War. Res. Vol. 27, No. 8, pp. 1339-1346,1993 0043-1354/93$6.00+ 0.

00
Printed in Great Britain, All rightsreserved Copyright © 1993PergamonPress Ltd

NON-FILAMENTOUS ACTIVATED SLUDGE BULKING IN

A LABORATORY SCALE SYSTEM
L. NOV~KI*O, L. L~atP.r.x2@, J. WANNERI@ and J. L. GARCIA-HERAS20
tDepartment of Water Technology and Environmental Engineering, Prague Institute of Chemical
Technology, Technick/~ 5, 166 28 Prague 6, Czech Republic and 2Department of Environmental
Engineering, CEIT, Apdo. 1555, 20080 San Sebastian, Spain

(First received January 1992; accepted in revised form January 1993)

Almraet--The paper describes the appearance of non-filamentous bulking in a lab-scale activated sludge
plant with nitrification-denitrification and reaeration of return sludge. The inlet part of the denitrification
zone was operated as an anoxic selector. Although filamentous microorganisms regularly o~urred, the
sludge settled properly. Deterioration of SVIs was observed after increasing the oleic acid concentration
in synthetic wastewater. The bulking period was related to the abundant growth of viscous zoogioeal
colonies. The intensive foam production resulted from the excessive appearance of exocellular polymers.
A structure of short intertwined fingered projections formed the zoogioeal flocs. After decreasing the oleic
acid proportion in the synthetic wastewater to the original level the zoogioeal riots gradually changed to
the well-settling sludge in 3 weeks.

Key words--activated sludge, bulking, filamentous microorganisms, foaming, selector, settling properties,
Zoogloea

INTRODUCTION which only settles slowly and compacts poorly be-

The activated sludge process works only as long as
the sludge settles and compacts in the secondary
settling tank to the extent that the return sludge
suspended solids concentration is high enough to
maintain the mixed liquor suspended solids concen-
tration (MISS) at a desired level.
Activated sludge solids separation problems can
be summarized (Jenkins, 1992) in several types. These
are, dispersed growth of microorganisms, non-
filamentous bulking, filamentous bulking, fungal
bulking, pin or pinpoint floe, blanket rising, foaming
and scum formation. Although the occurrence of any
above mentioned phenomena can negatively effect
the functioning of the secondary settling tank, sludge
bulking is one of the most significant.
Bulking definition
The term "bulking" can be analysed from different
viewpoints taking into consideration the reason
for sludge bulking development. Jenkins explains
filamentous bulking (Jenkins et al., 1986) as: " . . . a
macrostructure failure in which, in a sense, there is
too much macrostructure". This definition is based
on the fact that filamentous organisms that provide
the macrostructure are present in large numbers and
form the backbone of activated sludge floes (Sezgin
et al., 1978) and also interfere with the compaction
and settling of activated sludge. Eikelboom and van
Buijsen (1981) defined bulking sludge as: "Sludge
*Author to whom all correspondence should be addressed.
cause of an excessive growth of filamentous and/or
Zoogloea organisms". This definition also takes into
account a phenomenon of non-filamentous bulking.
Jenkins considers non-filamentous bulking separ-
ately. Whatever the approach, it seems that the term
"bulking" should be defined without any relation to
bulking reasons. It is a state when the sedimentation
rate of the act~mted sludge is low and the compaction
of activated sludge flocs is poor. This means the sludge
can be held up in the system but the clarifier function
is shifted to a considerably unstable state. The prob-
lem of compaction causes more negative effects. In a
case of insufficient compaction, the sludge can settle
well enough but cannot be thickened, the suspended
solids concentration at the bottom of the clarifier
becomes very low and a target value of MLSS
concentration in the aeration basin cannot be reached
or maintained. The compaction of activated sludge
plays a very important role especially in systems
with contact stabilization or separated regeneration
such as the regeneration-denitrification-nitrification
( R - D - N ) activated sludge process configuration.
Filamentous bulking
The most common sludge bulking problem is
caused by an excessive growth of filamentous micro-
organisms. The effect of filamentous organisms
on floc structure is the following: filaments stretch
out the flocs (diffused open structure) or form a
bridging structure. In both cases the filaments
mechanically prevent activated sludge floes from a
better compaction. This type of filamentous bulking

via ~ - - r 1339
1340 L. NovkK et al.
is well-known and described in detail in fiterature
(Albertson, 1991; Jenkins et al., 1986) along
with basic and common methods of filamentous
growth suppression based on a specific (selector
approach) and on a non-specific approach (the use
of selective toxicants) (Lakay et al., 1988). Neverthe-
less, the excessive growth of filamentous micro-
organisms need not always cause filamentous
bulking. It depends on the filament type, the shape
and the morphology of the floes (Pipes, 1979;
Wanner, 1992).
Non-filamentous bulking
The activated sludge bulking can also occur
when filamentous microorganisms are completely
missing. In this case we speak about the so-called
non-filamentous bulking; in the past termed viscous
bulking (Hale and Crarver, 1983; Jenkins eta/., 1986)
or zoogloeal bulking (Eikelboom and van Buijsen,
1981). The cause of non-filamentous bulking is pre-
dominantly an excessive growth of Zoogloea-like
microorganisms. In some cases other microorganisms
producing slime substances in excess can cause
similar irregularities (poly-P bacteria). Generally,
zoogloeal microorganisms form two types of
colonies: fingered and amorphous. If the zoogloeal
microorganisms are present in large amounts, the
exocellular slime causes the sludge floes to have
a voluminous character. In severe cases the slime
imparts a jelly-like consistency to the activated sludge
and causes foaming and scum formation. Polymeric
compounds produced by microorganisms which form
the exocellular slime usually contain a lot of water.
Very poor compaction of Zoogloea-like floes is
caused by a large amount of water absorbed by the
slime capsules, which prevents a rapid settling of
floes, and in the case of finger-like outgrowths for
mechanical reasons.
The deterioration of activated sludge settling prop-
erties in the case of non-filamentous bulking has
several different reasons:
--the composition of wastewater supports the
formation of zoogloeal growths, mainly the
high content of fatty and oleic compounds (van
Lecuwen, 1988);
--the high sludge loading and/or an influent contain-
ing an insufficient amount of certain nutrients. This
usually means a deficiency of nitrogen, phosphorus
and/or trace nutrients such as iron (Wood and
Tchobanoglous, 1974);
- - i n some cases selector systems significantly stimu-
late the growth of zoogloeal colonies (Gabb et al.,
1990;
- - i n anaerobic and/or anoxic systems which biologi-
cally remove phosphorus, an excessive growth of
poly-P bacteria, e.g. Acinetobacter spp which form
clusters of bacteria glued together by exocellular
polymers can cause some settling problems
(~tichov~t, 1990).
On the other hand almost no references to investi-
gations into the prevention and control of any bulk-
ing other than that caused by filamentous bacteria
could be found. Poor activated sludge settling proper-
ties due to viscous activated sludge, resulting from a
nutrient deficiency, cannot be satisfactorily con-
trolled by the addition of a polymer, chlorination
nor application of H202 (Jenkins et al., 1986). Suc-
cessful results for non-filamentous bulking control by
ozonation were achieved in South African activated
sludge plants treating fuel synthesis wastewater (van
Leeuwen, 1988).
Up to now the topic of non-filamentous bulking is
mentioned in literature very rarely. For that reason
the purpose of this paper is to describe a case of
zoogloeal bulking appearance in a bench-scale acti-
vated sludge plant with nitrification--denitrification,
in order to advance the knowledge of that type of
non-filamentous bulking.
EXPERIMENTALPROCEDURE
Bench-scale plant arrangement
The aim of the expenmentai programme of the plant was
to study the behaviour of the R-Ds-D-N process configur-
ation with two modifications, M I and M2, where the
internal recycle Rn was introduced into the denitrification
and regeneration zone respectively. Figure 1 depicts the flow
scheme. The experimental schedule was divided into two
periods:
Ist period---day 1-88, MI modification
2nd period---day 89-121, M2 modification.
Table 1 shows the nominal operational conditiom of the
system, Every tank of the plant was equipped with mechan-
icai stirring and a fine-bubble aeration in oxic tanks.
The composition of the synthetic wastewater is fisted in
Table 2. The wastewater was kept cool in a refrigerator to
prevent the decomposition of organic components.
Analytical methods
Samples were taken from the tanks during the transient
state and after reaching the steady state conditions which
were assumed to be achieved after at least three mild
retention times. Analyses of samples followed the pro-
cedures described in Standard Methods (APHA, 1985) and
An~lisis de las Aguas (Rodier, 1981). SVl-vaiues were
calculated from the 30 rain settled sludge volume determined
in unstirred 1000ml cylinders. Diluted DSVl-vaiues were
determined by measuring the volume occupied by the
diluted sludge after settling for 30 min. Dilution should
Fig. 1. Schematical outline of the R-Ds-D-N activated
sludge process configuration in MI and M2 modificationL
E, effluent; I, i n t u i t ; Ds, anoaic selector; D, denitrifl~tion
tank" N, nitrificationtank; It,regenerationtank;C, clarilk~,
R,, siudge r~'yc~; Rz(Ml), intmml r~'yc~ (Mi modifi-
cation); R~ (M2), internal recycle (M2 modification).
 Non-filamentous bulking 1341

Table I. Technological parameters of the MI and M2 modifications of the R - D s - D - N activated sludge process configuration
of the lab-scale model
MI modification M2 modification

Paranmer Unit R Ds D N R Ds D N

Volume I 10 3 7.5 30 10 3 7.5 30

HRT of mixed liquor in the individual reactors h -- 0.6 1.5 6 -- 0.6 1.5 6
HRT total h 10 10
Contact time of mixed liquor in the individual reactors h 4 0.4 0.6 2.4 1.3 0.2 0.6 2.4
Sludge recycle ratio (P~) 0.5 0.5
Internal recycle ratio (Rj) 1 1
SRT in the whole system d I1 11
Volumetric load on COD basis kg m -3 d - t 1.2 1.2
Flow rate I d -~ 121.1 121.2
Temperature °C 19 21
pH 7.5 7.5

R--regeneration tank; Ds---anoxic selector; N---nitrification tank; D--denitrification tank.

always be done to ensure that the sludge occupies a volume
of 20% or just less after 30 rain settling.
Microscopic ob~rvation of the biocenosls of activated
sludge was carried out according to Eikelboom and van
Buij~n (1981) and Jenkins et al. (1986). The exocellular
slime coat of zoogloeal formations was observed mivro-
scopically by reverse staining with India ink.
RESULTS AND DISCUSSION
From the beginning of the experimental pro-
gramme, an occurrence of Sphaerotilus natans, type
0041, and type 1701 was observed. These types of
filamentous microorganisms were present throughout
the whole period of the experiment and reached an
abundance of 4 and 5 (on a 6-point scale) (Jenkins
et al., 1986). See Fig. 2. They had no influence on the
deterioration of SVI. The sludge exhibited SVIs of
about 70 mi/g (Figs 3 and 4). From the 49th day up
to the 61st day of the experiment lecithin was not
available, and the composition of synthetic waste-
water was changed. The shortage of lecithin was
substituted by the same COD concentration of oleic
acid. After the 61st day the composition of synthetic
wastewater was the same as the original as listed in
Table 2.
The deterioration of SVI was observed after the
65th day of the experiment and it increased rapidly
during a few days reaching values above 300 mi/g
(Fig. 3). The increase of SVI was not connected to
an excessive growth of filamentous microorganisms
but with a very rarely observed non-filamentous
zoogloeal bulking. On the contrary, the number of
filamentous microorganisms significantly dropped
during the most excessive growth of zoogloeal micro-
organisms. From the initial state of an abundance of
5 before the zoogloeal excessive growth (day 49,
SVI---72 ml/g), the filaments reached a level of 3
during the most intensive period of zoogloeal bulking
(SVIs above 300 ml/g) (Fig. 2). The sludge had a
jelly-like viscous consistency with a great amount of
exoceilular slime causing viscous foaming in each
tank (Fig. 5) and scum formations on the claritier
surface (Fig. 6). The sludge became almost unfilter-
able. Microscopic examinations showed that neither
fmger-like nor amorphous-like colonies were formed
but a structure of short intertwined fingered projec-
tions appeared (Fig. 7). Under India ink reverse
staining large clear areas containing a low density
of cells and/or space-formations of zoogloeai out-
growths were observed (Fig. 8). Sedimentation prop-
erties were not bad but the thickening and
compaction were very poor. This resulted in a
decrease in suspended solids concentration at the
bottom of the clarifier and consequently in the regen-
eration tank but the overall performance of the
system was not influenced to a large extent. In order
to reduce the SVI every tank of the plant was cleaned
from the jelly cover and the D.O. concentration in
oxic zones was increased. From the 81st day SVI-
values began to decrease, the zoogloeal floes started
to change their consistency to normal flocs (Fig. 9).
The SVI decreased slowly in three weeks to values of
about 100 ml/g.

Table 2. Composition of synthetic wMtewater

Component In mg/I of COD Component In m8/I of weight
Glutami¢ acid 67 Na2CO 3 70
Yeast extract 57 NaHCO 3 400
Lecithin 71 CaCI=.6H20 54.3
Cellulose 22 FeEl 2 •61-12O 1.5
Casein 56 MsSO 4 •71-120 I 12.5
Oleic acid 95 K2 HPO4 I00
Lactose 44 KH2PO 4 40
Sucro~ 44 NH4CI 76.4
Staw~h 44 (NH4)=SO, 47. I

Total C O D , , 500 ms/I; TKN - 51 mg/I; N-NH4+ - 30 mll/I.

1342 L. NovkK et al.

U
t~
a
~3
<

2 S 15 24 31 49 56 63 70 77 84 91 98 105 113 119

Time (d)

Fig. 2. Abundance of filamentous microorganisms during the experiment.

The causes of all types of non-filamentous bulking
and the methods of its suppression are still unclear.
In the above described case it seems that the
zoogioeal growth was stimulated under steady state
conditions of the system by the change in synthetic
wastewater composition. The increased content of the
oleic component probably shifted the microorganism
population balance to a state where another form
of microorganism developed. Although it is not
clear if the real effect of the methods applied in
order to suppress the abundant Zoogloea-like micro-
organism growth was successful, it is supposed that
the decrease in the SVI was due to a delayed effect of
the original synthetic wastewater reconstitution after
the 61st day. Because the original wastewater still
contained oleic acid this could explain the slow
decrease of the SVI. A reduction in the SRT would
probably have increased the rate of the SVl-value
decline but it was not applied in order not to disturb
the experimental programme of the bench-scale
plant.
Considering that the combination of a regeneration
zone and a selector supports the growth of floe-form-
ing microorganisms (Grau et al., 1982), it is hypoth-
esized that the growth of zoogioeai microorganisms,
as a typical floe-former, was also supported to a large
extent and multiplied by the higher oleic component
content in the synthetic wastewater. In this context,

3SO
i mm •

300
Ill

Iiii •
250
• ,,, ",
oO
m 200 0 •
m o
II 0

rd'j 150 Oio 0

CZ:

II m
1oo O
0
.,,.. 9, "'." " %,, mmm •

5o

o I I I I I ! I
20 ,40 60 SO 100 120 14o
Time (d)

Fig. 3. Evolution of SVI- and DSVI-values.

 Non-filamentous bulking 1343

Fig. 4. Activated sludge from the lab-scale model, day 49, SVI = 72 ml/g (100 x , phase contrast).

Fig. 5. Foaming in the nitrification tank, day 74.

1344 L. NovkK et al.

Fig. 6. Formation of scum on the clarifier surface, day 74.

the decrease of filamentous microorganism abun-
dance during the period of non-filamentous bulking
could be explained by the competition between
filamentous and zoogloeal microorganism growth
as a result of the anoxic selector function, The
SVI-values reached a maximum above 300ml/g
and the DSVI-values slightly above 200 ml/g (Fig. 3).
This disproportion between SVI- and DSVI-values
along with the observation of sludge behaviour in
the sedimentation tank shows that there was no
problem with sludge settling (sludge formed large
flocs with a diameter of about 500#m exhibiting
good sedimentation properties) but the sludge did not
compact to higher MLSS-values than 5 g/1 at the
bottom of the clarifier. In the systems with separated
regeneration the decrease of MLSS concentration in

Fig. 7. Zoogloeal sludge flocs from the period of non-filamentous bulking, day 74, SVI ffi288 ml/g (100 x,
phase contrast).
 Non-filamentous bulking 1345

Fig. 8. Zoogloeal sludge flocs from the period of non-filamentous bulking, space maerostructure, day 74,
SVI = 288 ml/g (100 x , stained by India ink).

the regeneration tank can evoke a strong disturb- SUMMARY

ance in the regeneration and consequently the
accumulation function of the activated sludge. The
increase in the SVI-value above 300 ml/g took less
than 1 week, on the contrary it took more than 3
weeks to reach the SVI-value at a level of about
100 ml/g.
The lab-scale study with the R - D s - D - N process
configuration was performed with synthetic waste-
waters. The model of the activated sludge system
consisted of the sequence of denitrification-
nitrification reactors with the reaeration of return
1346 L. NOVAK et al.
sludge. The denitrification zone was equipped with an
anoxic selector. Besides the chemical analyses, the
settling properties were evaluated and microscopic
pictures of the activated sludge were examined.
Filamentous microorganisms were frequently pre-
sent in the biocenosis of activated sludge but their
presence did not worsen the settling properties. The
bulking period which was observed during the exper-
iment, was connected to the occurrence of viscous
jelly-like activated sludge. The enormous production
of exocellular polymers was related to the change in
wastewater composition when oleic acid was used
instead of lecithin. The non-filamentous bulking was
accompanied by an intensive foam production in
the system. The zoogloeal microorganisms formed
neither finger-like nor amorphous-like colonies but a
structure of short intertwined fingered projections.
The use of the original synthetic wastewater compo-
sition did not immediately stop the growth of
zoogloeal colonies but the biocenosis of the bulking
and foaming sludge gradually changed. The viscous
zoogloeal colonies were replaced by compact well-
settling flocs.
Acknowledgement--The authors wish to express their ac-
knowledgement to the Basque Government for its financial
support.
REFERENCES
Albertson O. E. (1991) Bulking sludge control--progress,
practice and problems. ;Vat. Sci. Technol. 23, Kyoto,
835-846.
APHA (1985) Standard Methods for the Examination of
Water and Wastewater, 16th edition. APHA, Washing-
ton, D.C.
Eikelboom D. H. and van Buijsen H. J. (1981) Microscopic
Sludge Investigation Manual. IMG-TNO Report A94a,
Delft.
Gabb D. M. D., Still D. A., Ekama G. A., Jenkins D. and
Marais D. v. R. (1991) The selector effect on filamentous
bulking in long sludge age activated sludge systems.
War. Sci. TechnoL 23, Kyoto, 867-877.
Grau P., Chudoba J. and Dohanyos M. (1982) Theory
and pract/ce of sccumulation-regeneration approach to
the control of activated sludge filamentous bulking. In
Bulking of Activated Sludge: Preoentativeand Remedial
Methods ~ t e d by Chambers B, and Tomlinson E. J.)
Ellis Horwood, Chichester.
Hale F. D. and Garver S. R. (1983) Viscous bulking
of activated sludge. Presented at the 56th Annual
Conf. Water Pollut. Control Fed., Atlanta, GA 2-7
October.
Jenkins D. (1992) Towards a comprehensive model of
activated sludge bulking and foaming. Wat. Sci. Technol.
25(6), 215-230.
Jenkins D., Richard M. G. and Dalgger G. T. (1986)
Manual on the Causes and Control of Activated Sludge
Bulking and Foaming. Water Research Commission,
Pretoria.
Lakay M. T., Wentzel M. C., Ekama G. A. and Marais
G. v. R. (1988) Bulking control with chlorination in a
nutrient removal activated sludge system. Water SA 14,
35-42.
van Leeuwen J. (1988) Ozonation for non-filamentous bulk-
ing control in an activated sludge plant treating fuel
synthesis waste water. Water SA 15, 127-132.
Pipes W. O. (1979) Bulking, deflocculation, and pinpoint
floc. J. War. Pollut. Control Fed. 51, 172-182.
Rodier J. (1981) An~lisis de las Aguas. Omega, Barcelona.
Sezgin M., Jenkins D. and Parker D. S. (1978) A unified
theory of filamentous activated sludge bulking. J. War.
Pollut. Control Fed. 50, 362-381.
~tichov~ O. (1990) Study of the mechanism of biological
enhanced phosphorus removal in SBRs. M.Sc. thesis,
Prague Institute of Chemical Technology, Prague (in
Czech).
Wanner J. (1992) Comparison of biocenoses from con-
tinuous and sequencing batch reactors. Wat. Sci. Technol.
25(6), 239-249.
Wood D. K. and Tchobanogious G. T. (1974) Trace
elements in biological waste treatment with specific refer-
ence to the activated sludge process. Presented at the 29th
Industrial Waste Conference, Pardue University, IN.