2363 © IWA Publishing 2013 Water Science & Technology | 67.

11 | 2013

Rethinking wastewater characterisation methods for

activated sludge systems – a position paper
Jean-Marc Choubert, Leiv Rieger, Andrew Shaw, John Copp,
Mathieu Spérandio, Kim Sørensen, Sabine Rönner-Holm,
Eberhard Morgenroth, Henryk Melcer and Sylvie Gillot

ABSTRACT
Jean-Marc Choubert (corresponding author)
Increasingly stringent effluent limits and an expanding scope of model system boundaries beyond
Irstea, UR MALY, 5 rue de la DOUA, CS70077,
activated sludge has led to new modelling objectives and consequently to new and often more 69626 VILLEURBANNE Cedex, France
E-mail: jean-marc.choubert@irstea.fr
detailed modelling concepts. Nearly three decades after the publication of Activated Sludge Model
Leiv Rieger
No1 (ASM1), the authors believe it is time to re-evaluate wastewater characterisation procedures and EnviroSim Associates Ltd,
McMaster Innovation Park,
targets. The present position paper gives a brief overview of state-of-the-art methods and discusses 175 Longwood Rd S, Suite 114A, Hamilton, Ontario,
L8P 0A1, Canada
newly developed measurement techniques on a conceptual level. Potential future paths are
Andrew Shaw
presented including on-line instrumentation, promising measuring techniques, and mathematical
Black & Veatch, 8400 Ward Parkway, Kansas City,
solutions to fractionation problems. This is accompanied by a discussion on standardisation needs to Missouri, 64114, USA

increase modelling efficiency in our industry. John Copp

Primodal, Inc., 122 Leland Street, Hamilton L8S
Key words | activated sludge, ASM, chemical oxygen demand fractionation, modelling, wastewater 3A4, Ontario, Canada
characterisation Mathieu Spérandio
Université de Toulouse; INSA,
UPS, INP; LISBP, 135 Avenue de Rangueil,
F-31077 Toulouse, France

Kim Sørensen
VA TECH WABAG GmbH,
Dresdner Strasse 87-91,
1200 Vienna, Austria

Sabine Rönner-Holm
LimnoTec Abwasseranlagen GmbH,
Ravensberger Straße 41, 32312 Lübbecke,
Germany

Eberhard Morgenroth
ETH Zürich, Institute of Environmental Engineering,
8093 Zürich, Switzerland
and
Eawag, Swiss Federal Institute of Aquatic Science
and Technology, 8600 Dübendorf, Switzerland

Henryk Melcer
Brown & Caldwell, 701, Pike Street, Suite 1200,
Seattle 98101, WA, USA

Sylvie Gillot
Irstea, UR HBAN, 1 rue Pierre-Gilles de Gennes, CS
10030, F-92761 Antony Cedex, France

INTRODUCTION

Wastewater characterisation has been an ongoing topic of
research (Spanjers et al. ; Henze et al. ) and is still
a major challenge for model application in engineering prac-
tice (Copp et al. ; Phillips et al. ). Fractionating the
variables that are traditionally measured by practitioners
(chemical oxygen demand (COD), nitrogen and phosphorus)
into model variables (different classes of biodegradability) is
an important input to the model. Calibration of the model
parameters by adopting an incorrect fractionation of the
wastewater composition may lead to compensation of the

doi: 10.2166/wst.2013.158

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 2364 J.-M. Choubert et al. | Rethinking wastewater characterisation methods for activated sludge systems
fractionation errors by introducing errors in the parameter
estimates (Rieger et al. ). Moreover, combined expertise
of modellers (Phillips et al. ) and sensitivity analysis
(Petersen et al. ) have shown profound effects of waste-
water characterisation on modelling outputs (Henze et al.
):
• Sludge production is influenced by the estimated inert
particulate COD.
• Oxygen demand is influenced by the estimated total bio-
degradable COD.
• Anoxic denitrification rate and anaerobic phosphorus
release are influenced by the estimated readily biodegrad-
able COD.
• Effluent COD is influenced by the estimated inert soluble
COD.
Although some milestone research has been published
(STOWA: Roeleveld & van Loosdrecht ; WERF:
Melcer et al. ; BIOMATH: Vanrolleghem et al. ),
and a recent attempt at guidelines has been proposed (Lu
et al. ), there is a lack of standardisation. Gillot &
Choubert () and Fall et al. () compared different
experimental COD fractionation approaches and found sig-
nificant differences in the results.
New measurement techniques allow for a more detailed
evaluation of the wastewater, but the techniques published
decades ago can provide significant insight into a particular
wastewater when performed properly and the results are ana-
lysed appropriately. In practice, questionable results are
commonplace due to a lack of standardisation and method
complexity (Rieger et al. ). In this context, this paper
gives a brief overview of state-of-the-art methods for waste-
water characterisation and discusses gaps in research. It
presents recently developed measurement techniques on a
conceptual level, and explains new insights into wastewater
characteristics and involved components. Potential future
paths are presented including on-line instrumentation, prom-
ising measuring techniques, and mathematical solutions to
fractionation problems. This is accompanied by a discussion
on standardisation needs to increase modelling efficiency in
the wastewater treatment field. The paper focuses on waste-
water characterisation methods for activated sludge systems
and does not intend to cover all aspects of wastewater charac-
terisation. Links are made to membrane systems, biofilm
systems and anaerobic digestion processes as they can
either provide ideas for improving, or taking advantage of
developments in, activated sludge wastewater characteris-
ation methods.
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Water Science & Technology | 67.11 | 2013
EXISTING WASTEWATER CHARACTERISATION
METHODS
In the Activated Sludge Model (ASM) framework (Henze
et al. ), the influent COD of a wastewater is split into
different fractions. For Activated Sludge Model No1
(ASM1), six fractions are considered (slowly biodegradable,
readily biodegradable, inert soluble and inert particulate,
heterotrophic and autotrophic biomass). In ASM No2d
(ASM2d), the readily biodegradable fraction was divided
into fermentable organic matter and fermentation products
(volatile fatty acids, VFA); moreover, storage compounds in
biomass and an additional biomass were considered. In
ASM No3 (ASM3), similar fractions to ASM1 are considered
with additional organics stored by heterotrophic biomass. In
the literature, most of the fractionation methods are dedicated
to the differentiation between inert and biodegradable frac-
tions, readily or slowly biodegradable variables, initially
defined for ASM1; however, most of the following protocols
and statements are valid by extension for ASM2 and ASM3.
This approach assumes the wastewater is devoid of biomass,
has no inert endogenous products and includes inert soluble
material. Although not ideal or detailed enough to account
for all constituents, these assumptions have proven to be effec-
tive in the past and are linked to the model structures.
Comparison of the most relevant protocols showed only
slight methodological differences for the characterisation of
the nitrogen and phosphorus fractions. However, for the
characterisation of the wastewater COD, there are different
approaches. Table 1 presents the methods and concepts. The
following discussion briefly presents the limitations caused
by the differences between conceptual meaning and exper-
imental results.
The existing methods of fractionating COD into the
different fractions of a given wastewater generally combined
the determination of the total biodegradable COD fraction
(CODtot,B) and the determination of the readily biodegrad-
able COD (noted SB).
Two types of biological assay have been proposed to
determine the total biodegradable COD fraction of a given
wastewater:
• Direct monitoring using the oxygen uptake versus time in
batch experiments. Various protocols exist, but typically
methodologies include respirometry monitoring the
modification of the oxygen uptake rate for a few hours
with a low substrate to biomass ratio (S/X ) (Spérandio
et al. ; Vanrolleghem et al. ), ultimate bio-
chemical oxygen demand (BOD) tests monitoring the
 2365 J.-M. Choubert et al. | Rethinking wastewater characterisation methods for activated sludge systems Water Science & Technology | 67.11 | 2013

Table 1 | Existing methods for wastewater COD fractionation

Conceptual meaning

Assessment of
biodegradable Assessment of readily
Experimental Targeted COD biodegradable Known limitations
protocol fractions (with respect to ASM1) References
Size of
organic
Direct Indirect Biodegradability molecules

Short-term bioassay SB, XCB XOHO • • Difficulty to distinguish UCT method (Ekama
at high and low endogenous et al. ) ASM Task
S/X (batch, pilot) respiration from XCB Group (Henze et al.
degradation (end of ) Spanjers et al.
test). Systematic low (); Spérandio
value of CODtot,B et al. (); WERF
(short test length) protocol (Melcer
et al. )
(respirometry)
10d bioassay at SB, XCB SU, XU,Inf • • Systematic low value of STOWA approach
high S/X CODtot,B (high S/X (Roeleveld & van
(batch) þ 0.1μm ratio) Loosdrecht )
filtration (ultimate BOD and
filtration)
Long-term bioassay SU, XU,Inf • • SU attributed to WERF protocol (Melcer
at low S/X (pilot, wastewater, whereas et al. ) UCT
batch) part is produced by method (Ekama et al.
biomass ) Stricker et al.
()
(biodegradation test)
0.45μm filtration of SU • Suitable only if removal UCT method (Ekama
treated effluent of biodegradable et al. )
COD is complete
Coagulation / SB, SU • • Link between size of Mamais et al. ()
flocculation þ organic molecules
filtration and biodegradability
not always valid

Notation according to Corominas et al. (2010) ¼ SB: readily biodegradable COD; XCB: slowly biodegradable COD; SU: inert soluble COD; XU,Inf: inert particulate COD; XOHO: ordinary hetero-
trophic organisms.

total oxygen uptake for 10 days with a high S/X ratio
(Roeleveld & van Loosdrecht ), or monitoring the
COD uptake versus time (total and soluble) for 30 days
with a high S/X ratio (Lesouef et al. ; Orhon et al.
; Stricker et al. ).
• Indirect assessment of CODtot,B by determination of the
particulate unbiodegradable COD (XU,Inf ) of the waste-
water. This fraction is deduced by using a simulation
model to match experimental sludge production in a bio-
reactor (Melcer et al. ).
Three distinct principles have been used to determine
the readily biodegradable COD (noted SB):
• Bioassay tests that are based on respirometric characteris-
ation methods where oxygen or nitrate utilisation rates
(OUR and NUR, respectively) are recorded (Ekama
et al. ; Naidoo et al. ; Spanjers et al. ).
• Physical–chemical methods that assume a direct link
between the size of organic molecules and their biodegrad-
ability. Filtrations with different pore sizes (Roeleveld &
van Loosdrecht ; Lu et al. ), and with or without
a flocculation step (Mamais et al. ) have been
proposed.
• VFAs are typically measured directly using gas chromato-
graphy method (Martin-Ruel et al. ).
COD fractions (and particularly readily biodegradable
COD) are measured (or estimated) with batch experiments
and first used for plant model set-up. Although wastewater
characterisation is considered as part of the data collection

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 2366 J.-M. Choubert et al. | Rethinking wastewater characterisation methods for activated sludge systems
step by the IWA Good Modelling Practice Task Group
(Rieger et al. ), there is a possibility to modify the obtained
fractions during one of the three main steps that compose the
calibration procedure. The latter starts with fixing the par-
ameters that represent hydrodynamics (number of tanks,
flow distribution), then continues with modifying the influent
wastewater or recycle streams’ characteristics, and finishes
with parameters’ estimation: settling parameters, aeration
parameters (oxygen mass transfer over time description)
and biokinetic parameters (stoichiometric, kinetic). Influent
wastewater characterisation and parameter assignment (also
called ‘calibration’ stricto sensu) are therefore related stages
when adjustments of the fractions are required (at least to
take into account measurement errors), for example modify-
ing the inert particulate COD (i.e. XU,Inf) to match the
sludge production. To avoid this fine-tuning, the industry
would have to develop methods to better represent the inert
particulate COD (i.e. XU,Inf).
ADAPTATION OF EXISTING METHODS
Changes to the influent fractions (i.e. percentage of total
COD) are not uncommon under wet-weather conditions or
due to temperature variations (Henze ). In plants with
long hydraulic buffering capacity (like those applied in con-
ventional activated sludge, 6–24 h) it is often sufficient to
separate the COD into biodegradable/unbiodegradable frac-
tions. However, with the migration to more stringent
effluent requirements, and the development of short hydrau-
lic retention time (HRT) processes (few tens of minutes), this
assumption may need to be revisited.
The following discussion briefly presents the limit-
ations caused by the differences between conceptual
meaning and experimental results, and suggests some
recommendations.
Impact of the experimental protocol on the
fractionation results
Even with defined approaches, the experimental methods
and resulting output still remain sensitive to practical inac-
curacies due to the complex ecosystem biology, the
biodegradable fractions that depend on protocol length
(few hours to a couple of days) and substrate to biomass
ratio (Gillot & Choubert ), or physical limitations such
as filter pore size (Fall et al. ). The wide variety of poss-
ible experimental conditions in the methods further
complicates the interpretation of results.
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Water Science & Technology | 67.11 | 2013
Even for apparently simple methods (e.g. physical–
chemical separation), differences are obtained (e.g. due to
differences in filter pore size 0.7, 0.45 or 0.1 μm according
to Fall et al. ()). Coagulation/flocculation prior to fil-
tration reduces the importance of the filter pore size;
however, if not standardised or reported it may lead to
further ambiguity in determining results.
A change in model structure or the introduction of
another state variable makes it necessary to adapt the exper-
imental methods also. It is important to use a fractionation
method that fits the model concepts. ASM1 is based on
the biodegradability of the fractions, but for practical
reasons the profession has turned to physical–chemical
methods (Roeleveld & van Loosdrecht ). Thus, several
concerns have been raised about the comparability of the
methods and how to link the physical–chemical methods
with the biodegradability of the different fractions.
The obtained fractionation based on measurements is
sometimes adapted during model calibration to be consist-
ent with other data collected on the plants (Rieger et al.
). During the calibration step, certain fractions can be
changed until simulation results match observed data; for
example, it is not unusual to modify the inert COD fraction
to fit the simulated sludge production to the observed
values. However, this indicates problems with the link
between what the modeller requires and what these exper-
imental procedures actually measure. As a result, the
value of expensive wastewater characterisation studies –
which are essential for good predictions – might be ques-
tioned if the results cannot be applied in all cases, and
might be critical for the application of modelling in
today’s engineering practice. Other sources of information
about plant behaviour are nevertheless always required
and can minimise the probability of a deficient wastewater
characterisation.
Changing COD fractions for long sludge retention time
processes
The plant type to which a wastewater is introduced deter-
mines how the wastewater should be characterised. Haider
et al. () and Spérandio et al. () clearly identified a
larger biodegradable fraction when the same wastewater
was subjected to a longer sludge age process. The definition
of specific fractions can be process specific. For example,
material considered inert in a short sludge retention time
(SRT) system (e.g. those applied in conventional activated
sludge) may not be inert in long SRT systems (e.g. membrane
bioreactor (MBR), sludge reduction techniques, constructed
 2367 J.-M. Choubert et al. | Rethinking wastewater characterisation methods for activated sludge systems
wetlands). New model concepts that improve the understand-
ing and models incorporating hydrolysis are suggested below.
Changing COD fractions versus time
Under dry-weather conditions, a basic assumption in today’s
modelling practice is that influent fractions are stable over
time during a 24 h period (Henze ). However, it may
change with temperature (Henze ). Also in the case of
wet-weather treatment, significant changes in the fraction
of XU,Inf and XCB due to rain may have to be considered.
Improving the determination of the colloidal COD fraction
is important for a better prediction of sludge production
and optimisation studies (Petersen et al. ; Rönner-
Holm et al. ).
Variations in the estimated COD fractions through the
day are another potential concern as clearly shown in
Figures 1(a) and 1(b). These results were obtained with the
STOWA protocol on 2 h composite samples (Barry et al.
; Morvannou ). One would expect more stable
ratios but the measurements of lumped COD show that
the CODfiltered/CODtot ratio varied between 20 and 60%
(Figure 2(a)) and the SB fraction varied between 5 and 25%
of the total COD over a period of a few hours (Figure 2(b))
(Hess ). Large variations in the COD fractions over
the course of a single day were also noted in Germany
(Hintermaier & Thiele ; Affeld ; Roog ).
Assuming that the previous variations are not plant
specific and are not due to measurement artefacts, it is
clear that the plant behaviour may well vary throughout the
day as a direct result of the variability in wastewater fractions.
The impact of these fraction variations is even more critical
when modelling systems with HRTs of only a few tens of min-
utes (e.g. of A-stage of an A/B plant, moving-bed biofilm
bioreactor (MBBR)), so it is reasoned that a higher temporal
resolution of the fractionation would then improve
Figure 1 | Evolution of COD fractions (XU,Inf, XCB, SB, SU) versus time at two French wastewater treatment plants (WWTPs).
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Water Science & Technology | 67.11 | 2013
simulation results (Haider et al. ). Future research
needs to address the magnitude and the impact of the vari-
ations. However, in the short term, it would seem prudent
to estimate the variations in the COD fractions using a time
series of total, filtered and flocculated-filtered COD, instead
of using incorrectly measured SB and XCB values.
NEW AND ALTERNATIVE METHODS
The development of new treatment processes and the need
to predict very low nutrient effluent concentrations requires
model extensions and consequently adapted influent charac-
terisation methods. Still, models and fractionation methods
have to be complementary, and the model objective should
be taken into consideration when selecting a fractionation
method. To meet this next challenge, and to meet the
needs of different modelling objectives, modellers will
have to consider new measurements, experiments/pro-
cedures, checks, data resolution and also model changes.
Table 2 provides expected adaptations of fractionation con-
cepts published in the literature that are then explained in
the subsequent paragraphs.
Biomass determination
Most characterisation protocols assume that wastewater is
devoid of biomass (XOHO ≈ 0), including the related COD
in XU,Inf and XCB. These assumptions have proven to be
effective in the past as it did not impact model predictions
in activated sludge systems (Melcer et al. ) and the
assumptions were linked to the model structure. However,
active heterotrophic biomass has been estimated to be up
to 25% of the raw wastewater COD (Henze ; Spérandio
et al. ). Compensating for this assumption (i.e. consider-
ing this COD to be slowly biodegradable XCB) may require
 2368 J.-M. Choubert et al. | Rethinking wastewater characterisation methods for activated sludge systems
Figure 2 | COD versus time at Morgenthal plant (Switzerland, 28 Nov.–8 Dec. 2000) (Hess
2001).
that the yield coefficient be increased by approximately 10%
(Henze ).
This is observed when modelling systems operated at
high loading rates, or when modelling downstream of a pre-
liminary stage of biological treatment, as the influent
biomass should not be incorrectly assumed to be zero, other-
wise the behaviour of the models may have to be incorrectly
altered (by changing parameters) to compensate. Thus a
more systematic measurement of active biomass content in
influents might reduce calibration efforts. Respirometry
can be used for active biomass determination and it is still
the recommended method for modelling application. Pro-
gress towards biomass determination is expected with the
development of quantitative biomolecular techniques (e.g.
quantitative real-time polymerase chain reaction) but until
now these methods have not demonstrated their superiority
for model validation or calibration.
Standardisation of biological tests
Respirometric biological tests are subjected to uncertainty
(Gatti et al. ), for example the endogenous state can
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Water Science & Technology | 67.11 | 2013
be confounded by slow hydrolysis, or sludge that is not
adapted (unacclimatised) to the wastewater. Two sugges-
tions are made to improve the results: the first one is to
standardise the conditions that define when a respirometric
test enters the endogenous respiration phase (i.e. when all
substrate is consumed). Experience indicates that most of
the error is introduced because of the uncertainty in identi-
fying this transition.
The second option is to improve our understanding
of biomass development inside the bioreactors (e.g.
compliance to biomass growth and decay concepts used
to model OUR), and to develop ways to grow the biologi-
cal inoculums for seeding bioreactors, so that the results
are not dependent on the level of acclimation of the
biomass.
New probes
On-line sensors for measuring COD, NH4, PO4, NOx and
total suspended solids (TSS) have shown the potential for
on-line analysis and control of biological systems. Newer
types of probes offer even more potential for collecting
more detailed wastewater characterisation data. The data
from on-line respirometer or UV-vis probes (with appropri-
ate correlations) allow for a continuous estimation of
wastewater fractions. It is expected that significant strides
towards ensuring the quality of these data will have to be
made in the future. However, the benefits gained with
respect to parameter uncertainty and model predictive
power have been shown by Cierkens et al. ().
Inorganic compounds
The use of inorganic suspended solids (ISS) (Melcer et al.
; Ekama & Wentzel ) has improved the predictions
of sludge production in terms of suspended solids (total, TSS).
As the requirement for more detailed models increases, there
is a clear need for an improvement in the prediction of inor-
ganics, and especially those involved in precipitation/
dissolution (sulphide, magnesium, calcium, iron, aluminium,
phosphates and nitrogen). This requirement may include a
procedure to distinguish between different fractions of min-
eral suspended solids in raw wastewater and in biological
sludge (e.g. inert, bound to organic matter, etc.). There will
be a requirement for more detailed influent information
including pH, and salt/metal concentrations. It is also poss-
ible that the use of models for warm climate applications
may require sulphur modelling, and the measurement of the
different sulphur fractions (Sharma et al. ).
 2369 J.-M. Choubert et al. | Rethinking wastewater characterisation methods for activated sludge systems
Table 2 | Summary of published wastewater characterisation approaches
Expected evolution of wastewater
characterisation
Improved / new Measurement of biomass content of
experimental wastewater (XOHO) instead of
procedures lumping it in XCB and XU,Inf
Measurement of time-varying
fractions of COD for specific
operating conditions
Reduced uncertainties
Continuous estimation of WW
fractions. Set-up of correlations
with lumped parameters
Systematic use of inorganic
suspended solids (ISS) and
chemical compounds involved in
dissolution/precipitation (e.g.
sulphide, Mg, Ca, Fe, Al)
Comprehensive data treatment,
including uncertainty of sampling,
lab-analysis, data reconciliation
New or extended model New state variables and fractions
components/concepts
Consider the biochemical forms
(like lipids, proteins, sugars) and
also particle size
Very slowly biodegradable organic
nitrogen fraction (soluble) Refined
nitrogen fractionation
(Measurement of aminoacids and
new substances)
Mathematical estimation/verification
With more and often contradictory data, new methods would
be helpful to facilitate data reconciliation in wastewater frac-
tionation analysis. Automated techniques based on statistical
analyses could improve fractionation quality and conse-
quently reduce calibration needs of kinetic parameters in
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Water Science & Technology | 67.11 | 2013
Examples/objectives References
Active biomass determination Wentzel et al. ()
(wastewater, intermediate point Spérandio et al. ()
like between separate reactors)
Fraction variations influence This paper
modelling systems with short
hydraulic residence times
Standardisation or reporting of –
biological tests
New probes (on-line respirometers, Cierkens et al. ()
UV-vis probes with correlations)
TSS calculation/prediction of sludge Ekama et al. ()
production using inert suspended
solids and chemical compounds
Mathematical estimation/ Grau et al. ()
verification
New variable of interest like Ni et al. ()
greenhouse gas (GHG), Barret et al. ()
micropollutants, exo- Naessens et al. ()
polysaccharides (EPS), sulphur Sharma et al. ()
production
New process units: membrane Naessens et al. ()
bioreactor (MBR), moving-bed Barry et al. ()
biofilm bioreactor (MBBR), sludge
reduction tanks
New operating conditions: long Ménniti et al. ()
sludge retention time (SRT)/short Spérandio et al. ()
hydraulic retention time (HRT)
e.g. hydrolysis of particulate
undegradable endogenous
products (XU,E) and particulate
undegradable organics from the
influent (XU,inf ) at long SRT
Better hydrolysis description Sophonsiri and
Morgenroth ()
Low nitrogen effluent limits Pehlivanoglumantas-
Mantas & Sedlak ()
biological reactors. Grau et al. () proposed a relationship
between the model component and the most common
analytical measurements in the wastewater, combining a
priori knowledge of the expected wastewater composition
and experimental information. An optimisation algorithm
uses an extended list of non-redundant model components
with definition of their elemental mass balance fractions in
 2370 J.-M. Choubert et al. | Rethinking wastewater characterisation methods for activated sludge systems
terms of C, H, N, O, P and charge. This approach might be
applied more systematically to improve the estimation of
COD fractions.
HOW MODEL EXTENSIONS INFLUENCE FUTURE
WASTEWATER CHARACTERISATION
The need to account for other processes is obvious when
dealing with the modelling of sulphur production (Sharma
et al. ), modelling of the fate of micropollutants (Plósz
et al. ; Pomiès et al. ), modelling of MBR clogging
(Naessens et al. ) or modelling the emission of green-
house gas (Ni et al. ).
Future wastewater characterisation may need to account
for new model components such as a very slowly biodegrad-
able COD fraction, detailed organic matter characterisation
(e.g. molecular level variables and/or particle size distri-
bution), or fractionation for nutrients.
Very slowly biodegradable COD fraction
Recent analyses (e.g. Menniti et al. ; Spérandio et al.
) has demonstrated that hydrolysis of particulate unde-
gradable endogenous products (XU,E) and particulate
undegradable organics from the influent (XU,inf ) – using a
slow hydrolysis rate (Ramdani et al. ) – might be suitable
to deal with measured behaviour in long SRT systems such
as processes operated under very low organic loading rate
(AS or MBR plants designed with long time capacity). This
approach might also be introduced to model sludge
reduction where a portion of the inert particulate COD
might become biodegradable (e.g. Cannibal process,
mycelium hydrolysing). A state variable for ‘very slowly bio-
degradable matter’ might increase the range of applicable
SRTs. However, a dedicated procedure is still needed to
accurately assess this fraction in the wastewater.
Molecular level variables/particle size distribution
The hydrolysis rate is a function of the influent characteris-
ation. Simulations of biological nutrient removal (BNR)
plants with ASM1 suppose a conversion of XCB into SB,
and use the instantaneous XCB/XOHO ratio in a bioreactor
to predict the hydrolysis rate (Henze et al. ). However,
in some cases, more detailed organic matter characteris-
ation (e.g. molecular level variables, particle size
distribution) will benefit description of hydrolysis phenom-
ena. More accurately predicted hydrolysis rates would
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Water Science & Technology | 67.11 | 2013
then be obtained for denitrification or anaerobic phos-
phorus release (Drewnowski & Makinia ). Also, the
assumption that compounds of a certain class (colloidal or
particulate) are hydrolysed at an equal rate irrespective of
their source (i.e. centrate or influent) is potentially proble-
matic for modelling. More detailed organic matter
characterisation becomes also relevant for wastewater
characterisation when wastewater contains industrial dis-
charges, whereas it is probably not necessary for standard
applications and objectives for domestic wastewater.
Besides using substrate availability, a few protocols have
been proposed to help with the calibration of hydrolysis at
macro-scale (Naidoo et al. ; Martin-Ruel et al. ).
Recently, papers reported that a deeper characterisation of
the organic matter should be considered to improve the
hydrolysis prediction:
• Fractionating the influent at a molecular level (i.e. lipids,
proteins and carbohydrates) because hydrolysis rates are
likely to be component specific. Indeed, systematic
measurements of these components may provide added
clarity (Gorini et al. ). The correlation between
these components and the COD fractions is still insuffi-
ciently investigated. In comparison, those components
were included in Anaerobic Digestion Model No 1
(ADM1) for the prediction of gas production with
wasted sludge (Batstone et al. ; Henze et al. )
and were shown to be essential for the prediction of the
behaviour of some micropollutants (Barret et al. ).
• Particle size distribution, in association with a surface-
based kinetics, may lead to a better description of
hydrolysis, assuming that particles breaking up into smal-
ler ones will allow for an increasing hydrolysis rate over
time (Dimock & Morgenroth ). The molecular
weight and the nature of chemical bonds may also pro-
vide information about the transformation pathways
(Sophonsiri & Morgenroth ).
This elucidation not only gives a better understanding of
hydrolysis but it also requires model extensions to accom-
modate improved characterisation of wastewaters (e.g.
known phenomenon of soluble slowly biodegradable
material exiting a primary clarifier (Ginestet et al. )).
Also, in MBRs, a better characterisation of the colloidal frac-
tion could improve prediction of fouling in membrane
bioreactors (Naessens et al. ), and it is useful to account
for particle sizes.
 2371 J.-M. Choubert et al. | Rethinking wastewater characterisation methods for activated sludge systems
Fractionation for nutrients
Plants facing very low effluent nutrient limits (one tenth to
one hundredth milligram per litre, Neethling et al. (),
may need to consider several things including refined frac-
tionation methods as well as model extensions. Nutrient
sources and fractionation becomes important (e.g. treating
return liquors from digesters) and the modelling of munici-
pal wastewaters containing some industrial discharges
with unbiodegradable nitrogen may become significant.
Urea and ammonia are the main sources of nitrogen in
raw municipal wastewater (SNHx ∼ 60–75% of total Kjeldahl
nitrogen) while the rest is typically organic nitrogen, split
into slowly biodegradable (XCB,N ∼ 22%), readily biodegrad-
able (SB,N ∼ 2%) and inert soluble (SU,N ∼ 1%) (Henze ;
Rieger et al. ). Future experimental methods should be
developed to characterise the dissolved organic nitrogen in
a discharged effluent. As nutrient limits decrease, this frac-
tion becomes increasingly important and could account for
0.7–2.1 mgN/L (Pehlivanoglumantas-Mantas & Sedlak
). Often, this nitrogen is incorrectly believed to be inert
whereas it may be composed of very slowly biodegradable
substances. Baiyang et al. () found degradation of up to
30% of this nitrogen in 5-day degradation tests. Arnaldos &
Pagilla () reported that amino acids account for less
than 30%, with the rest being heterogeneous in nature, and
composed of low-molecular weight nitrogenous substances.
CONCLUSIONS
This paper has discussed different challenges to consider
during the process of wastewater characterisation. These
issues are becoming and will become more important for mod-
elling more stringent effluent limits and the widening of model
system boundaries beyond activated sludge systems. The dis-
cussion included new variables of interest, new fractions for
new process units, and new operating conditions, as well as
time-varying influent fractionation. To address these require-
ments, the use of new experimental measurement or
estimation techniques have been proposed like particle size
distribution, molecular level fractionation, active biomass
determination and increased measurement frequency. These,
along with method standardisation and new data treatment
techniques will become increasingly important as the require-
ments for model predictive capabilities increase. However, it
should be understood that not all of the concepts are appli-
cable to every modelling project and to every wastewater.
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Water Science & Technology | 67.11 | 2013
Some of these concepts are research focused, while others
are dedicated to engineering application.
ACKNOWLEDGEMENT
This paper was first presented at WWTmod2012 and the
fruitful discussions are kindly acknowledged.
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