ARTICLE IN PRESS

Journal of Environmental Management 81 (2006) 1–18

www.elsevier.com/locate/jenvman

Review

Modelling anaerobic biofilm reactors—A review

V. Saravanan, T.R. Sreekrishnan
Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology Delhi, New Delhi-110 016, India
Received 7 October 2004; received in revised form 4 October 2005; accepted 5 October 2005
Available online 6 March 2006

Abstract

Anaerobic treatment has become a technically as well as economically feasible option for treatment of liquid effluents after the
development of reactors such as the upflow anaerobic sludge blanket (UASB) reactor, expanded granular sludge bed (EGSB) reactor,
anaerobic biofilter and anaerobic fluidized bed reactor (AFBR). Considerable effort has gone into developing mathematical models for
these reactors in order to optimize their design, design the process control systems used in their operation and enhance their operational
efficiency. This article presents a critical review of the different mathematical models available for these reactors. The unified anaerobic
digestion model (ADM1) and its application to anaerobic biofilm reactors are also outlined.
r 2006 Elsevier Ltd. All rights reserved.

Keywords: Mathematical model; UASB; AFBR; EGSB; Biofilter; Biofilm; Anaerobic

Contents
1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2. Models for UASB reactors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2.1. Flow model . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
2.2. Reactor model . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
3. Proposed models for the structure of the biofilm . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
3.1. Multi-layer model. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
3.2. Syntrophic microcolony model . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
3.3. Non-layered structure model . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3.4. Granule cluster structure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
4. Models for AFBR . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
4.1. Bed fluidization model . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
4.1.1. Terminal settling velocity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
4.1.2. Fluidization mechanics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
4.1.3. Effect of gas production on hydrodynamics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
4.1.4. Bed stratification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
4.2. Kinetic and reactor sub-models . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
4.2.1. Stratified biofilm models . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
5. The EGSB reactor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
6. The anaerobic biofilter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
7. The unified model for anaerobic digestion (ADM1) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
7.1. The extension of ADM1 to biofilm reactors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
8. Summary and conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15

Corresponding author. Tel.: +91 11 26591014; fax: +91 11 26582282.

E-mail address: sreekrishnan_t_r@hotmail.com (T.R. Sreekrishnan).

0301-4797/$ - see front matter r 2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.jenvman.2005.10.002
 ARTICLE IN PRESS
2 V. Saravanan, T.R. Sreekrishnan / Journal of Environmental Management 81 (2006) 1–18

1. Introduction Biogas

The term ‘‘biofilm reactor’’ refers to that class of Effluent

bioreactors where the biocatalyst exists in an anchored
form, either on the surface of an inert ‘‘carrier’’ or attached
to one another. The carrier could be the wall of the reactor,
baffles provided for this purpose or particles of some inert
material. Biocatalysts such as microorganisms could also
grow attached to one another, giving rise to a ‘‘biogra-
nule’’. The carrier or the biogranule could be stationary as
in a packed-bed or expanded bed system or mobile as in the
case of a fluidized bed system. Typically, in such reactors,
the rate of substrate conversion is limited by the rate of
transport of substrate into the biofilm. In an anaerobic
biofilm reactor, the biocatalyst will include all the different
bacterial species responsible for the break down of complex
organic molecules to a final end product consisting of
methane and carbon dioxide. The anaerobic treatment, as
the main biological step in wastewater treatment systems,
was scarce until the development of the upflow anaerobic
sludge blanket (UASB) reactor in the early 1970s (Lettinga
et al., 1980). UASB processes are based on the develop-
ment of dense granules (1–4 mm) formed by the natural
self-immobilization of the anaerobic bacteria. This kind of
immobilization does not employ any support material such
as Raschig rings or clay in the reactor (Nicolella et al.,
2000). A schematic of a typical UASB reactor is shown in
Fig. 1. Wastewater enters the bottom of the reactor
through the inlet liquid distribution system and passes
upward through the dense anaerobic sludge bed. Because
of the high biomass concentration, it was demonstrated
that volumetric organic loading rates as high as 50 kg
chemical oxygen demand (COD) per m3 per day could be
employed (Hulshoff Pol, 1989). The liquid velocity inside
the reactor is usually in the range of 0.5–1.0 m/h. This
reactor consists of a sludge bed, a sludge blanket and a
clarifier zone supplemented with a physical device called
the gas–solid separator.
In anaerobic fluidized bed reactor (AFBR), the liquid to
be treated is pumped through a bed of inert particles
(typically sand with a particle size range of 0.2–0.8 mm) at
a velocity sufficient enough (10–20 m/h) to cause fluidiza-
tion (Nicolella et al., 2000). In the fluidized state, the media
provides a large surface for attached biological growth and
allows biomass concentrations to develop in the range of
10–40 kg/m3 (Cooper and Sutton, 1983). A typical flow
diagram of an AFBR is shown in Fig. 2. Compared to
other high rate anaerobic reactors such as UASB, the
fluidized bed system is claimed to have the following
advantages: higher purification capacity, no clogging of
the reactor (as in filters), no problem of sludge washout
(as in UASB systems if granular sludge is not obtained),
and small volume and land area requirements (Heijnen
et al., 1989).
To study the sensitivity of the process to various
operating parameters and to optimize the design of these
reactors, it is necessary to have mathematical models which
Weir
Settler
Baffles
Sludge Blanket
Sludge granules
Sludge Bed
Influent
Fig. 1. Schematic diagram of UASB reactor.
Biogas
Treated water
Recycle line
Biofilm
Carrier
Wastewater feed
Fig. 2. Anaerobic fluidized bed reactor (AFBR).
are simple in concept and close to the physical situation
existing in these reactors. During the past few decades there
have been a number of studies on modelling of UASB and
AFBR reactors. An effort is made in this article to present
the gist of different approaches used in developing
mathematical models for these reactors. This article also
reviews different proposals available for modelling the
structure of the biological granules and biofilm-covered
particles in these reactors.
2. Models for UASB reactors
The formation of anaerobic granular sludge is consid-
ered to be a necessary condition for the successful
operation of a UASB reactor. The efficiency of the reactor
depends mainly on active biomass concentration and the
influent flow rate. As the reactor reaches steady state, a
dense bed composed of granulated sludge develops at the
bottom. Immediately above the sludge bed, a zone
consisting of finely suspended particles called the sludge
blanket forms. A clear zone over this sludge blanket
constitutes the settling zone. To keep the sludge granules in
suspended condition inside the reactor, the inlet flow rate
 ARTICLE IN PRESS
V. Saravanan, T.R. Sreekrishnan / Journal of Environmental Management 81 (2006) 1–18
should be below a threshold limit (normally below 1 m/h).
This low influent flow rate causes channelling problems in
the sludge bed zone, resulting in lowered efficiency of the
reactor. Hence to develop mathematical models for this
reactor, it is important to analyse the flow pattern inside
the reactor and reaction kinetics within the biological
granules. In general, models for UASB reactors consist of
two parts:
1. fluid flow model;
2. reactor model.
2.1. Flow model
Fluid flow models for UASB reactors differ considerably
in their approach. The different zones of a UASB reactor
are modelled as continuos stirred tank reactors (CSTR) or
plug flow reactors (PFR) having dead volume with bypass
flow between the zones (Bolle et al., 1986; Van der Meer,
1979; Wu and Hickey, 1997). The flow behaviour within a
zone mainly depends on the concentration and character-
istics of the biomass (Ojha and Singh, 2002).
Wu and Hickey (1997) have developed a flow model to
describe the flow pattern in a UASB reactor. They modelled
the sludge bed and blanket as a non-ideal CSTR by using a
combination of an ideal CSTR along with a dead zone and
a bypass flow. This CSTR is in series with a dispersed plug
flow reactor (PFR) (non-ideal PFR) that represents the
clarification zone above the sludge blanket (Fig. 3).
The equation of the flow model is given by
dC
Vb ¼ V b EðtÞ
Qf CðtÞ,
dt
EðtÞ ¼ VM f
b T in
if 0ptpT in ;
¼0 if T in p0;
where C(t) is the tracer concentration within the CSTR, Vb
the CSTR working volume, E(t) the input function for
tracer impulse, Qf the low fraction that enters the working
volume, Mf the fraction of mass input that go through
reactor’s working volume, and Tin the tracer injection time.
Ojha and Singh (2002) analysed the flow distribution in
different zones of the reactor in order to simulate the
UASB reactor performance. They used the flow resistance
approach to represent flow distribution. It was found that
with an increase in flow resistance in the UASB reactor
system, the magnitude of short-circuiting flows at the
reactor bed increased. The flow distribution at the blanket
and clarifier was found to have an influence on the flow
resistance. But no generalized model could be obtained.
Bolle et al. (1986) divided the reactor into three
compartments (Fig. 4): sludge bed, sludge blanket and
settler. The liquid flow in the sludge bed and the sludge
blanket were described by completely stirred tank reactor
systems. Liquid flow in the internal settler was described by
a plug flow model.
3
By-pass flow
CSTR Dispersed
plug flow
Dead volume
Fig. 3. Representation of hydraulic model (Wu and Hickey, 1997).
Effluent
Settler
Sludge blanket
Short circuit
Dead space Sludge bed
Influent
Fig. 4. Block diagram of fluid flow pattern (Bolle et al., 1986).
(2.1)
Narnoli and Indu (1997) developed a model for the
sludge blanket of a UASB reactor. According to them, the
(2.2) maximum organic load which a reactor can assimilate
depends on proportioning of the reactor height into the bed
and the blanket sections. A condition of force equilibrium
was considered between the rising gas bubbles from the
sludge bed and the adjacent water mass. The negative
pressure behind the bubble attracts the water mass and
leads to the formation of a wake. Using diffusion concepts,
solid particles moving along the wake due to the
concentration gradient were equated to those settling down
under the influence of gravity at steady state. The solid
concentration along the blanket height was computed and
found to match the experimental observations. This model
could be used to optimize the reactor dimensions and the
desludging schedule.
Singhal et al. (1998) reported that a simple two-zone
axial dispersion model adequately describes the fluid flow
characteristics of UASB reactors. They found that the fluid
flow behaviour is very sensitive to the volume of zones and
degree of bypassing between them but less sensitive to the
dead volume. The model assumed that some of the liquid
bypasses the first zone and enters directly into the second
zone. The schematic representation is shown in Fig. 5.
 ARTICLE IN PRESS
4 V. Saravanan, T.R. Sreekrishnan / Journal of Environmental Management 81 (2006) 1–18
Sludge Clarifier
Blanket
Zone-1 Zone-2
Fig. 5. Schematic of the two-compartment axial dispersion model
(Singhal et al., 1998).
They had verified the model efficacy through tracer
studies. The step response of an axially dispersed tubular
flow reactor for an inert tracer is described by the following
partial differential equation:
1 q2 C qC qC

¼ , (2.3)
Pe qZ2 qZ qy
where C is the dimensionless concentration of the tracer
(c/c0), Pe is the Peclet number (uL/D), Z is the dimension-
less distance (z/L), y is the dimensionless time (t/t), u is the
linear average velocity of the liquid in the reactor, L is the
length of the reactor, and D is the axial dispersion
coefficient. The simulation and experimental results gave
a very good fit validating this flow model.
All the above multi-compartment models were capable
of fitting laboratory scale experimental data well. In the
case of full-scale reactors, the feed distribution is very
different from that in the laboratory scale reactor. Distinct
zones of sludge bed, sludge blanket and clarifier may not be
observed due to the large volume of the reactor and uneven
flow distribution. This will make the task of quantifying the
extent of non-ideality in the flow patterns difficult. It may
not be correct to predict the fluid flow pattern based on lab
scale studies alone. Hence, further investigations are
required to test the models for full-scale reactors.
2.2. Reactor model
Substrate degradation in the bioreactor is dependant on
the observed reaction rate. In a biofilm type of reactor set-
up, the observed reaction rate is normally less than the rate
predicted by the reaction kinetics. The reason for this is
that the substrate concentration actually available to the
microorganisms is less than the substrate concentration
present in the bulk liquid. This is due to the control exerted
by the mechanism of molecular diffusion on the penetra-
tion of substrate into the biofilm. It is, therefore, important
to analyse the rate-determining step while developing
models. Hence, the reactor model consists of 3 parts:
substrate utilization within the biofilm (granules), mass
transfer and transport in the granule bed and blanket, and
mass transport within the clarifier.
Kinetic models are based on the relationship between
limiting substrate concentration and growth rate as
proposed by Monod (1950). James (1961) pointed out that
in biological processes a wide variety of substances might
act as limiting substrates. Stewart (1956) and Agardy et al.
(1963) have used COD as the limiting substrate in the
development of their models for the anaerobic digestion
process. Lawrence and McCarty (1967) used volatile acids
concentration as the limiting substrate, since it is generally
believed that the rate-limiting step in the anaerobic
decomposition of complex organics is the conversion of
volatile acids to methane. Andrews (1969) has presented a
dynamic model for the anaerobic digestion process, which
considers only the methanogenesis step. It is commonly
observed in the field that a high volatile acids concentration
and low pH value leads to digester failure. To represent this
phenomenon in a quantifiable way an inhibition function
(Haldane type) was used in the model. The un-ionized form
of volatile acids was considered as the rate limiting
substrate since it is a function of both pH and the total
volatile acids concentration.
Rittman and co-workers have presented a biofilm model
describing the biofilm growth and substrate flux under
steady-state conditions (Rittmann, 1982; Rittmann and
McCarty, 1980a). UASB reactor models using methano-
genic biofilm models have focused on mass transfer
limitations and single as well as multiple limiting substrates
(Alphenaar et al., 1993; Atkinson and Davies, 1974;
Atkinson and How, 1974; Buffiere and Steyer, 1995; de
Beer et al., 1992; Lens et al., 1993; Lin, 1991).
Thick biofilms may give rise to mass-transfer limitations
for the substrate within the biofilm, resulting in an overall
reduction in the substrate conversions achieved. Under this
condition, the transport of substrate into the biofilm or
transport of products out of the biofilm could become the
rate-determining step. Contradictory results have been
reported on the impact of internal (within the biofilm) and
external (in the bulk liquid phase) mass transport. While
some reports indicate that both internal and external
diffusion limitations influence the rate of substrate utiliza-
tion (Dolfing, 1985; Wu et al., 1995), others indicate that
mass transport limitations are not observed in anaerobic
biofilms (de Beer et al., 1992; Schmidt and Ahring, 1991)
even at film thicknesses as high as 2.6 mm (Droste and
Kennedy, 1986). In addition, it has been reported that pH
profiles inside anaerobic granules may affect the conversion
rate more than mass transport limitations (de Beer et al.,
1992).
Gonzalez-Gil et al. (2001a) studied the influence of
external and internal mass transport in anaerobic granular
sludge. For this, kinetic properties of acetate degrading
methanogenic sludge granules of different mean diameters
were assessed at different up-flow velocities. It was
experimentally found that external mass transport resis-
tance can be neglected and that biogas formation does not
influence the diffusion rates. The substrate uptake could be
explained by biofilm (internal) diffusion.
The general approach to the modelling problem starts
with development of the biofilm model. These are then
used to find the substrate flux at the surface of each
granule. The biofilm model is then tailored with the flow
 ARTICLE IN PRESS
V. Saravanan, T.R. Sreekrishnan / Journal of Environmental Management 81 (2006) 1–18 5

model to predict the overall performance of the reactor. In where Ar is the total granule surface area; Tin the substrate
general, the following assumptions were made in develop- injection time for the acetate impulse; kl Ar ðsb
sð0; tÞÞ
ing anaerobic biofilm models: describes the substrate transferring from the bulk solution
through the liquid boundary layer into the granules. Data
1. Granules are spherical. from an acetate impulse experiment was simulated using
2. Substrate utilization is described by the Monod model. these hydraulic- reaction–diffusion models with a good fit.
3. External mass transport is negligible. The results indicated that diffusion inside the granules,
4. Density of the biofilm is constant throughout the reaction kinetics and hydraulic behaviour play important
granule. roles in the performance of a UASB reactor.
5. The relative concentrations of the acidogenic and Bolle et al. (1986) developed an integrated dynamic
methanogenic bacteria are constant throughout an model for the UASB reactor. Their flow model has been
anaerobic granule. This remains true irrespective of described in Section 2.1. Substrate and biomass balance
the location in the reactor from where the granule was were done over the sludge bed and sludge blanket. Thus the
taken. model was developed by integrating the fluid flow pattern
in the reactor, the bacterial growth and substrate utiliza-
tion kinetics, and the mass transport between different
Many of the reported studies deal with steady-state
compartments and different phases. This model was able to
conditions. Unsteady state is the most critical situation for
predict the sludge bed height, the biomass concentration in
modelling, associated with real-time control strategies. Wu
the sludge blanket, the short-circuiting flows over the bed
and Hickey (1997) developed a dynamic model to describe
and the blanket, and the effluent COD concentrations as a
UASB reactors with methanogenic anaerobic granules.
function of the hydrodynamic load, COD load, pH and
They assumed that substrate diffuses into the granules’
settler efficiency.
outer layer upto a thickness of 100 mm. At the inner edge of
Skiadas and Ahring (2002) proposed a model for
this layer, the substrate gradient reaches zero. Growth of
UASB reactors based on the cellular automata (CA) concept.
acetate utilizing methanogens is neglected due to their low
A cellular automation is a simulation, which is discrete in
growth rate.
time, space and state. A CA model usually consists of an
By applying an acetate mass balance on anaerobic
array of compartments similar to the spaces in a game of
granules, the granular bed and the clarifier, respectively,
naughts and crosses. The CA theory has been applied to
the dynamic model equations are obtained.
predict the layer structure of the granules, which is high
For anaerobic granules:
acidogen and low methanogen concentrations at the outer

2 
qs qs 2 @s km xm s granule layers and the reverse at the inner granule layers. It
¼D

. (2.4) has also predicted the granule diameter and granule
qt qx2 R
x @x ks þ s
microbial compositions as functions of the operational
Boundary conditions: parameters. The other models do not consider the effect of
operational parameters on granule size and composition.
1.
D qs þ kl sb ¼ kl sb ; x ¼ 0, Details on automation models can be found in the excellent
qx review paper by Wimpenny and Colasanti (1997).
Even though different authors have presented good
2. qs ¼ 0; x ¼ d, experimental fit for their models, a unified approach is
qx lacking. The above kinetic models, in general, assume
external mass transfer to be negligible. Some of the
where R is the granule radius; km the specific substrate assumptions may lead to poor predictions. For example,
utilization rate; ks the half velocity constant; D the effective the assumption of Monod kinetics, spherical shape and
diffusion coefficient; kl the mass transfer coefficient; xm the uniform density of the granules may not be true in actual
active acetate utilizer biomass within the outer layer d; s the reactors. The lack of versatile models in the literature
acetate concentration within the biofilm. shows that a lot of improvements can be made in future
A mass balance on a granule bed involves three terms: models. Some of them could be
substrate entering the reactor, substrate leaving the reactor,
and substrate being taken up by the granules and
subsequently being utilized: 1. Incorporation of the variation of density within the
ds biofilm/granule.
Vb ¼ V b EðtÞ
Qf sðtÞ
kl Ar ðsb
sð0; tÞÞ. (2.5) 2. Kinetic expressions, which include inhibition terms.
dt
3. Kinetic expressions, which include the non-uniform pH
Initial condition: sb(0) ¼ sb0 profile inside the biofilm.
4. Diversity in the bacterial population distribution
EðtÞ ¼ VM f
b T in
if 0ptpT in for impulse;
(2.6) inside the granule in terms of the predominant species/
¼0 if T in p0; groups.
 ARTICLE IN PRESS
6 V. Saravanan, T.R. Sreekrishnan / Journal of Environmental Management 81 (2006) 1–18
3. Proposed models for the structure of the biofilm
In an anaerobic reactor, under favourable conditions,
bacterial cells can attach to each other and grow as a
granule. This granule has a 3-D structure. When some inert
carrier particles or surfaces are introduced in the system,
cells grow on the carrier particles/surfaces forming a
biofilm. Depending upon the structure of the carrier
material the resulting biofilm will have a 2-D or 3-D
structure. Microbial composition and their distribution
inside the granules play an important role in substrate
conversion. Conversion could be limited by the process of
diffusion of the substrate within the biofilm as well as the
concentration of biomass bringing about the conversion.
This information is very essential to develop the biofilm
model. Many authors have reported that the structure of
the granule/biofilm is mainly determined by the substrate
degradation kinetics rather than the geometry with which it
is formed (Batstone et al., 2004; Fang et al., 1995; Guiot et
al., 1992). Batstone et al. (2004) have developed a biofilm
structure model which predicts the structure of a 2-D
biofilm. This model has been formulated based on
substrate degradation and diffusion kinetics. They exam-
ined four different types of granules obtained from UASB
reactors treating wastewaters from a cannery, a slaughter-
house, and two breweries. The microbial structures of these
granules were assessed by fluorescence in situ hybridization
probing with 16S rRNA-directed oligonucleotide probes,
scanning electron microscope and transmission electron
microscope. The biofilm model could exactly predict the
structures of different types of granules observed through
the experiments when the model was simulated for the
same operating conditions. This proves that the structure
of the biofilm is influenced by the substrate kinetics and not
by the geometry of the biofilm. Van Loosdrecht et al.
(2002) has shown that a 2-D model is sufficient to represent
a 3-D structure. Picioreanu et al. (2001) also has supported
the same. Hence, the different models proposed for the
biofilm structure in this section are applicable to both
granules and 2-D biofilms.
3.1. Multi-layer model
MacLeod et al. (1990) and Guiot et al. (1992) proposed a
three-layered structure for the bacterial aggregates treating
carbohydrates in a UASB reactor. According to this
model, the microbiological composition of granules is
different in each layer. The inner layer mainly consists of
methanogens that may act as nucleation centres necessary
for the initiation of granule development. H2-producing
and H2-utilizing bacteria are dominant species in the
middle layer, and a mixed species including rods, cocci and
filamentous bacteria takes the predominant position in the
outermost layer (Fig. 6).
To convert a target organic compound to methane, the
spatial organization of methanogens and other microbial
species in the granules (such as those in a UASB reactor) is
Acidogens
Methanothrix
H2 producing Acetogens and
H2 consuming organisms
Fig. 6. Three-layered structure of the bacterial aggregates (MacLeod
et al., 1990).
essential. The layered structure of UASB granules is
supported by the works of Arching et al. (1993) and Lens
et al. (1995) with immunological and histological methods.
The dynamic model proposed by Arcand et al. (1994) also
supports this structure. Similar support for the layered
structure is found in reports from Santegoeds et al. (1999)
using microelectrodes. Sekiguchi et al. (1998, 1999) and
Tagawa et al. (2000) also confirmed this structure by
fluorescence in situ hybridization using 16S rRNA targeted
oligonucleotides. A distinct layered structure was also
found in the methanogenic–sulfidogenic aggregates, with
sulfate-reducing bacteria in the outer 50–100 mm and
methanogens in the inner part (Sekiguchi et al., 1998).
Unlike the initial multi-layer model proposed by MacLeod
et al. (1990) recent research showed that UASB granules
have large, dark, non-staining centres, in which neither
archaeal nor bacterial signals could be found (Rocheleau
et al., 1999). In fact, the non-staining centre in the UASB
granules might be the result of the accumulation of
metabolically inactive, decaying biomass and inorganic
materials (Sekiguchi et al., 1998).
The importance of ECP in anaerobic granulation has
been observed by Schmidt and Ahring (1994, 1996). They
conclude that ECP may play an important role in building
spatial structure and maintaining the stability of UASB
granules, but are unsure of its contribution to the initiation
of anaerobic granulation. A large quantity of ECP is
unnecessary for making up active granules since it
decreases the porosity and thereby increases the diffusional
resistance for substrate to penetrate the granule. Too much
ECP could even cause deterioration of floc formation
(Schmidt and Ahring, 1996).
3.2. Syntrophic microcolony model
According to the syntrophic microcolony model, a close
synergistic relationship among different microbial groups is
essential for efficiently breaking down the complex organic
compounds. In fact, the syntrophic microcolonies provide
the kinetic and thermodynamic requirements for inter-
mediate transference and therefore efficient substrate
conversion (Schink and Thauer, 1988). They state that
the synergistic requirements would drive bacteria to form
granules, in which different species function in a synergistic
way and can easily survive.
 ARTICLE IN PRESS
V. Saravanan, T.R. Sreekrishnan / Journal of Environmental Management 81 (2006) 1–18
3.3. Non-layered structure model
Contrary to the multi-layer model, anaerobic granules
with non-layered structure have also been reported (Fang et
al., 1995; Grotenhuis et al., 1991; Wu et al., 2001).
There is evidence that a layered structure of the UASB
granules would be developed with carbohydrates and a non-
layered one using substrates having a rate-limiting hydrolytic
or fermentative step (e.g. proteins) (Fang, 2000; Fang et al.,
1995). This is probably due to different initial steps in the
degradation of carbohydrate as compared
to that of proteins. Degradation of carbohydrates to
small molecules is faster than the subsequent degradation
of the intermediates, whereas the initial step in protein
degradation is a rate-limiting step. That is, the protein
degradation step is slower than the steps which follow it.
Results from fluorescence in situ hybridization com-
bined with confocal scanning laser microscopy clearly
showed that protein-fed granules possess non-layered
structure with a random distribution of Methanosaeta concilii
(Rocheleau et al., 1999). However, granules, which differ in
their composition in terms of the predominant microbial
species, can still be formed from the same substrate
(Daffonchio et al., 1995; Schmidt and Ahring, 1996).
Based on microscopic examination of the UASB
granules, recently Fang (2000) proposed that the microbial
distribution of the UASB granules strongly depends on the
degradation thermodynamics and kinetics of individual
substrate. Therefore, it appears that different dominating
catabolic pathways may give rise to granules, which are
different in their structure. Spontaneous and sudden
washout of the established granular sludge bed, as a result
of a change in wastewater composition, is a common
problem encountered in the operation of UASB systems.
So far, none of the individual models for the granule
structure can explain this phenomenon. If a factor that is
independent of the wastewater composition can initiate the
formation of UASB granules, a change in the wastewater
composition should not lead to the washout of the entire
granular sludge bed. Thus, it is a reasonable speculation
that there should be a substrate composition-associated
factor that highly contributes to the formation of UASB
granules, but is not yet included in the present models of
granule structure (Liu et al., 2003).
Liu et al. (2003) proposed a general model for anaerobic
granulation in UASB reactors. This model consists of four
steps for granulation.
1. Physical movement to initiate bacterium-to-bacterium
contact or bacterial attachment onto nuclei.
2. Initial attractive forces to keep stable multi-cellular
contacts, e.g. physical, chemical and biochemical forces.
3. Microbial forces to make cell aggregation mature, e.g.
production of ECP.
4. Hydrodynamic shear force shaping 3-D structure of
microbial aggregates.
7
3.4. Granule cluster structure
Recently the structure of anaerobic granules of an
expanded granular sludge bed (EGSB) reactor was studied
by Gonzalez-Gil et al. (2001b). They found black spherical
granules having numerous whitish spots on their surfaces.
Cross-sectioning these aggregates revealed that the whitish
spots appeared to be white clusters embedded in a black
matrix (Fig. 7). High magnification electron microscopy
showed that the white clusters mainly consisted of acetate
utilizing methanogens (Mathanosaeta spp.) and the black
matrix consisted of syntrophic species and hydrogenotrophic
methanogens (Methanobacterium like and Methaospirillum
like organisms). Fluorescent in situ hybridization using 16 s
rRNA probes of crushed granules showed that 70% of the
cells belonged to the archaebacterial domain and 30% to
eubacterial domain. The authors have provided a very logical
explanation as to why a cluster structure, as observed, is
advantageous over a layered structure. Acetate produced in
the black zone is transported by random diffusion in all
directions and thus penetrates the Methanosaeta clusters from
all sides. Hence, substrate depleted zones are circumvented,
which allows the growth of more active biomass per unit area
of aggregate.
Batstone et al. (2004) studied the influence of substrate
degradation kinetics on the microbial community structure
in granular anaerobic biomass. The granules, which were
grown in the effluent containing soluble as well as
particulate protein, had homogeneous structure. The
primary cause of this structure was assessed through
biofilm modelling. They postulate that the particulate
nature of the wastewater and the slow rate of particulate
hydrolysis, rather than the presence of proteins in the
wastewater, was responsible for the homogeneous structure
of the granules. Because solids hydrolysis was rate limiting,
soluble substrate concentrations were very low (below
Monod half-saturation concentration), which caused low
growth rates.
From the above information it can be concluded that
there are two key factors which determine the structure of a
granule (i.e. the organization of the microbial community
within a granule). They are
1. The nature of organic compounds present in the
wastewater.
2. The kinetics of substrate degradation.
Methanosaeta clusters
Zone with syntrophic
eubacteria and
hydrogenotrophic
methanogens
Seed aggregate
Fig. 7. Schematic representation of the architecture of anaerobic
aggregates (Gonzalez-Gil et al., 2001b).
 ARTICLE IN PRESS
8 V. Saravanan, T.R. Sreekrishnan / Journal of Environmental Management 81 (2006) 1–18

The relative concentrations of different microbial species
within a granule is another important factor required in
developing a biofilm model. This seems to depend on the
concentration of the substrate. Since there could be
concentration gradients with respect to substrate concen-
tration within the same reactor, the composition of the
granule in terms of the microbial species may vary from
granule to granule obtained from different parts of the
same reactor. Hence a robust model, capable of incorpor-
ating the structure as well as composition of the granules in
terms of the different microbial communities present, is
required. Such a model for the biofilm/granule can be used
in developing reactor models which are more versatile as
well as reproducible with a higher level of accuracy.
4. Models for AFBR
The AFBR uses inert carrier particles to provide
mechanical support for growth of the biofilm. These
biogranules are maintained in a fluidized state by using
the energy of the influent liquid. Such a fluidized bed
system is free from the channelling problems encountered
in other biofilm reactors. A serious operational problem
associated with the AFBR is that when the biofilm grows
on the carrier surface, the composite density of the film-
covered particle decreases, ultimately resulting in the carry
over of the film-covered particles out of the reactor. Full-
scale application of AFBRs is not common due to lack of
sound design principles. Many attempts have been made to
study the process kinetics and the factors affecting process
performance.
In general, models for AFBRs include the following
elements:
Di Felice, 1995), have to be modified to take into account
the effect of the biofilm layer on the rigid particles
(Nicolella et al., 2000).
4.1.1. Terminal settling velocity
The terminal settling velocity of a single spherical
particle in an infinite expanse of fluid is
 
4gðrs
rl Þ 0:5
ut ¼ . (4.1)
3C D rl
Depending on the biofilm thickness and on the carrier
type, values for the equivalent density of biofilm particles
(rs ) range typically from 1100 to 1500 kg/m3. The drag
coefficient CD is a function of the particle Reynolds
number defined as
rl d s ut
Ret ¼ . (4.2)
ml
Biofilm particles are in the intermediate flow regime
(1oReto100) for the vast majority of cases, obtained
when sand (0.5–1 mm) or carrier materials with densities in
the same range are used as the inert support. In the
intermediate flow regime, the drag coefficient for a smooth,
rigid sphere is (Perry and Green, 1997)
C D ¼ 18:5Ret
0:6 . (4.3)
This correlation cannot be used for carrier-supported
granules, since they are neither smooth nor rigid. For this
reason, empirical correlations have been suggested for the
estimation of CD for biofilm particles:
C D ¼ 17:1Ret
0:47 ; 50oRet o100 (4.4)
Hermanovicz and Ganczarczyk (1983),

1. A bed fluidization model which describes the size and C D ¼ 36:66Ret
0:67 ; 40oRet o90 (4.5)
number of bioparticles per unit fluidized bed volume. Mulcahy and Shieh (1987),
2. A biofilm model which describes the rate of substrate
24
conversion per individual granule. CD ¼ þ 21:55Ret
0:518 ; 15oRet o87 (4.6)
3. A reactor flow model, which links the biofilm and bed Ret
fluidization models to yield substrate concentration as a Ro and Neethling (1990),
function of axial position within the AFBR. 24
CD ¼ þ 14:55Ret
0:48 ; 40oRet o90 (4.7)
Ret
4.1. Bed fluidization model Yu and Rittmann (1997),
C D ¼ 29:6Ret
0:6 ; 7oRet o90 (4.8)
Hydrodynamic behaviour of carrier supported biogra-
nules plays an important role in designing AFBRs. When Nicolella et al. (1999).
the granules grow, their size, shape and composite density The drag coefficient of the biofilm covered carrier
change. This has an impact on the hydrodynamic particle is generally found to be more than that of smooth,
behaviour of the granules. Information on settling and rigid spheres. Surface roughness has generally been
fluidization characteristics, such as fluidized bed-height, as considered as the reason for the increase in the drag
a function of liquid velocity is required for the designing of coefficient of biofilm covered carrier particles (Hermano-
AFBRs. Information on fluidized-bed height is important vicz and Ganczarczyk, 1983; Mulcahy and Shieh, 1987).
because it establishes the solids residence time and the The above correlations are all empirical since they were
specific biofilm surface area in the biologically active zone. obtained by fitting experimental data generated by
The relationships valid for fluidization of rigid particles, different groups, who employed fluidized bed systems,
readily available in the chemical engineering literature (e.g. which differed from one another in one or more aspects.
 ARTICLE IN PRESS
V. Saravanan, T.R. Sreekrishnan / Journal of Environmental Management 81 (2006) 1–18
The deformable nature and surface roughness of the
biofilms and the type of carrier particle used play a major
role in determining the hydrodynamic nature of the biofilm
covered particle. Hence, bed fluidization models should be
able to incorporate those parameters which have an
influence on the hydrodynamic behaviour of the biofilm
covered particles. It may not be feasible to have an
analytical expression for this purpose, given the complexity
and variability inherent to the system. While an empirical
relationship is acceptable, it should be able to cater to a
wider range of operational parameters normally encoun-
tered rather than a narrow set of operating conditions.
4.1.2. Fluidization mechanics
In a fluidized bed anaerobic reactor, the film-covered
particles are kept in a fluidized state by the incoming liquid.
The bed porosity and biomass concentration in the bed are
determined by the mechanics of fluidization. Hence, a
realistic mathematical expression for the bed fluidization is
necessary.
For a bed of uniform spherical particles, the following
relation was proposed (Richardson and Zaki, 1954):
us ¼ ui ðeÞn , (4.9)
where us is the superficial liquid velocity, e ¼ porosity and
ui ¼ ut 10
d=D , (4.10)
where ut is the terminal settling velocity of particle, d the
particle diameter and D the diameter of bed, n is a constant
given by
n ¼ 4:65 þ 20d=DðRet o0:2Þ, (4.11)
n ¼ ð4:4 þ 18d=DÞðRet Þ
0:03 ð0:2oRet o1Þ, (4.12)
n ¼ ð4:4 þ 18d=DÞðRet Þ
0:1 ð1oRet o200Þ, (4.13)
n ¼ 4:4ðRet Þ
0:1 ð200oRet o500Þ, (4.14)
n ¼ 2:4ð500oRet Þ. (4.15)
Whether this equation can be applied directly and in its
entirety to a biological fluidized bed is a controversial
subject. Richardson and Zaki have derived the equation for
a bed of uniform spherical particles, which are hard and
have a smooth surface. But the biological film covered
particles seldom have these characteristics.
Andrews and Tien (1979) related the bed height to the
amount of biomass in the bed. The fluidized bed tends to
stratify vertically based on the settling velocity of the
bioparticles. If no stratification is assumed, the bed height
is related to average biofilm thickness x̄ by
H ð1
ec Þð1 þ x̄Þ
¼ . (4.16)
H c 1
ec ½ð1 þ x̄Þ1=3 =ð1 þ Ax̄Þ1=n
In the case of complete stratification
Z xmax
H ð1 þ xÞ
¼ ð1
ec Þ gðxÞ dx. (4.17)
Hc 0 1
e
9
A practical working expression was derived based on the
above two expressions by Andrews and Tien (1979) as
follows:
H
¼ 1 þ ð1 þ BÞx̄, (4.18)
Hc
where


 
ec D 1 þ ec 1
3A2 1
3A
B¼ 3 þ x0 2 þ D
D¼ ,
3ð1
ec Þ 1
ec 1
3A 3n
where H is the bed height, e the bed porosity, x the film
volume/clean particle volume, x̄ the mean value of
distribution function of bacterial film gðxÞ, A the buoyant
density of bacterial film/buoyant density of clean particle, n
the exponent in the Richardson–Zaki correlation. The
subscript ‘c’ denotes the bed of clean particles.
This expression was found to predict the experimental
data well.
Mulcahy and La Motta (1978) have developed specific
correlations to determine ut and n for the case of
bioparticles in a fluidized bed as follows:
" #0:75
ðrs
rl Þgd 1:67
ut ¼
p
, (4.19)
27:5r0:33
l m0:67
n ¼ 10:35 Re
0:18
t ð40oRet o90Þ, (4.20)
where rs is the density of bioparticles; rl the liquid density;
m the liquid viscosity.
Ngian and Martin (1980) found that the Richardson and
Zaki correlations gave a satisfactory estimate for ui for
small support particles whereas ui was 30–70% below the
experimentally determined value for larger support parti-
cles. Nicolella et al. (1999) found ui to be only 80% of the
unhindered settling velocity. They recommend that the
correlation should be used with caution while determining
the constant ui.
In general, the Richardson and Zaki correlation is found
to describe the bed expansion characteristics of a fluidized
bed. But the values of the constants n and ui seem to be a
function of the property of the biofilm covered particles.
The application of these correlations to a full-scale reactor
containing a wide size distribution of biofilm covered
particles needs to be verified.
4.1.3. Effect of gas production on hydrodynamics
The effect of gas production on hydrodynamics of
fluidized beds is an important factor to be studied for
design and scale-up of AFBRs. Many investigations on the
flow pattern in an AFBR suggested that an axially
dispersed plug flow model can be used for the flow model
(Hirata et al., 2000; Seok and Komisar, 2003). In these
studies the effect of gas production on the flow pattern was
not considered. Diez-Blanco et al. (1995) have studied the
effect of gas production on the hydrodynamic behaviour of
an AFBR. In this study, the bed contraction due to biogas
production in a fluidized bed of 6 m height was estimated to
 ARTICLE IN PRESS
10 V. Saravanan, T.R. Sreekrishnan / Journal of Environmental Management 81 (2006) 1–18
be less than 6%. Based on this, the investigators considered
the effect of biogas on the hydrodynamic behaviour to be
negligible. In contrast, Buffiere et al. (1998a) reported that
the effect of biogas on bed height could be negligible. But
the biogas effervescence affects the phase hold-ups (which
affects the liquid solid contact time) and liquid mixing of
the reactor system. The experimental study of Buffiere et al.
(1998b) compared the hydrodynamic behaviour of a gas
producing fluidized bed with a classical gas injected three
phase fluidized bed reactor. The overall gas hold up was
found to be more in the case of the gas producing fluidized
bed than the gas injected one. They also observed axial
variation of phase hold-up.
4.1.4. Bed stratification
Schreyer and Coughlin (1999) reported a stratification of
biofilm coated sand particles in a fluidized bed reactor.
Stratification can be attributed to the influence of a biofilm
on a particle’s settling velocity. The presence of a biofilm
cover decreases a particle’s overall density, thereby
increasing its buoyancy. The biofilm also increases the
particle’s size thereby increasing the drag force exerted on
it by the liquid flowing upward. The particles in a fluidized
bed are expected to segregate according to size and mean
density (Ro and Neethling, 1994; Safferman and Bishop,
1996; Trinet et al., 1991).
Bed stratification has many negative effects on the
performance of the reactor. Thicker biofilms pose diffusion
limitation and wash out problems. Hence, to prevent
stratification and to maintain uniform particle size, efforts
have been made to remove excess biofilm from larger
particles (Ruggeri et al., 1994; Safferman and Bishop, 1996;
Shieh et al., 1981; Trinet et al., 1991). Examples of such
efforts include external sand-biomass separators (screens),
operation of an impeller at the top of the bed and internal
screen cleaning devices. Many researchers have examined
the effect of shear on biofilm and biofilm detachment rate,
mainly as a tool to control biofilm thickness (Chang and
Rittmann, 1988; Chang et al., 1991; Gjaltema et al., 1997;
Peyton and Characklis, 1992; Rittmann, 1982; Safferman
and Bishop, 1996; Stewart, 1993; Trinet et al., 1991).
Biofilm detachment rate appears to depend on turbulence
and particle concentration; an increase in either increases
detachment rate. Biofilm has been observed to be relatively
smoother and more homogeneous under conditions of high
shear (i.e., high liquid velocity) than under low shear
conditions (Lau, 1995; Zhang and Bishop, 1994). The study
by Schreyer and Coughlin (1999) showed that the
introduction of a thinner to increase the shear resulted in
a non-stratified bed.
Bed stratification could occur due to differences in the
biofilm thickness or differences in the carrier particle size or
both. But bed stratification is most common when carrier
particles are not of uniform size. A completely mixed bed
(i.e. no stratification) was observed by Andrews and Tien
(1979) when uniform carrier particles were used.
4.2. Kinetic and reactor sub-models
Substrate removal within a biological film of uniform
thickness attached to a spherical particle is described by


D d 2 ds
r ¼ Rt , (4.21)
r2 dr dr
where D is the effective diffusion coefficient of substrate
within the biological film; r the radial coordinate measured
from the centre of the support particle; s the substrate
concentration within the biofilm and Rt the intrinsic rate of
substrate consumption per unit volume of biological film.
It is generally accepted by many authors that substrate
removal kinetics can be modelled using the microbial
growth model proposed by Monod. Many researchers
(Grady, 1982; Henze and Harremoes, 1983; Iwai and
Kitao, 1994) have suggested that depending on the values
of the model constants, reaction rate can be represented by
first- or zero-order kinetics. For AFBRs, many authors
demonstrated that zero-order kinetics provide an adequate
description of substrate consumption (i.e. considering the
acidogenesis and methanogenesis phases together) (La
Motta and Patricio, 1996; Mulcahy and La Motta, 1978;
Mulcahy et al., 1980; Shieh et al., 1982).
For zero-order reaction, the reaction rate term is
Rt ¼ rk0 . (4.22)
The solution of Eq. (4.21) for complete substrate
penetration inside the biofilm and for the partial penetra-
tion has been presented by Mulcahy et al. (1980).
For fully penetrated biofilm:
Observed rate ¼ 43prk0 ðr3p
r3m Þ. (4.23)
For partial substrate penetration:
ðr3p
r3m Þ1:9
Observed rate ¼ 1:76ðrk0 Þ1:45 ðr3p
r3m Þ1:9 s0:45
b 0:45
.
r1:8
p D
(4.24)
rm is the radius of support particle; rp the radius of
biological particle; sb the concentration of substrate in the
bulk of the liquid within the fluidized bed.
A simple plug flow model was used to describe dissolved
substrate transport in the axial direction in a fluidized bed
reactor (Mulcahy and La Motta, 1978):
dsb
U þ Rv ¼ 0. (4.25)
dz
With the boundary condition
z ¼ 0; sb ¼ s0
where U is the average liquid velocity in the longitudinal
direction; sb the substrate concentration in the bulk of the
liquid; s0 the influent substrate concentration; Rv the rate
of reaction.
For fully substrate penetrated biofilm, the concen-
tration profile along the reactor was given by Mulcahy
 ARTICLE IN PRESS
V. Saravanan, T.R. Sreekrishnan / Journal of Environmental Management 81 (2006) 1–18 11

et al. (1980) as Eq. (4.29) gave

"
3 #

Vm rp sb sss
se ¼ s0
rk0
1 , (4.26) Rt ¼ K , (4.32)
Q rm km þ sss
where K ¼ ðrb dmmax =Y x=s Þ:
where se is the concentration of substrate in the effluent
Transforming the above equation using Lineweaver–
stream. La Motta and Patricio (1996) tested this model by
Burke linearization,
conducting experiments. This test showed a reasonable
agreement between the zero-order kinetic model with sb km 1 1
¼ þ . (4.33)
complete substrate penetration and the experimental data. Rt K sss K
Hirata et al. (2000) used a different approach to evaluate
Plotting sb/Rt versus 1/sss gave a straight line with slope
the kinetic parameters of the biochemical reactions taking
km/K and intercept 1/K. Using the above plot for the
place in a three phase fluidized bed reactor. Using the
known value of inlet substrate concentration, the steady-
substrate balance at steady state and assuming Monod
state substrate concentration could be found.
kinetics, an equation relating the substrate consumption
Buffiere et al. (1998c) studied the biofilm activity along
rate to substrate concentration (expressed as Biochemical
the reactor height. They developed a biofilm model
Oxygen Demand, BOD5) and total biofilm surface area was
assuming
established. The following assumptions were made in
formulating the model:
1. Homogeneous biofilm of uniform thickness.
2. Spherical support media of uniform size.
1. Reactor system is completely mixed.
3. Internal mass transfer described by Fick’s law.
2. Total organic carbon (TOC), which is expressed in terms
4. Liquid phase perfectly mixed with homogeneous con-
of BOD5, is the only rate-limiting substrate. Other
centration.
substrates are in excess.
5. No external mass transfer limitation.
3. The reaction follows Monod kinetics and substrate
inhibition is negligible.
4. Reaction occurs at constant volume. The mass balance for a substrate s in the biofilm is
expressed by

 X
Performing a substrate balance at steady state yielded D d 2 ds
r ¼ Vsi . (4.34)
1 r2 dr dr i
F ðsin
sss Þ ¼ rx V , (4.27)
Y x=s The boundary conditions are
where F is the inlet flow rate, V the reactor volume, rx the r ¼ rp ; s ¼ s0 ,
biofilm growth rate, Yx/s the yield coefficient ¼ mass of
biomass formed/mass of substrate consumed, Sss the ds
r ¼ rc ; ¼ 0,
steady-state value of the rate limiting substrate concentra- dr
tion inside the reactor, Sin the inlet substrate concentration.

If the reaction followed Monod kinetics, then the proposed xs mmax s
Vs ¼ . (4.35)
rate equation was Y x=s ks þ s


mmax sss The right-hand side of (4.34) is the sum of all substrate
rx ¼ mx ¼ x, (4.28)
km þ sss uptake rates minus the sum of all substrate production
rates.
where m is the specific growth rate, mmax is the maximum
In the liquid phase, the mass balance for one substrate s
specific growth rate and km the Monod constant.
is given by
Substituting rx into the substrate balance equation

  !
1 m max sss ds ds
F ðsin
sss Þ ¼ V x ¼ Rt , (4.29) V L ¼ Qðsin
sÞ þ DAp
 , (4.36)
Y x=s km þ sss dt dr r¼rp

where s is the concentration of component s; sin the inlet

V x ¼ rb dsb ,
where rb is the biomass dry density, d the effective biofilm
thickness and sb the total biofilm surface area. The total
surface area was computed as follows:
sb ¼ pðDave Þ2 N,
where N is the total number of particles inside the reactor
and Dave is the average particle diameter. Modifying
(4.30)
concentration of s; Q the input flow rate; rp the bioparticle
radius; r the radial distance measured from bioparticle
centre; D the diffusivity of component s in the biofilm; mmax
the maximum specific growth rate for s-utilizing bacteria;
Vsi the reaction rate of s through reaction I; Ap the
(4.31)
exchange area of the bioparticles; VL the liquid phase
volume; xs the s-utilizing bacteria concentration; Y x=s the
biomass yield factor for s-utilizing bacteria.
 ARTICLE IN PRESS
12 V. Saravanan, T.R. Sreekrishnan / Journal of Environmental Management 81 (2006) 1–18

Table 1 Bed contraction was found to reduce the liquid–solid

Biomass composition of an anaerobic granule contact by 10–25%:
Type of bacteria Substrates for the %Distribution in the es
¼ 1 þ 0:045U 0:4
g . (4.38)
bacteria granule es0
Acidogens Glucose 65 Experimental results of gas hold-up in the reactor gave
Acetogens Butyrate 2.5 the following correlation:
Propionate 2.0
Methanogens Acetate 7.0 eg ¼ ð13  1:2Þd 0:168
p U 0:7
g . (4.39)
H2 utilizing bacteria Hydrogen 23.5
An axially dispersed plug flow model was found to
describe the liquid mixing in the reactor. The mass balance
The substrate gradient at the surface of a bioparticle in of the reactor was given by the equation
Eq (4.36) is taken from the biofilm model. The biomass 1 d2 s ds s
composition used in this simulation was averaged from 2
¼ þ Da (4.40)
Pe dx dx lþs
several studies such as the modelling work of Mosey (1983)
with the following notations:
and the experimental investigation of Sanchez et al. (1994)
(Table 1). z s rmax Hel U lH
x¼ ; s ¼ ; Da ¼ ; and Pe ¼ .
A set of batch experiments was conducted using H ks ks U l el E zl
bioparticles taken from different heights. Glucose, acetate The axial dispersion co-efficient was found experimen-
and propionate were used as substrates. Substrate con- tally using the tracer injection method. The experimental
sumption with time was monitored. Simulation results were results were found to fit the correlation of Muroyama et al.
found to give reasonable fit for experimentally observed (from Fan, 1989) very well:
values. The biomass composition (i.e. the relative concen-
tration of acidogens and methanogens) within the biofilm Dc U l
¼ 1:01U 0:738
l U
0:167
g D
0:583
c , (4.41)
was manipulated to fit the experimental results. This el z
indicated the crucial role of biomass composition in where dp is the particle diameter; Dc the column diameter;
substrate kinetics. The specific activity of the biofilm was Ezl the axial dispersion coefficient; g the gravitational
also measured. The following were the findings: acceleration; H the bed height; ks the half-saturation
concentration in Monod expression; rmax the maximal
1. Thicker film bioparticles were found on the top of the reaction rate in Monod expression; s the substrate
reactor and thinner in the bottom. concentration; Ug the gas superficial velocity; Ul the liquid
2. Glucotrophic activity decreased from bottom to top and superficial velocity; Ut the terminal velocity of particles; x
methanogenic activity increased from bottom to top. the reduced bed height; es the solid hold up; es0 the solid
3. This indicates the change in biomass composition with hold-up in the liquid–solid fluidized bed; eg the gas hold-up.
biofilm size. Thus by knowing hold-ups (from Eq. (4.38) and (4.39))
and Ez (from Eq. (4.41)), the performance Eq. (4.40) was
Buffiere et al. (1998a) developed a model for AFBRs solved. The model results were found to give a more
based on total carbon removal kinetics. They considered realistic picture.
the effects of gas production in their model which were of
two kinds: 4.2.1. Stratified biofilm models
A stratified biofilm model was presented by Canovas-
1. Gas production modified the degree of axial mixing, Diaz and Howell (1988). The anaerobic biofilm was
which is responsible for the establishment of a concen- modelled as two distinct layers with the inner layer
tration gradient in the reactor. consisting of methanogens and the outer layer consisting
2. Gas production is responsible for bed contraction, of acidogens. The substrate is converted to acids in the
which reduces the contact between the liquid and outer layers, and is subsequently converted to methane by
bioparticles. the bacteria in the inner layer.
The differential equations for substrate uptake in the two
The TOC removal kinetics were found to be in good layers are:
agreement with the Monod model. The TOC uptake rate
d2 G
can be expressed by D1 ¼ k1 x1 , (4.42)
dz2
S
rTOC ¼ rmax , (4.37) d2 F k2 x2
ks þ S D2 ¼
ak1 x1 þ , (4.43)
where S is the TOC concentration in the reactor (the dz2 1 þ F =ki
reactor is assumed to be perfectly mixed). Parameters rmax where D1, D2 is the diffusivities of substrate through the
and ks were found by plotting the inverse of rTOC and 1/S. acidogenic and methanogenic layer. G, F the concentration
 ARTICLE IN PRESS
V. Saravanan, T.R. Sreekrishnan / Journal of Environmental Management 81 (2006) 1–18
of glucose and fatty acids, k1 the zero-order kinetic
constant with respect to sugar uptake and k1 the zero-
order rate constant k2 the kinetic constant for VFA uptake;
ki the inhibition constant; z the distance through biofilm;
x1, x2 the mass volume densities of acidogens and
methanogens.
The authors also explain why a stratified biofilm is
advantageous as compared to an un-stratified biofilm.
When the bulk VFA concentration is high, in the case of an
unstratified film, methanogens will face VFA inhibition. In
the case of stratified biofilm, the inhibition level is
minimized by the presence of the outer layer.
Droste and Kennedy (1986) have given a model for
sequential substrate utilization in a biofilm. This model
assumes that no interactions occur between the two groups
of microorganisms that will cause kinetic or diffusion
parameters to change from values associated with indivi-
dual cultures of each group. Unstratified biofilm and
Monod-type kinetics were assumed.
The governing differential equations are:
d 2 s1 k1 x1 s1
D1 2
¼ , (4.44)
dx K 1 þ s1
d 2 s2 k 2 x 2 s2 Yk1 x1 s1
D2 2

¼
, (4.45)
dx K 2 þ s2 K 1 þ s1
where D is the diffusivity; k the maximum specific reaction
velocity; K the half-velocity constant; s the substrate
concentration; x the distance; x the active microorganism
concentration; Y the acetate yield coefficient (g acetate/g
primary substrate).
From numerical analysis of the governing equation, it
was found that the production of intermediate substrate in
the biofilm increased the conversion of primary substrate to
ultimate product. The increase was not always significant.
To summarize, two ways of approaching the problem of
modelling substrate uptake kinetics are explained in this
section. In the first one, TOC is considered as the rate-
limiting substrate. In this case biomass composition,
individual reaction steps and substrate diffusion limitations
are not considered. The parameters involved in the model
are evaluated by fitting the model to the experimental
results. Even though this approach seems to be simple, it
does not have a sound theoretical explanation and remains
empirical. In the second approach, the kinetic model is
developed considering the individual substrate kinetics,
biofilm composition and diffusion limitations. This seems
to be a more realistic approach. Ultimately the validity of
these models has to be verified for large-scale reactors.
5. The EGSB reactor
The EGSB reactor comes under the family of UASB
reactors. The use of effluent recirculation in a UASB (or a
high height/diameter ratio), resulted in the EGSB reactor
(Seghezzo et al., 1998). Here also, the biomass is present in
a granular form. The higher upflow liquid velocity keeps
13
the granular sludge bed in an expanded condition
(Zoutberg and Frankin, 1996). Reports on models for
EGSB reactors are very scarce. But based on the knowl-
edge of UASB reactors and AFBR models, modelling of
EGSB reactor can be attempted.
1. The biofilm model is similar to the UASB reactor
biofilm model. The composition and structure of the
biofilm is expected to be influenced by the nature as well
as concentration of the substrate. However, there is no
reason to believe that this influence will be significantly
different from what has been observed for biogranules
in the UASB reactor (Section 3.4). The only significant
difference will be the reduced level of the substrate
concentration gradient along the height of the reactor
due to the effect of recirculation.
2. The liquid flow pattern could be expected to be
somewhere between completely mixed and dispersed
plug flow, the exact pattern depending on the recycle
ratio employed. However, any flow model employed will
need validation through tracer studies or any other
suitable experimental studies.
3. A fluidization model which can predict the variation of
bed height with upflow liquid velocity has to be
developed based on experimentation.
Tailoring of the above three models could result in a
complete model for an EGSB reactor.
6. The anaerobic biofilter
The anaerobic filter is an anaerobic packed-bed biofilm
reactor. Though both upflow as well as down flow
configurations are possible, the upflow mode of operation
is more common. The biomass forms a film on the surface of
the packing media. The kinetic model of such a biofilm
reactor has been studied quite extensively (Chang and
Rittmann, 1987; Hamoda and Kennedy, 1987; Meunier
and Williamson, 1981; Rittmann and McCarty, 1980a;
Rittmann and McCarty, 1980b). Fig. 8 illustrates the ideal
biofilm having uniform microbial density (Xf) and uniform
thickness (Lf). When the substrate utilization follows the
Monod relationship, the biofilm model incorporating the
external mass transfer and internal simultaneous diffusion
and reaction can be derived as follows (Huang and Jih, 1997):
d2 S f kX f S f
¼ . (4.46)
dZ 2 ½Df ðK s þ S f Þ
Boundary conditions:
dS f Ds

Df ¼ ðS b
S s Þ ¼ J at Z ¼ 0,
dZ L
dS f
¼ 0 at Z ¼ Lf ,
dZ
where S is the biofilm substrate concentration, Z is the
distance normal to the biofilm surface, D is the diffusivity, Ds
 ARTICLE IN PRESS
14 V. Saravanan, T.R. Sreekrishnan / Journal of Environmental Management 81 (2006) 1–18
Outlet
Sampling port
Gas phase
Inlet
Fig. 8. Schematic diagram of an anaerobic filter showing the differential section.
is the axial dispersion coefficient, Xf is the microbial density
in the biofilm, Ks is the Monod constant, k is the maximum
specific substrate utilization rate, J is the substrate flux.
Here, the subscripts ‘f’ and ‘b’ denote biofilm and bulk
liquid, respectively.
The greatest difficulty in applying the model is to
measure the biofilm thickness (Lf). Rittmann and McCarty
(1978) and Suidan (1986) imposed another boundary
condition to the model, i.e.S f ¼ 0 at Z ¼ Lf
This model is called the deep-biofilm model. Another
reported approach for indirectly estimating the steady-state
biofilm thickness was by assuming that the biomass growth
equals the biomass decay and/or shear losses in biofilm
reactors. For instance, Rittmann and McCarty (1980a)
computed the steady-state biofilm thickness using para-
meters of the substrate flux, microbial growth, biomass
decay and sloughing rate (Eq. (4.47)). The model obtained,
called the steady-state biofilm model, is given by
JY
Lf ¼ , (4.47)
bX f
where Y is the biomass yield coefficient and b is the
sloughing rate.
The models mentioned above have been experimentally
verified or simulated using the results reported in the
literature (Chang and Rittmann, 1987; Liu et al., 1991;
Meunier and Williamson, 1981; Rittmann and McCarty,
1980b, Wang et al., 1987). The kinetic parameters included
in the models were either determined by independent
experiments or obtained from published papers. The liquid
flow regime was assumed to be completely mixed or plug-
flow with axial dispersion (dispersion coefficients were
calculated using empirical equations). An axial dispersion
model coupled with deep-biofilm kinetics was used by
Huang and Jih (1997). The experimental results and the
calculated values showed good agreement. The tracer study
Ss Biofilm
Liquid
Phase Sb Carrier
material
Lf
of Huang and Jih (1997) confirmed that the flow regime is
close to completely mixed.
7. The unified model for anaerobic digestion (ADM1)
The IWA Anaerobic Digestion Modeling Task Group
developed a generalized anaerobic digestion model (Bat-
stone et al., 2002). The biochemical as well as physico-
chemical processes were included in the model. The
biochemical steps include (i) disintegration from homo-
geneous particulates to carbohydrates, proteins and lipids,
(ii) the extracellular hydrolysis of these particulate sub-
strates to sugars, amino acids, and long chain fatty acids
(LCFA), respectively, (iii) acidogenesis from sugars and
amino acids to volatile fatty acids (VFAs) and hydrogen
(iv) acetogenesis of LCFA and VFAs to acetate and (v)
separate methanogenesis steps from acetate and hydrogen/
CO2. The physico-chemical equations describe ion associa-
tion and dissociation, and gas–liquid transfer. The inhibi-
tion kinetics have been incorporated in the biochemical
process.
7.1. The extension of ADM1 to biofilm reactors
The ADM1 model gives a unified approach to anaerobic
digestion. This model was successfully implemented for a
CSTR (Batstone et al., 2002). ADM1 was developed for a
suspended cell system, where there is no mass transfer
limitation for movement of the substrate from the bulk
liquid phase to the cells. On the other hand, in biofilm
reactors, the rate of transport of substrate from the bulk
liquid to the microbial population is controlled by diffusion
of substrate within the biofilm. Hence to extend the ADM1
for biofilm systems, the substrate utilization kinetics of the
single cell system must be replaced with a biofilm model.
The biofilm models for different biofilm reactor systems
have been extensively reported in the previous sections. In
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V. Saravanan, T.R. Sreekrishnan / Journal of Environmental Management 81 (2006) 1–18
a similar way the reactor flow system of ADM1 (i.e. CSTR
system) also has to be modified. In the case of a UASB
reactor, the reactor flow system will have a combination of
CSTRs and a plug flow system as explained in Section 2.1.
AFBR and EGSB reactors will have axially dispersed plug
flow systems as shown in Sections 4.2 and 5. A biofilter
flow system is similar to that of ADM1. The physico-
chemical process system of ADM1 can be directly extended
to the biofilm reactor systems. Thus, the modification of
the biochemical processes and the reactor flow system of
ADM1 with that of the biofilm system could result in a
unified model for biofilm reactors.
8. Summary and conclusions
Development of biofilm reactors has made anaerobic
treatment an attractive option to treat wastewaters. For
optimum design and scale-up of these reactors, mathema-
tical models are required. In this paper, various parameters
affecting the performance of anaerobic biofilm reactors
were reviewed. The important parameters are:
1. the effect of hydrodynamics/flow pattern on reactor
performance;
2. the mass transfer within granules/biofilms;
3. the kinetic effects;
4. the structure and composition of biogranules.
In UASB reactors the different zones are idealized with
different flow patterns. Sludge blankets and sludge beds are
mostly described by a CSTR flow pattern with bypass. The
clarifier zone is described by the axially dispersed plug flow
model. The application of these models for actual full-scale
reactors is yet to be studied because these models do not
consider the existence of non-ideal conditions in full-scale
reactors. These non-ideal conditions may include non-
existence of different zones, improper flow distribution and
dead zones.
Reactor models for UASB reactors consider many
questionable assumptions such as spherical granules,
steady-state operation, description of substrate degrada-
tion by simple Monod kinetics, no substrate/product
inhibition and the effect of pH. Further studies are
required to develop models which do not depend too
much on these assumptions for their development.
In developing kinetic models for both UASBs and
AFBRs, granule structure plays an important role. Studies
show that the structure of the granules and bacterial
composition depends on the type of effluent being treated.
Various theories are provided to support the layered and
un-layered structures of the granules. The variation of
granule structure within the same reactor remains un-
explained as of today. Incorporation of this variation in the
models poses a challenge for the modellers.
The hydrodynamics and bed expansion characteristics of
AFBRs have been reported in the literature. It is generally
found that the drag co-efficient of a biofilm covered
15
granule is more than that of a rigid particle of the same
size. The experimental evidence published so far (Andrews
and Tien, 1979; Hermanovicz and Cheng, 1990; Mulcahy
and Shieh, 1987; Ngian and Martin, 1980; Nicolella et al.,
1999) suggests that the fluidized bed expansion pattern is
observed to follow the Richardson and Zaki correlation.
The studies show that the expansion index can be
calculated through the Richardson and Zaki correlation
(Nicolella et al., 1999). But the parameter ui was found to
be 30–80% of the experimentally determined ut value.
All these studies were conducted with uniform biogranule
sizes.
Many authors (Ro and Neethling, 1994; Safferman and
Bishop, 1996; Schreyer and Coughlin, 1999; Trinet et al.,
1991) observed that the bed becomes stratified due to the
presence of granules with different biofilm thicknesses. In
such a context, the application of the Richardson and Zaki
correlation remains dubious. Hence models considering
expansion characteristics of stratified beds are necessary
for proper process design.
Ideal plug flow and CSTR models were used to describe
the flow pattern inside a laboratory scale AFBR. The use
of this assumption in large scale reactors has to be
investigated. Similar to UASB reactors, while developing
kinetic models for AFBRs, the influence of pH, substrate
and product inhibition, validity of Monod kinetics,
microbial composition and location and sphericity of the
granules have to be studied. In the case of anaerobic filters,
the deep-biofilm model seems to represent the laboratory
scale reactors. But, the assumption of the substrate
concentration reaching zero at the filter media has to be
verified in the large-scale reactor. A realistic model
considering all the above-mentioned short comings is
necessary for proper design and scale up of such reactors.
The integration of the flow model and biofilm model for
these biofilm reactors with ADM1 can result in a robust
model, which can be a tool for design purposes.
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