Biochemical Engineering Journal 32 (2006) 127134

A novel technology for bulking control in biological wastewater

treatment plant for pulp and paper making industry
Y.F. Tsang

, H. Chua, S.N. Sin, C.Y. Tam

Department of Civil and Structural Engineering, The Hong Kong Polytechnic University, Hung Horn, Hong Kong, China
Received 5 May 2006; received in revised form 3 August 2006; accepted 4 August 2006
Abstract
Filamentous foaming and bulking in activated sludge process are common problems in pulp and paper mill wastewater treatment processes.
In this study, the predominant foaming and bulking causing lamentous microorganism in the pulp and paper mill efuent treatment system was
identied. Its growth kinetics was studied and used as a theoretical basis to develop an optimal operational strategy for foaming and bulking
preventive control. Filamentous actinomycetes, Nocardia amarae was found to be the predominant lamentous microorganisms. The kinetics
study revealed that the Food to Microorganism (F/M) ratio was a critical factor affecting the excessive growth of N. amarae. Accordingly,
a novel Feast-Fast Operation (FFO) strategy was developed, in which the sludge microbes underwent a repetitive switch between high and
low F/M ratio during the course of wastewater treatment. Feast-to-Fast Loading Ratio (FFLR) was proposed and used as the parameter for
optimization of FFO performance. At FFLR of 4 and 3.5, the sludge volume index (SVI) rapidly reduced to a healthy level of 70 ml g
1
and the
systems were free from stable foam, while the biochemical oxygen demand (BOD) removal efciency remained above 96.2%. The FFO strategy
effectively suppressed lamentous overgrowth and improved the settleability of activated sludge without adversely affecting the BOD removal
efciency.
2006 Elsevier B.V. All rights reserved.
Keywords: Bulking control; Pulp and paper mill efuent; Waste-water treatment; Filamentous bacteria; Bioreactors; Optimization
1. Introduction
Filamentous overgrowth is a ubiquitous operational problem
in activated sludge process for wastewater treatment, especially
in pulp and paper mill efuent treatment systems [1]. The excess
growth of lamentous microorganisms causes bulking that dete-
riorates sludge settleability, resulting in decreased sludge set-
tling rate and incompact sludge blanket, usually indicated by an
increased SVI [2]. Filamentous species attach to and stabilize air
bubbles to become thick, stable, persistent and scum-like foam.
Foaminterferes level sensors and hinders sequencing batch reac-
tor (SBR) operations. Spillage of foam from aeration tanks to
walkways causes safety problems. Dried foam results in air-
borne pathogens that cause public health problems. Thick foam
blocks scumremoval systems andreduces oxygensupply. Above

Corresponding author at: W614a, Department of Civil and Structural Engi-

neering, The Hong Kong Polytechnic University, Hung Horn, Hong Kong,
China. Tel.: +852 2766 4486; fax: +852 2334 6389.
E-mail address: 04900146r@polyu.edu.hk (Y.F. Tsang).
all, both bulking and foaming increase efuent BOD and sus-
pended solids (SS) levels due to the sludge or the foam escaping
from clariers.
Bulking and foaming problems have been alleviated by addi-
tion of toxic chemicals such as chlorine or hydrogen peroxide
to the aeration tank or the return sludge line, to kill the l-
amentous microorganisms selectively [3]. Metal ions such as
calcium, magnesium, iron [1,4], synthetic polymer [5], and
multi-component additive [6] are observed to control bulking
effectively. Application of mineral talc also is effective in sludge
settleability improvement in paper mill efuent treatment plant
[7]. However, these chemical treatment methods are costly and,
most important, they only offer short-term solution as bulking
and foaming will resume when chemical additions are stopped.
Kinetic selection is an alternative to control lamentous over-
growth specically under low F/M condition, it was rst pub-
lished by Chudoba et al. [8]. The theory stated that substrate
concentration gradient favors the growth of oe-formers, instead
of lamentous bacteria, among the activated sludge microbes.
Successful applications have been accomplished by installation
of selector upstream to the aeration tank [9], intermittently fed
1369-703X/$ see front matter 2006 Elsevier B.V. All rights reserved.
doi:10.1016/j.bej.2006.08.014
128 Y.F. Tsang et al. / Biochemical Engineering Journal 32 (2006) 127134
Fig. 1. The activated sludge process treating pulp and paper mill efuent.
operation [10,11], or implementation of plug ow conditions
[12]. Although many studies have been carried out, the root
causes of lamentous overgrowth are still not fully understood
and contradicting views remain [13]. Since universal lamen-
tous control strategy is still unavailable, a specic analysis is
necessary to determine the control method for each problem
case.
The objectives of this study were: (1) to identify the pre-
dominant lamentous microorganism in bulking sludge; (2) to
develop a novel activated sludge process (FFO) based on the
growth kinetics study of the lamentous microorganisms; (3) to
evaluate the effectiveness of the modied process on lamentous
control.
2. Materials and methods
2.1. Identication of predominant lamentous
microorganism
The microorganisms in activated sludge samples collected
from the paper mill wastewater treatment plant were isolated
by plate cultures with Czapeks agar supplemented with 0.4%
yeast extract, incubated at 28

C for 3 weeks. The microbes

were identied according to the criteria adopted by Blackall et
al. [14] and Jenkins et al. [2]. The isolated bacteria were cultured
in 500 ml asks with 100 ml sterilized LBmedium, on an orbital
shaker at 28

C and 200 rpm for 120 h.

2.2. Kinetic selection studies
The predominant lamentous microorganism(later identied
as Nocardia amarae, ATCC 27810) and Pseduomonas aerugi-
nosa (CRCC 10261) were used to represent the lamentous and
non-lamentous oc-forming bacteria in activated sludge. The
cultures were maintained on yeast-extract-glucose agar slants at
24

C.
The inocula of N. amarae and P. aeruginosa used for the
kinetics studies were derived fromthe agar slants and maintained
at 30

C for 72 and 24 h, respectively. Minimal salt medium

(MSM) was prepared according to the method proposed by Chua
et al. [15]. The MSMwas adjusted to pH6.8 with NaOHsolution
and autoclaved. Paper mill efuent, which acted as the carbon
source, was ltered through a 0.45 mmembrane lter and ster-
ilized. Ninety milliliters of MSM, 10 ml of paper mill efuent
and 5 ml of inoculum were added into a 500 ml bafed conical
ask. Both types of bacteria were cultured at 30

C and 200 rpm

for a period required to enter the stationary phase.
2.3. The activated sludge simulator system
The simulator system was comprised of two 3-l aeration
columns and a settling tank (Fig. 1). Both aeration columns
received the paper mill wastewater and return activated sludge
(RAS) at pre-determined ow rates. The system was operated
under conventional conditions for 36 days to attain stable oper-
ation before switching to FFO mode, during which the two
aeration columns were operated at different hydraulic reten-
tion times (HRT), return sludge rates and MLSS concentrations,
resulting in relatively high and low F/M ratios. The simulator
systemwas operated in an air-conditioned laboratory at the tem-
perature of 25 1

C. All operating conditions of conventional

and FFO mode are summarized in Table 1.
2.4. Seed and inuent
Activated sludge with excessive lamentous growth fromthe
paper mill wastewater treatment plant was used as seed for the
lab-scale bioreactor. The collected sludge was screened with a
2-mm sieve to remove coarse particles before transferring to the
simulator system for start-up. The efuent from the dissolved
air otation unit, used as the inuent of the treatment system,
was collected and immediately refrigerated at 4

C. The char-
acteristics of the inuent of system are shown in Table 2. Prior
to the experiment, the inuent was supplemented with nutrients
(N: urea and P: phosphoric acid) to maintain a BOD:N:P ratio
of 100: 5:1.
Y.F. Tsang et al. / Biochemical Engineering Journal 32 (2006) 127134 129
Table 1
Operational conditions in the conventional operation and FFO programmes
System condition Conventional operation during start-up FFO 1 FFO 2 FFO 3
Feasting Fasting Feasting Fasting Feasting Fasting
Effective volume (l) 6 3 3 3 3 3 3
Inuent ow rate (l h
1
) 0.6 0.4 0.2 0.35 0.25 0.3 0.3
HRT (h) 10.0 7.5 15.0 8.6 12.0 10.0 10.0
pH 67 67 67 67 67 67 67
Dissolved oxygen (mg l
1
) 24 24 24 24 24 24 24
Inuent BOD (mg l
1
) 500 500 500 500 500 500 500
Sludge wastage (l day
1
) 0.4 0.2 0.2 0.2 0.2 0.2 0.2
SRT (d) 12 12 12 12 12 12 12
MLSS (mg l
1
) 3200 2100 4200 1800 4500 1500 4800
F/M (mgBODmgMLSS
1
day
1
) 0.38 0.76 0.19 0.78 0.22 0.80 0.25
FFLR N.A. 4.0 3.5 3.2
Table 2
Characteristics of the inuent of the activated sludge simulator
Parameter Concentration
Dissolved COD (mg l
1
) 905 52.3
BOD (mg l
1
) 496 15.4
Suspended solid (mg l
1
) 30.2 3.2
pH 6.5 0.32
2.5. Analytical methods
All parameters of mixed liquor and wastewater quality were
determined according to standard methods [16]. Biomass was
quantied in terms of MLSS because MLVSS/MLSS was often
close to 1.0 in this system, which had very low mineral content.
A foam measurement method was applied to assess the
effectiveness of the foam control strategy. A 500 ml sample of
mixed liquor was placed in a 11 graduated cylinder and aer-
ated, from the bottom, at an air ow rate of 21 min
1
through a
sintered-sand diffuser for 60 s. The mixture was then left under
quiescent conditions for another 60 s. The instantaneous foam
height, as indicated by an inverted conical polyethylene marker,
was recorded at 15-s intervals throughout the 120-s period.
The degree of foaming was measured by the maximum vol-
ume of foam produced in the aeration period, where 6080 and
020 ml of foam were typical values for severely foaming and
healthysludge, respectively. The degree of foamingcouldalsobe
expressed by dividing the maximum volume of foam produced
by the sample volume. These results could then be used to fore-
cast the actual depth of the foam or scum layer in the treatment
plant. The foam stability was quantied by dividing the mini-
mum volume of foam in the quiescent period by the maximum
volume of foam in the aeration period, where 0 and 1 are the
normalized minimum and maximum, respectively, of foam sta-
bility. This procedure was rst developed and successfully used,
both in the laboratory and sewage treatment plants, for a wide
range of activated sludge samples with different grease, pro-
tein and lamentous bacterial contents producing reliable and
reproducible results [17].
3. Results and discussion
3.1. The predominant lamentous bacteria
The micrograph of the sludge sample of bulk oe from
paper mill efuent treatment system is illustrated in Fig. 2,
which shows an overgrowth of lamentous bacteria with
right-angled branching, and the lament dimensions were
0.51.0 m80160 m. The colonies on plate cultures
demonstrated aerial hyphae that gave a supercially matte and
chalky appearance. The generation time of the isolated bacteria
in shake ask cultures at 28

C was 10.5 h. The isolated bacteria

were found Gram-positive and Neisser-negative, and could not
Fig. 2. The predominant N. amarae in the sludge sample of bulk oe from paper mill efuent treatment system (left: 200, right: 400).
130 Y.F. Tsang et al. / Biochemical Engineering Journal 32 (2006) 127134
Fig. 3. LineweaverBurk plot of N. amarae.
survive after an8-hincubationat 50

Cunder heat resistance test.

These morphological and physiological properties identied the
bacteria as N. amarae [14,2]. This lamentous actinomycetes is
commonly found in biological systems treating paper mill efu-
ent [18]. The growth of N. amarae was supported as the efuent
that contains organic additives, oil-based defoamers and wood
extractives [15].
3.2. Kinetic selection studies of lamentous and
oc-forming bacteria
The maximumspecic growth rate (
m
) and the half-velocity
constant (K
s
) of the Monod kinetics model were obtained from
the batch culture growth data, followed by processing with
LineweaverBurk equations and linear regression techniques.
The double reciprocal plots for lamentous and oc-forming
bacteria are demonstrated in Figs. 3 and 4. The
m
and K
s
of
N. amarae were 0.073 h
1
and 546 mg l
1
, respectively. The
m
and K
s
of N. aeruginosa were 1.25 h
1
and 2148 mg l
1
, respec-
tively. According to the results, both
m
and K
s
of N. amarea
were signicantly lower than that of P. aeruginosa. These obser-
vations were in agreement with the study by Chudoba et al. [8]
that lamentous bacteria are slow-growing microorganisms that
possess
m
and K
s
lower than non-lamentous oc-formers.
The specic growth rates of lamentous N. amarae and oc-
forming P. aeruginosa under different F/M ratios are shown
in Fig. 5. The cut-off value of F/M ratio was observed at
0.52 mgBODmgMLSS
1
day
1
, below which N. amarae grew
faster than P. aeruginosa. While at F/M ratio above the cut-
Fig. 4. LineweaverBurk plot of P. aeruginosa.
Fig. 5. Growth kinetics of N. amarae and P. aeruginosa.
off value, the growth rate was higher for P. aeruginosa. These
results showed that lamentous N. amarae grew faster and
had a stronger afnity towards the substrate, and turned out
to be the predominant strain at low substrate concentrations,
while the oc-forming P. aeruginosa required higher organ-
ics concentrations to grow, and dominated at higher substrate
levels [15]. Such growth behaviors implied that lamentous
overgrowth was inevitable in most activated sludge processes
that are continually exposed to the F/M ratio between 0.2 and
0.5 mgBODmgMLSS
1
day
1
.
3.3. Development of the feast fast operation strategy
For the FFO strategy, the simulator system was used as
the realization technique and FFLR was used as the con-
trol parameter. The kinetics selection results revealed that
activated sludge process operating at F/M ratio of above
0.52 mgBODmgMLSS
1
day
1
rendered lamentous bacte-
ria to be at a competitive disadvantage relative to the non-
lamentous microbes. Hence, high F/M ratio was regarded
as a prerequisite for effective selection of the desired non-
lamentous bacteria from the sludge population. Moreover,
other studies found that the sludge bacteria, after exposing to
a high F/M environment, required sufcient time for oxidation
of adsorbed/accumulated substrates [19,20]. It was also inves-
tigated that activated sludge with good settleability was best
achieved when the microorganisms were periodically exposed
to high substrate concentration and long-lasting near-starvation
conditions [21]. This information provided the theoretical basis
for FFO development in this study. In the dual-reactor sys-
tem, the feasting column was operated at a higher F/M ratio of
0.8 mgBODmgMLSS
1
day
1
, while the fasting column was
run at a lower F/M ratio of 0.2 mgBODmgMLSS
1
day
1
. The
FFO mode allowed an overall process F/M ratio to maintain
within the normal range while avoiding prolonged exposure of
the activated sludge to an F/M ratio below the cut-off value of
0.52 mgBODmgMLSS
1
day
1
.
3.4. Start-up of the treatment process
The simulator systems were operated with conditions as
shown in Table 1. Each dual-reactor system was started up with
Y.F. Tsang et al. / Biochemical Engineering Journal 32 (2006) 127134 131
Table 3
Sludge production and return sludge rate at different FFLRs during FFO programmes
Module System condition
FFLR=4.0 FFLR=3.5 FFLR=3.2
Feasting Fasting Feasting Fasting Feasting Fasting
Inuent ow rate (l h
1
) 0.4 0.2 0.35 0.25 0.3 0.3
MLSS (mg l
1
) 2100 4200 1800 4500 1500 4800
Sludge production rate (mg h
1
) 840 840 630 1125 450 1440
Proportion of sludge production (%) 50 50 36 64 24 76
Return sludge rate (l h
1
) 0.05 0.05 0.04 0.08 0.11 2.20
Proportion of sludge replenishment (%) 50 50 33 67 5 95
conventional operation for microbial acclimatization. The sys-
tems were fed with paper mill efuent and seeded with bulking
and foaming sludge obtained from the paper mill wastewa-
ter treatment plant. In each system, the inuent ow and the
return sludge rate were maintained at 0.6 and 0.8 1 h
1
, respec-
tively, which resulted in MLSS of 3200 mg l
1
and F/M ratio
of 0.38 mgBODmgMLSS
1
day
1
. The initial values of SVI,
degree of foaming and foam stability, at 235 ml g
1
, 70 ml
and 0.28, respectively, have increased and became steady at
275 ml g
1
, 75 ml and 0.35, respectively. At the same time, the
BOD removal efciency increased from 86% to 97.5%. These
observations indicated that the quantity of lamentous bacteria
within the sludge ecosystemincreased under conventional oper-
ation with a typical F/Mratio of 0.4 mgBODmgMLSS
1
day
1
during the 36-day start-up period. A sludge sample of bulk oe
at this point showed an overgrowth and overwhelming domi-
nance of lamentous N. amarae (Fig. 6). These ndings also
provided further support that paper mill wastewater treatment
plants with conventional F/Mratio were subjected to lamentous
overgrowth. Fromday 37, the systems were switched to the FFO
programme with predetermined conditions (Tables 1 and 3). The
target MLSS of each column was attained by adjusting the return
sludge rate according to the inuent ow rate, the MLSS of the
aeration column and the suspended solid concentration of the
settled sludge.
3.5. Process performances under FFO strategy
Three simulator systems were operated under different con-
ditions in order to study the treatment performance of FFO
Fig. 6. Filamentous dominance during the system start-up period (200).
strategy for foaming and bulking control in paper mill efu-
ent treatment plant. These operating conditions were main-
tained at the same overall process F/M ratio of 0.38 and total
inuent ow rate of 0.6 1 h
1
as in the conventional opera-
tion during start-up. However, the relative F/M ratios within
each system were varied in order to optimize the effective-
ness in lamentous control. The F/M ratios were allocated
through manipulating the inuent ow rate and MLSS in both
feasting and fasting columns within each system. Hence, the
Feast-to-Fast Loading Ratio (FFLR), which is dened as
the proportion of the F/M ratios between the feasting column
and the fasting column, was used as the parameter for process
optimization.
The performances of FFO programme after the start-up
period are shown in Figs. 710. Figs. 7 and 8 show the his-
tory of F/M ratios and MLSS during the FFO operation. The
ne-tuning of the F/M ratios allowed the exposure of the acti-
vated sludge microbes to different degree of feasting and fasting
conditions. The prole of degree of foaming, foam stability, and
SVI are illustrated in Figs. 9 and 10. Generally, the FFO strat-
egy gave a quick response in lamentous control as indicated by
the improved settleability and reduced foaming capacity of the
systems.
At FFLR of 4.0, the degree of foaming dropped from 75 to
20 ml, while foam stability was below 0.05 within the 24-day
operation of FFO programme (Fig. 9). The foam measurements
in two columns were very similar and the sludge was almost free
from stable foam from day 60 onwards. The sludge settleability
also responded rapidly to the FFOoperation. After the simulator
was switched to the FFO mode, the SVI returned rapidly to a
healthy level of 70 ml g
1
within 22 days. The SVI dropped
further, and gradually stabilized at around 60 ml g
1
during the
next 30-day operation (Fig. 10). At the steady state condition,
SVI values of around 55 and 65 ml g
1
were observed in the
feasting and fasting columns, respectively.
At FFLR of 3.5, the degree of foaming and foam stability
dropped to 20 ml and below 0.05, respectively, within the 33-
day FFO programme. After the day 69, the system was almost
free from stable foam (Fig. 9). For the sludge settleability, the
SVI returned rapidly to a healthy level of 70 ml g
1
within 33
days and became stabilized at SVI of around 65 ml g
1
on day
70 (Fig. 10). When the feasting and fasting columns were at
steady state, the SVI were found at 60 and 75 ml g
1
, respec-
tively. Although the process response at FFLR of 3.5 was not as
132 Y.F. Tsang et al. / Biochemical Engineering Journal 32 (2006) 127134
Fig. 7. The MLSS prole during operation.
Fig. 8. The F/M prole during operation.
Fig. 9. The prole of degree of foaming and foam stability during operation.
Y.F. Tsang et al. / Biochemical Engineering Journal 32 (2006) 127134 133
Fig. 10. The SVI prole during operation.
fast as that at FFLR of 4.0, the performance in terms of lamen-
tous control was still satisfactory.
For the system operated at FFLR of 3.2, the degree of foam-
ing and foam stability slightly dropped to 60 and maintained at
around 2.8, respectively (Fig. 9) and stable foam was frequently
observed. The SVI reduced to a stable value of 220 ml g
1
(Fig. 10). The system failed in controlling of foaming and bulk-
ing under this FFLR value.
The process conguration under the FFO strategy enabled
the settled activated sludge fromthe settling tank to be randomly
recycledbetweenthe feastingandfastingcolumns, thus avoiding
a prolonged exposure of the sludge to a low F/M condition.
This created a favorable environment for the non-lamentous
oc-forming bacteria to gain dominance in the ecosystem. In
addition, the effectiveness of lamentous control was enhanced
by increasing the FFLR (Figs. 9 and 10), which controlled the
specic F/Mratios towhichthe activatedsludge microorganisms
were exposed.
FFLR was affected by many operating parameters, including
inuent ow rate, MLSS concentration and return sludge rate
(Table 3). FFLR also correlated to the sludge production rate
from the feasting and fasting column, and the proportion of the
return sludge that replenished to each aeration columns. There-
fore, a narrowchange in FFLRresulted in a signicant change in
various operating conditions (Table 3). Lower FFLR decreased
the rate of sludge production and the return sludge replenishment
in the feasting column. However, it increased the same in fasting
column (Table 3). Consequently, a large quantity of sludge was
forced to expose to a fasting condition that resulted in higher
degree of sludge starvation. For the case at FFLR of 3.2, as
high as 76% of the sludge production of the system was derived
from the fasting column, and about 95% of the return sludge
owed back into the fasting column (Table 3), in which the
F/M ratio was maintained at 0.25 mgBODmgMLSS
1
day
1
(Table 1). In this condition, excess growth of lamentous bac-
teria was promoted, resulting in unsolved bulking and foaming
problems.
The ultimate function of the wastewater treatment system is
to remove the organic matter to meet the corresponding dis-
charge standard (BOD20 mg l
1
) set by the local government
[22]. Table 4 indicates that the BOD removal efciencies during
the process of FFOprogramme were not substantially lower than
that during conventional operation. The systemshowed the BOD
removal efciency of 98.1% with treated efuent BOD level
at 9.45 0.34 mg l
1
(n =19) during start-up operation. After
switching to FFO strategy, the BOD removal efciencies were
found to be 96.2% (n =20), 97.0% (n =20) and 97.4% (n =20),
for FFLRs of 4.0, 3.5 and 3.2, respectively. All the FFO operat-
ing conditions were able to meet the BOD discharge standard.
However, the BOD concentration of treated efuent was found
very close to the upper limit of the standard at FFLR of 4.0.
The slightly lowered BOD removal efciency was attributed to
the shorter HRT in the feasting column, and the lower degree of
sludge starvation in the system. Higher FFLR responded more
rapidly to bulking and foaming control, whereas, lower FFLR
enhanced the BOD removal. Hence, the FFLR was an appro-
priate tool for optimizing the FFO strategy between lamentous
control and BOD removal performance.
Table 4
BOD removal performance at different operating conditions
System condition Conventional operation
during start-up (n =19)
FFO 1 FFLR=4.0 (n =20) FFO 2 FFLR=3.5 (n =20) FFO 3 FFLR=3.2 (n =20)
Inuent BOD (mg l
1
) 500 16.1 493 13.7 493 13.7 493 13.7
Efuent BOD (mg l
1
) 9.5 0.34 19.2 1.77 15.1 0.34 12.8 0.27
BOD removal efciency (%) 98.1 96.2 97.0 97.4
134 Y.F. Tsang et al. / Biochemical Engineering Journal 32 (2006) 127134
4. Conclusions
N. amarae, which was induced by low F/M ratio, was found
to be the predominant lamentous species in the activated sludge
process treating paper mill efuent. The excess growth of this
actinomycetes was effectively controlled by the implementa-
tion of the novel FFO strategy. The FFLR proposed in this study
allowed ne-tuning of the relative F/Mratio between the feasting
and fasting columns, and hence can be used as the parameter for
optimization of FFO performance on foaming and bulking con-
trol. The SVI was rapidly reduced from 275 ml g
1
to a healthy
level of 70 ml g
1
and stable foamwas extinguished after imple-
mentation of FFO at FFLR of 4 and 3.5. The FFO strategy
successfully suppressed lamentous overgrowth and improved
the settleability of activated sludge without adversely affecting
the performance of BOD removal.
Acknowledgement
The authors wish to thank the Hong Kong Polytechnic Uni-
versity Research grants for the nancial support.
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