Journal of Biotechnology 80 (2000) 1 – 33

www.elsevier.com/locate/jbiotec

Review article

Wastewater treatment with particulate biofilm reactors

C. Nicolella a,1, M.C.M. van Loosdrecht b,*, J.J. Heijnen b
a
Department of Food Science and Technology, The Uni6ersity of Reading, P.O. Box 226, Reading RG6 6AP, UK
b
Department of Biochemical Engineering, Kluy6er Institute for Biotechnology, Delft Uni6ersity of Technology, Julianalaan 67,
2628 BC Delft, The Netherlands
Received 22 November 1999; received in revised form 21 February 2000; accepted 25 February 2000

Abstract

The review presented in this paper focuses on applications of particulate biofilm reactors (e.g. Upflow Sludge
Blanket, Biofilm Fluidized Bed, Expanded Granular Sludge Blanket, Biofilm Airlift Suspension, Internal Circulation
reactors). Several full-scale applications for municipal and industrial wastewater treatment are presented and
illustrated, and their most important design and operation aspects (e.g. biofilm formation, hydrodynamics, mass
transfer, mixing) are analysed and discussed. It is clear from the review that this technology can be considered a
grown up technology for which good design and scale-up guidelines are available. © 2000 Elsevier Science B.V. All
rights reserved.

Keywords: Wastewater treatment; Biofilm reactor; Internal circulation; USB; EGSB; Fluidized bed; Airlift

1. Nomenclature De diffusion coefficient (m s−2)

DH dilution rate (day−1)
a specific surface area (m2 m−3) FC organic loading rate (kg day−1)
A area (m2) F/M food to microorganism ratio
C substrate concentration (kg (kgCOD kg−1SS )
m−3) Fr Froude number
CD drag coefficient g acceleration of gravity (m s−2)
d diameter (m) h height (m)
k0 zeroth order kinetic rate con-
* Corresponding author. Tel.: +31-152-781618; fax: + 31- stant (kg m−3 s−1)
152-782355.
kLa volumetric gas–liquid mass
E-mail addresses: c.nicolella@afnovell.reading.ac.uk (C.
Nicolella), mark.vanloosdrecht@stm.tudelft.nl (M.C.M. van transfer coefficient (h−1)
Loosdrecht) kLS liquid–solid mass transfer coeffi-
1
Fax: +44-118-9310080. cient (m h−1)

0168-1656/00/$ - see front matter © 2000 Elsevier Science B.V. All rights reserved.
PII: S 0 1 6 8 - 1 6 5 6 ( 0 0 ) 0 0 2 2 9 - 7
2 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33

n single cells, in downstream processing, by facili-

N substrate flux (kg m−2 h−1) tating cell–liquid separation by sedimentation or
OTR oxygen transfer rate (kg m−3 filtration. The term floc is used to refer to an
day−1) assemblage of individual cells and micro-colonies
q overflow rate (m3 m−2 h−1) occurring under specific reactor conditions or af-
Q flow rate (m3 day−1) ter addition of various agents to the medium
Re Reynolds number (Boonaert et al., 1999). A biofilm can be defined
Sc Schmidt number as a complex coherent structure of cells and cellu-
Sh Sherwood number lar products, like extra-cellular polymers
u velocity (m s−1) (Characklis, 1990), which either form sponta-
V volume (m3) neously as large, dense granules (Lettinga et al.,
X biomass concentration (kg m−3) 1980), or grow attached on a static solid surface
Y biomass yield coefficient (static biofilms) or on suspended carriers (particle-
YO oxygen yield coefficient supported biofilms) (Heijnen, 1984).
Greek letters Microbial aggregates (either in the form of
a height to diameter ratio biofilms, granules or flocs) and the bulk culture
b reaction order transition charac- medium constitutes two distinct phases. This key
teristic [(Eqs. (12) and (13)] feature has three major consequences.
d biofilm thickness (mm) 1. Biomass retention can be used to improve
g settler to reactor volume ratio reactor volumetric conversion capacity when
o hold-up the conversion is limited by the amount of
m viscosity (kg m−1 s−1) biomass present. If no biomass retention is
mmax maximum specific growth rate applied (e.g. in the standard chemostat), the
(day−1) biomass concentration depends only on the
n viscosity (m s−2) substrate concentration in the feed, and conse-
r density (kg m−3) quently large retention times are required in
rS particulate biofilm density (kg the presence of diluted feeds. Depending on
m−3) the settling characteristics of the aggregates,
tH retention time (days) biomass can be readily separated (e.g. by sedi-
mentation) from the bulk liquid and retained
Subscripts
in the bioreactor. In this respect, granules and
c reactor
particle-supported biofilms have an extra ad-
d downcomer
vantage in that they can be more easily sepa-
f biofilm
rated than flocs (i.e. higher biomass
G gas
concentration possible) and have a high reac-
L liquid
r riser tor specific surface area (i.e. a large mass
S solid transfer area than static biofilms).
t terminal 2. The substrates (e.g. oxygen, carbon and nitro-
gen sources) have to cross the aggregate–liq-
uid interface and be transported through the
aggregate to reach the microbial cells and be
consumed. This transport is in general by dif-
2. Introduction fusion and results in a concentration gradient
within the aggregate. The penetration depth of
Microbial cell aggregates, such as flocs and substrates in biofilms mainly depends on the
biofilms, are of great interest in biotechnology. porosity of the biofilm, substrate concentra-
They offer advantages, with respect to suspended tion in the bulk liquid, mass transfer at the
 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33
biofilm–liquid interface and reaction rate in
the biofilm. For poorly soluble substrate (e.g.
oxygen) the penetration depth is shallow
(typically 100–150 mm for oxygen) (Denac et
al., 1983).
3. Due to diffusional substrate concentration
gradients, a growth rate gradient also exists
within the aggregate. In multi-species biofilm
systems this will lead to a biofilm with a
layered structure, where the organisms with
the highest growth rate will be found at the
outside of the biofilm, whereas slower grow-
ing organisms will be found inside (Heijnen
et al., 1989). As a result of this organization,
slower-growing organisms will be protected
from external shear forces, and are less likely
to be lost due to detachment and wash-out.
In this case not the absolute maximum
growth rate of the organisms should be con-
sidered but the maximum growth rate under
conditions in the reactor (e.g. in the presence
of an inhibitor).
The extent to which these features are rele-
vant for a specific system depends, among other
factors, on the physical and structural properties
(e.g. size, density, porosity, settling velocity, etc.)
of the aggregates. Due to a smaller size (typi-
cally between 10 and 150 mm) and higher poros-
ity, diffusional transport is generally faster in
flocs than in granules or biofilm (whose size is
usually in the range 0.5 – 3 mm). The substrate
concentration and biomass distribution gradients
are, therefore, less important in flocs than in
biofilms. On the other hand, biofilms and gran-
ules present better settling properties than flocs
(terminal settling velocity of about 40 and 5 m
h − 1 for particle-supported biofilms and flocs, re-
spectively), and can be more easily retained in
the bioreactors. The physical and structural
properties of particle-supported biofilms and
granules are similar, and so are their hydrody-
namic, mass transfer and reaction characteris-
tics. For this reason, in the following
particle-supported biofilms and granules will be
considered as a single category, the particulate
biofilms (Fig. 1).
When the substrate concentration in the feed
is high ( \10 gCOD l − 1) and rapidly growing
3
organisms (growth rate\ 0.1 h − 1) are used,
there is no advantage in biomass retention.
Sufficient biomass will be formed to metabolize
the substrate within relatively short residence
times. For the majority of industrial fermenta-
tion processes, where high substrate concentra-
tions are used, biofilm formation is either
unnecessary or even disadvantageous, and the
range of applications of immobilized-cell systems
in industry is limited. Several applications of
biofilm reactors in various biotechnological pro-
cesses (e.g. fermentation, production of enzymes,
production of primary and secondary metabo-
lites, production of antibiotics and bioconver-
sions), have been reviewed by Furusaki (1988),
Schugerl (1989, 1997), and Godia and Sola’
(1995). However, with the exception of wastewa-
ter treatment processes, the application of bio-
film reactors at full industrial scale is scarce
(Godia and Sola’, 1995).
An extensive use of biofilms is made within
the field of environmental biotechnology for
three main reasons:
1. compared with most other industrial biopro-
cesses, large volumes of dilute aqueous solu-
tions have to be treated;
2. natural, mixed populations of microorgan-
isms, which readily form biofilms, are used;
3. the process can be operated at high biomass
concentration in the reactor, without the
need for settlers for biomass retention and
recirculation. A polishing step of the effluent
is usually needed to remove remaining sus-
pended (detached) biomass.
Trickling filters have been in use in wastewa-
ter treatment processes for over a century, and
their design and operation are well established
in the practice of wastewater engineering (Met-
calf and Eddy, 1991). The application of partic-
ulate biofilm reactors is relatively recent, and,
though in most cases the principles of their de-
sign and operation are known or can be derived
from other disciplines, there is a lack of system-
atic information. This article covers relevant en-
gineering aspects of particulate biofilm reactors
and reviews several examples of full-scale appli-
cations.
4 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33

2.1. Application range static biofilms (e.g. in trickling filters), particulate

biofilms (e.g. in biofilm fluidized bed reactors,
Wastewater treatment processes are based on upflow anaerobic sludge blanket reactors and bio-
the use of three types of microbial aggregates: film airlift suspension reactors), and flocs (in acti-

Fig. 1. Particulate biofilms. (a) Aerobic granules (average diameter: 1.2 mm); (b) particle supported biofilms (average diameter: 1.7
mm).
 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33
Fig. 2. Concentration-flow rate phase diagram for application of floc and biofilm reactors.
vated sludge processes). Basic design criteria can
be used to identify the range of operating condi-
tions suitable for the application of these aggre-
gates in wastewater treatment processes. Fig. 2
shows, in the concentration-flow rate plane, the
lines corresponding to design criteria applicable
to different reactor configurations for the case
of industrial aerobic processes (details are given
in Appendix A). These lines define different re-
gions of applicability in the C – Q phase dia-
gram:
A. Retention time is so long that microorgan-
isms grow in suspension.
B. At high flow rates, particulate biofilms and
flocs are washed-out and only static biofilms
can be retained in the reactors, or the reac-
tors get a very flat and extended shape.
C. Flow and loading conditions are suitable for
application of particulate biofilm reactors.
This range of applications is covered in the
present review.
D. Flow and loading conditions are suitable for
applications of flocs, provided that separa-
tion and biomass recycle are used (e.g. acti-
vated sludge processes). This region partially
overlaps the particulate biofilm region.
E. For high strength and low flow wastewater,
upflow sludge blanket reactors can be used.
5
The sludge is retained in the reactor without
need for external separation and recycle.
3. Applications of particulate biofilms in
environmental biotechnology
The main reactor types applicable for the sus-
pension of particulate biofilms in wastewater
treatment processes are Biofilm Upflow Sludge
Blanket (USB), Fluidized Bed (BFB), Expanded
Granular Sludge Blanket (EGSB), Biofilm Airlift
Suspension (BAS), and Internal Circulation (IC)
reactors (Fig. 4). In USB, BFB and EGSB reac-
tors (Fig. 4a,b,c, respectively), particles are kept
fluidized by the up-flowing influent. In BAS re-
actors (Fig. 4d) an airlift suspension is obtained
by pumping air into the system, whilst in IC
reactors (Fig. 4e) the gas produced in the system
drives the circulation and mixing of liquid and
solids in the reactor. The advantages and disad-
vantages of particulate biofilm reactors are pre-
sented in Table 1 (Heijnen et al., 1989; Bryers
and Characklis, 1990; Harremoes and Henze,
1995; Wright and Raper, 1996). The basic de-
sign characteristics and some examples of indus-
trial applications of particulate biofilm reactors
are given in Tables 2 and 3, respectively.
6 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33
3.1. Historical o6er6iew
Although the development of water and
wastewater biological reaction systems using a
fluidized bed of biomass can be traced back to
1940 in the UK (Mishra and Sutton, 1991), devel-
opment of particle-supported biofilm reactors did
not occur until the early 1970s. Researchers at
Manhattan College in New York were granted a
US patent in 1976 (Jeris et al., 1976) for the
application of Biofilm Fluidized Bed (BFB) pro-
cess configuration to denitrifying wastewater. The
first commercial-scale application for the BFB
process configuration occurred at Pensacola
(USA) in the mid-1970s, with the installation of a
system for the denitrification of nitrified munici-
pal wastewater (Mishra and Sutton, 1991). The
first commercial-scale application of the BFB
technology to industrial treatment occurred in the
late 1970s when an Ecolotrol HY-FLO system
was installed at a soft drink bottling plant in
Birmingham (USA) (Jeris, 1983). Since the early
1980s, the BFB process has been used on an
industrial scale for most conventional-sewage
treatment processes including carbonaceous oxi-
dation (Jeris et al., 1977; Cooper and Wheeldon,
1982), nitrification (Dunn et al., 1983; Nutt et al.,
1984), denitrification (Melcer et al., 1984; Nutt et
al., 1984), and anaerobic treatment (Hickey and
Owens, 1981; Jewell et al., 1981; Jeris, 1983).
Table 1
Advantages and disadvantages of particulate biofilm reactors
Advantages
High terminal settling velocity of solids (50 m h−1; for
flocculated sludge: 5 m h−1), leading to possible
elimination of external clarification/separation stages
High reactor concentration (30 kg m−3; for flocculated sludge
systems: 3 kg m−3)
High biofilm surface area (3000 m2 m−3; in trickling filters:
300 m2 m−3)
High biomass concentration and mass transfer area result in
high conversion capacities (for oxygen, 20 kg m−3 day−1;
in activated sludge and trickling filter processes: 3 kg m−3
day−1)
Compact reactor with small area requirements
High biomass age (several weeks); and minimization of excess
sludge production
Perhaps the most significant recent develop-
ment at the commercial-scale level is the use of
upflow fluidized bed reactors containing granules
and no bed media. The USB reactor was devel-
oped in The Netherlands in the late 1970s for
anaerobic treatment of low-strength wastes, and
the first full-scale plant (200 m3) was installed in
1978 at a sugar-beet factory in Halfweg (The
Netherlands) (Lettinga et al., 1980). The EGSB
and the IC reactors are the most recent evolutions
of the USB concept. Biothane Biobed EGSB
(Lourens and Zoetemeyer, 1992) and Paques IC
(Vellinga, 1986) systems operate at much higher
liquid upflow velocities than the conventional
USB.
Fundamental and applied research on Biofilm
Airlift Suspension (BAS) reactors in the late 1980s
(Heijnen, 1985; Heijnen et al., 1991, 1993), includ-
ing research at Gist-Brocades, TNO (Dutch Or-
ganisation for Applied Scientific Research) and
Delft University of Technology (The Nether-
lands), led to the concept of CIRCOX airlift
reactor, developed and patented by Gist-Brocades
and commercialized by Paques.
High rate anaerobic pre-treatment followed by
aerobic post-treatment using biofilm reactors
(Heijnen et al., 1991) has been shown on numer-
ous occasions to be a very cost effective way to
remove organic compounds from wastewaters
(Fig. 3). The IC anaerobic reactor followed by the
Disadvantages
Biofilm formation on carriers poses problems leading to long
start-up times
Control of biofilm thickness is difficult
Overgrowth of biofilms leads to elutriation of particles
Liquid distributors for fluidized systems are costly for
large-scale reactors and pose problems with respect to clogging
and uniform fluidization
 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33
Table 2
Some design characteristics of particle biofilm reactors

Reactor type Commercial names Flow pattern Liquid velocity H/D Mixing References

BFB HY-FLO, Ecolotrol (USA); Up-flow 10–30 m h−1 (upward) 2–5 Liquid Heijnen et al. (1989)
ANAFLUX, Degremont
(France); OXYTRON, ANY-
TRON, Dorr-Oliver (USA)
UASB BIOPAQ, Paques (The Nether- Up-flow 0.5–1 m h−1 (upward) 0.2–0.5 Gas produced Lettinga et al. (1980),
lands) Pereboom and Vereijken
(1994), Austermann et al.
(1999)
EGSB BIOBED, Biothane (USA) Up-flow 10–15 m h−1 (upward) 4–5 Liquid, gas pro- Zoutberg and Frankin (1996)
duced
IC IC, Paques (The Netherlands) Mixed Bottom: 10–30 m h−1; top: 4–8 3–6 Gas produced Pereboom and Vereijken
m h−1 (circulation) (1994), Gorur et al. (1995)
BAS CIRCOX, Paques (The Mixed 0.4–0.8 m s−1 (circulation) 4–5 Gas introduced Heijnen et al. (1993), Gorur et
Netherlands) al. (1995)

7
 8
Table 3
Examples of full-scale applications of particle biofilm reactors for environmental control

System Development topics Examples of full-scale Design parametersa References

applications

BFB ANAFLUX, Degremont, Upflow anaerobic FBF using a
France mineral support (BIOLITE
R280) as fluid bed media to
treat a variety of brewery,
food-processing and paper
Starch factory, Habourdin, N: 1; D: 6 m; OLR: 12 000 Holst et al. (1997)
France (1993) kgCOD day−1; RC: 60 kgCOD
m−3 day−1; hCOD: 86%

C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33

industry wastewater
BFB OXYTRON, ANYTRON, Carbonaceous oxidation, By-product coke plant, N: 2; D: 9 m; Hc 8.5 m; Sutton et al. (1999)
Dorr-Oliver, USA nitrification, denitrification and Ont., Canada (1996) OLR: 6690 kgCOD day−1; RC:
anaerobic reduction of 10.5 kgCOD m−3 day−1; Qw:
municipal and industrial 1104 m3 day−1
(automotive industry,
coke-making operations)
wastewater using sand and
activated carbon as fluid bed
media
UASB BIOPAQ, Paques, The Processes incorporating the Distillery, Wellington, N: 1; V: 450 m3; OLR: 6750 Wolmarans and Driessen
Netherlands sludge granulation concept for South Africa (1996) kgCOD day−1; RC: 15 kgCOD (1996), Wolmarans and Nell
the anaerobic treatment of high m−3 day−1; hCOD: \90% (1996)
strength wastewaters
EGSB BIOBED, Biothane, Processes combining the best Chemical factory N: 1; V: 220 m3; H: 17.3 m; Zoutberg and Frankin (1996)
USA features of the UASB and BFB (production of OLR: 2880 kgCOD day−1; RC:
technologies for the anaerobic formaldehyde from 13.1 kgCOD m−3 day−1; Qw:
treatment of high strength methanol), Rotterdam, The 144 m3 day−1; Qr: 2400 m3
wastewaters (wastewater from Netherlands (1993) day−1; uL: 6.3 m h−1; uG:
food, brewery, yeast, 0.65 m h−1; HRT: 2.7 h
pharmaceuticals, paper, chemical
industry)
IC Paques, The Netherlands System consisting of two UASB Inuline and fructose N: 1; V: 1100 m3; H: 22 m; Habets et al. (1997a,b)
reactor compartments on top of production from chicory OLR: 35 000 kgCOD day−1;
each other, one high loaded and beet, Rosendaal, The RC: 31 kgCOD m−3 day−1; X:
one low loaded. The gas Netherlands (1995) 31 kg m−3; hCOD: 70–80%
collected in the first stage drives
a gas lift and an internal
circulation. The system is used
for the anaerobic treatment of
high strength wastewaters
 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33
Table 3 (Continued)

System Development topics Examples of full-scale Design parametersa References

applications

BAS CIRCOX, Paques, The
Netherlands
Airlift technology with biomass
on carrier (sand or basalt) app-
lied to pharmaceutical and
brewery wastewaters which are
anaerobically pre-treated, and to
municipal wastewater
Brewery, Enschede, The
Netherlands (1994)
N: 1; V: 230 m3; H: 19 m; Gorur et al. (1995)
HRT: 1.3 h; Qw: 2245 m3
day−1; F/M: 0.2 kgBOD kg−1
MLSS
day−1; RC: 4–14 kgCOD m−3
day−1

a
N, number of units; D, column diameter; H, column height; V, reactor volume; OLR, organic loading rate; RC, reactor capacity; h, removal efficiency; HRT,
hydraulic retention time; uL, liquid velocity; uG, gas velocity; F/M, food to microorganism ratio; X, volatile suspended solid concentration; Qw, wastewater flow rate;
Qr, recirculation flow rate.

9
10 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33
Fig. 3. Full scale Paques CIRCOX (front, 140 m3) and IC
(back, 385 m3) reactors at a brewery in Brazil.
CIRCOX airlift aerobic technology has been re-
cently selected to treat the effluent from a large
brewery in The Netherlands (Gorur et al., 1995).
After a year of operation, this innovative combi-
nation was reported to have overall total and
soluble COD removals of 80 and 93.5%, respec-
tively. Current volumetric loading rates have aver-
aged 14 kg m − 3 day − 1 in the IC and 10 kg m − 3
day − 1 in the airlift system.
3.2. Upflow Sludge Blanket (USB) reactor
The principle of operation of the Upflow
Anaerobic Sludge Blanket (USB) reactor (Fig. 4a)
(Lettinga et al., 1980) is similar to that of biofilm
fluidized bed reactors, but hydrodynamic condi-
tions are more quiescent due to lower superficial
liquid velocities. USB processes are based on the
development of dense granules (1 – 4 mm) in the
reactor. Wastewater enters the bottom of the reac-
tor vessel through the inlet distribution system
and passes upward through the dense anaerobic
sludge bed (biomass concentration: 60–70 kg
m − 3). Soluble COD is readily converted to
biogas, which is rich in methane, and an upward
circulation of water and gasborne sludge is estab-
lished. A settler section allows effective degassifi-
cation to occur. The dense, granular sludge
particles now devoid of attached gas bubbles, sink
back to the bottom, establishing a return down-
ward circulation. The upward flow of gasborne
sludge through the blanket combines with the
return downward flow of degassed sludge and
creates continuous convection. This insures effec-
tive sludge to wastewater contact. The design of
the reactor allows a highly active biomass concen-
tration and thereby high loading rate (10–15 kg
COD m − 3 day − 1), i.e. short hydraulic retention
time (less than 48 h for most applications).
Suspended solids in the influent, which accumu-
late in the reactors, pose a major problem to the
operation of USB and reduce the reactor capacity.
To overcome these limitations, the BFB and,
later, the EGSB and the IC concepts were
developed.
3.3. Biofilm Fluidized Bed (BFB) reactor
In Biofilm Fluidized Bed (BFB) reactors (Fig.
4b), the liquid to be treated is pumped through a
bed of small media (typically sand with a particle
size range of 0.2–0.8 mm) at a sufficient velocity
to cause fluidization. In the fluidized state the
media provide a large specific surface for attached
biological growth and allows biomass concentra-
tions in the range 10–40 kg m − 3 to develop
(Cooper and Sutton, 1983). For aerobic treatment
processes the reactor is aerated. This is done by
recirculating the liquid from the reactor to an
oxygenator where air, or possibly oxygen, is bub-
bled (Cooper, 1981). To overcome problems re-
lated to high recirculation rates, needed when
there is an high oxygen demand in the reactor, the
reactor might be aerated directly (three-phase
BFB reactor) (Fan et al., 1986; Tang and Fan,
1987; Trinet et al., 1991).
The BFB technology is typically most useful for
treatment of streams contaminated with organic
or inorganic compounds (e.g. ammonia) requiring
 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33 11

long solids residence time conditions (longer than OXYTRON (Sutton et al., 1980; Hoyland and
15 days) for biological oxidation or reduction and Robinson, 1983) and ANYTRON (Sutton and
low (less than 100 mg l − 1) concentrations of Mishra, 1990, 1994) are the commercial embodi-
suspended solids (Sutton and Mishra, 1990). ments of, respectively, the aerobic and anaerobic

Fig. 4. Biofilm reactor configurations. (a) USB; (b) BFB; (c) EGSB; (d) BAS; (e) IC.
12 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33
BFB process configuration developed by Dorr-
Oliver, Inc. Twelve aerobic BFB reactors were
installed at General Motors manufacturing facili-
ties in the 1980s (Mishra and Sutton, 1991), and
four anaerobic BFB reactors were started-up in
1982 at a soy protein factory in Muscatine, IA
(Heijnen et al., 1989). Most recently, the BFB
technology provided by Dorr Oliver was selected
for the treatment of the wastewater from by-
product coking operations with a phenolic load of
1120 kg day − 1 (Sutton et al., 1999). This BFB
system consists of two towers of 9 m diameter and
8.5 m height. The system includes provisions for
controlled pre-dissolution of high purity oxygen
to the influent using an oxygen contactor and a
means for monitoring and controlling expansions
of the fluidized bed because of biofilm growth on
the sand media.
Biothane B.V. has built several anaerobic two-
stage BFB plants for purification of their fermen-
tation process wastewater (Heijnen et al., 1989)
treating wastes at COD volumetric loading rates
in excess of 20 kg m − 3 day − 1. The first two
plants were built in the mid-1980s at Delft in The
Netherlands and Prouvy in France.
Degremont S.A. developed the ANAFLUX
process (Durot and Prevot, 1987; Bernard and
Rovel, 1989; Oliva et al., 1990), an upflow anaero-
bic reactor using a mineral bacterial support, in
the mid-1980s. The first industrial reactor was
built in 1986 for the treatment of brewery waste-
water. Since then, Degremont S.A. has con-
structed over 25 full-scale ANAFLUX reactors
treating, world-wide, a variety of industrial waste-
waters (Holst et al., 1997). The system is highly
efficient due to the large reactor biomass concen-
tration (30–90 kg m − 3) and to the intimate con-
tact between biomass and substrate created by
high upflow liquid velocities (5 – 10 m h − 1).
3.4. Expanded Granular Sludge Bed (EGSB)
reactor
The Expanded Granular Sludge Bed (EGSB)
reactor (Fig. 4c) (Frankin et al., 1992; Zoutberg
and Frankin, 1996) combines both characteristics
of USB and BFB processes. Biomass is present in
a granular form, but conditions with respect to
the upflow velocities for liquid (10 m h − 1) and
gas (7 m h − 1) approach those of the BFB system.
EGSB reactors can be operated as ultra high
loaded anaerobic reactors (up to 30 kg COD m − 3
day − 1) to treat effluents from chemical, biochemi-
cal and biotechnological industries (Zoutberg and
de Been, 1997). Biothane B.V. have built dozens
of Biobed EGSB units for various types of
wastewater (food, chemical, pharmaceutical in-
dustry) in all parts of the world.
3.5. Biofilm Airlift Suspension (BAS) reactor
Airlift reactors (Chisti, 1989) consist of two
connected sections, a riser and a downcomer.
Different configurations are possible, including
internal loop and external loop reactors. The prin-
ciple of operation is the same for both configura-
tions. A gas is sparged at the bottom, moves
upward and exits at the top of the riser section. In
internal-loop airlift reactors, air may recirculate
through the downcomer section and provide aera-
tion throughout the reactor. The difference in
density between riser and downcomer, due to the
difference in gas hold-up, drives the liquid to
circulate between the two sections. When the liq-
uid velocity is sufficiently high, small particles will
be suspended and recirculated with the liquid.
This results in a thorough mixing of both particles
and liquid throughout the reactor. The airlift
technique has found two major applications in
wastewater treatment processes, the Biofilm Air-
lift Suspension (BAS) reactor (Fig. 4d) for aerobic
treatment and the gas-lift reactor for anaerobic
treatment. The BAS technology was originally
developed for aerobic purification of anaerobi-
cally treated industrial wastewaters (Heijnen,
1984; Heijnen et al., 1990, 1993). In anaerobic
processes, the circulation in the reactor is induced
by recirculating the gas produced by the biochem-
ical reactions (gas-lift reactors; van Houten et al.,
1994, 1997).
CIRCOX, a commercial embodiment of the
BAS reactor, is an aerobic system which ensures
high biological load (4–10 kg m − 3), short liquid
residence time (0.5–4 h), high biomass settling
velocities (50 m h − 1) and high biomass concentra-
tion (15–30 kg m − 3). The CIRCOX airlift tech-
 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33
nology has been applied to treat municipal
wastewater at a sewage plant in The Netherlands
(Frijters et al., 1997). Two CIRCOX reactors, an
airlift reactor and an airlift reactor with integrated
denitrification, were used to treat pre-settled influ-
ent. High BOD and nitrogen removal rates could
be obtained resulting in a good effluent quality.
Approximately 15 such systems are presently in
operation.
TURBOFLO is a biofilm reactor for secondary
and tertiary wastewater treatment developed by
Cie Lyonnaise des Eaux (France) using the con-
cept of an internal circulating airlift reactor
(Lazarova and Manem, 1996; Lazarova et al.,
1997; Mousseau et al., 1998). The reactor com-
prises a rectangular column filled with high den-
sity polyethylene granules (size: 0.5 – 2.5 mm;
density 860 kg m − 3). An industrial-scale reactor
was operated at a wastewater treatment plant at
Evry (France).
An industrial prototype of an external loop
airlift reactor, the BIOLIFT process, was devel-
oped by OTV S.A. (France) and installed at the
Maxeville (France) wastewater treatment plant
(Badot et al., 1994).
3.6. Internal Circulation (IC) reactor
The IC reactor (Fig. 4e) (Vellinga, 1986) con-
sists of two USB-like reactor compartments on
top of each other, one highly loaded and one low
loaded. The first reactor compartment contains an
expanded granular sludge bed, where most of the
COD is converted to biogas. The biogas produced
in this compartment contains an expanded bed, is
collected by the lower level phase separator and is
used to generate a gas lift by which water and
sludge are carried upward via the riser pipe to the
gas/liquid separator on the top of the reactor.
Here the biogas is separated from the water/
sludge mixture and leaves the system. The water/
sludge mixture is directed downwards to the
bottom of the reactor via the concentric downer
pipe, resulting in the internal circulation flow. In
the lower section, the upward liquid velocity
varies from 10 to 20 m h − 1. The effluent from the
first compartment is post-treated in the second,
low loaded compartment, where remaining
13
biodegradable COD is removed. The liquid veloc-
ity in this compartment normally varies from 2 to
10 m h − 1. The IC system consists of a slender
vertical reactor with a height of up to 25 m and a
relative small surface area.
Paques has several full-scale IC systems in oper-
ation in Europe. One of the largest systems is
located at a brewery in The Netherlands
(Pereboom and Vereijken, 1994) and consists of
six IC reactors each 162 m3 in volume. The reac-
tors are designed at a COD loading rate of 24 kg
m − 3 day − 1. The largest reactors (22 m high, 9.5
m in diameter and approximately 1500 m3 in
volume) are presently at ADM in Ireland and at
Zhongya Chemicals in China.
4. Engineering aspects
4.1. De6elopment of particulate biofilms
For a reliable operation of biofilm reactors,
high biomass concentrations, present in stable
granular biofilms, are essential. For this reason,
the carriers should be completely covered with
biofilms. However, these biofilms should be
smooth, otherwise the biofilm particles will be
easily washed out of the reactor.
Biofilm development is the difference between
biofilm growth and attachment on one hand, and
detachment processes on the other. Biofilm devel-
opment is influenced by various processes, includ-
ing adsorption and desorption of microorganisms
to and from the solid surface, attachment of
microorganisms to the surface, biofilm growth
and biofilm detachment (Characklis, 1990). In
steady state, the balance between biofilm growth
and detachment determines the physical structure
of the biofilm (van Loosdrecht et al., 1995), and
thereby the settling and fluidization characteris-
tics.
4.1.1. Biofilm formation
Heijnen (1984) postulated the hypothesis that,
in ideally mixed reactors, formation of biofilm
covered carriers only takes place if the hydraulic
retention time is less than the inverse of the
maximum growth rate (the dilution rate is larger
14 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33
than the maximum growth rate). Experimental
evidence for this hypothesis was produced by
Tijhuis et al. (1994) for the formation of hetero-
trophic aerobic biofilms on small suspended
basalt particles in airlift reactors. These authors
also showed that, during biofilm development in
the start-up of BAS reactors, more than 95% of
the attached biomass production is detached to
the liquid phase and washed out of the reactor.
The amount of detached biomass, and therefore,
biofilm accumulation, is strongly influenced by
abrasion processes in the reactor, which are
mainly due to particle interactions. It was found
for the BAS reactor that the rate of biofilm spe-
cific surface detachment rate is a function of the
bare carrier concentration, independent of the
surface substrate loading rate and biofilm thick-
ness. Because the bare carrier concentration de-
creases during the start-up, abrasion and
therewith detachment rate decreases. This results
in a strong non-linear increase of the biofilm
thickness with time.
The influence of carrier type on adhesion and
biofilm formation of pure and mixed cultures was
studied by Gjaltema et al. (1997a) using sus-
pended carriers (standard, roughened, hydropho-
bic and positively charged glass beads, sand and
basalt grains) in laboratory airlift reactors. The
results clearly show that in airlift reactors hydro-
dynamic conditions and particle collisions control
biofilm formation. Increased surface roughness of
the carriers promoted biofilm accumulation on
suspended carriers, whilst the physico-chemical
characteristics of the carrier surface proved to be
less important.
It was shown that the biofilm morphology is
influenced to a large extent by the surface sub-
strate loading and applied detachment forces (Ti-
jhuis et al., 1995a; Kwok et al., 1998). A moderate
surface substrate loading and a high detachment
force yielded smooth and strong biofilms, whilst
the combination of a high surface substrate load-
ing and low detachment forces led to rough bio-
films. The strength of biofilms also appeared to be
related to the detachment forces applied during
biofilm formation, in combination with the sub-
strate loading.
4.1.2. Biofilm detachment
Biofilm detachment, the interphase transport of
biomass from an attached microbial film to the
bulk liquid phase, has generally been attributed to
four different processes (Characklis, 1990), includ-
ing grazing (the consuming of bacteria from the
outer surface of the biofilm by protozoa), slough-
ing (the periodic loss of large patches of biofilm),
erosion (the continuous removal of small particles
from the surface of the biofilm, primarily caused
by liquid shear stress), and abrasion (analogous to
erosion, but caused by collisions of particles).
Among the mechanisms controlling biofilm reac-
tor performance, biofilm detachment is one of the
least studied and understood. The biofilm detach-
ment rate is a complicated function of many
variables, including hydrodynamics of the liquid
flow, biofilm morphology, and support
characteristics.
The mechanisms controlling biomass loss have
been investigated for different biofilm reactors,
including solid/liquid BFB reactors (Chang et al.,
1991; Nicolella et al., 1996), three-phase BFB
reactors (Rittmann et al., 1992), and BAS reactors
(Tijhuis et al., 1995b; Gjaltema et al., 1995,
1997b,c).
Chang et al. (1991) made direct measurements
for the specific biofilm loss rate coefficient and the
total biofilm accumulation in a laboratory BFB
reactor with varying liquid velocities and support
particle concentrations. Multiple regression analy-
sis of the results showed that increased particle-to-
particle attrition and increased turbulence caused
the biofilm to be denser and thinner. The specific
detachment rate coefficient increased as inert par-
ticle concentration and particle Reynolds number
(i.e. turbulence) increased, and the turbulence and
attrition of bed fluidization appeared to be domi-
nant detachment mechanisms. Similar results are
reported by Gjaltema et al. (1995) for the detach-
ment of biomass from suspended biofilm pellets in
the presence of bare carrier particles in BAS
reactors under non-growth conditions. The de-
tachment rate was dominated by collisions be-
tween bare carrier particles and biofilm pellets,
whilst hydrodynamic conditions and substrate
loading rates have a minor effect (Tijhuis et al.,
1995b). The collisions between biofilm pellets and
 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33
bare carrier particles cause an on-going abrasion
of the biofilm pellets, leading to a reduction in
pellet volume, whilst the breakage of the biofilm
pellets is negligible (Gjaltema et al., 1997b). These
findings were confirmed by Kwok et al. (1998) for
growing biofilm suspensions.
The experimental data on biofilm detachment
rate in BFB reactors reported by Nicolella et al.
(1996) showed that the specific detachment rate
coefficient strongly increases with increasing liq-
uid velocity. Other parameters, such as particle
concentration and liquid shear stress, were found
to be less significant. Nicolella et al. (1996) found
that the biofilm detachment rate is very low under
the flow conditions used in their experiments,
which are comparable to those of many industrial
applications of fluidized bed technology in
wastewater treatment processes (e.g. liquid veloc-
ities in the range 1– 10 mm s − 1).
The high detachment rate caused by bare carri-
ers has some consequences for BAS reactor opera-
tion: while bare carrier are necessary for biofilm
formation, they can at the same time severely
hinder or delay biofilm formation, especially dur-
ing the start-up phase. On the other hand, bare
carrier particles can be used to prevent excessive
biofilm formation, which can lead to reduced
reactor performance. Contrary to airlift reactors
where solids are completely mixed in the reactor,
in BFB, bare particles will accumulate at the
bottom of the bed, whilst thick fluffy biofilms will
accumulate at the top of the bed (Di Felice et al.,
1997). In BFB, control of biofilm development by
particle shear is therefore not as easy as in an
airlift reactor.
4.2. Hydrodynamic characteristics of particulate
biofilms
A sound knowledge of the hydrodynamic be-
haviour of biofilm-coated particles is an essential
step for the correct design of particle-supported
biofilm reactors. In these systems, biofilm growth
alters particle size, apparent density, shape and
roughness, and all these factors have a strong
influence on the hydrodynamic behaviour of the
reactors.
15
Successful design and operation of particle type
biofilm reactors rely on information on settling
and fluidization characteristics, such as fluidized
bed-height as a function of liquid velocity. Infor-
mation on fluidized-bed height is important be-
cause it establishes the solid’s residence time and
the specific biofilm surface area in the biologically
active zone. The relationships valid for fluidiza-
tion of rigid particles, readily available in the
chemical engineering literature (e.g. Di Felice,
1995), have to be modified to take into account
the effect of the biofilm layer.
4.2.1. Terminal settling 6elocity
The terminal settling velocity of a single spheri-
cal particle in an infinite expanse of fluid is:
ut =
4g(rS − rL) n
0.5
(1)
3CDrL
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 m − 3. The drag coefficient CD is a func-
tion of the particle Reynolds number
rLdSut
Ret =
mL
Biofilm particles are in the intermediate flow
regime (1B Ret B 100) for the vast majority of
cases, invariably obtained when sand (0.5–1 mm)
or similar material is used as inert support. In the
intermediate flow regime, the drag coefficient for
a smooth, rigid sphere is (Perry and Green, 1997):
CD = 18.5Ret− 0.6 (2)
Biofilm particles are of nearly spherical shape,
but they are not smooth or rigid, and therefore
Eq. (2) cannot be used to calculate their settling
velocity. For this reason, other empirical correla-
tions have been suggested for the estimation of
CD for biofilm particles. A selection of these
correlations is presented in Table 4 and plotted in
Fig. 5. The drag coefficient of the biofilm particles
is clearly higher than that of smooth, rigid
spheres, also reported in Fig. 5, over the whole
range of Ret investigated.
Nicolella et al. (1999b) could estimate the ter-
minal settling velocity of particles used in their
16 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33

Table 4
Selected correlations for CD, n and ui for biofilm particles

Reference Ret CD n ui

Ngian and Martin (1980) 4.4Re−0.1

t ut
Hermanovicz and Ganczarczyk (1983) 50–100 17.1Re−0.47
t
Thomas and Yates (1985) 20–100 30Re−0.505
t ut
Mulcahy and Shieh (1987) 40–90 36.66Re−0.67
t 10.35Re−0.18
t ut
Harada et al. (1987) 10–50 8.733Re−0.341
t ut
−0.21
Hermanovicz and Cheng (1990) 50–100 9.11Ret ut
24
Ro and Neethling (1990) 15–87 +21.55Re−0.518
t
Ret
24
Yu and Rittmann (1997) 40–90 +14.55Re−0.48
t ut
Ret 4.526Re−0.0126
t
Nicolella et al. (1999b) 7–90 29.6Re−0.6
t 4.4Re−0.1
t 0.8ut

work and those reported by other authors with an Richardson and Zaki (1954) showed that the
average error of 10% by: extrapolation of the velocity to o= 0, ui, and the
single particle settling velocity, ut, are virtually
(CD)biocovered coincident for systems where the particle diameter
=1.6 (3)
(CD)clean is considerably smaller than the column diameter.
The expansion index n was found to be a function
Surface roughness has generally been invoked
of the flow regime only, and, for the range of
as the reason for the increase in the drag coeffi-
Reynolds number of interest in the case of biofilm
cient of biofilm particles (Hermanovicz and
particles (1BReB 500), Richardson and Zaki
Ganczarczyk, 1983; Mulcahy and Shieh, 1987).
(1954) proposed:
Ro and Neethling (1990) introduced a form factor
and a shape factor. The experimental value of
both parameters, however, was very close to one, n= 4.4Ret− 0.1 (5)
so that their effective contribution was not tested.
Nicolella et al. (1999b) found that the ratio of
drag coefficient for biofilm particles to drag coeffi-
cient for smooth rigid solid is independent of
biofilm thickness, and concluded that particle de-
formability has a negligible effect on CD. They
also showed that as the Reynolds number de-
creases (up to 0.001), the experimental measure-
ments for CD become closer to the correlation for
rigid smooth particles, thereby indicating that the
surface roughness indeed plays an dominant role
in the determination of CD.

4.2.2. Expansion characteristics

The empirical equation of Richardson and Zaki
(1954) is widely used to describe the bed expan-
sion characteristics of concentrated suspensions of
rigid spherical particles:
uL
o nL = (4) Fig. 5. Selected correlations for drag coefficient of particulate
ui biofilms.
 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33
Fig. 6. Selected correlations for the expansion index of partic-
ulate biofilms.
For BFB, the experimental evidence published
so far (Andrews and Tien, 1979; Ngian and Mar-
tin, 1980; Mulcahy and Shieh, 1987; Hermanovicz
and Cheng, 1990; Csizkor et al., 1994; Nicolella et
al., 1999b) suggests that bed expansion character-
istics can still be represented by an equation like
that of Richardson and Zaki (i.e. a linear relation-
ship in logarithmic co-ordinate between voidage
and liquid velocity). However, the dependence of
the parameters n and ui on the system’s physical
characteristics has yet to be fully established. Fig.
6 reports the estimates of the expansion index n as
a function of the terminal Reynolds number as
suggested by different authors for biofilm parti-
cles. The values obtained by Nicolella et al.
(1999b) are well in line with those found by
Richardson and Zaki (1954) for liquid beds of
smooth rigid particles but much smaller than the
estimates reported for biological beds (e.g.
Thomas and Yates, 1985; Mulcahy and Shieh,
1987; Hermanovicz and Cheng, 1990; Yu and
Rittmann, 1997). Nicolella et al. (1999b) found
that the ratio between ui and ut for the particle
considered in their experimental work, is around
0.8, in contrast with the work of Richardson and
Zaki (1954), but in agreement with a more re-
cently published study (Di Felice, 1995). Using
17
their findings for the two numerical parameters
characterising the expansion law (Eq. (4)),
Nicolella et al. (1999b) were able to predict the
overall bed height of the biological fluidised bed
as a function of the fluidising velocity for systems
whose physical parameters are reported in litera-
ture (Ngian and Martin, 1980; Mulcahy and
Shieh, 1987).
Experimental data on bed expansion character-
istics can be used in combination with the empiri-
cal equation of Richardson and Zaki (1954) (Eq.
(4)) to give an estimate of biofilm thickness
(Nicolella et al., 1995) and reactor biomass con-
centration (Nicolella et al., 1997) in BFB reactors
through a non-linear parameter estimation rou-
tine. This unique estimation does not require any
measurement other than bed height versus liquid
velocity, and constitutes a feature of BFB sys-
tems, as in other biofilm systems it is impossible
to give an estimation of biofilm thickness and
reactor biomass concentration without measuring
these variables experimentally.
4.2.3. Effects of biofilm o6ergrowth
A problem with particulate biofilm reactors is
that the biofilm has a tendency to change in
density, thickness, and shape over the cycle of
operating life. The major result of this change is
that the mass transfer, reaction and hydrody-
namic characteristics of the bed alter over time.
All these effects can dramatically influence the
operation of the unit. In steady state operation,
biomass would tend to grow continuously around
the inert support, and this growth would be only
partially balanced by loss due to liquid shear and
particle attrition. Since the biomass has a density
usually lower than the support (typical value for
the wet density range around 1100 kg m − 3), the
increase in biofilm thickness due to biological
growth modifies the mixing and fluidization char-
acteristics, resulting in an expansion of the bed.
The overall particle density reduction can cause
carry over of the particles, leading to problems
such as loss of particles and active biomass into
pumps (Wright and Raper, 1996). Hence particles
need to be filtered out, the biofilm removed and
the clean particles returned to the reactor. This
practice requires additional capital expenditure on
18 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33
particle separation equipment such as vibrating
screens (Shieh et al., 1981; Fan, 1989). In response
to biofilm overgrowth, a number of different reac-
tor configurations have been proposed to enable
the removal of the excess biofilm within the
fluidized bed (Wright and Raper, 1996). This is
done by creating areas of high turbulence on the
top of the reactor to shear the biofilm off the
support particles. As a result, two solid phases are
present at the same time in the reactor: one fully
covered with the biofilm and the other where the
biofilm is thin or even absent. Due to different
hydrodynamic characteristics, these two solid
phases tend to segregate within the bed.
4.3. Flow regimes in airlift suspension reactors
In BAS reactors, gas sparging is used to keep
the biofilm particles in suspension. In most full-
scale applications, this is achieved by using an
internal draught tube (riser). The riser is sparged
with gas, and the resulting gas hold-up difference
between the gas-sparged riser and the unsparged
downcomer leads to a difference in the bulk densi-
ties of the fluids in the two zones and hence an
induced fluid circulation (upflow in the riser,
downflow in the downcomer) is realised. The
magnitude of liquid circulation is one of the most
important design and scale-up parameters for air-
lift reactors: liquid circulation influences the gas
hold-up in the vessel, mass transfer coefficients,
the extent of mixing in the reactor and the pre-
vailing flow regimes for gas, liquid and solid
phases (Chisti et al., 1988; Heijnen et al., 1997).
Depending on liquid circulation velocity, different
flow regimes exist with respect to gas bubble
entrainment into the downcomer. For aerobic sys-
tems, the liquid velocity should be sufficient to
ensure entrainment of bubbles into the down-
comer and from the downcomer into the riser
again. This gas recirculation is desired to achieve
maximal aeration (gas hold-up). In addition, there
is a minimal gas supply rate below which the
liquid circulation velocity is insufficient to main-
tain a stable particle suspension. The liquid circu-
lation rate should at least be well above the
settling velocity of the particles. Knowledge of the
liquid circulation rate is therefore of obvious im-
portance. Reactor geometry (height, diameter,
draught tube dimensions), gas supply rate and
solids loading and properties will all determine
the liquid circulation velocity and therewith mix-
ing, gas hold-up and solid suspension.
4.3.1. Liquid circulation
Several hydrodynamic models have been devel-
oped to derive theoretical relationships between
the gas hold-up, circulation pressure drop, and
liquid velocity in two-phase airlift reactors (Bello
et al., 1985; Chisti et al., 1988; Siegel and
Robinson, 1992; Ayazi Shamlou et al., 1994).
These models were derived using either energy
balances on the gas input or macroscopic momen-
tum balances. A key parameter is the total fric-
tional resistance to fluid flow in the circulating
liquid loop, including the flow reversal at the top
and bottoms of the columns. Friction coefficients
have been estimated by applying standard rela-
tionships for single-phase flow and two-phase
flow in pipes and bends, or by experimental fitting
(Siegel and Robinson, 1992).
Most of the hydrodynamic models of three-
phase airlift reactors concern external loop sys-
tems (Douek et al., 1994), systems with small solid
particles of density close to that of water (whose
flow behaviour is similar to that of two-phase
systems, Lu et al., 1995; Hwang and Cheng,
1997), or lab-scale systems (where the hydrody-
namic regime 1 and 2 predominate, Livingston
and Zhang, 1993). Heijnen et al. (1997) have
developed a model based on a momentum balance
to predict data measured in large scale internal-
loop airlift reactors (pilot-scale, 0.4 m3, and full-
scale, 300 m3) with high gas recirculation through
the downcomer (regime 3). Garcia-Calvo et al.
(1999) presented a similar model for the descrip-
tion of flow behaviour in three-phase airlift reac-
tors, which is based on an energy balance. This
model predicts liquid and solid circulation veloc-
ity, gas hold-up and solid distribution within the
reactor, and describes the three solid flow regimes
(packed bed, fluidized bed, and complete recircu-
lation). Experimental data obtained by various
authors for reactors of different shape (internal
and external loops) and size (0.02–284 m3), differ-
ent solid particles (sizes from 0.1 to 3 mm and
 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33
densities from 2550 to 3700 kg m − 3) and different
experimental conditions are simulated
satisfactorily.
van Benthum (1998) has presented a new reac-
tor concept, the biofilm airlift suspension exten-
sion, for simultaneous nitrification and
denitrification of wastewater. The reactor consists
of a three-phase (biofilm particles, water and air)
internal-loop airlift reactor for nitrification, which
is extended with a two-phase (biofilm particles
and water) external concentric downflowing bed,
the extension for denitrification. Due to the spe-
cial design of the reactor, the liquid velocity in the
extension can be manipulated by the overpressure
in the headspace of the aerobic compartment,
therewith controlling the liquid recirculation ratio
between aerobic compartment and extension (van
Benthum et al., 1999a, 2000).
4.3.2. Solid flow regimes
Three solid regimes are reported in the litera-
ture (Douek et al., 1994) for three phase airlift
systems. These are, with increasing gas flow rate,
packed bed (settled solid), fluidized bed (no solids
recirculation) and circulated bed (complete sus-
pension regime). In the packed bed regime, the
riser may be assumed as a gas – liquid bubble
column, while the downcomer contains static liq-
uid. This condition is undesirable for operation of
biofilm reactors. The condition to suspend the
solid settled in the bottom may be written as
(Garcia-Calvo et al., 1999):
rS
oG − − 1 o 0S ]0 (6)
rL
The initial solid distribution is decisive in the
change from packed to fluidized or circulating bed
regimes. When the gas velocity, uG, is increased
from zero to the minimum value needed for
fluidization, uGmf, o 0S corresponds to the solid ini-
tially settled in the bottom of the riser. If uG
decreases from high values to reach uGmf, the
condition of Eq. (6) is fulfilled when the apparent
densities in the riser and downcomer are equal,
and o 0S = oSr −oSd. The different o 0S values corre-
sponding to these two alternatives are related to
the hysteresis effect (Heck and Onken, 1988).
19
The behaviour of BAS reactors in the fluidized
regime is similar to that of three-phase BFB reac-
tors. The boundary between the fluidized bed and
the circulating bed regimes is established by the
condition:
uLr ] ut (7)
Riser liquid velocities higher than the particle
settling velocity result in solid recirculation
through the downcomer. As the circulating bed
regime is the normal operation mode, the opera-
tional conditions to reach this regime are among
the main design criteria for BAS reactors.
4.3.3. Gas flow regimes
Based on observations at different scales
(10 − 3 –102 m3), three flow regimes have been
noted for three-phase airlift reactors when gas
velocity increases (Heijnen et al., 1997):
1. No gas entrainment in the downcomer. This
occurs at low gas velocities when the liquid
circulation velocity is insufficient to entrain
bubbles in the downcomer. The downcomer
liquid velocity is lower than the bubble swarm
velocity.
2. Gas entrainment in the downcomer, but no
gas circulation. When the liquid velocity in the
downcomer becomes about equal to the bub-
ble swarm velocity, gas is entrained into the
downcomer. Often an axial bubble distribution
is present in the downcomer. Complete solids
suspension is commonly but not always
present.
3. Complete gas circulation. All the gas bubbles
that are entrained into the downcomer are
carried all the way down the downcomer and
into the riser again. There is no axial bubble
distribution in the downcomer. The liquid ve-
locity in the downcomer is significantly higher
than the bubble swarm velocity, which leads to
a large downward gas flow in the downcomer.
The riser gas flow rate, combining the sparged
gas and the recirculated gas, is significantly
higher than the injection flow rate. Complete
solids suspension is common, but incomplete
suspension cannot be excluded.
Fig. 7 compares the gas hold-up measured for a
two-phase lab-scale airlift reactor by Nicolella et
20 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33
al. (1998a) and Bakker et al. (1993). Both sets of
data show the same trend. The slope of the graph
of oG versus uG changes drastically on two occa-
sions, and the curves present a platform, corre-
sponding to the transition between the
hydrodynamic regime of no gas entrainment in
the downcomer (regime 1) and complete gas recir-
culation (regime 3). Such transitions are indicated
by vertical lines in Fig. 7. Data on liquid circula-
tion velocity reported by Bakker et al. (1993) are
also plotted in Fig. 7, where it can be observed
that the liquid circulation velocity changes drasti-
cally when the flow regimes changes from 2 to 3,
remaining almost constant in regime 2 and rapidly
increasing with increasing gas velocity in regime 3.
Similar results are reported by van Benthum et al.
(1999b) for a pilot-scale (1 m3) internal-loop airlift
reactor with two phases (water and air) and three
phases (water, air and polystyrene particles).
In general, the importance of regimes 1 and 2
decreases with increasing scale. In large-scale re-
actors regime 1 is hardly ever realised, and the
transition from regime 2 to regime 3 depends
strongly on sparger design and reactor geometry
(Heijnen et al., 1997).
Fig. 7. Gas hold-up and circulation velocities for different flow
regimes in airlift reactors.
4.4. Mass transfer
Particle-supported biofilm reactors can be
classified as three-phase gas–liquid–solid reac-
tors. The solid phase is constituted by biofilm
particles suspended in the bulk liquid. The gas
phase is constituted by air (or oxygen) in aerobic
systems or biochemical reaction products (CH4,
CO2, H2S, and N2) in anaerobic systems, and
moves in the form of dispersed bubble through
the bulk liquid. In aerobic systems, oxygen is
transferred from the air bubbles through the bulk
liquid to the biofilm particles, where it is con-
sumed by biochemical reactions (Fig. 2). Gas–liq-
uid (G–L) and liquid solid (L–S) mass transfer
coefficients (MTCs) are, together with reaction
kinetic parameters, important design parameters
of gas–liquid–solid bioreactors for biochemical
applications.
Reliable techniques exist for the estimation of
the volumetric G–L MTC (kLa) and kinetic
parameters in biofilm reactors. The well known
transient gassing technique (Fuchs et al., 1971) is
often used to measure kLa; this technique was
recently applied specifically to BAS reactors
(Nicolella et al., 1998b). The biofilm kinetics can
be measured by means of various techniques such
as the measurement of the oxygen uptake rate
(Tijhuis, 1994). A new method has been recently
presented for the measurement of L–S MTC in
biofilm systems (Nicolella et al., 1998b, 1999a).
This method is based on the analysis of oxygen
consumption rate data in biofilm reactors and
biological oxygen monitoring systems (Tijhuis,
1994), and allows the independent estimation of
G–L MTC, L–S MTC and biofilm reaction rate
parameters.
4.4.1. Gas hold-up and gas–liquid mass transfer
The determination of the gas hold-up and volu-
metric G–L MTC is an important factor in the
design and operation of three-phase biofilm reac-
tors and has been the subject of much research
interest. Extensive data are available for the esti-
mation of these design parameters in two-phase
G–L systems (see for example Chisti, 1989 and
Bello et al., 1985 for airlift reactors; Linek et al.,
1987 for aerated agitated vessels; Shah et al.,
 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33
1982, Heijnen and van’t Riet, 1984, and Deckwer
and Schumpe, 1993 for bubble columns) and in
G – L slurry reactors (Chaudari and Ramachan-
dran, 1980; Pandit and Joshi, 1986; Siegel and
Robinson, 1992; Beenackers and van Swaaij,
1993), where the liquid and solid phases are con-
sidered as one pseudo-homogeneous phase. For
the specific case of three-phase gas – liquid – solid
reactors, there is still no universally applicable
relation describing the influence of all types of
particles in any weight fraction in any liquid and
for any reactor scale (Beenackers and van Swaaij,
1993).
The volume fraction of gas in gas – liquid – solid
dispersions (gas hold-up, oG) has a strong influ-
ence on the performance of pneumatic biofilm
reactors, such as the BAS and the IC reactors.
The residence time of the gas in the liquid, the
gas – liquid contact area for mass transfer and the
design volume of the reactor depend on the gas
hold-up which occurs under given operating con-
ditions. In addition, the gas hold-up in conjunc-
tion with the knowledge of mean bubble diameter
allows the determination of the interfacial area
and thus leads to the mass transfer rate between
the gas and the liquid phase. In pneumatic reac-
tors, the principal operating influence on gas
hold-up is the gas velocity in the vessel. Conse-
quently the effect of gas velocity on gas hold-up
and G–L MTC has been much investigated (Shah
et al., 1982; Heijnen and van’t Riet, 1984; Chisti
and Moo-Young, 1987, 1988; Fan et al., 1987).
Hydrodynamics and mass transfer have been
studied extensively for gas – liquid – solid fluidiza-
tion (Muroyama and Fan, 1985), and to a less
extent for three-phase pneumatic reactors. Most
of the papers on the hydrodynamics and mass
transfer of three-phase gas – liquid – solid reactors
have dealt with systems of relatively small scale.
The influence of the presence of solids has been
reported for various types of particles including
glass beads (Koide et al., 1985), plastic beads
(Miyahara and Kawate, 1993), polystyrene cylin-
ders (Hwang and Lu, 1997), activated carbon
particles (Muroyama et al., 1985), Raney nickel
particles (Gavroy et al., 1995), calcium alginate
beads (Lu et al., 1995), and basalt (Nicolella et al.,
1998a). For the case of biofilm coated particles,
21
Ryhner et al. (1988) reported that the gas–liquid
volumetric mass transfer coefficient in a three-
phase biofilm fluidized sand bed reactor decreased
(in the range 0.02–0.04 s − 1) with increasing
amounts of clean sand and was almost indepen-
dent of the sand fraction with biofilm-covered
sand. Direct information on biofilm airlift suspen-
sion reactors is provided by Nicolella et al.
(1998a). The gas hold-up in aerated suspensions
of solid particles is generally reported to decrease
significantly with increasing solids loading. Miya-
hara and Kawate (1993) showed that for low-den-
sity particles the gas hold-up decreased
significantly when the solid hold-up was larger
than about 0.2. Some of the available information
on gas hold-up and G–L mass transfer in three-
phase sparged reactors is reported in Table 5.
Empirical (Koide et al., 1985) and theoretical
(Livingston and Zhang, 1993; Lu et al., 1995)
models have also been proposed to describe the
hydrodynamics of small-scale systems (0.001–0.01
m3). Only recently, Heijnen et al. (1997) proposed
a model for the hydrodynamic behaviour of large
scale three-phase airlift reactors (0.1–500 m3).
The effect of solid size should be mainly related
to the effect of the particle terminal settling veloc-
ity (ut). The particle terminal settling velocity has
a direct influence on the difference in solid hold-
up between the riser and the downcomer (DoS). In
turn, the difference DoS strongly influences the
hydrodynamics of the system: if the solid hold-up
in the riser is larger than in the downcomer, the
presence of solids lowers the driving head of the
system, and thereby the liquid circulation rate and
the gas recirculation, i.e. the gas residence time
(Heijnen et al., 1997). Nicolella et al. (1998a)
measured the gas hold-up in a concentric tube
airlift reactors in the presence of basalt and bio-
film particles of different sizes. The terminal set-
tling velocities of basalt particles varied in a wider
range (19–129 mm s − 1) than those of biofilm-
coated particles (30–49 mm s − 1), and the influ-
ence of particle size was more noticeable in the
first case than in the latter. In both cases, the gas
hold-up decreases with increasing particle settling
velocity. The observed influence of solid size on
the hydrodynamics is consistent with that found
by other authors for similar systems (Koide
 22
Table 5
Selected gas hold-up and G–L mass transfer correlations for three-phase (gas–liquid–solid) sparged reactors

System Range of variables Gas hold-up correlationa G–L mass transfer correlation(*) Reference

AL. L, water, electrolyte, glycerol dS =0.08–0.2 mm;  n, n

oG oG
  
CS 0.069
Koide et al. (1985)

C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33

rS =4680–8770 kg m−3;
       n
and ethylene glycol aqueous (1−oG)4 (1−oG)4 [kLa]/[kLa]0 = 1+0.099
0
rS
solutions. G, air; S, glass and ut =0.165–2.44 m s−1; CS 0.091 rLs 3L 0.023
ut 0.046 −1
bronze spheres uG =0.03–0.15 m s−1; hc = = 1+0.17

    n
rS gm 4L uG
1.5 m; dc =100–300 mm;
dr =60–190 mm rLs 3L 0.043
ut 0.067 −1

gm 4L uG

  n
DTFB. L, water; G, air; S, glass dS =0.3–0.7 mm; [kLa]/[kLa]0
Fan et al. (1987)
beads, activated carbon rS =1500–2500 kg m−3; oSRet
particles ut =0.023–0.0935 m s−1; = 0.143 1−31.5

 
ReS
uG =0–17.069 m s−1; hc =1.22
oSRet 0.002

 
m; dc =760 mm; dr =500 mm × (1−oS)10.31
ReS
FB. L, water; G, air; S, glass dS =0.05–8 mm; rS =2510–2950 oS
kLa= 0.39 1− u 0.67 Nguyen-Tien et al.
spheres kg m−3; oS =0.005–0.53; 0.58
g
(1985)
uL =0.046–0.116 m s−1; hc =m;
dc =140 mm

'  
AL. L, water; G, air; S, plastic dS =2.6–12.7 mm; 0.4
Fr% Miyahara and
oG =
beads rS =1046–1135 kg m−3; uL Kawate (1993)
uG =0.03–0.15 m s−1; uL =2–9 1+0.4 Fr% 1+
uG
m s−1; hc =1 m; dc =148 mm;
dr =60–100 mm
FB. L, water; G, air; S, glass dS =0.53–0.755 mm; rS =2338 kLa= 0.99u 0.5 0.97 −0.27
G uL dS (1−oS)0.53 Zheng et al. (1995)
beads, activated carbon kg m−3; uG =0.01–0.1 m s−1; (1−0.89
h+0.2h)
particles uL =0.05–0.5 m s−1; hc =4.1 m;
dc =285 mm; oS =0.02–0.165

a
Subscript 0 refers to the two-phase gas–liquid system.
 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33
Fig. 8. Selected correlations for liquid–solid mass transfer
coefficient in three-phase sparged reactors.
et al., 1985) and that predicted by the model of
Livingston and Zhang (1993).
The effect of the ratio of downcomer to riser
cross-section area (Ad/Ar) on hydrodynamic and
mass transfer in airlift reactors was studied by
Bello et al. (1985). They measured gas hold-up
and G–L MTC in two-phase internal and exter-
nal loop reactors with Ad/Ar varying in the range
0.11 –0.69, and proposed unifying generalised cor-
relations for both parameters.
4.4.2. Liquid solid mass transfer
Mass transfer to particles submerged in a fluid
is generally described by equations based on the
boundary layer theory, which leads to a correlat-
ing equation of the form:
Sh = 2.0+ CRenScm
Correlations of this form involve calculating the
particle Reynolds number. In problems involving
liquid–particle mass transfer in three-phase sus-
pension the Reynolds number is frequently
defined according to Kolmorgoff’s theory of
turbulence:
od 4S
Re =
n3
23
where o is the energy dissipation rate.
One approach to analysing the mass transfer
data is to assume that the biofilm reactor may be
treated as having essentially uniform energy dissi-
pation rates throughout the entire volume. The
energy dissipation rate so calculated is:
o= uGg (10)
Kolmorgoff’s theory has been used extensively
in correlating mass transfer data for bubble
columns (Sano et al., 1974; Sanger and Deckwer,
1981), fluidized beds (Arters and Fan, 1986; Liv-
ingston and Chase, 1990) and airlift reactors
(Nicolella et al., 1998b). An exponent of 1/3 in
Eq. (8) is often chosen for the Schmidt number.
Published correlations valid for different types of
three-phase reactors are compared in Fig. 8. The
characteristics of the systems used by different
authors are reported in Table 6. For the case of
mass transfer to biofilm particles in a BAS reac-
tor, Nicolella et al. (1998b) correlated their exper-
imental data with the following relationship:
od 4S 0.241
Sh= 2.0+ 0.265 Sc1/3 (11)
n3
The L–S MTC measured for particle-supported
biofilms was found to be smaller (by a factor of
approximately 15%) than the values reported for
rigid particles, and it was therefore concluded that
L–S mass transfer should be regarded as critical
process in biofilm systems.
4.4.3. Intraparticle mass transfer and reaction
Particle-supported biofilm reactors are charac-
terised by microorganisms being attached to a
solid surface in the form of a biofilm. Biofilms are
dense layers through which substrates have to be
(8) transported for biochemical reaction to occur.
This transport takes place by molecular diffusion,
which is a slow process. In practice it proves that
the general rule is that the removal is limited by
diffusion, and one of the major disadvantages of
biofilm reactors is the low efficiency of thick
biofilms (Harremoes and Henze, 1995).
The substrate concentration in a biofilm is de-
termined by the substrate conversion process and
diffusion. The latter can be described by Fick’s
(9)
law with an effective diffusion coefficient. In gen-
 24
C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33
Table 6
Selected liquid–solid mass transfer correlations in aerated suspensions

Reference System Particles uG (mm s−1) Sc Re= od 4/n 3 Sh

Sano et al. BC Ion-exchange resin, benzoic acid, KMnO4, b-naphthole; 0–170 217–1410 1–107 2+0.4Re1/4Sc1/3
(1974) oS =0.03–0.07; dS =0.06–0.833 mm
Arters and Fan FB Benzoic acid; rS =1298 kg m−3; dS =1.8–4.4 mm 0–260 1960–3500 2.5×105 2+0.695Re0.2Sc1/3
(1986) –4.8×109
Goto et al. BCDT Ion-exchange resin; oS =0–0.015; dS =0.55–0.92 mm 1–10 143 103–106 2+1.01Re0.173Sc1/3
(1989)
Livingston and BCDT Ion-exchange resin; oS =0.04–0.1; dS =0.655–1.119 mm 0–60 400 1.5×104 2+0.275Re0.274Sc1/3
Chase (1990) –6.77×105
Kikuchi et al. BC, FB Ion-exchange resin; oS =0–0.4; dS =0.32–2.59 mm 0–100 300–600 10–104 2+0.47Re0.21Sc1/3
(1995)
Nicolella et al. BAS Biofilms; oS =0.05–0.15; dS =0.47–1.95 mm 2–27 400 103–2×106 2+0.275Re0.274Sc1/3
(1998a)
 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33
eral the effective diffusion coefficient is 80 – 90%
of the diffusion coefficient in water. The substrate
flux at the biofilm surface (if the concentration is
well above the substrate affinity coefficient, which
is often the case for biofilm processes) can be
calculated by considering two distinct reaction
regions with abrupt transition in the order of the
reaction (Harremoes, 1978). This transition is
characterised by:
b=
' 2DeC i
f
(12)
k0d 2
For b \ 1, the biofilm is fully penetrated by the
substrate and the substrate flux is zero order with
respect to the substrate concentration at the bio-
film surface:
b \ 1 [ NS =k0d (13)
For b B1, the biofilm is partially penetrated by
the substrate and the substrate flux is half order
with respect to the substrate concentration at the
biofilm surface:
b B1 [ NS =
2Dek0C if (14) and biofilm specific activity changes along the
The zero-order reaction rate constant is defined
as (Harremoes, 1978)
k0 = mmaxXf/Y (15)
4.5. Mixing and scale-up effects
Liquid mixing in particulate biofilm reactors
affects the interphase mass transfer, the reactant
concentration distribution, and ultimately reac-
tant conversions. Information on axial liquid mix-
ing is therefore crucial to reactor design and
optimisation. Fluidized bed systems and airlift
suspension systems present different mixing char-
acteristics. Airlift suspension reactors are usually
considered ideally mixed for liquid and solids
(Heijnen et al., 1993). Axial liquid mixing in liq-
uid – solid and gas– liquid – solid fluidized beds is
often analysed using the axial dispersion model
(Levenspiel, 1972; Davidson et al., 1985; Fan,
1989). In these systems, a distribution of solid size
and/or density results in a distribution of terminal
settling velocities leading to a classification within
the fluidized bed. This classification is caused by a
25
sorting effect where particles with higher settling
velocity are found at the bottom of the bed (Di
Felice, 1995).
The most common technique to study the mix-
ing characteristics of fluidized bed and airlift sus-
pension systems is the injection and response
method (Levenspiel, 1972).
4.5.1. Fluidized bed systems
In BFB reactors, solid stratification occurs ow-
ing to the difference in size and density among the
biofilm particles. Thicker biofilms are usually
found at the top of the bed, whilst bare carriers
and thin biofilms remain in the bottom region of
the column (Shieh et al., 1981; Hermanovicz and
Ganczarczyk, 1983; Boaventura and Rodrigues,
1988). Particle stratification has been attributed to
differences in drag and buoyancy that affect parti-
cle terminal settling velocity (Di Felice, 1995).
Simple models have been developed to simulate
solid stratification in BFBs (Di Felice et al., 1997).
As a consequence of stratification and size distri-
bution, biodegradation rate, biofilm composition
height of the bed. For example, in anaerobic
BFBs, the glucotrophic activity decreases along
the bed from the bottom to the top, whilst the
methanogenic activity increases and is maximum
at the top of the bed (thickest biofilms) (Buffiere
et al., 1998a). In the upper portion of the bed,
where the thickest biofilms are present, diffusional
mass transfer limitations are particularly impor-
tant and reduce the conversion capacity of the
reactor (Schreyer and Coughlin, 1999).
Axial liquid mixing in liquid–solid fluidized
beds and three-phase fluidized beds have been
extensively studied for systems with particles hav-
ing densities significantly greater than that of the
liquid medium (e.g. Davidson et al., 1985;
Nguyen-Tien et al., 1984). Only a few studies
(Kikuchi et al., 1984; Tang and Fan, 1990; Zanin
et al., 1993) focused on axial liquid mixing in
fluidized beds containing particles akin to particu-
late biofilms. In particular, Tang and Fan (1990)
measured axial dispersion coefficients in liquid
fluidized beds of low density particles (1.05–1.3
kg m − 3). The axial dispersion coefficient was
found to be independent of bed height and to
26 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33
increase monotonically with increasing liquid su-
perficial velocity. The extent of axial liquid mixing
in beds of light particles is significantly less than
that in beds of heavy particles, and therefore the
particle density effect on axial liquid mixing is
important. Gas production affects the hydrody-
namics and mixing characteristics of BFBs
(Buffiere et al., 1998b), resulting in a bed contrac-
tion and in an increase of dispersion. These effects
have in turn an influence on reactor performance,
which, at high reactor loading, can be reduced by
10 – 15% (Buffiere et al., 1998c).
The axial dispersion coefficient in liquid
fluidized bed reactors decreases with increasing
ratio of height to diameter ratio (Thommes et al.,
1995). This results in different mixing characteris-
tics in lab-scale reactors and full-scale applica-
tions. As the H/D ratio is larger in lab-scale
reactors (H/D \10) than in pilot-scale and full-
scale applications (H/D B5, see Table 2), the
liquid axial mixing is normally greater in indus-
trial columns than it is in small bench reactors.
This effect considerably affects the scale-up of
BFBs.
Though USB and EGSB present some hydro-
dynamic similarities with fluidized systems, the
simple axial dispersion model is insufficient to
describe their mixing characteristics. Based on
experience with a model with all dimensions 1/20
of a full-scale USB plant, Bolle et al. (1986)
observed that in USB reactors the sludge bed and
the sludge blanket behave like mixed tank reac-
tors with short-circuiting flows, whilst the settling
volume can be described as a plug flow reactor.
The short-circuiting flows over the sludge bed and
the sludge blanket (i.e. the mixing characteristics
of these compartments) were affected by the su-
perficial gas velocity. An optimal bed height of
3.5 – 4 m was found corresponding to minimal
short-circuiting flows.
4.5.2. Airlift suspension systems
The airlift reactor can be considered as a system
consisting of interrelated zones: riser, downcomer,
bottom section, top section and settler. Mixing
efficiency is due to the circulation of liquid and
solid particles around the airlift column, and to
the backmixing in the settler (Obradovic et al.,
1994). The degree of mixing in the riser is re-
ported to be higher than that in the downcomer,
and mixing in the two-phase systems is higher
than that in three-phase systems (Lu et al.,
1994a).
The axial dispersion model is generally used to
describe the overall mixing phenomena in airlift
reactors. The overall axial dispersion coefficient
increases as aeration rate increase (Lu et al.,
1994b). This result is expected since a higher
aeration rate corresponds to a higher circulation
velocity, and, consequently, to a higher degree of
turbulence.
Axial dispersion in airlift reactors is higher than
in fluidized beds. For lab-scale reactors of com-
parable size, Schoutens et al. (1986) reports axial
dispersion coefficients of 4–16× 10 − 4 and 3–
10× 10 − 3 m2 s − 1 for fluidized beds and airlift
columns, respectively. The degree of mixing is
therefore higher in airlift reactors than in fluidized
beds.
Heijnen et al. (1993) report that a pilot-scale
(0.25 m3) and a full-scale (280 m3) BAS reactor
have nearly the same gas hold-up. This implies
that in airlift reactors there is a negligible effect of
scale on mass transfer and mixing characteristics.
The effect of reactor scale on biomass structure,
biofilm detachment, liquid mixing, gas–liquid
mass transfer and liquid–solid mass transfer in
two- and three-phase particulate biofilm reactors
has been analysed extensively by Heijnen (1996).
In particular:
1. The importance of undesired biomass struc-
tures (e.g. growth in suspension, growth on
static reactor surface growth) is clearly scale-
dependent. In particular the concentration of
suspended organisms and debris in the reactor
depends on their settling velocity relative to
the wash-out liquid velocity in the three-phase
separator. It is important to maintain identical
settler liquid velocities at different reactor
scales to eliminate undesired biomass
structures.
2. As discussed above, most biofilm detachment
in particulate biofilm reactors is due to parti-
cle–particle interactions in collisions. In two-
phase systems, abrasion only occurs between
biofilm particles, because bare carriers are con-
 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33
centrated at the bottom of the reactor due to
stratification. The rate of abrasion in two-
phase systems increases at increasing particle
concentrations, i.e. at decreasing liquid veloc-
ity. Liquid velocity should therefore be main-
tained constant in scaling-up a two-phase
reactor. For three-phase systems it is noted
that, contrary to two-phase systems, biofilm
particles are mixed up with bare carriers. If a
heterogeneous flow regime is maintained,
scale-up effects are not significant.
3. In three-phase BAS reactors, there is an im-
portant effect of scale on the oxygen transport,
which is related to the circulation of gas bub-
bles in the downcomer. As discussed above,
gas regime 1 (no bubble circulation) and 2
(presence of bubbles in the downcomer but no
bubble circulation) only occur in small reac-
tors. Full-scale reactors always operate with
full bubble recirculation (gas regime 3), which
is obviously the most favourable for oxygen
transfer due to the presence of gas flow
everywhere.
5. Conclusions
A number of particulate-biofilm reactor tech-
niques (USB, BFB, EGSB, IC, BAS) have been
developed in the last two decades for applications
to industrial and municipal wastewater treatment.
Several full-scale applications have been presented
and illustrated in this review, and the engineering
aspects of their design and operation have been
analysed and discussed. Most fundamentals of
biofilm formation, hydrodynamics, mixing, mass
transfer and chemical kinetics of these systems are
well understood, and empirical correlations and
mathematical models are available for the predic-
tions of relevant parameters and variables needed
in the design and operation of full-scale reactors.
One of the least understood aspects in particu-
late-biofilm reactors is detachment of biomass.
Although the mechanism seems to be particle –
particle collisions, there is no design rule for rates
of detachment. This is an important lack of
knowledge, which requires further research.
27
Appendix A. Design criteria for particle-biofilm
reactors
A.1. Condition for biofilm de6elopment
Biofilms will form in a bioreactor when the
dilution rate (DH = 1/tH = Q/V) is larger than the
maximum growth rate (mmax) (Heijnen, 1984; Ti-
jhuis, 1994):
1 Q
= \mmax (A1)
tH V
In some cases biofilm formation is already ob-
served at a dilution rate below the maximum
growth rate. Upon analysis of the system it will be
generally observed that despite the biofilm accu-
mulation the majority of the conversion still takes
place in suspension.
For an aerobic system where the rate limiting
factor is the gas–liquid oxygen mass transfer, the
reactor size can be designed as a function of
organic loading rate FC = Q/C, oxygen yield (YO)
and oxygen transfer rate (OTR):
FC Y QC
V= = O (A2)
OTR OTR
By substituting Eq. (A2) and rearranging terms
in Eq. (A1), the condition for biofilm develop-
ment (Eq. (A1)) can be rewritten as:
OTR
CB (A3)
YOmmax
The line in Fig. 1 delimiting the region of
applications of particle-biofilms and flocs (line 1)
is defined assuming a maximum oxygen transfer
rate of 10 kg m − 3 day − 1, an oxygen yield coeffi-
cient of 1 and a maximum specific growth rate of
1 day − 1.
A.2. Condition for particle retention
For particle retention in a bioreactor, the su-
perficial liquid velocity u=Q/A must be lower
than the particle settling velocity (ut):
Q
u= B ut (A4)
A
28 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33
Using Eq. (A4), and assuming a cylindrical
geometry for the bioreactor, the reactor volume
can be expressed as a function of the liquid veloc-
ity and the height to diameter ratio a = H/D:
'
Q 4Q
V=a (A5)
u pu
By combining Eq. (A5) with Eq. (A2), the
condition for particle retention in the biofilm re-
actor (Eq. (A4)) can be rewritten as:
pu 3t C 2Y 2O
QB (A6)
4a 2OTR2
The lines in the diagram C – Q of Fig. 1 delimit-
ing the regions of applications of particle-biofilm
suspensions and floc suspension (line 2 and 3,
respectively) are obtained by assuming settling
velocities of 30 and 5 m h − 1 for particle biofilms
and flocs, respectively, and an aspect ratio a =5
(the maximum oxygen transfer rate is fixed at 10
kg m − 3 day − 1 as in the previous section).
A.3. Liquid– solid mass transfer limitations
In biofilm reactors, the process rate may be
limited by liquid–solid mass transfer. For aerobic
processes, assuming for the sake of simplicity that
oxygen concentration at the biofilm interface is
negligible, the oxygen flux (N) from the bulk
liquid to the biofilm surface is given by:
N =kLSCL (A7)
The volumetric oxygen transfer rate (OTR) is
the product of flux (N) and biofilm specific sur-
face area (a).
OTR = Na (A8)
For systems with low values of biofilm specific
surface area (e.g. static biofilm reactors), the liq-
uid – solid mass transfer may become the rate lim-
iting process. Assuming values of 1 m day − 1 for
kLS and 8 g m − 3 for CL (dissolved oxygen concen-
tration in water at equilibrium using air at ambi-
ent temperature and pressure), the maximum
oxygen transfer rate in a static biofilm reactor
with a specific surface area of 200 m2 m − 3 is 2 kg
m − 3 day − 1. Introducing this value in Eq. (3), a
vertical line in the diagram C – Q (line 4 in Fig. 1)
is obtained which limits the region of applicability
of static biofilm reactors.
A.4. Limitations on biomass separation
The separation step (usually by sedimentation)
is the main limitation in processes based on the
use of flocs (e.g. activated sludge processes). Floc
settling characteristics are rather poor (ut B 5 m
h − 1) and limits the maximum volatile suspended
solid concentration in the liquors to about 3 kg
m − 3 (Metcalf and Eddy, 1991). This in turn
results in a limitation of the reactor capacity,
which can be expressed as a function of the
reactor biomass concentration by considering the
food to microorganism ratio (defined as the ratio
between the organic load to the reactor and the
reactor biomass concentration: F/M=QC/VX):
OTR= (F/M)XYO (A9)
The food to microorganism ratio is an empiri-
cal parameter frequently used for the design of
activated sludge processes. Typical values for con-
ventional systems range from 0.3 to 1.5 kgCOD
−1
kgSS day − 1 (Metcalf and Eddy, 1991). Assuming
−1
a value of 1 kgCOD kgSS day − 1, and taking YO as
1, the maximum oxygen transfer rate in an acti-
vated sludge process can be estimated as 3 kg
m − 3 day − 1, which, using Eq. (A3) results in a
vertical line in the diagram C–Q (line 5 in Fig. 1).
When the solid loading to the settler increases,
the settling area required to achieve a desirable
effluent quality in a reasonable time may become
unrealistic. There is no general rule, since in prin-
ciple the settling volume could be chosen as large
as required, but a sensible design would probably
not consider settling volumes much larger than
the oxidation volumes. If, for example, it is as-
sumed that the settler to reactor volume ratio is
limited to a given value (let g be this ratio), then
it can be shown (with calculations similar to those
performed in the previous section) that the region
of applicability of activated sludge processes is
delimited in the plane C–Q by the line:
pq 3C 2Y 2O
Q= (A10)
4(a/g)2OTR2
 C. Nicolella et al. / Journal of Biotechnology 80 (2000) 1–33
where q is the overflow rate to the settler. Values
for q and a in Eq. (A10) can be obtained from the
practice of wastewater engineering (Metcalf and
Eddy, 1991). The line 6 of Fig. 1 is plotted
assuming q = 2 m h − 1, a =0.5, g =5.
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