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Journal of Hazardous Materials 106B (2004) 83–92

Advanced sludge treatment affects extracellular polymeric

substances to improve activated sludge dewatering
Elisabeth Neyens a,1 , Jan Baeyens b , Raf Dewil b,∗ , Bart De heyder a
a Aquafin n.v., Dijkstraat 8, B-2630 Aartselaar, Belgium
b Department of Applied Biological Sciences, University of Antwerp, Middelheim, Groenenborgerlaan 171-U537, B-2020 Antwerp, Belgium

Received 30 December 2002; received in revised form 23 May 2003; accepted 26 November 2003


The management of wastewater sludge, now often referred to as biosolids, accounts for a major portion of the cost of the wastewater treatment
process and represents significant technical challenges. In many wastewater treatment facilities, the bottleneck of the sludge handling system
is the dewatering operation. Advanced sludge treatment (AST) processes have been developed in order to improve sludge dewatering and
to facilitate handling and ultimate disposal. The authors have extensively reported lab-scale, semi-pilot and pilot investigations on either
thermal and thermochemical processes, or chemical oxidation using hydrogen peroxide. To understand the action of these advanced sludge
technologies, the essential role played by extracellular polymeric substances (EPS) needs to be understood. EPS form a highly hydrated biofilm
matrix, in which the micro-organisms are embedded. Hence they are of considerable importance in the removal of pollutants from wastewater,
in bioflocculation, in settling and in dewatering of activated sludge. The present paper reviews the characteristics of EPS and the influence of
thermochemical and oxidation mechanisms on degradation and flocculation of EPS. Experimental investigations on waste activated sludge
are conducted by the authors to evaluate the various literature findings. From the experiments, it is concluded that AST methods enhance
cake dewaterability in two ways: (i) they degrade EPS proteins and polysaccharides reducing the EPS water retention properties; and (ii) they
promote flocculation which reduces the amount of fine flocs.
© 2003 Elsevier B.V. All rights reserved.

Keywords: Sludge; Advanced treatment; Extracellular polymeric substances; Dewaterability; Flocculation

1. Introduction minor fraction of the design and construction budget re-

lated to sludge management. Unfortunately this approach
Although biological processes are an effective way of does not represent the reality and today it is recognised that
treating wastewater and ensuring minimum residual impact the management of sludge, or biosolids, represents major
on the aquatic environment, they have the serious drawback investment and operating costs and forms an important tech-
of producing huge amounts of excess sludge: the more nical challenge. The associated capital and operating costs
effective the process is in removing contaminants from may be as high as 25–50% of the total cost of the wastew-
the wastewater, the more sludge is produced. Historically, ater treatment process (Karr and Keinath [1]; Baeyens
it was common to see schematics that showed the water et al. [2]).
treatment scheme in detail with all of the appropriate unit The bottleneck of the sludge handling system is the
processes and an arrow at the end that simply said “sludge dewatering operation. Advanced sludge treatment (AST)
to disposal”. The assumption that the sludge would some- processes have been developed and investigated in order
how be dealt with was supported by the fact that only a to improve sludge dewatering, and facilitate the ultimate
disposal. Different processes using thermal hydrolysis (neu-
tral, acid, alkaline) or chemical oxidation (H2 O2 , O3 , O2 ,
∗ Corresponding author. . . . ) have been proposed in literature. The authors (Neyens
E-mail addresses: elisa (E. Neyens), and Baeyens [3,4]; Neyens et al. [5–9]) have recently (J. Baeyens), (R. Dewil).
1 Present address: United Oil Products (UOP) n.v., Noorderlaan 147, summarized the literature findings and added extensive
B-2030 Antwerpen, Belgium. Tel.: +32-3-265-32-35; experimental results concerning the optimum treatment
fax: +32-3-265-32-25. conditions of thermal and thermochemical processes, as

0304-3894/$ – see front matter © 2003 Elsevier B.V. All rights reserved.
84 E. Neyens et al. / Journal of Hazardous Materials 106B (2004) 83–92

Polysaccharides have often been assumed to be the most

Nomenclature abundant components of EPS in early biofilm research (ap-
AST advanced sludge treatment proximately 60% of the dry solids content). However, pro-
COD chemical oxygen demand teins, nucleic acids and amphiphilic compounds including
CST capillary suction time (phospho)-lipids have also been shown to appear in signif-
d daily icant amounts (up to 40% of the DS content) (Wingender
DS dry solids et al. [10]).
EPS extracellular polymeric substances EPS are involved in both detrimental and beneficial char-
FTIR-ATR Fourier transform infrared attenuated acteristics of microbial aggregates (Flemming and Wingen-
total reflection der [13]). In biofouling they are responsible for the increase
IE equivalent-inhabitant of friction resistance, change of surface properties such
ODS organic dry solids as hydrophobicity, roughness, colour, etc. In biocorrosion
ORD oxidative reductive oxidation of metals they are involved by their ability to bind metal
P pressure ions. In bioweathering, they contribute by their complexing
T temperature properties to the dissolution of minerals. The EPS present
WWTP wastewater treatment plant adsorption sites for pollutants such as heavy metal ions and
organic molecules hence contributing to the water purifica-
tion, but possibly increasing the hazardous properties of the
well as peroxidation (H2 O2 ) for thickened sludge (5–6% sludge produced. Other biotechnological uses of EPS ex-
DS) from a municipal sewage treatment plant. ploit their contribution to viscosity, e.g. in food, paints and
Effective dewatering requires the attack of the micro- oil-drilling ‘muds’, whereas their hydrating properties find
organisms. The vast majority of these micro-organisms live applications in cosmetics and pharmaceuticals (Flemming
in aggregates such as films, flocs and sludges. A common and Wingender [13]). EPS may furthermore have potential
feature is that the micro-organisms are embedded in a matrix uses as biosurfactants, e.g. in tertiary oil production, and as
of extracellular polymeric substances (EPS). To understand biological glue (Flemming and Wingender [13]).
the action of the AST technologies in the sludge dewater-
ing process, the essential role played by the EPS must be
assessed and understood. 3. EPS in activated sludge
The present paper reviews the characteristics of EPS and
assesses the mechanisms in, and results of, AST processes. EPS are present in varying quantities in sewage sludge,
occurring both as a highly hydrated capsule surrounding the
bacterial cell wall and loose in solution as slimy polymers.
2. Extracellular polymeric substances: generalities Extracellular polymers are thought to be of considerable
importance in the removal of pollutants from wastewater, in
The production of EPS is a general property of micro- bioflocculation and settling (Eriksson and Alm [14]; Bruus
organisms in natural environments (Wingender et al. [10]). et al. [15]; Urbain et al. [16]), and in the dewatering of acti-
The abbreviation EPS is used as a more general and compre- vated sludge (Urbain et al. [16]; Pere et al. [17]; Forster and
hensive term for different classes of organic macromolecules Lewin [18]; Li and Ganczarczyk [19]; Sanin and Vesilind
such as polysaccharides, proteins, nucleic acids, lipids, and [20]). Li and Ganczarczyk [19] found that EPS form the
other polymeric compounds which have been found to oc- third biggest component in an activated sludge floc after the
cur in the intracellular space of microbial aggregates, more cells and water.
specifically at or outside the cell surface (Flemming and
Wingender [11]). 3.1. Characterization of the organic matter in EPS
Films, flocs and sludges are accumulations of micro-
organisms, EPS, multivalent cations, biogenic and inorganic In activated sludge, between 70 and 80% of the ex-
particles as well as colloidal and dissolved compounds. tracellular organic carbon could be attributed to proteins
EPS are mainly responsible for the structural and functional and saccharides (Dignac et al. [21]). Some researchers
integrity of the aggregates and are considered as the key have suggested that polysaccharides play the major role
to the physicochemical and biological properties (Flem- in flocculation: Bruus et al. [15] indicated that divalent
ming and Wingender [11]). EPS form a three-dimensional, cations bridge negatively charged groups on alginate-like
gel-like, highly hydrated and often charged biofilm matrix, polysaccharides within bioflocs. Forster and Dallas-Newton
in which the micro-organisms are embedded and more or [22] also suggested cations may bridge between negatively
less immobilized (Wingender et al. [10]). The total mass charged carboxyl groups on uronic acids. Most research on
of EPS (EPS and water held within the EPS-structure) has the characterization of EPS from activated sludge has fo-
been found to represent up to 80% of the mass of activated cused on exocellular polysaccharides (Forster [23]; Horan
sludge (Frolund et al. [12]). and Eccles [24]; Jorand et al. [25]). However, many studies
E. Neyens et al. / Journal of Hazardous Materials 106B (2004) 83–92 85

have reported that the exocellular protein concentration in ter within the sludge matrix, which counteracts the bene-
activated sludge systems exceeded the exocellular polysac- fit of flocculation. The water in the EPS will shield many
charide concentration (Tenney and Verhoff [26]; Brown and of the potential binding sites which could otherwise bridge
Lester [27]; Barber and Veenstra [28]; Eriksson and Alm the macromolecules by electrostatic interactions, hydrogen
[14]; Urbain et al. [16]; Jorand et al. [25]; Frolund et al. bonds and London dispersion forces (Flemming et al. [35]).
[12]; Higgins and Novak [29]). The predominance of pro-
teins in activated sludge EPS could be due to the presence 3.2.2. Water binding mechanisms
of large quantities of exoenzymes in the floc (Frolund et al. The water retained in the EPS-structure is bound mainly
[30]). The extracellular proteins could also originate from by the polysaccharides and proteins of the EPS in the acti-
wastewater compounds or from cell lysis compounds. vated sludge. The molecular mechanisms of water binding
On the basis of their relatively high content of negatively are of crucial importance for a rational basis of the improve-
charged amino acids, proteins are supposed to be more in- ment of dewatering techniques. Schmitt et al. [36] inves-
volved than sugars in electrostatic bonds with multivalent tigated water binding by measuring the exchange of D2 O
cations, underlining their key role in the floc structure (Frol- against H2 O in an FTIR-ATR study. Their results showed
und et al. [30] and Sutherland [31]). The difference in protein that treatment with chlorine clearly facilitated the exchange,
content between activated sludge and pure cultures is pos- indicating that chlorine may have destroyed water binding
sibly the result of the complexity of the substrate present in structures, probably in the EPS.
wastewater which necessitates the presence of exoenzymes According to Flemming [37], two types of binding mech-
around bacteria in activated sludge, while the synthetic sub- anisms between water molecules and the EPS-structure must
strate added to pure cultures is much more simple and easy be considered, i.e. electrostatic interactions and hydrogen
to degrade. bonds:
The remaining 20–30% uncharacterised organic carbon
(i) Electrostatic interactions are active between the per-
of EPS are possibly composed of humic compounds, uronic
manent dipole of the water molecules and ions or per-
acids, nucleic acids and lipids.
manent/induced dipoles of the functional groups of the
EPS-structure. Considering the strength of the potential
3.2. EPS and sludge dewatering
interactions, carboxylate groups and hydroxyl groups
are expected to play the most important roles among the
3.2.1. Generalities
functional groups present in EPS.
The fact that EPS are highly hydrated and are able to
(ii) Hydrogen bonds are mainly active between the EPS
bind a large volume of water was discovered by Forster
hydroxyl groups (particularly frequent in polysaccha-
[32]. EPS therefore play an important ecological role in
rides) and water molecules. They also support the ter-
protecting biofilm organisms from desiccation (Flemming
tiary structure of proteins.
and Wingender [13]): it maintains an environment in which
microbial life is possible. Both types of binding forces contribute to the overall bind-
The technical impact of this water retention affects the ing energy, although possibly to different extents. The indi-
dewaterability of activated (biological) sludge: EPS water vidual binding force of any type of weak interaction is very
needs to be removed. This is a significant economical draw- small compared to a covalent C–C bond (Flemming [37]).
back of the water retention by EPS, considering the energy However, the large number of functional groups of a macro-
required for dewatering and the millions of tons of sewage molecule may result in an overall binding energy which is
sludge which have to be dewatered every year. well in the range of several covalent C–C bonds. These inter-
The influence of EPS on dewatering has been investigated actions are affected by the water content and increase with
in various studies. Kang et al. [33] found that extracellu- increasing polymer concentration which is another argument
lar polymer substance extracted from one activated sludge to degrade the EPS to improve dewaterability.
and then added to another activated sludge has a detrimen-
tal effect on the dewatering process of the latter sludge. Ac-
cording to Houghton et al. [34] however, there appears to 4. Advanced sludge treatment and EPS
be a level of EPS at which the sludge should be easiest
to dewater. Increasing relatively low levels of EPS can aid The authors have previously summarized the literature
sludge dewaterability by improving the level of sludge floc- findings concerning the optimum treatment conditions
culation (Sanin and Vesilind [20]). This reduces the num- of thermal and thermochemical pre-treatment processes
ber of small particles present in the sludge, a factor that has (Neyens and Baeyens [3]) and chemical oxidation using hy-
been shown previously to make a sludge easier to dewater drogen peroxide (Neyens and Baeyens [4]). These reviews
(Karr and Keinath [1]). Once optimum sludge flocculation were complemented by extensive lab-scale (Neyens et al.
has been attained, further increases in EPS become detri- [5–7]), semi-pilot and pilot-scale investigations (Neyens
mental to sludge dewaterability. It is envisaged that this is et al. [8,9]) for treating thickened sludge (5–6% DS) from a
due to the highly hydrated nature of the EPS retaining wa- municipal sewage treatment plant. From these experimental
86 E. Neyens et al. / Journal of Hazardous Materials 106B (2004) 83–92

results and reviews, we suspect that the AST methods im- of temperature cause them to loose their natural shapes (de-
prove the dewaterability of the sludge by affecting the extra- naturation) and sometimes to precipitate irreversibly out of
cellular polymeric substances in two ways: (i) they have the solution in an inactive form.
potential to degrade the EPS and; (ii) they affect the multi- Results of Hatfield [39], Everett [41] and Neyens et al. [5]
functional groups of the EPS and promote their participation revealed that heat treatment alters the structure of the sludge,
in several interactions. Both effects are studied below. reduces the specific resistance and transforms some of the
suspended organic solids into soluble compounds. Everett
4.1. Degradation of EPS [41] confirmed the increase in COD of the water phase as
a result of thermal hydrolysis of organic macromolecules:
Considering the high water binding properties of EPS amino acids, volatile acids, and simple sugars are produced.
(Forster [32]; Roberson and Firestone [38]; Wingender [10]; Brooks [40,44] treated sludge at 180 ◦ C and found that hy-
Flemming and Wingender [13]), together with the finding drolysis of the exocellular and intracellular organic fraction
that the mass of total EPS represents up to 80% of the mass of the sludge, i.e. proteins, carbohydrates and lipids, leads
of activated sludge (Frolund et al. [12]), it is concluded that to destruction of the colloidal properties of these macro-
the dewatering efficiency of activated (biological) sludge can molecules with a resulting release of the water originally
be increased by degrading the EPS. The susceptibility to- bound to the particles.
wards degradation depends both upon the agent which leads Edelhoch and Osborne [45] also concluded that the heat
to the breaking of polymer chains, and upon the chemical treatment reduces or denatures the substances with high
nature of the polymer (Wingender et al. [10]). We there- affinity towards water such as proteins, carbohydrates and
fore review the literature on EPS degradation and present nucleic acids.
some experimental results concerning the influence of the
proposed AST methods on degradation of EPS. Acid/alkaline thermochemical treatment. Acids
4.1.1. Literature review of degradation of EPS and alkalis act as catalysts in the thermal hydrolysis of or-
by AST methods ganic macromolecules. Everett [41] already noted that their
addition to the sludge prior to thermal conditioning resulted Heat treatment. Thermal conditioning signifi- in an additional improvement of the rates of mechanical de-
cantly improves the dewatering properties of waste activated watering and increased percentages of DS in the filter cake.
and primary sludge (Hatfield [39]; Brooks [40]; Everett [41]; The results of the sensitivity analysis by the authors
Marshall [42]). Semi-pilot-scale experiments by the authors (Neyens et al. [9]) concerning optimum treatment condi-
(Neyens et al. [9]) demonstrated that thermal hydrolysis tions are summarized in Table 1. Both acid thermal and
most effectively reduces the residual sludge amounts and alkaline thermal hydrolysis increase the rate of filtration
enhances the dewaterability when performed at tempera- (as measured from CST-values), the amount of DS to be
tures of 120 ◦ C for a 60 min reaction time. The rate of fil- dewatered is reduced to 45% of the initial untreated amount
tration, as measured from CST-values, decreases compared for acid hydrolysis and to 62% for alkaline hydrolysis; the
to the initial untreated sludge sample. The amount of DS DS-solid content of the dewatered cake increases from 25%
to be dewatered is reduced to 58% of the initial untreated DS (initial untreated) to approximately 45% DS for acid
amount and the DS-solid content of the dewatered cake hydrolysis and from 28 to 48% DS for alkaline hydrolysis.
increases from 30% DS (initial untreated) to approximately Extremes of pH (acidity or alkalinity) cause EPS proteins
43% DS (Table 1). to loose their natural shapes (Watson et al. [43]).
Watson et al. [43] found that protein, the main component Polysaccharides, another component of EPS, are gener-
of EPS in activated sludge, is degraded by heating. Extremes ally unstable in strong acids, which leads to acid hydrolysis

Table 1
Comparison between acid and alkaline thermal hydrolysis: semi-pilot-scale experiments
Thermal Acid thermal Alkaline thermal
hydrolysis hydrolysis hydrolysis
Reagent H2 SO4
Optimum conditions
T (◦ C) 120 120 100
pH 7 3 10
Time of reaction (min) 60 60 60
Rate of mechanical dewatering after neutralisation, as measured by CST Decreased Increased Increased
Amount of DS to be dewatered as a fraction of the initial untreated amount (%) 58 45 62
% DS in filter cake 43 45 48
% DS initial untreated 30 25 28
E. Neyens et al. / Journal of Hazardous Materials 106B (2004) 83–92 87

of the glycosidic linkages (Wingender et al. [10]). They Extracellular polymers are susceptible to oxidative-
are however quite stable towards degradation in alkali, reductive depolymerization (ORD) which results in cleavage
especially at high temperature (Wingender et al. [10]). of linkages in the polymeric backbone (Herp [50]; Chris-
Strong alkali may solubilize gels not only because of tensen et al. [51]; Wingender et al. [10]). The degradation
chemical degradation, but also because of the ionization of of alginate by ORD has been studied in detail (Smidsrod
the hydroxyl groups (–OH → –O− ) which leads to exten- et al. [52–54]). ORD involves a series of free radical re-
sive swelling and subsequent solubilization (Wingender et al. actions which ultimately lead to chain scission. Molecular
[10]). oxygen and transition metal ions (e.g. Fe2+ /Fe3+ ) are ef-
At extremes of pH and after the solubilization of gels ficient catalysts. The Fenton reagent effectively degrades
(EPS), the cell looses its viability, it cannot maintain an polymers (Christensen et al. [55]). ORD has the advantage
appropriate turgor pressure and disrupts. Acids/alkali added that relatively high depolymerization rates may be obtained
to the cell suspension react with the cell walls in several at ambient temperature due to the low activation energies,
ways, including the saponification of lipids in the cell walls, whereas acid or alkaline hydrolysis require high tempera-
which leads to solubilization of membrane (Erdincler and tures. Acids or alkalis can be employed, but the degradative
Vesilind [46]). Disruption of sludge cells leads to leakage of effects are non-specific and may severely affect the cells.
intracellular material and water out of the cell which further Most cells have an intracellular defense system against toxic
improves dewaterability. radicals. This means that peroxides may operate within a
biofilm, but that they degrade only extracellular material Advanced oxidation. The oxidation processes uti- such as the matrix (Christensen et al. [55]).
lizing activation of H2 O2 by Fe(II)-salts are referred to as
Fenton’s reactions. The Fenton reaction causes the disso- 4.1.2. Experimental investigations
ciation of the oxidant and the formation of highly reactive In the following, the Advanced Sludge Treatment methods
hydroxyl radicals that attack and destroy organic pollutants. are studied from the point of view of degrading the EPS.
Fenton’s reagent is a mixture of H2 O2 and ferrous iron, Sludge samples were obtained from the underflow of the
which catalyses the formation of hydroxyl radicals according thickener (approximately 6% DS) at the municipal sewage
to the reaction (Kitis et al. [47]; Yoon et al. [48], Lu et al. treatment plant of Kessel-Lo (Belgium).
[49]): To get a clear indication on how the proposed AST meth-
ods influence the EPS-content of the activated sludge, the
Fe2+ + H2 O2 → Fe3+ + OH• + OH− presence of the main components of EPS, i.e. proteins and
polysaccharides, in the untreated and treated sludges is mea-
Advanced sewage sludge treatment by peroxidation was sured. Proteins and polysaccharides are also the most water
studied by the authors. Both lab-scale (Neyens et al. [6]) retaining components, hence influencing the dewaterability
and pilot-scale experiments (Neyens et al. [8]) were con- characteristics of the sludge. Since the mass of total EPS
ducted to investigate the efficiency of the process in function (EPS and water held within its structure) represents up to
of the operating conditions (temperature, pH, Fe2+ ). The 80% of the mass of the activated sludge (Frolund et al.
same properties as in the case of hydrolysis were measured. [12]), the measuring of sludge protein and polysaccharide
The optimum results of these experiments are summarized content instead of EPS protein and polysaccharide content
in Table 2. should give a reasonable indication of the degradation of
EPS-components by the AST methods.
The content of protein compounds was determined by the
Table 2
Comparison between lab-scale and pilot-scale peroxidation modified Lowry method described by Frolund et al. [30]
with bicinchoninic acid (BCA) as a standard. The polysac-
charide content was determined by the anthrone method
peroxidation (1000 l)
modified by Raunkjer et al. [56] and glucose was used as a
Optimum conditions
T (◦ C) Ambient
P (atm) Ambient The experimental results for the untreated sludge and the
pH 3 sludges treated with the various AST methods are indicated
H2 O2 (g/kg DS) 25 in Table 3. The protein and polysaccharide content are given
Fe2+ (g/kg DS) 1.67 as percentages of the dry solids content. All samples are
Time of reaction (min) 60–90
treated at the optimum conditions (given in Table 1) derived
Reduction of DS and ODS during peroxidation 30% from experiments reported in Neyens [9].
Rate of mechanical dewatering, as measured Increased
The results of Table 3 illustrate that proteins are more
by CST
Amount of DS to be dewatered as a fraction 40% prevalent than polysaccharides and probably are the most
of the initial untreated amount abundant component of the sludge (EPS). Over one-third of
% DS in filter cake 45% (27% DS the dry solids and over one half of the organic dry solids
initial untreated) of the untreated sludge consists of proteins (Table 3). Over
88 E. Neyens et al. / Journal of Hazardous Materials 106B (2004) 83–92

Table 3 4.2.1. The effects of pH on surface charge and stability of

Protein and polysaccharide content of untreated and AST-treated sludges sludge flocs (electrostatic interactions)
originating from WWTP Leuven (ODS/DS = 0.68)
Blanco Thermal Acid thermal Peroxidation Low pH. Everett [41], Karr and Keinath [1] and
hydrolysis hydrolysis Neyens et al. [5] showed that the dewaterability of acti-
Proteins (% of 36.2 30.1 25.7 24.3 vated sludge improved as the pH decreased. The presence
of acids seemed to alter the particle size distribution as the
Polysaccharides 22.4 20.7 19.3 18.5
(% of DS) concentration of supracolloidal solids was noted to have de-
creased with pH (Karr and Keinath [1]). It has furthermore
been shown that the amount of supracolloidal particles in
85% of the organic carbon in the untreated sludge could be the size range 1–100 ␮m seems to be an important factor
attributed to proteins and polysaccharides (Table 3). for the filterability of activated sludge (Karr and Keinath
It is clear from the results that the thermochemical [1]; Barber and Veenstra [28]). The reason for this is gen-
pre-treatment methods cause a destruction of proteins and erally believed to be blinding of the sludge cake or filter
polysaccharides. It has to be remarked that the degradative medium due to clogging with fine particles. Given this back-
effect caused by heating the sludge is strongly improved by ground, it can be assumed that the amount of fine particles
the addition of acid. In case of thermal hydrolysis 13% of in a sludge will be of importance for both clarification and
total mass of polysaccharides and proteins is destroyed, in filterability.
the case of acid thermal hydrolysis 23%. However, peroxi- Liao et al. [58] illustrate that the sludge flocs were gen-
dation under ambient conditions is more efficient in degrad- erally more stable (smaller dissociation constant) in the pH
ing proteins and polysaccharides (27% destroyed). Each of range of 2.5–9.5 than those in the pH range of less than
the three tested AST methods caused a larger destruction 2. A minimum dissociation constant occurred in the pH
for proteins than of polysaccharides. range of 2.6–3.6, which is the pH range of the isoelectric
Experimental investigations done by the authors (Neyens point. The presence of a net charge on sludge surfaces cre-
et al., [57]) moreover revealed that the degradation of EPS ates electrostatic repulsion that prevents close contact of
by the AST methods caused the release of a significant sludge micro-organisms (Liao et al. [58]). The variation in
amount of heavy metals into the water phase. Therefore, dissociation constants in terms of pH indicates that repul-
the water phase (filtrate) will be subject to metal precipita- sive electrostatic interactions were involved in disrupting
tion and separation prior to being recycled to the WWTP the stability of sludge flocs. The variation in the dissocia-
influent. tion constants with pH may be explained by the presence,
on sludge surfaces, of different functional groups, such as
4.2. The role of EPS in flocculation carboxyl, amino, and phosphate groups with different ion-
ization constants (pKa ). The major source of surface charge
The stability of sludge flocs or floc strength is important was the ionisation of carboxyl and amino groups from
in determining the efficiency of biosolids–liquid separation proteins, the main component of EPS (Liao et al. [58]).
in the activated sludge process. The presence of a large The repulsive electrostatic interactions were minimized
number of fine flocs significantly reduces not only the ef- in the pH range of 2.6–3.6, near the isoelectric point, so
fluent quality but also the dewatering properties of excess that fine flocs could approach each other closely, and the
sludge. These fine flocs can result in the clogging of the dissociation constants of sludge flocs were also at a min-
pore structure in sludge cakes and may also lead to a higher imum as mentioned earlier. The sharp increase in the dis-
level of bound water content of the sludge. Consequently, sociation constants at a pH less than the isoelectric point
the minimization of fine flocs in the activated sludge pro- could be related to the adsorption of H+ by amino groups
cess is essential for better effluent quality and for improved (R–NH2 ) to form R–NH3 + under low pH conditions. The
dewatering properties of sludge. relatively stable dissociation constants, in the pH range of
A number of mechanisms, such as charge neutralization, 4.5–9.5, could be attributed to the combined influence of
hydrophobic interactions, hydrogen bonds, ionic interac- the amino and carboxyl groups on sludge surfaces (pKa val-
tions, and physical enmeshment have been recognized as ues of 1.71–3.0 and 8.2–11, respectively (Smith and Martell
important in controlling the formation of the sludge floc and [59]; Nelson and Cox [60]). The naturally occurring amino
its stability. Since EPS (present at or near sludge surfaces) groups might form a stable five member ring chelated with
are composed of diverse high-molecular-weight biopoly- divalent cations, such as Ca2+ and Mg2+ , and therefore neu-
mers (e.g. polysaccharides, proteins, DNA, and lipids) that tralize the amino groups (Huang et al. [61]). At least 99%
contain many functional groups capable of participating of the carboxyl groups of amino acids would ionise into
in interactions, it is likely that the stability of sludge flocs COO− groups when pH = pKa + 2 i.e. pH 4–5), so that the
is related to the interparticle interactions at or near sludge magnitude of the surface charge was relatively stable, which
surfaces. The effects of AST methods on interparticle inter- further explains the stability of the dissociation constants in
actions are now reviewed. the pH range of 4.5–9.5.
E. Neyens et al. / Journal of Hazardous Materials 106B (2004) 83–92 89 High pH. Neyens et al. [7] showed that the treat- 4.2.3. Experimental investigations
ment of activated sludge at high pH improved the dewat- Most information on floc size diameter in the literature
erability. This improved dewaterability is also illustrated in concerns activated sludge. Andreadakis [65]; Higgins and
Table 1. Novak [66]; Biggs et al. [67] found that for well mixed
Katsiris and Kouzeli-Katsiri [62] stated that when the pH activated sludge systems the typical range of floc sizes was
of sludge samples increased, the bacterial surfaces become between 10 and 70 ␮m. No information was found on size
increasingly negatively charged. This creates high electro- distribution of thickened biological sludge. To assess the
static repulsion which causes desorption of some part of ex- possible influence of AST on the biosolids size distribu-
tracellular polymers. These dissolved extracellular polymers tion, an identical sample of 6% DS-thickened sludge was
and intracellular material released by cell disruption are all treated by the different thermal and chemical AST presented
high molecular weight polymeric materials that possibly act above. The experimental set-up is described in Neyens
as polyelectrolytes and thereby promote flocculation (Val- et al. [9].
lom and McLoughlin [63]) and dewaterability. Untreated and treated samples were analysed for size dis-
Since cells are disrupted both at low and high pH, the tribution using a Malvern Mastersizer laser diffractometer.
water-floc separation properties of sludge are improved by This is a laserbeam scattering instrument that operates on
the release of interstitial water trapped inside the sludge the principle of Fraunhofer Diffraction Theory. The parti-
cells and by the promotion of the flocculation, reducing the cle size detection range is between 5 and 300 ␮m. This size
amount of small flocs. range is within the range indicated to represent most of the
activated sludge flocs (Jorand et al. [25]). The particle diam-
4.2.2. The effects of cations on flocculation (ionic eters were calculated assuming spherical particles. Samples
interactions) were diluted before analysis to obtain particle counts within
Several researchers have suggested that cations in general, the concentration limits of the analyser.
and bi-valent ones specifically, aid flocculation and dewa- The results are illustrated in Table 4, and expressed as the
tering by bridging negative sites on exocellular biopolymers respective particle sizes corresponding with 10, 50 and 90%
which promotes an increase in floc size, floc density, floc of the size histogram i.e. 10% of the flocs have sizes in the
resistance to shear and dewaterability (Tezuka [64]; Bruus range [0 ␮m-d10 ], 50% in the range [0 ␮m-d50 ] and 90% in
et al. [15]; Eriksson and Alm [14]; Andreadakis [65]). Hig- the range [0 ␮m-d90 ].
gins and Novak [66] showed that the settling and dewatering Since larger flocs are easier to dewater than smaller flocs,
properties of the activated sludge systems were dependent and since the presence of smaller flocs tend to clog the
on both the concentrations and ratios of cations added with cake during filtration thus reducing the dewaterability, the
high concentration ratios of divalent/monovalent cations sig- evolution of the d10 , d50 and d90 sizes give a clear indication
nificantly improving settling and dewatering. This improve- of the positive or negative effects.
ment correlates with an increase in the bound exocellular With the exception of thermal hydrolysis, all techniques
biopolymer content involved in the aggregation process. Di- tested increase the size of the smaller flocs being present.
valent cations act to bind protein and not polysaccharide Thermal hydrolysis moreover significantly reduces the other
within the floc in these systems (Higgins and Novak [66]). characteristic particle sizes, thus pointing towards an over-
As a result, the improvements in settling and dewatering are all reduction of flocculation performance. However, this
well correlated to the bound protein content in addition to reduced flocculation behaviour does not significantly cause
the increase in divalent cation concentration. These results reduced dewatering properties (Table 1), since the reduced
suggest that protein is an important biopolymer component flocculation is offset by the degradation of the hygroscopic
in the flocculation process. It is likely that carboxyl groups EPS and subsequent disruption of the cells as explained
from amino acids in proteins and phosphate groups from in Section 4.1.2. This release of bound water improves the
DNA were the dominant functional groups involved in salt dewaterability irrespective of the decreased flocculation
bridging (Liao et al. [58]). behaviour.

Table 4
Particle sizes and standard deviation after applying the different AST methods
Treatment applied d10 (␮m) σ 10 (␮m) d50 (␮m) σ 50 (␮m) d90 (␮m) σ 90 (␮m)

Untreated (blanco) sludge sample 32.82 6.11 106.64 7.12 169.47 2.71
Thermal hydrolysis 34.92 7.23 65.99 7.55 117.36 12.78
Acid thermal hydrolysis 54.88 9.84 106.93 8.10 169.57 3.11
Alkaline thermal hydrolysis 58.46 1.88 105.24 4.97 209.16 4.05
Acid treatment at ambient conditions 64.06 1.26 121.36 0.92 193.48 0.41
Alkaline treatment at ambient conditions 66.07 2.33 125.13 12.43 226.77 7.93
Peroxidation 47.38 3.84 91.83 7.53 161.47 10.59
90 E. Neyens et al. / Journal of Hazardous Materials 106B (2004) 83–92

Acid thermal hydrolysis causes the smaller flocs to ag- technique, there is hence a saving of over 800 000 EUR per
gregate (Table 4). This explains that the breakdown of flocs year on the disposal cost of the annual amount of 6570 ton
caused by thermal hydrolysis is compensated by adding acid DS. Considering the costs and the savings, there is a net
(H2 SO4 ), which is indicated in the results of acid treatment saving of 800 000 − 460 000 EUR per year = 340 000 EUR
(at ambient conditions) in Table 4. This illustrates that at per year or 52 EUR/ton DS for a WWTP of 300 000 IE.
pH 3 the repulsive interactions and the resulting dissociation
constants are minimized as stated above.
A similar compensation effect is noticed in the case of 6. Conclusion
alkaline thermal hydrolysis: the thermal breakdown of flocs
is compensated by the alkaline effect. The results in Table 4 Advanced sludge treatment processes such as thermal hy-
also stress the importance of divalent Ca2+ -ions as bridg- drolysis and peroxidation have been developed to improve
ing agents: divalent ions aid flocculation and dewatering by sludge dewatering and to facilitate ultimate disposal. Imple-
bridging negative sites on exocellular biopolymers which menting these methods in a wastewater treatment plant: (i)
promotes an increase in floc size, floc density, floc resistance slightly reduces or increases the filtration rate; (ii) decreases
to shear and dewaterability (Tezuka [64]; Bruus et al. [15]; the amounts of DS to be dewatered; and (iii) increases the
Eriksson and Alm [14]). DS content of the dewatered cake. The present paper made
Peroxidation slightly reduces the value of d50 and d90 , an attempt to explain these improvements in terms of the
effects counteracted by the marked increase of the d10 -size. current knowledge concerning EPS which constitute up to
This improved flocculation performance in combination 80% of the mass of activated sludge. It is concluded that
with the capability of the hydroxyl radicals to degrade EPS AST methods degrade the EPS and influence the dewater-
by ORD, turns peroxidation into a promising AST method ability of the sludge in two ways:
(Table 1). (i) Degradation of the EPS reduces their water retention
properties thereby releasing the EPS-bound water and
increasing the dewatering efficiency of activated sludge.
5. The economy of using AST methods The advanced methods degrade proteins and polysac-
charides of activated sludge EPS, which constitute al-
Complete and detailed cost–benefit analysis of the AST most 60% of EPS. The extent of the degradation varies
methods are reported in Neyens [68], the most important among the AST methods from 23% for acid thermal
results are summarized below. hydrolysis to 27% for peroxidation.
Peroxidation gave the best results with respect to reducing (ii) Degradation of the EPS reduces cell stability but in-
the amounts of sludge and improving the product quality. creases the size of sludge flocs. The presence of a
An economic assessment is therefore made for a WWTP of net surface charge on sludge surfaces creates electro-
300 000 equivalent-inhabitant (IE) with sludge dewatering static repulsion that prevents close contact of sludge
and incineration. For the case of a daily (d) sludge produc- micro-organisms. These repulsive interactions are min-
tion of 60 g DS/IE d, a total annual amount of 6570 ton DS imized in the pH range of 2.6–3.6, near the isoelectric
has to be dealt with. The annualised cost of operating a tra- point, so that the dissociation constants of sludge flocs
ditional 300 000 IE WWTP where the dewatered sludge is are also at a minimum. For this reason the flocculation
incinerated is 1 900 000 EUR. of activated sludge improved as the pH decreased. The
The peroxidation process requires the use of corrosion- improved flocculation phenomena (bridging, ionic ac-
resistant construction materials. The fixed equipment costs tion) moreover reduce the amount of fine flocs, thus
are estimated to be 50 000 EUR per year when written off facilitating cake dewatering. This statement was under-
over 10 years. The maintenance costs are approximately lined by conducting experiments on the size distribution
10 000 EUR per year. Variable costs consist of chemical of sludge samples treated by the different AST methods
costs and electricity. Chemical costs involved are rather high. (Table 4).
Electricity is only necessary for the pumps. The sum of these
variable costs is estimated to be 400 000 EUR per year. The
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