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Strength characteristics of aerobic granular sludge

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DOI: 10.2166/wst.2012.837 · Source: PubMed

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1 © IWA Publishing 2011 Water Science & Technology | in press | 2011

Strength characteristics of aerobic granular sludge

A. Nor-Anuar, Z. Ujang, M. C. M. van Loosdrecht, M. K. de Kreuk
and G. Olsson

ABSTRACT

Aerobic granular sludge has a number of advantages over conventional activated sludge flocs, such A. Nor-Anuar (corresponding author)
Z. Ujang
as cohesive and strong matrix, fast settling characteristic, high biomass retention and ability to Institute of Environmental and Water Resource
Management (IPASA),
withstand high organic loadings, all aspects leading towards a compact reactor system. Still there are Universiti Teknologi Malaysia,
81310 Johor,
very few studies on the strength of aerobic granules. A procedure that has been used previously for Malaysia
E-mail: aznah@utm.my
anaerobic granular sludge strength analysis was adapted and used in this study. A new coefficient
M. C. M. van Loosdrecht
was introduced, called a stability coefficient (S), to quantify the strength of the aerobic granules.
M. K. de Kreuk
Indicators were also developed based on the strength analysis results, in order to categorize aerobic Department of Biotechnology,
Delft University of Technology,
granules into three levels of strength, i.e. very strong (very stable), strong (stable) and not strong Julianaalan 67,
NL-2628 BC,
(not stable). The results indicate that aerobic granules grown on acetate were stronger (high Delft,
density: >150 g T SSL
1 and low S value: 5%) than granules developed on sewage as influent. A lower The Netherlands

value of S indicates a higher stability of the granules. G. Olsson

Department of Industrial Electrical Engineering
Key words | aerobic granular sludge, granule density, stability, strength and Automation,
Lund University,
S-221 00 Lund,
Sweden

INTRODUCTION

Aerobic granular sludge is a recent innovation in
biological wastewater treatment. Granulation is a
self-immobilization process in which microorganisms
agglomerate and develop to dense and compact biomass
granules. Aerobic granular sludge has a number of advan-
tages over conventional activated sludge flocs, such as
regular and strong structure, good settling ability (sludge
volume index (SVI) <50 mL g
1), high biomass retention
(up to 20 g SSL
1), and ability to withstand high organic
loadings. Furthermore, they are able to convert organic
substrates, nitrogen compounds and phosphate simul-
taneously at high removal efficiencies (Dulekgurgen
Q1 et al. ; De Kreuk & Van Loosdrecht ; Gao
et al. ). In order to be able to design a robust system
with the aerobic granular sludge technology, knowledge
about the strength and stability of the granules is essential.
Sufficient shear stress is needed for the formation of stable
and dense granular sludge (Beun et al. ; Liu & Tay
; Liu et al. ). However, extensive shear stress in
the reactor (e.g. by mechanical mixing, aerated mixing
or design of the airlift) affects the aerobic granular
sludge settling characteristics or worse, might damage
the granules. In addition, sludge handling (processing
surplus sludge, inoculation of new reactors etc.) often
requires pumping of a water/granule mixture, which also
leads to high stress on the granules and thus to breakage.
Damaging aerobic granules during reactor operation or
sludge handling can lead to decreased settling ability
and subsequent washout of the biomass. Changes in gran-
ule diameter might influence the efficiency of
simultaneous nitrification and denitrification, because of
a changed anoxic volume inside the granules (De Kreuk &
Van Loosdrecht ). In order to understand the capabi- Q2
lity of granules to withstand shear, more knowledge
about the physical strength of aerobic granular sludge is
needed. The granule strength is defined as the resistance
to attrition and/or breaking by a mechanical force or the
liquid shear stress. A procedure that has been used pre-
viously for anaerobic granular sludge strength analysis
was adapted and used in this study (Pereboom ). This
study aimed to develop a new aerobic granule strength
characterisation test and to be able to predict the stability
of aerobic granule in full scale systems. A new coefficient
was introduced, called a stability coefficient (S), to quantify

doi: 10.2166/wst.2011.837
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2 A. Nor-Anuar et al. | Aerobic granules strength
the strength of the aerobic granules. Indicators were also
developed based on the strength analysis results, in order
to categorize aerobic granules into three levels of strength,
i.e. very strong, strong and not strong.
METHODS
Aerobic granular sludge samples
Samples of aerobic granules from two different temperatures
have been studied, i.e. aerobic granules developed at 20 and
W W
30 C. Granules developed at temperature 20 C were
sampled from two different sources, (1) from a Nereda
pilot plant, operated by DHV (www.dhv.com) at the waste-
water treatment plant and (2) from a 3-L laboratory-scale
reactor at Delft University of Technology, The Netherlands.
Figure 1 shows the picture of pilot plant and sampled
granules.
W
Meanwhile, granules developed at 30 C were sampled
from a 3-L laboratory-scale reactor at Universiti Teknologi
Malaysia. All reactors were operated in Sequencing Batch
(SBR) mode and description of the operating conditions of
the aerobic granules samples is given in Table 1. During
start-up, biomass concentration in the reactor was main-
tained around 8–10 g TSS/L. Aerobic granules was
collected during steady state operation of the reactor and
sieved, granules with the size >2 mm were analysed as for
this research.
Figure 1 | (a) Photograph of Granular Sludge Bioreactor (GSBR): Nereda™ Pilot Plant. (b) Picture of aerobic granular sludge sampled from GSBR. Granules were observed using inverted
microscope with magnification of 40× and photo were taken using LeicaQWin Image Analyser.
Water Science & Technology | in press | 2011
Experimental set-up: strength measurement
The strength was measured by shear experiments in a 2 L
standard geometry vessel (Tv ¼ 0.133 m) at 200 RPM stirrer
(D ¼ 0.075 m) speed for 10 min. The vessel equipped with
a lid and metal turbine type stirrer. The rotational speed
was measured by an optical revolution meter. The position
of the turbine stirrer in the vessel is 0.015 cm from the
bottom. Before conducting the shear experiments, physical
characteristics of the aerobic granules were analysed. Size
distributions of aerobic granules were determined by sieving
the granules using 0.2, 0.4 and 0.6 mm sieves. The total sus-
pended solid (TSS) concentration of the total sample and
the dry weight of each fraction were determined by drying
the samples for 24 h at 105 C (APHA ). Solid mass of
W
granule or solid density (ρ) was determined with a dextran
blue method (Beun et al. ). Changes in diameter of
the aerobic granules (φ) before and after the shear test
were observed using an Image Analyser. This observation
is also important for over viewing the ruptured granules.
Determination of strength of aerobic granular sludge
Strength of the aerobic granules was measured based on the
stability of aerobic granules against shear stress during reactor
operation. Therefore, a procedure was developed as
described in Table 2, to evaluate the strength of aerobic gran-
ules. This test procedure was based on the premise that if the
aerobic granules were subjected to fluid shear stress beyond a
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3 A. Nor-Anuar et al. | Aerobic granules strength Water Science & Technology | in press | 2011

certain limit, the quantity of sludge released in the surround-

Pre-treated sewage was taken directly

ing fluid would be a function of shear strength of the aerobic

stored at 4 C for a maximum of

after the 6 mm screen and were

Activated sludge was taken from an aeration tank of

Method of aeration used is diffused aeration system and oxygen in the reactor was controlled between 2 and 3 mg/L during aeration. pH was not
granules. As mentioned before, a procedure that has been

3 h (60 min anaerobic feeding, aeration 110 min

used previously for anaerobic granular sludge strength analy-

aeration, 5 min settling and 5 min effluent

sis was adapted and used in this study (Pereboom ).
Therefore, it was found that a 10 min time limit is adequate.
The shear stress was introduced indirectly and approximately

5.8 h & 70 days

W

by mixing a well-defined aerobic granules sample in a small

10 days

impeller-stirred vessel as described in the experimental set-

Granules developed at 30 C

up. In study conducted by Pereboom, granules were tested

W

Extended Aeration

at constant shear rate 1,600 RPM. In this study, stirring

3 L – laboratory scale

rates of impeller-stirred vessel used can only be set from mini-

5.8 h & 70 days

withdrawal)
et al. 2005)

mum 200 RPM up to maximum 1,200 RPM. The minimum

(de Kreuk

value of 200 RPM was applied to give a good distinction of

Acetate

stability of the different granules tested.

3–4 h (60–70 min simultaneous feeding and effluent

RESULTS AND DISCUSSION

0–10 min denitrification (no aeration), 10–25 min
Pre-treated sewage was fed from the bottom of the

withdrawal, 5 min drain, 100–165 min aeration,

reactor after passing an aerated sand and fat

Physical characteristics of aerobic granular sludge

At the beginning of the experiment, the solid density and the

size distribution of the aerobic granules sample based on
remover and a 6 mm screen

total suspended solids (TSS) concentration was determined,

in order to analyse the differences between all types of gran-
ules. The results are summarized in Table 3. More than 50%
of each sample had diameter more than 0.6 mm. The fraction
Activated sludge was taken from an aeration tank of SBR

2.8 h & 30 days

between 0.4 and 0.6 mm differed slightly; more than 10% of

1.4 m3 pilot plant

controlled but was observed to vary from 6.0 to 8.0

the aerobic granules from the laboratory scale reactor were

settling

found in this fraction compared to only 4% for aerobic granules

pilot plant. The remaining fraction passed the 0.2 mm sieve
was not considered as granule and therefore not taken into
account in the shear experiment. The solid density of aerobic
aeration 112 min aeration, 3 min

W
granules (ρ) acetate-fed developed at 20 C was 5 times
3 h (60 min anaerobic feeding,
Acetate (de Kreuk et al. 2005)

higher compared to aerobic granules sewage-fed and 2 times

settling and 5 min effluent

W
higher for 30 C-aerobic granules. This was mainly due to the
Granules developed at 20 C
W

different type of feeding materials; sewage contains many

types of COD (readily biodegradable COD as well as colloidal
3 L – laboratory scale

5.6 h & 65 days

Description of aerobic granules samples

and particulate COD), leading to more irregular and less dense

withdrawal)

structures (Beun et al. ). On the other hand, synthetic influ-

ent fed into laboratory scale reactor contains only sodium
Reactor operating conditions

acetate as organic source for microorganism growth. A temp-

erature difference as well as different sewage used also
Operation successive
Sequencing batch airlift

contributes as a factor that affects the results.

concentration &
Dissolved oxygen
Inoculums/seed

Strength of aerobic granular sludge

Q5 Influent feed

SRT & HRT

sludge
|

cycle
Samples

reactors
Table 1

The strength of aerobic granules is presented in the form of

pH

stability coefficient (S) and percentage of change (Σ) in

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4 A. Nor-Anuar et al. | Aerobic granules strength Water Science & Technology | in press | 2011

Table 2 | Procedures for evaluation of aerobic granules strength and determination of stability coefficient (S) and percentage of change granules diameter, (Σ)

No Procedure Remark

Step 1 For each one of the samples, prepare four fractions Samples used in this study are shown in Table 1.
of granule samples and label as (a), (b), (c) and (d).
Each fraction contains approximately 1,000
granules (≈100 ml) with diameter (φ) greater than
0.2 mm
Step 2 Sample (a) will be used to determine the total dry X ¼ total dry weight of granules (g TSS L
1)
weight (TSS concentration) using Standard This TSS concentration value also represents the total dry
Method (APHA ) weight of other samples
Step 3 Analyze the average diameter of granules sample A ¼ mean granule diameter before shear test
through microscopic examination Microscopic examination conducted using Image Analyser
Step 4 Pour sample (b) into a small impeller-stirred vessel The strength was measured by shear experiments in a 2,000 ml
and fill with water until the volume reaches standard geometry vessel (Tv ¼ 0.133 m) at 200 RPM stirrer
300 ml. Then, stir the sample at 200 RPM for (D ¼ 0.075 m) speed. The shear rate applied is calculated
10 min according to Equation (1). This equation is only valid for
Similar ratio as in reactor was used, in which 1/3 fully filled reactor
and 2/3 from total working volume is granules and
water. Figure below shows the schematic diagram
of experimental set up to study the shear
sensitivity of aerobic granules

Step 5 Collect the sample after stirring and sieve over a A particle with size less than 0.2 mm is considered as detached
0.2 mm sieve material
Step 6 Determine the dry weight of detached material X0 ¼ dry weight of detached material (g TSS L
1)
Detached material was considered as a broken/damage
granule during shear test
Step 7 For particles larger than 0.2 mm, re-analyze the size B ¼ mean granule diameter after shear test
with IA A particle with size larger than 0.2 mm is considered as granule
Step 8 Sample (c) and (d) will be used in a repeat test for Average value is reported in results and discussion
accuracy A repeat test should be minimum 3 times. As proposed in Step
1, four fractions of granule samples need to be prepared: (a)
for initial measurement of dry weight and (b) (c) (d) for 3
times repetition
Step 9 The fraction of detached material used for evaluation S is related to the stability of aerobic granules against shear
of granule strength. The results are expressed in stress, which can thus be interpreted as an indicator for
term of stability coefficient, S defined herein as the stability of aerobic granules. The lower the value of S, the
ratio of detached material to the total weight of greater is aerobic granules strength. In other words, the
granules, expressed in percent. S is defined as better is the stability of aerobic granules. S is not an accurate
follows: tool to measure the exact shear strength in the bioprocess;
  however it is reasonable and logical to demonstrate the
X
X0
S¼ × 100% index indicative of strength of the aerobic granules against
X
shear stress

(continued)
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5 A. Nor-Anuar et al. | Aerobic granules strength Water Science & Technology | in press | 2011

Table 2 | continued

No Procedure Remark

S ¼ Stability Coefficient (%), X ¼ Total dry weight of

granules (g TSS L
1), X0 ¼ Total dry weight of
detached material (g TSS L
1)
Step 10 An equation to calculate the Σ is determined as The stability of aerobic granules has also been evaluated by
follows: analysing the changes of granule size before and after shear
  stress was applied. The percentage of change (Σ) was
A
B
Σ¼ × 100% calculated based on the differences of mean granule
A
diameter before and after the shear test The lower the value
Σ ¼ Percentage of change of granule diameter (%), of Σ, the greater is the aerobic granules strength
A ¼ granule diameter before shear test (mm),
B ¼ granule diameter after shear test (mm)

Table 3 | W
Physical characteristics of aerobic granules developed at 20 and 30 C Table 4 | Strength characteristics of aerobic granules developed at 20 and 30 C
W

Granules developed Granules developed Granules developed at Granules developed at

W W W W
at 20 C at 30 C 20 C 30 C
Physical characteristics Acetate Sewage Acetate Sewage Acetate Sewage Acetate Sewage

1
Size distribution (% of TSS) TSStotal (g L ) 39.5 30.2 35.0 32.5
>0.6 mm 63 72 65 52 TSSfines (g L
1) 1.3 3.5 1.8 2.6
0.4–0.6 mm 12 4 13 11 S (%) 4 12 5 8
0.2–0.4 mm 4 4 6 12 φ before (mm) 0.9–1.0 0.9–1.1 1.0–1.1 0.9–1.0
<0.2 mm (≠granules) 21 20 16 25 φ after (mm) 0.8–0.9 0.3–0.5 0.7–0.9 0.5–0.7
ρ (g TSS L
1 of granules) 150 30 135 75 Σ (%) 9 40 10 24

granule diameter before and after shear test as shown in

Table 4. The lower the value of S and Σ, the higher the stab-
ility of aerobic granules. S value of the aerobic granules fed
with acetate is similar even though it was developed at
different temperature. For aerobic granules fed with
W
sewage, the S value of aerobic granules developed at 20 C
W
is greater than the aerobic granules developed at 30 C.
Meanwhile, if compared between S value of the granules
fed with acetate and sewage, the granules fed with acetate
stronger and stable. This is due to the sewage contains
many types of COD (readily biodegradable COD as well
as colloidal and particulate COD), leading to more irregular
and less dense structures (Beun et al. ). A similar trend
was also observed in the Σ value. The result showed that the
percentage is 10% for granules fed with acetate developed
W
at 20 and 30 C approximately. As Table 4 illustrates, for Figure 2 | Correlation between stability coefficient (S) and aerobic granule density (ρ).
(◊) granule: acetate-fed, developed at 20 C (Δ) sewage-20 C (O) acetate-
W W

granules fed with sewage, the Σ value of aerobic granules W W

W
30 C (□) sewage-30 C.
developed at 20 C is greater than that of the aerobic gran-
W
ules developed at 30 C. Again, overall results indicated
that the different type of feeding materials and temperature This study also observed that there is a correlation
difference also contributes as a factor that affects the between the strength and the solid density of aerobic gran-
strength and stability of granules. ules as shown in Figure 2.
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6 A. Nor-Anuar et al. | Aerobic granules strength Water Science & Technology | in press | 2011

Table 5 | Statistical results of correlation between stability coefficient (S), density (ρ) and percentage of change of granules size (Σ) with temperature (T )

Acetate-fed Sewage-fed
Correlation Correlation equation ρ-valuea p-levelb Correlation equation ρ-valuea p-levelb

ρ vs T 191e
0.01T
0.99 0.00006 145e
0.02T
0.99 0.00002

0.02T
S vs T 2.5e 0.98 0.0003 5.6e0.01T 0.97 0.0011
Σ% vs T 7.5e0.01T 0.93 0.0061 21e0.01T 0.97 0.0013
a
Pearson correlation coefficient: þ1.00: perfect positive correlation;
1.00: perfect negative correlation.
b
Statistical significance: p  0.5: less significant; p  0.05: significant; p  0.005 or 0.001: highly significant.

It shows that S decreases for increasing ρ. For a higher

ρ, the value of S will be lower, i.e. a better strength. Simi-
lar relations between shear stress, strength and density are
Q3 also reported for biofilm studies (Gjaltema et al. ;
Pereboom ; Kwok et al. ; Beun et al. ,
Q4 Villaseñor et al. ; Verschuren & van den Heuvel
; De Bruin et al. ). The local shear stress in the
well mixed laboratory scale airlift reactor and the pilot
plant bubble column are expected to be different as well
(Tay et al. ), resulting in lower strength for the pilot
W
plant granules (sewage-fed, 20 C). Correlations between
S, ρ and Σ with temperature (T ) were also studied. Statisti-
cal results of correlation obtained are summarized in
Table 5.
From the correlation curve observed, the results were Figure 4 | The stability coefficient (S) as a function of the percentage of change of
granule diameter (Σ) for different temperatures (T ). The functions are derived
then re-calculated, whereas the value is fixed with the from the data.
equation and thus, overall conclusion on correlation
between S and ρ at different T is illustrated in Figure 3. In
addition, Figure 4 illustrates the relationship between S higher growth rate of microorganism as suggested in the pre-
and Σ. The results shows that the higher the temperature, vious studies (Kwok et al. ; Villaseñor et al. ).
the lower the stability of granule. This may be related to The growth rate will increase with the increase of
temperature, while the stability or strength can be
improved by selection of slow growing microorganism in
a total granule (De Kreuk & Van Loosdrecht ).
The functions in Figures 2 and 3 can be expressed by fol-
lowing simple models:

S ¼ So0  exp(
α0  ρ) (2)

S ¼ So0  exp(þ α0  Σ) (3)

The values of So0 , So0 and α0 , α0 respectively for the exper-

Figure 3 | The stability coefficient (S) as a function of the granule solid density (ρ) for
different temperatures (T ). The functions are derived from the data.
iments are shown in Table 6.
Throughout these experimental results, a guide was
developed for ease of reference to evaluate the physical
strength of aerobic granules, as described in Table 7.
The illustrations on how the granule detached according
to their strength is also given in Table 7. The range of
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7 A. Nor-Anuar et al. | Aerobic granules strength Water Science & Technology | in press | 2011

Table 6 | Correlation function parameters of the experiment CONCLUSIONS

S vs ρ S vs Σ
Temperature ( C)
W
So0 α0 So0 α0 A new coefficient was introduced, called a stability coeffi-
cient (S), to quantify the strength of the aerobic granules.
10 20 0.010 2.2 0.05
Indicators were also developed based on the strength analy-
15 20 0.010 2.2 0.05
sis results, in order to categorize aerobic granules into three
20 16 0.009 2.8 0.04
levels of strength, i.e. very strong (very stable), strong (stable)
25 5.5 0.008 3.0 0.04
and not strong (not stable). The results indicate that aerobic
30 14 0.007 3.5 0.03 granules grown on acetate were stronger (high solid density:
>150 g T SSL
1 and low S value: 5%) than granules devel-
oped on sewage as influent. A lower value of S indicates a
higher stability of the granules. The reason is the different
Table 7 | Guide for evaluation of aerobic granule strength
hydrodynamic shear stress encountered in the pilot and
Indicator Strength
lab-scale reactors and also the different substrates. Other
Very strong Strong Not strong factors that could be included are the operating variables
Stability Very stable Stable Not stable in the laboratory scale reactor. They were easier to control,
S (%) S<5 5  S  20 S > 20
having fewer uncertainties.

ρ (g T SSL
1) ρAGS > 120 10  ρAGS  120 ρAGS < 10

Σ (%) Σ < 10 10  Σ  40 Σ > 40
Detachment ACKNOWLEDGEMENT

The authors would like to express thanks to the Universiti

Teknologi Malaysia (UTM) collaboration with Delft Univer-
sity of Technology (TUDelft), The Netherlands for the
support of this study, and Ministry of Science, Technology
and Innovation for financial grant (Vot:79004).

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Author Queries
Journal: Water Science & Technology
Manuscript: WST-WSTWS-EM11755R1

Q1 Please confirm the change of year from citation Dulekgurgen et al. (2000) to Dulekgurgen et al. (2003) as per
the reference list
Q2 Please confirm the change of citation from De Kreuk et al. (2004) to De Kreuk & Van Loosdrecht (2004) as per
the reference list
Q3 Please confirm the change of spelling from citation Gjatelma et al. (1997) to Gjaltema et al. (1997) as per the
reference list
Q4 Please confirm the change of spelling from citation Villasenor et al. (2000) to Villaseñor et al. (2000) as per the
reference list
Q5 de Kreuk et al. (2005) is not listed in the reference list. Please provide publication details to insert in the list
Q6 Please provide all editors name.

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