentrations of 3.5 mg NH3±N/l. Many other
researchers found out similar results. Ford et al.
(1980) reported total inhibition of nitri®cation ac-
tivity at FA levels of 24 mg NH3±N/l, but noted
that system recovery was possible, even at levels as
high as 56 mg NH3±N/l. Cecen and Gonenc (1994)
noted, in the batch start-up phase, the combined
e€ect of high ammonia and high pH (8.5) inhibited
Nitrobacter and led to accumulation of nitrite.
Mauret et al. (1996) showed that high FA concen-
tration inhibits Nitrobacter, in the range of 6.6 and
8.9 mg NH3±N/l. Balmelle et al. (1992) reported
that the inhibition e€ect of nitratation could be
observed at concentrations as low as 1 mg NH3±N/l.
Anthonisen (1976) found out values of 0.1±1.0 mg
NH3±N/l.
Abeling and Seyfried (1992) stated that in order
to attain the highest nitritation rate, it was decisive
to prevent the inhibition of the Nitrosomonas
caused by FA. The FA concentration necessary for
the inhibition of Nitrobacter must be kept low
enough to ensure that the inhibition of nitritation
does not also take place. At pH = 8.5 and
T = 208C, the optimal FA concentration for maxi-
mum nitritation and minimum nitratation was
found to be around 5 mg NH3/l (Abeling and
Seyfried, 1992). On and after approximately 7 mg
NH3/l an inhibition on nitritation could be noticed.
At concentrations of around 20 mg NH3/l there was
very little nitri®cation activity. Limits to prevent
nitritation inhibition had been found out in batch
tests which ranged between 10 and 150 mg NH3/l
(Anthonisen, 1976). Also Neufeld et al. (1980) con-
®rmed the beginning of nitritation inhibition at
10 mg NH3/l.
Temperature
Balmelle et al. (1992) showed that in spite of con-
centration of FA normally inhibiting for the
Nitrobacter of between 2 and 5 mg NH3±N/l, the
Nitrobacter was active over a range of temperature
between about 10 and 208C. Under these con-
ditions, nitrite build-up remained low, since the
e€ect of the Nitrobacter activation by temperature
prevailed over its inhibition by FA. On the other
hand, beyond a temperature of 20±258C, a slowing
of the nitratating activity was observed together
with an activation of the nitritating activity, which
passes through a maximum at 258C. These results
were in agreement with the data of Quinlan (1986),
that showed high Nitrobacter activity for tempera-
ture below 158C. With respect to Nitrosomonas
 Nitri®cation and dentri®cation via nitrite
under these conditions, the inhibiting e€ect of FA
was preponderant for temperature higher than
258C. This result con®rmed that of Anthonisen
(1976), but fairly disparate observations had been
found in the literature, notably those of Ford et al.
(1980) which suggested the optimal temperature
range between 30 and 368C.
Lag-time when changing from anoxic to aerobic con-
dition
Turk and Mavinic (1986) observed a lag time in
nitratation. Even with a bacterial population con-
taining Nitrobacter acclimated to FA, nitritation
rates lagged nitratation rates following the exit
from the anoxic cell and entering the aerobic cell,
resulting in a temporary, but signi®cant, accumu-
lation of nitrite which lasted for several hours in
the aerobic cell. Senanyake (1982), and Turk and
Mavinic (1986) have reported that the extension of
aeration time alleviates stress on the nitrite oxidizers
and results in the loss of nitrite build-up. Duration
of aeration time was found to be inversely related
to the degree of nitrite build-up.
Low DO concentration during aeration
Cecen and Gonenc (1994) reported that nitrite ac-
cumulation reached a considerable degree at DO to
FA concentration ratios lower than 5 during nitri®-
cation, and the formation of nitrate was inhibited.
No nitrite occurrence was encountered when this
ratio exceeded 5, which implied that oxygen limi-
tation lead to nitrite accumulation. Therefore, e€ec-
tive nitritation might be achieved by supplying less
air during nitri®cation.
Free-hydroxylamine (FH) concentration
Yang and Alleman (1992) concluded that the DO
level alone did not appear to be the dominant fac-
tor behind nitrite build-up, and its correlation with
free ammonia (FA) concentration alone was also
erratic. On the other hand, hydroxylamine
(NH2OH/NH3OH+), an intermediate in nitri®ca-
tion by Nitrosomonas, was found to inhibit nitrata-
tion by the Nitrobacter. Indeed, hydroxylamine
appeared likely to accumulate in a nitrifying system
with high NH3/NH+ 4 concentration, de®cient oxy-
gen, and high pH. This research showed that the
presence of unionized or the so called free hydroxy-
lamine (FH) fraction (as opposed to ``total'')
appeared to have a consistent correlation with low
nitratation activity. FH was believed to be a major,
if not principal, factor behind nitrite build-up in the
batch nitrifying systems.
Process guidelines for suppressed nitratation
Therefore, to achieve e€ective SND via nitrite, it
is necessary to (1) use simultaneous and/or alternat-
ing nitri®cation/denitri®cation process in the same
reactor (factors 1, 3 and 4 above); (2) maintain low
DO during aeration (factors 1, 4, and 5); (3) keep
147
the microorganisms in direct contact with the in¯u-
ent wastewater in oxygen-de®cient condition to
induce contact with high concentration of FA and/
or FH (factors1 and 5); (4) raise the pH (factors 1
and 5); (5) add hydroxylamine to the reactor (factor
5); and (6) maintain the reactor temperature near
258C (factor 2).
PROPOSED PROCESS
The following cyclic activated sludge process con-
sisting of three di€erent periods (Fig. 1) was pro-
posed. It is a modi®ed form of an intermittently
decanted extended aeration (IDEA) process (®rst
developed by the Department of Public Works and
Services, NWS, Australia in 1965).
``Anoxic'' is de®ned, in this process, as the state
or condition where denitrifying bacteria carry out
denitri®cation utilizing nitrite and/or nitrate as elec-
tron acceptors, independent of the DO concen-
tration in the surrounding environment. Such
condition may take place in the entire water body
of the reactor during the initial aeration phase, and
in the settling/settled sludge layer during the settling
phase. ``Anaerobic'' condition in this process takes
place inside the settled sludge layer after all the
nitrite and nitrite are used up as electron acceptors
inside the layer. ``Aerobic'' condition takes place in
the reactor after some time has elapsed into the
aeration period-denitri®cation is suppressed due to
the rising DO concentration and nitri®cation takes
place.
As shown in Fig. 1, the in¯uent wastewater is fed
constantly from the bottom throughout the cycle.
During the settling and decanting periods oxygen is
de®cient, and the Nitrobacter are in direct contact
with the FA in the in¯uent wastewater, which
theoretically has a high probability to be converted
to FH by the Nitrosomonas species. The settling/
settled sludge acts as an anoxic reactor where deni-
tri®cation takes place. When the aeration period of
the subsequent cycle starts, the condition in the
reactor is anoxic even with the added air; it takes
some time for the DO level to rise high enough to
suppress denitri®cation. Denitrifying activity is not
completely suppressed, and the converted nitrite
and nitrate from nitri®cation with the added oxygen
will be denitri®ed-thus the simultaneous nitri®cation
and denitri®cation. Therefore, this modi®ed IDEA
process takes advantage of the non-instantaneous
transition from anoxic/anaerobic to aerobic con-
dition of the microorganisms. Such alternating
between anoxic/anaerobic and aerobic condition is
necessary to take advantage of the lag-time of the
nitrite-oxidizers behind the ammonium-oxidizers.
Such lag time may be di€erent for each species of
denitrifying bacteria, and certain species may carry
out denitri®cation throughout the entire cycle. The
length of the aeration period should be long enough
for e€ective COD removal, nitri®cation and denitri-
148 Hyungseok Yoo et al.

Fig. 1. Modi®ed IDEA process description and operation strategy: (a) three periods of the process; and
(b) length ratios of the three periods, corresponding conditions and the ideal DO pattern (determined
in this experiment).

®cation, but short enough to prevent the adaptation Experimental set-up and reactor operation
of nitrite-oxidizers to the aerobic environment. The two modi®ed IDEA reactors were variable-volume,
Ideal dissolved oxygen (DO) level curve for one and seeded with a grab sample of mixed liquor from the
cycle is shown in Fig. 1(b). Minimum DO concen- aerobic basin of the Kwangdongli Sewage Treatment
tration of the aeration period is achieved several Plant (Kyung-Gi-Do, Korea). They were operated for
122 days. Sample analysis commenced after approximately
minutes into the period. Intuitively, optimal average 22 days of operation and continued for the next 100 days.
and maximum DO concentration levels exist for The reactors had circular bottom with diameter of 20 cm,
nitrogen removal in this process. DO levels above and their maximum working volume was 17 l. Discharge
these values will yield too much nitrite/nitrate and sampling ports were located at di€erent levels. The
top of the reactors was open to air. The reactors were
(NOx) in the e‚uent, whereas levels too low will operated in parallel, with varying hydraulic retention times
induce a high level of NH3/NH+ 4 . It was one of the (HRT's), and sludge retention times (SRT's). The in¯uent
purposes of this experiment to determine such opti- wastewater was kept in a refrigerator (at 48C). The reactor
mal DO levels. This proposed process does not temperature was maintained at 22±278C.
require a separate mixing period, since the settling Cyclic operation as described in Fig. 1 was controlled
by a programmable logic controller (PLC). Aeration
and decanting periods, and the initial portion of the period was set at 72 min, settling period at 48 min, and
aeration period are anoxic/anaerobic treatment decanting period at 24 min for the reactors; the duration
phases. The amount of air supplied is such that the of each cycle was 144 min (2.4 h) i.e. 10 cycles per day.
DO level does not rise too quickly into the aeration 10 l of wastewater was fed to each reactor everyday, and
period.
Table 1. Composition of the two types of in¯uent wastewater
Added compounds In¯uent wastewater 1 In¯uent wastewater 2
(per l in¯uent) (mg) (mg)
MATERIALS AND METHODS
CH3COONa ÿ 256.41
Composition of the synthetic wastewater CH3COONH3 240.88
KH2PO4 43.94
Two types of synthetic wastewater, made accord- NaHCO3 125.00
ing to Table 1, were fed to reactors 1 and 2. CaCl2 10.00
FeCl2 0.375
The di€erence between in¯uent wastewater 1 and MnSO4 0.038
2 was the COD/NH4±N ratio Ð approximately 5:1 ZnSO4 0.035
and 10:1 Ð and COD/phosphate ratio-approxi- MgSO4 25.00
Yeast extract 50.00
mately 10:1 and 20:1, respectively.
 Nitri®cation and dentri®cation via nitrite 149

Table 2. In¯uent wastewater characteristics

In¯uent 1 In¯uent 2
Parameter average std. dev. average std. dev.

COD (TCOD = SCOD) 226.2 10.0 401.1 21.3

(mg/l)
NH4±N (mg/l) 41.3 1.8 41.4 0.8
HPO4±P (mg/l) 13.5 7.5 11.5 6.5
NO3 ±N (mg/l) 1.1 0.3 1.1 0.6
COD/NH4±N ratio 5.5 9.7

all in¯uent, e‚uent, and waste activated sludge (WAS)

streams were controlled using variable-speed Master¯ex
pumps. When changing the HRT, the amount of in¯uent
feed was kept constant and the reactor volume was chan- Fig. 2. MLSS concentration for reactor 1 (W) and reactor
ged. All water and WAS tubes were cleaned once a week. 2 (w).
To control the SRT, WAS was wasted during the last min-
ute of the aeration period (not collected). Stirrers, operat-
ing at 30 rpm, provided mixing during the aeration period
RESULTS AND DISCUSSION
in addition to the compressed air. Air was di€used by
aquarium-type di€user-stone, and the supply rate was con-
trolled using ¯owmeter. Supplied air was in the range of Mixed liquor suspended solids (MLSS) and COD
0.3±1.0 l/min. DO level control system was purposely not removal
used to observe nitrogen removal at various DO concen-
tration and pattern, and to obtain the optimal DO level. As shown in Fig. 2, MLSS concentration for
DO concentration was measured 5 min into (minimum both reactors dropped relatively quickly from day 1
DO) and at the end of (maximum DO) the aeration until day 30, and continued to drop until day 60;
period, using Series 5905 DO probe and Model 58 DO
however such drop did not a€ect the two reactors'
meter (YSI, OH); the membrane of the DO electrode was
changed every 2 weeks. The DO probe was inserted into COD removal which was very high (Figs 3 and 4).
the water surface about 5 cm for measurement. Therefore, the initial HRT's were believed to be too
high, and on day 61, the HRT's were nearly
reduced by half (Table 3). The HRT reduction did
Sampling methods not a€ect the reactors' COD removal, and resulted
Grab in¯uent wastewater samples were analyzed once a in stable MLSS concentration in reactor 1 from day
week. E‚uent samples were collected as the entire dec- 61, but in reactor 2 MLSS concentration increased
anted e‚uent from the same cycle of everyday, two to slightly. SRT for both reactors was increased to
four times a week and analyzed. Cyclic studies were per- 45 days on day 26 to slow down the dropping rate
formed several times throughout the operation with
of MLSS concentration. The SRT increase had no
sampling intervals between 1 and 15 min. Samples for the
determination of soluble components were immediately ®l- e€ect on the reactors' performance (Figs 3 and 4).
tered using 0.45 mm ®lter paper and cooled (48C) in order COD removal rate for both reactors was very
to prevent further reaction after sampling. Sample analysis high throughout the operation, una€ected by the
was performed according to the standard methods changes in the MLSS concentration, HRT and
(APHA, 1992).
SRT. Based on the average incoming soluble COD
(SCOD) concentration of 226.2 mg/l (reactor 1) and
401.1 mg/l (reactor 2), the removal eciency was
General reactor operation
between 89% and 98% with the average of 95%
Table 2 shows the characteristics of the two types of (Std. Dev. = 2.0) for reactor 1, and between 85%
in¯uent wastewater.
and 99% with the average of 97% (Std.
Since this process combined anoxic, aerobic and settling
phases in a single reactor, and aimed for high perform-
ance, initial HRT's were set at relatively high values.
HRT's were to be reduced if the reactors showed high per-
formance with continued decrease in the MLSS concen-
tration. Table 3 describes change in the HRT's and SRT's.
(All calculation was based on the reactors' volume at the
bottom water level (BWL) at the end of the decanting
period.) HRT's for both reactors were lowered nearly by
half on day 61. (``Day'' will be referred according to the
sample analysis dates.)
The sludge settleability for both reactors was good, with
the sludge volume index (SVI60) between 78 and 165 ml/g.
SVI60 was not a€ected by changes in HRT and SRT. pH
was measured at the end of aeration, settling and decant-
ing periods; pH values for both reactors were in the neu-
tral range between 6.9 and 7.8. Average pH values were
7.2 and 7.5 for reactors 1 and 2 respectively. Although pH Fig. 3. E‚uent SS concentration of reactor 1 (W) and
bu€er solution was added to the in¯uent wastewater, SND reactor 2 (r), and SCOD concentration of reactor 1 (.)
aided in maintaining the neutral pH. and reactor 2 (w).
150 Hyungseok Yoo et al.

Fig. 4. E‚uent nitrogen compounds and DO concentration of reactor 1 (a) and reactor 2 (b). (``NH3 ±
N'' in this Figure is technically the sum of NH3±N and NH+
4 ±N.)

Dev. = 2.4) for reactor 2, respectively. Figure 3
shows the e‚uent suspended solids (SS) and SCOD
concentration.
Scum ¯oated on the water surface of both reac-
tors; an attempt was not made to remove the scum.
In full-scale operation, decanting system that separ-
ates the scum from the e‚uent should be installed.
In this experiment, e‚uent samples contained the
scum and exhibited relatively high SS concen-
tration.
Nitrogen removal, and changes in DO, HRT and
SRT
Figure 4 shows the e‚uent nitrogen (N) com-
pounds' concentration with the minimum and maxi-
mum DO concentration for reactors 1 and 2.
Surprisingly, nitrite accumulation was very success-
ful in both reactors from day 1. For many days,
high concentration of nitrite was observed in the
e‚uent, but low nitrate concentration. A corre-
lation between the e‚uent nitrite concentration and
DO levels was observed.
According to Fig. 4, high concentration of nitrite
resulted when the maximum and minimum DO con-
centration in the reactors was relatively high. High
NH3/NH+ 4 concentration was observed when the
maximum and minimum DO concentration was
relatively low. Generally, the minimum DO concen-
tration was below 0.4 mg/l. From Fig. 4, SND via
nitrite seemed very successful. Also, the initial guess
of the existence of optimal DO levels for N-removal
proved to be correct. pH was never intentionally
raised or the addition of hydroxylamine made to
suppress nitratation; initially it was thought that
these steps would be necessary. To con®rm the
SND via nitrite, NOx (sum of NOÿ ÿ
2 and NO3 ) con-

Table 3. HRT's and SRT's of the two reactors

Reactor 1 Reactor 2
Day HRT (h) SRT (days) HRT (h) SRT (days)

1±25 25 39 31 25
Fig. 5. Nitrite concentration for reactor 1 (W) and reactor
26±60 25 45 31 45
61±100 13 45 17 45 2 (.), and nitrate concentration for reactor 1 (r) and reac-
tor 2 (w), at the end of aeration period.
 Nitri®cation and dentri®cation via nitrite 151

Fig. 6. Cyclic study of reactor 1 on day 87. (``NH3±N'' is the sum of NH3±N and NH+
4 ±N.)

centration was measured at the end of the aeration
period.
Figure 5 shows high concentration of nitrite at
the end of aeration period in both reactors. Nitrite
oxidation was e€ectively suppressed in this pro-
posed process.
Cyclic study
Figure 6 shows data from one of the cyclic stu-
dies, which showed ideal DO pattern described in
Fig. 1. All samples for the cyclic studies were col-
lected by immersing 50 ml beaker slightly into the
water surface and extracting about 20 ml. The
samples were then immediately ®ltered using
0.45 mm ®lter paper. DO level changed sensitively in
all cyclic studies.
In Fig. 6, DO concentration rose in 2nd-order
fashion after about 5 min into the aeration period,
reaching 2.5 mg/l by the end. The median DO level,
de®ned as the mid-value between the minimum and
maximum DO values in a given aeration period,
was 1.3 mg/l in this cyclic study. To obtain the true
average DO concentration, integration must be per-
formed for the DO curve during the aeration period
and then division by the time; however, such inte-
gration was not performed in this experiment.
Using the average DO concentration together with
the minimum and maximum concentration would
be the accurate way to describe the pattern of DO
level. During settling and decanting periods, DO
level in the upper region of the reactor dropped
quickly reaching 0.4 mg/l by the end. Figure 6
showed the ideal pattern of DO concentration; it
dropped and then started to rise several minutes
into the aeration period, and passed 1.0 mg/l at
minute 38, about half-way into the period. Thus,
during the initial phase, mostly denitri®cation took
place indicated by the drop and low concentration
of nitrite, and slight increase in pH. NH3/NH+ 4 ±N
concentration stayed constant at around 6.5 mg/l
during the initial phase, but then dropped as the
DO concentration rose, and disappeared when the
DO concentration was between 1.5±2.0 mg/l. On
the contrary, nitrite concentration rose with the DO
level, indicating nitri®cation was active in the later
portion of the aeration period. Nitrite appeared as
the DO level passed through 1.0 mg/l and its con-
centration stayed high until the end. Due to nitri®-
cation, pH dropped slightly. Nitrite concentration
rose steadily until the end of aeration. After several
minutes into the settling period, nitrite concen-
tration started to drop. During the decanting
period, NH3/NH+ 4 appeared again due to di€usion
of the in¯uent wastewater's NH3/NH+ 4 . About
1.3 mg/l of nitrite and 0.4 mg/l of NH3/NH+ 4 was

Fig. 7. E‚uent NO3±N (*), NOx±N (R), and (NH3+NOx)±N (w) concentrations of reactors 1 (a) and
2 (b) versus median DO levels (data from entire operation). (``NH3±N'' in this Figure is the sum of
NH3±N and NH+ 4 ±N. ``NOx ±N'' is the sum of NO2 ±N and NO3 ±N.)
152 Hyungseok Yoo et al.
Fig. 8. The e€ect of DO level on speci®c nitri®cation and denitri®cation rates for reactor 1 during days
61±100. (Dotted lines show the intuitive pattern.)
observed in the e‚uent of this speci®c cycle indicat-
ing e€ective N-removal.
From the cyclic studies, the importance of DO
level was realized. DO level should be controlled
two-dimensionally: (1) the maximum level of DO
during the aeration period, and (2) the increase rate
of the DO level-aerobic condition should not be
created too quickly or slowly. Therefore, aeration
system capable of such DO control must be im-
plemented in the full-scale plant of this proposed
process. The aeration system should be easy to
install since there are four control parameters of air
supply: (1) DO level sensor, (2) ¯owmeter installed
at the air pump, (3) air-supply valve, and (4) ¯exi-
bility in the length of the aeration period.
Determining the optimal DO concentration for nitro-
gen-removal
The e€ect of maximum and minimum DO con-
centration on the e‚uent N compounds is shown in
Fig. 7; the median DO concentration is the x-axis.
According to Fig. 7, when the median DO level
was high, NOx concentration was high and NH3/
NH+ 4 was mostly removed from the e‚uent, and
Fig. 9. Speci®c N-removal rates for reactor 1 during day
61±100. (Dotted line shows the intuitive pattern).
the opposite trend occurred when the DO value was
low. The median DO value of 1.0±1.5 mg/l seemed
to be optimal, implying that if the minimum DO
level was in the range of 0.0±0.4 mg/l, the maximum
DO level should be in the range of 1.6 and 3.0 mg/l.
It is generally known that DO concentration above
1 mg/l is essential for nitri®cation; if the DO level is
lower, oxygen becomes the limiting factor and nitri-
®cation slows or ceases. On the contrary in denitri®-
cation, high DO level will suppress the necessary
enzyme system. Thus, when SND via nitrite
becomes the focus, controlling the DO level is criti-
cal to balance the degrees of nitri®cation and deni-
tri®cation, and the resulting levels of N compounds
in the e‚uent.
With certain portion of the data, speci®c nitri®ca-
tion, denitri®cation and N-removal rates were cal-
culated to study the e€ect of di€erent DO levels on
the relative degrees of nitri®cation and denitri®ca-
tion. For such study to be valid, MLSS concen-
tration must be constant. Therefore, data from
reactor 1 during days 61±100 (Fig. 2) were used.
Figure 8 shows that nitri®cation rate increased
with the DO concentration, whereas the denitri®ca-
tion rate decreased. Figure 9 shows the speci®c N-
removal rate at various DO levels. The optimal me-
dian DO level for nitrogen removal was 1.3 mg/l.
Review of factors for nitratation suppression
Concentration of FA (NH3) was calculated
throughout the operation. For reactor 1, the maxi-
mum NH3/NH+ 4 ±N in the e‚uent was 23.1 mg/l
obtained on day 71; pH was 6.9 during aeration
and 7.2 during decanting which corresponded to
0.10 and 0.20 mg/l NH3±N at 258C. For reactor 2,
the maximum NH3/NH+ 4 ±N in the e‚uent was
27.6 mg/l obtained on day 64; pH was 7.1 during
aeration and 7.5 during decanting which corre-
sponded to 0.19 and 0.48 mg/l NH3±N at 258C. If
no NH3/NH+ 4 ±N removal is assumed for reactor 1,
 Nitri®cation and dentri®cation via nitrite
Fig. 10. Extension of the aeration period in reactor 1 on day 96.
the in¯uent 41.3 mg/l NH3/NH+ 4 ±N at 258C should
yield 0.07 mg/l NH3±N at pH = 6.5, 0.23 mg/l at
pH = 7.0, and 0.71 mg/l at pH = 7.5; so in reactor
1, NH3±N concentration should not exceed 1.0 mg/l
at any time unless the pH reaches 7.65; such pH
was never recorded. For reactor 2, the in¯uent
41.4 mg/l NH3/NH+ 4 ±N at 258C should yield
0.71 mg/l NH3±N at pH = 7.5, and 1.40 mg/l at
pH = 7.8 which was the maximum pH recorded.
Such result implies that the reported values by
Abeling and Seyfried (1992), Turk and Mavinic
(1986, 1987), Wong-Chong and Loehr (1978), Ford
et al. (1980), and Mauret et al. (1996) for the FA
level inhibiting nitratation was not reached in either
of the reactors. Only the values reported by
Balmelle et al. (1992) at 1 mg NH3±N/l, and
Anthonisen (1976) at 0.1±1.0 mg NH3/l were close
to the levels observed in our experiment. Inhibition
e€ect of FA was maximum in the region of in¯uent
feed at the bottom of the reactor, especially during
settling and decanting periods. Hydroxylamine was
not measured in this experiment. Thus, correlation
between hydroxylamine concentration and nitrita-
tion/nitratation activity cannot be stated in numeri-
cal terms. However, hydroxylamine was never
added to the in¯uent or to the reactors, as initially
thought necessary to achieve SND via nitrite.
Theoretically, hydroxylamine had high probability
of build-up (by the Nitrosomonas) in the settled
sludge during settling and decanting periods due to
de®cient oxygen, and could have suppressed nitrata-
tion irreversibly.
Temperature was in the range of 22±278C during
the entire operation, inducing maximum nitritation
activity. Maximum nitratation activity is expected
in the temperature range below 158C.
DO to FA ratio was calculated. Cecen and
Gonenc (1994) suggested that below the ratio of 5,
nitrite accumulation was high and the formation of
nitrate inhibited. For reactor 1, maximum FA con-
centration in the e‚uent was 0.20 mg/l NH3±N
(calculated previously), which corresponded to
0.24 mg/l NH3. For the DO to FA (NH3) ratio to
be below 5, the DO level must be lower than
153
1.2 mg/l. For reactor 2, maximum FA concentration
was 0.58 mg/l NH3 (0.48 mg/l NH3±N) in the e‚u-
ent, and the DO level must be lower than 2.9 mg/l
for the ratio to be below 5. DO level in both reac-
tors changed throughout the cycle as well as from
day to day. The calculated DO levels (as reference
points) suggest that DO:FA ratio was below 5 more
than half of the time in both reactors.
The most in¯uential factor was believed to be the
lag-time of nitrite-oxidizers behind ammonia-oxidi-
zers during the transition from anoxic/anaerobic to
aerobic condition. Turk and Mavinic (1986) stated
such lag-time to be ``several hours'' but did not pro-
vide a speci®c numerical value. To study the e€ect
of the lag-time, on day 96, aeration period in reac-
tor 1 was extended for 330 min (the normal length
had been 72). DO level was controlled at approxi-
mately 2.5 mg/l by adjusting the air ¯owmeter.
In¯uent wastewater was fed continuously, but the
e‚uent was not decanted; the water surface level
continued to rise. All other experimental settings
stayed the same.
72 min, the normal length of aeration, was ade-
quate for e€ective COD and N-removal (Figs 3 and
4). In Fig. 10, normally the aeration period should
have ended at minute 77. After minute 77, pH con-
tinued to drop but never fell below 6.5; ammonia
did not appear and nitrite concentration increased
indicating on-going nitri®cation. Nitrate concen-
tration rose as the aeration continued indicating
that (1) the nitrite-oxidizers were present in the
reactor, and (2) they were adjusting to the aerobic
environment. Substantial portion of nitrite-oxidizers
could not adapt to the aerobic environment in the
72 min of normal aeration.
It was concluded that the active nitritation and
suppressed nitratation was caused by the combined
e€ect of all the factors reviewed, and the most im-
portant one was the lag-time of nitrite-oxidizers
when transitioning from the anoxic/anaerobic to
aerobic environment. The investigation to determine
exactly to what degree each of the factors was in¯u-
ential would be the subject of future research.
154 Hyungseok Yoo et al.
CONCLUSION
This investigation showed that N-removal from
synthetic wastewater utilizing SND via nitrite was
successful in this proposed intermittently-aerated
cyclic activated-sludge single-reactor process con-
sisting of 72 min aeration, 48 min settling and
24 min decanting, which should be advantageous in
treating wastewater with low COD:N ratio. SND
via nitrite was sustained for 122 days. However, for
e€ective SND via nitrite to take place in this pro-
posed process, careful control of parameters such as
the DO level, pH, FA and FH concentration, tem-
perature and aeration length is necessary. Also, the
continuous feeding of the in¯uent wastewater from
the bottom of the reactor was critical for e€ective
SND via nitrite.
Aeration time per cycle should not be too long or
short. Aeration of 72 min per cycle, and 12 h per
day was adequate and e€ective for this proposed
process. However, the data showed that longer
aeration time per cycle may be possible once the
sludge in the reactor is acclimated to nitratation-
suppressed condition (Fig. 10).
DO concentration during aeration should be con-
trolled two-dimensionally to e€ectively suppress
nitratation: control of the maximum DO concen-
tration reached at the end of aeration period, and
the DO concentration increase rate. The minimum
DO level was generally below 0.4 mg/l. Ideal maxi-
mum DO concentration should be 2.0±2.5 mg/l.
Ideal median DO level was 1.3 mg/l. Active denitri-
®cation took place in the initial phase of the aera-
tion period when the DO concentration was below
1.0 mg/l. However, during the later portion of the
period when the DO concentration exceeded
1.0 mg/l, nitri®cation was more active. Active deni-
tri®cation commenced again once the settling period
started and continued until the end of the cycle.
It is important to create the environment where
oxygen is de®cient and the sludge is in close contact
with the high level of FA in the in¯uent wastewater.
Under such condition, build-up of hydroxylamine
by the Nitrosomona has high probability to take
place. In this proposed process, continuous feeding
of the in¯uent from the bottom of the reactor
during settling and decanting periods was e€ective
in creating such environment.
AcknowledgementsÐThis research was funded jointly by
the Ministry of Science and Technology (MOST), and the
Kumho Construction Company, Ltd., of the Republic of
Korea.
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