Bioresource Technology 111 (2012) 111–121

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Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Development of a calibration protocol and identification of the most

sensitive parameters for the particulate biofilm models used in biological
wastewater treatment
Ahmed Eldyasti a, George Nakhla a,b,⇑, Jesse Zhu b
a
Department of Civil and Environmental Engineering, The University of Western Ontario, London, Ontario, Canada N6A 5B9
b
Department of Chemical and Biochemical Engineering, The University of Western Ontario, London, Ontario, Canada N6A 5B9

a r t i c l e i n f o a b s t r a c t

Article history: Biofilm models are valuable tools for process engineers to simulate biological wastewater treatment. In
Received 7 December 2011 order to enhance the use of biofilm models implemented in contemporary simulation software, model
Received in revised form 2 February 2012 calibration is both necessary and helpful. The aim of this work was to develop a calibration protocol of
Accepted 6 February 2012
the particulate biofilm model with a help of the sensitivity analysis of the most important parameters
Available online 14 February 2012
in the biofilm model implemented in BioWinÒ and verify the predictability of the calibration protocol.
A case study of a circulating fluidized bed bioreactor (CFBBR) system used for biological nutrient removal
Keywords:
(BNR) with a fluidized bed respirometric study of the biofilm stoichiometry and kinetics was used to
Biofilm model
Activated sludge models
verify and validate the proposed calibration protocol. Applying the five stages of the biofilm calibration
Nitrification procedures enhanced the applicability of BioWinÒ, which was capable of predicting most of the perfor-
Denitrification mance parameters with an average percentage error (APE) of 0–20%.
Calibration protocol Ó 2012 Elsevier Ltd. All rights reserved.

1. Introduction many), AQUASIMÒ (EAWAG, Switzerland), EFORÒ (DHI Inc., Den-

mark), BioWinÒ (Envirosim Associates Ltd., Burlington, ON), GPS-
During the last few years, there has been renewed interest in XÒ (Hydromantis Inc., Hamilton, ON), AQUIFASÒ (Aquaregen,
attached growth biological treatment processes, which prove to Mountain View, CA), Pro-2DÒ (CH2M HILL, Inc., Colorado, US),
be economic and efficient. Along with the growing interest in STOATÒ (WRc, Wiltshire, England), WESTÒ (Mostforwater, Bel-
attached (biofilm) treatment processes, there have been also gium), and TRIFLÒ (Penn State University, Pennsylvania, USA).
numerous efforts towards their numerical analysis and biofilm Amongst them, Pro-2DÒ, WESTÒ, and TRIFLÒ model the biofilm as
modeling studies. Five mathematical classes, characterized by a a homogeneous layer while the other models the biofilm as a het-
mixed culture and biofilm models, have been published and pre- erogeneous biofilm layers, which gives the biofilm model a higher
sented as: analytical (A), pseudo analytical (PA) (Wanner and Rei- level of accuracy (Boltz et al., 2010; Eldyasti et al., 2011).
chart, 1996), one-dimensional numerical (1-D) (Wanner et al., In order to successfully apply these simulation packages to a
2006), two-dimensional numerical (2-D) (Boltz et al., 2010), and biological system, calibration of the model is absolutely necessary.
three-dimensional numerical (3-D) (Xavier et al., 2005). These Petersen et al. (2002) defined the model calibration as the adapta-
models vary in complexity from simple analytical models to mul- tion of the model to fit a certain set of information obtained from a
ti-dimensional dynamic models. For engineering design and analy- practical wastewater treatment plant. A methodology of the
sis, a balance between the simplified and complex mechanistic activated sludge models (ASMs) calibration procedures has been
approach is required. Hence, one-dimensional fully dynamic and defined by a number of researchers: ‘‘ASMs calibration protocol’’
steady-state simulation models are widely used and implemented developed by Petersen et al. (2002); ‘‘STOWA calibration protocol’’
in contemporary simulation software (Wanner and Gujer, 1984; developed by Dutch Foundation of Applied Water Research (Huls-
Wanner, 1986; Wanner and Reichart, 1996; Xavier et al., 2005; beek et al., 2002); ‘‘BioMath-calibration protocol’’ developed by
Boltz et al., 2010). The biofilm models are available in several Vanrolleghem et al. (2003); and ‘‘Hochschulgruppe (HSG) proto-
user-friendly forms such as SimbaÒ (Ifak GmbH, Magdeburg, Ger- col’’ developed by Langergraber et al. (2004). Thus, calibration of
ASMs have been successfully applied both in research and practice,
and serves as the benchmark for new or expanded activated sludge
⇑ Corresponding author at: Department of Civil and Environmental Engineering,
models (Boltz et al., 2010).
The University of Western Ontario, London, Ontario, Canada N6A 5B9. Tel.: +1 519
661 2111x85470; fax: +1 519 850 2921. Attached growth reactor (AGR) mathematical modeling is more
E-mail address: [email protected] (G. Nakhla). complicated than suspended growth reactor (SGR) due to: (a)

0960-8524/$ - see front matter Ó 2012 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2012.02.021
112 A. Eldyasti et al. / Bioresource Technology 111 (2012) 111–121

Nomenclature

A cross sectional area MTBL mass transfer boundary layers

AGR attached growth reactor MWW municipal wastewater
AOBs ammonia oxidizing bacteria NOBs nitrite oxidizing bacteria
APE average percentage error OHOs ordinary heterotrophic organisms
ASM activated sludge models PAOs phosphorus-accumulating organisms
b endogenous decay coefficient SGR suspended growth reactor
BNRb Biological nutrient removal endogenous decay coeffi- Si,j normalized sensitivity coefficient
cient SRT solids retention time
CFBBR circulating fluidized bed bioreactor SSAmax maximum possible surface area
COD chemical oxygen demand SSV specific surface volume
D diffusion coefficient TBOD total biochemical oxygen demand
dm media diameter TCOD total chemical oxygen demand
dp particle diameter TSA total surface area
EBCTs empty bed contact times TSS total suspended solids
Fup nonbiodegradable particulate chemical oxygen demand VSS violate suspended solids
fraction WWTP wastewater treatment plant
Fxsp particulate biodegradable fraction of slowly biodegrad- xi input variable
able COD YH true biomass yield
g acceleration of gravity yi output variable
HRT hydraulic retention time dmsqr
j
mean square sensitivity measure
KS monod half saturation coefficient DO2 oxygen consumption
L length of the section DpS additional pressure drop
LL mass transfer boundary layer thickness e axial void fraction
M/AL solids concentrations eS solids hold-up
MBBR moving bed bioreactor lmax maximum specific growth rate

increased complexity in describing the pollutant biodegradation BioWinÒ. The pilot-scale facility was developed based on the labo-
rate in the biofilm, which in turn depends on the intrinsic micro- ratory-scale experience reported by Chowdhury et al. (2008). The
bial degradation and growth kinetics in the biofilm; (b) the biore- pilot-scale CFBBR consists of an anoxic compartment (riser) fol-
actor hydrodynamic characteristics including the flow pattern and lowed by aerobic compartment (downer) and recirculation lines
the impact of bulk-liquid hydrodynamics, voidage, pressure and between downer and riser as shown in Fig. 1a and b. Table 1a illus-
filling factor; (c) liquid–solid mixing states; (d) varying detach- trates the municipal wastewater, characterized predominantly by
ment and attrition rates; and (e) boundary layer mass transfer, carbon to nitrogen ratio of 8:1, total chemical oxygen demand to
and biofilm diffusional resistances and mass transfer (Nicolella violate suspended solids (TCOD/VSS) ratio of 2:1 and total Bio-
et al., 2000; Boltz et al., 2010). Therefore, calibration of the biofilm chemical oxygen demand to total chemical oxygen demand
models is significantly more intricate than SGR. A comprehensive (TBOD/TCOD) of 0.65. Table 1b shows the detailed operational con-
literature review of biofilm modeling, calibration protocol, and cal- ditions and reactor design parameters of the CFBBR; further details
ibration approach demonstrated that there are no readily available of the reactor and operational conditions are presented elsewhere
complete calibration protocols for biofilm reactors. The existing (Chowdhury et al., 2008; Eldyasti et al., 2010).
calibration protocols have been developed for the ASM model only
and cannot be applied for biofilm model (Boltz et al., 2010; WEF, 2.1.2. Biofilm support media (bioparticles)
2011). Lava rock particles with an average diameter of 600 lm (300–
Thus, the primary goal of this work was to develop a calibration 850 lm) were used as the carrier media (bioparticles) for biofilm
protocol for the particulate biofilm model with the help of the sen- attachment in the CFBBR. The particle porosity was about 33% and
sitivity analysis in BioWinÒ software and verify the predictability the total porosity (particle porosity and voids between particles)
of the calibration protocol. A case study of the patented circulating was 61%. The bulk density (considering packed media filled with
fluidized bed bioreactor (CFBBR) system used for biological nutri- water) of particles was approximately 1720 kg/m3, with true den-
ent removal (BNR) from municipal wastewater (Nakhla et al. sity (the ratio of sample mass to its true volume) of 2560 kg/m3
2004; Chowdhury et al., 2010), with biofilm stoichiometry and and a high specific surface area of 10,950 m2/m3. The CFBBR was
kinetics derived from a fluidized bed respirometric study (Chow- started with 125 and 421 kg of fresh lava rock particles with the cor-
dhury et al., in press) was adopted for validation of the proposed responding compact bed volumes of 80 and 277 L in the riser and
calibration protocol. the downer respectively. The amount of bioparticles was deter-
mined considering the observed nitrification–denitrification rates
2. Methods of 0.14 g N/(g VSSd) and 0.24 g N/(g VSSd) respectively and at-
tached biomass of 5–20 mg VSS/g lava rock in the lab-study (Chow-
2.1. Experimental data dhury et al., 2008, 2010). The observed attached biofilm thicknesses
on the aerobic and anoxic bioparticles in the pilot-study were 60–
2.1.1. Pilot-scale of circulating fluidized bed bioreactor 190 and 500–700 lm, respectively. Biofilm-coated particles were
The experimental data obtained from a pilot-scale CFBBR oper- periodically taken from sampling ports along the columns for the
ated for 255 days to treat municipal wastewater (MWW) at the purpose of measuring the biofilm thickness. The sampling was done
Adelaide Pollution Control Plant, London, Ontario, Canada was by using a syringe at the same pressure inside each column to min-
used to calibrate and validate the biofilm model implemented in imize disturbances to the biofilm structure; further details of the
 A. Eldyasti et al. / Bioresource Technology 111 (2012) 111–121 113

Fig. 1. (a) Schematic diagram of CFBBR. (b) 2-D view of the pilot-scale CFBBR. (c) Bioparticles distribution in the CFBBR system. (d) BioWinÒ schematic flow diagram of CFBBR
model.

Table 1a
Influent and effluent characteristics at different phases.

Parameter Phase I Phase II Phase III Phase IV

Influent⁄ Effluent⁄ Influent⁄ Effluent⁄ Influent⁄ Effluent⁄ Influent⁄ Effluent⁄
pH 6.6–7.1 6.9–7.3 6.6–7.2 6.9–7.2 6.6–7.1 7.0–7.4 6.5–7.1 6.8–7.2
Alkalinity⁄⁄ 223 ± 17 89 ± 23 245 ± 29 116 ± 12 292 ± 39 75 ± 38 253 ± 19 79 ± 24
COD (mg/L) 332 ± 42 26 ± 3 349 ± 38 39 ± 8 578 ± 39 41 ± 14 496 ± 152 45 ± 7
SCOD (mg/L) 71 ± 14 13 ± 4 100 ± 16 15 ± 4 192 ± 82 20 ± 8 117 ± 23 23 ± 5
NH4–N (mg/L) 22.1 ± 5.2 0.6 ± 0.5 24.6 ± 2.9 0.9 ± 0.3 35.2 ± 8 0.9 ± 0.6 25.8 ± 1.1 3.9 ± 0.9
NO3–N (mg/L) 0.4 ± 0.6 3.6 ± 1.2 0.4 ± 0.1 4.7 ± 1.3 <0.06 5.4 ± 1.3 0.4 ± 0.1 2.8 ± 0.6
TKN (mg/L) 39.7 ± 10.5 2.5 ± 0.4 40.5 ± 3.5 3 ± 0.6 60.2 ± 11 2.3 ± 0.6 54.9 ± 9.1 6.7 ± 1.2
TN (mg/L) 40.3 ± 10.5 6.2 ± 1.1 42.1 ± 9.5 7.6 ± 1.3 61.5 ± 14 7.7 ± 0.8 55.8 ± 10.3 9.8 ± 1.2
PO4–P (mg/L) 2.2 ± 0.9 0.7 ± 0.1 1.9 ± 0.4 0.5 ± 0.1 3.7 ± 2.9 1.3 ± 0.4 2.3 ± 0.2 0.6 ± 0.2
TP (mg/L) 4.9 ± 1.0 1.0 ± 0.1 4.2 ± 0.8 1.2 ± 0.2 5.2 ± 4 0.7 ± 0.2 5.9 ± 0.6 1.2 ± 0.4
TSS (mg/L) 217 ± 27 11 ± 2 219 ± 26 22 ± 6 443 ± 11 41 ± 20 443 ± 17 27 ± 6
VSS (mg/L) 174 ± 28 9±2 171 ± 23 16 ± 5 339 ± 19 22 ± 11 315 ± 11 21 ± 6
BOD (mg/L) 217 ± 18 16 ± 5 211 ± 28 19 ± 3 321 ± 26 21 ± 8 352 ± 89 23 ± 3
SBOD (mg/L) 51 ± 5 3±1 69 ± 11 4±2 97 ± 55 7±2 88 ± 15 7±2
C:N:P 8.2:1:0.1 8.3:1:0.1 9.6:1:0.2 9.8:1:0.1
⁄ ⁄⁄
Average ± SD (number of samples, 20) with a frequency of a sample every 3 days. (mg CaCO3/L).

bioparticles sampling and measurement are presented elsewhere compact bed was 170 and 1070 m2, respectively calculated as
(Chowdhury et al., 2008; Eldyasti et al., 2010). The maximum possi- shown in Eq. (1).
ble surface area (SSAmax) in the anoxic and aerobic reactors was cal-
Total surface area ðTSAÞ ¼ SSAmax  ð1  eÞ  EBCT  Q ð1Þ
culated considering zero void ratio and biofilm thickness of 500 and
120 lm diameter and a bare lava rock particles of 600 lm diameter
as 3750 and 7060 m2/m3, respectively. Considering bed voidage (e), 2.1.3. Respirometry study
spherical lava rock particles occupy 44% of the total reactor volume, A new respirometric approach has been developed by Nakhla
translating into a possible surface area for the anoxic and aerobic and coworkers (Chowdhury et al., in press) to determine heterotro-
reactors of 2100–3113 m2/m3 and 3318–6989 m2/m3, respectively phic biofilm kinetics of particulate bioreactors employing liquid
as shown in Fig. 1c. Thus, the total surface area (TSA) of the carrier recirculation for particle fluidization were used to determine the
media for the entire anoxic and aerobic reactors considering the biofilm kinetics and stoichiometry. The modified respirometry
114 A. Eldyasti et al. / Bioresource Technology 111 (2012) 111–121

Table 1b
Operating conditions.

Phase I Phase II Phase III Phase IV

Influent flow, Qin (L/d) 2880 ± 140 4320 ± 140 5000 ± 140 5800 ± 140
Average organic loading (kg COD/(m3 d)) 1.55 2.26 4.33 4.12
Average nitrogen loading (kg N/(m3 d)) 0.17 0.26 0.45 0.49
Average phosphorus loading (kg P/m3 d)) 0.018 0.024 0.09 0.051
R–R recirculation ratio (Qr–r/Qin) 15.5 10.3 8.9 7.7
D–R recirculation ratio (Qd–r/Qin) 7.7 5.2 4.4 3.9
D–D recirculation ratio (Qd–d/Qin) 17.3 11.5 10 8.7
EBCT (h)⁄ Anoxic 0.67 0.45 0.39 0.34
Aerobic 2.3 1.4 1.30 1.2
Nominal HRT (h)⁄⁄ Anoxic 0.45 0.31 0.27 0.23
Aerobic 1.58 1.05 0.99 0.79
Avg. attached biomass (mg VSS/g lava rock) Anoxic 15.11 15.51 16.01 16.12
Aerobic 7.55 7.62 7.77 7.85
Biomass (g VSS) Anoxic 1828.3 1876.7 1879.4 1886.4
Aerobic 3201.2 3230.9 3236.4 3243.6
Food/microorganisms ratio (g COD/(g VSSd)) 0.26 0.30 0.46 0.55
Detachment rates (1/d) Anoxic 0.129a 0.149 0.152 0.168
Aerobic 0.081a 0.093 0.092 0.091
Estimated SRT (d) Anoxic 15 12 9 8
Aerobic 24 20 14 12
Overall 39 32 23 20
Run time (d) 44 58 120 33
⁄
EBCT = Vcompact/Q; ⁄⁄Nominal HRT = EBCT  (1  compact bed porosity).
a 0 QX 1
Detachment rates ðb Þ ¼ MX m
.
b
SRTanoxic ¼ SRTTotal Maerobic XMaerobic
anoxic X anoxic
þM anoxic X anoxic .

SRTTotal ¼ Maerobic
c X aerobic þM anoxic X anoxic
Q effluent VSSeff X wastage .

successfully estimated in-situ biofilm kinetics of the circulating example, the unbiodegradable particulate (Fup) and noncolloidal
fluidized bed bioreactors (CFBBR) as presented elsewhere (Chow- slowly biodegradable (Fxsp) were adjusted to 0.1 g COD/g TCOD
dhury et al., in press). The observed maximum specific growth rate and 0.8 g COD/g sbCOD, respectively. Table S1b illustrating the
(lmax) of 3.69 ± 0.44 1/d, biomass true yield (YH) of 0.36 ± 0.03 g simulated characterization compared to the experimental influent
COD/g COD, and endogenous decay coefficient (b) of 0.26 1/d in characterization for calibrated phase (phase III) confirms the valid-
the fluidized bed respirometers reported by this study were used ity of the specified of various organic and nutrient fractions.
in this calibration protocol (Chowdhury et al., 2010). The lmax
was determined using the application of Eq. (2) (Kappeler and Gu- 2.3. Biofilm calibration protocol
jer, 1992) where b is endogenous decay coefficient, while YH was
obtained using Eq. (3) (Orhon et al., 1995), estimated from oxygen Biofilm calibration for particulate biofilm systems is composed
consumption (DO2) vs. SCOD reduction (DSCOD). of five stages as shown in Fig. 2, including definition of objectives,
layout and data collection, data analysis, model running and cali-
OUR
ln ¼ ðlmax  bÞt ð2Þ bration, and sensitivity analysis. It is noteworthy that some of
OURinitial
the biofilm calibration protocol stages proposed in this work have
been used for the ASMs calibration protocols proposed by Petersen
DO 2
YH ¼ 1  ð3Þ et al. (2002), Hulsbeek et al. (2002), Vanrolleghem et al. (2003),
DSCOD Langergraber et al. (2004). The details of the proposed calibration
protocol for three thematic applications are summarized in the fol-
2.2. Biofilm model lowing sections.

BioWinÒ (3.0) developed by Envirosim Associates Ltd. (Burling- 2.3.1. Stage I: Definition of objectives
ton, ON, Canada) was used to apply the proposed calibration proto- Three main objectives are used for this purpose: educational
col for the CFBBR system as an example of particulate biofilm (research), simulation/optimization, and retrofitting purposes as
reactors. shown in Fig. 2. This stage is very important for calibration and de-
The influent characteristics of the municipal wastewater, simu- fines the main theme. For educational and research purposes, the
lated using the influent specifier associated with BioWinÒ, revealed biofilm model is used to understand and design a research plan
the carbonaceous and nutrient fractions according to the biode- for the biofilm lab-scale system. Different scenarios can be evalu-
gradable and nonbiodegradable organic matter in the influent ated using the biofilm model (e.g. study the effect of solids reten-
wastewater. Some of these fractions are expansive and cannot be tion time (SRT), temperature, biofilm kinetics/stoichiometry, and
measured frequently and are calibrated as a function of carbona- mass transfer effect on the BNR performance) in order to establish
ceous and filtered BOD, total suspended solids (TSS), and VSS con- a biofilm system and validate the model predictions.
centrations using nonbiodegradable particulate chemical oxygen For simulation/optimization theme, a biofilm model is devel-
demand (COD) fraction (Fup) with a range of (0–1) along with the oped to simulate an experimental biofilm system data and used
particulate biodegradable fraction of slowly biodegradable COD to optimize and control a biofilm system with dynamic changes
(Fxsp) with a range of (0–1) and its relationships estimated in the of the influent characteristics to simulate shock loadings while
influent wastewater as summarized in Table S1a. In phase III for for the design/retrofitting theme, the biofilm model is used to
 A. Eldyasti et al. / Bioresource Technology 111 (2012) 111–121 115

Fig. 2. Calibration protocol for the particulate biofilm reactors.

study the viability of applying the particulate biofilm approach in a for this purpose can be obtained from the literature along with an
new/existing activated sludge reactors and estimate the bioparticle evaluation step of the biofilm reactor performance. For example,
system requirements i.e. size of reactors, aeration requirement, the influent and effluent characteristics, bioreactor details, and
weight of media, nitrate recycle, and external carbon source. sludge wastage data of a moving bed bioreactor (MBBR) can be used
to establish a research plan of such a system.
2.3.2. Stage II: Layout and data collection In simulation and optimization theme, the biofilm model con-
The layout and data collection is the most important step in a figuration; simulating the existing treatment processes; is cali-
biofilm model. Layout information, as defined in the ‘‘BIOMATH’’ brated using operational data. Generally, a detailed layout of the
calibration protocol for SGR, consists of a general plant layout biofilm system is a crucial step of a simulation study. Identification
and configuration, compartments/process units, volume/area/ of these basic configurations is carried out by gathering informa-
depth, water and sludge lines, installed capacities, pumps, aerators, tion on: operational data, hydrodynamic data, biofilm kinetics
and hydraulic layout (Vanrolleghem et al., 2003). Additionally in a and stoichiometry, and performance data. Operational data is de-
biofilm model, bioparticles and media properties are included in fined as main and recycle flow rate, reactor volume and hydraulic
the main layout information. retention time (HRT). Particulate biofilm attached growth systems
For the educational purpose, the typical layout of the biofilm are dependent on solids holdup and mixing regimes. Thus, infor-
system i.e. influent, unaerated media bioreactor, aerated media bio- mation on hydrodynamic data (i.e. voidage, pressure, fill factor,
reactor, nitrate recirculation, clarifiers, effluent, and sludge wastage SSA, and specific surface volume (SSV)) and axial solid–liquid dis-
effluent, are used to understand the biofilm model. Calibration data tribution is crucial for reactor design and optimization (Nicolella
116 A. Eldyasti et al. / Bioresource Technology 111 (2012) 111–121

et al., 2000). In this calibration protocol, a hydrodynamic profile of properties. In biofilm models, the biofilm thickness is predomi-
biofilm reactor is defined by determination of the pressure gradi- nantly governed by the detachment rate and the kinetics and
ent, using on-line pressure transducers data along the bioparticles stoichiometry do not significantly affect the biofilm thickness.
reactors in order to calculate the fill factor of the bioparticles using Hence, the attachment/detachment approach is applied first fol-
axial void fraction (e), solids hold-up (eS), and solids concentrations lowed by kinetics/stoichiometry and biofilm properties ap-
(M/AL) as shown in Eqs. (4)–(6) (Epstein, 2003). proaches. According to Takács et al. (2007), the turbulence and
the mean velocity gradient (G-value) is not included explicitly
Dps ¼ L  ð1  eÞ  ðqp  qÞ  g ð4Þ
in the attachment and detachment expressions in BioWinÒ bio-
film model. Therefore, the detachment rate coefficient was used
eS ¼ 1  e ð5Þ
initially to fit the experimental biofilm thickness. A higher shear
force is implemented in the aerated biofilm reactor to simulate
M
¼ qp  ð1  eÞ ð6Þ the effect of the diffused aeration and the three phase fluidized
AL
bed bioreactor.
where, L, A, g, DpS, q, and qp are length of the section (m), cross sec- The second approach is to calibrate the kinetic and stoichiome-
tional area (m2), acceleration of gravity (m/s2), additional pressure tric parameters. It is interesting to note that the kinetics and stoichi-
drop (kPa) due to the presence of solids, liquid density (kg/m3), ometric parameters implemented in BioWinÒ are limited for the
and particle density (kg/m3) respectively. suspended growth bioreactor and do not account for the effect of
Biofilm kinetics and stoichiometry information is also a critical the oxygen diffusional limitations inside the biofilm and the reduc-
parameter in biofilm modeling in order to simulate biofilm reactor tion of external mass transfer resistance by fluidization. Therefore,
performance. Recent developments in instrumentation have ren- calibration of the ammonia oxidizing bacteria (AOBs), nitrite oxidiz-
dered respirometric techniques a useful tool to measure biokinetics ing bacteria (NOBs), ordinary heterotrophic organisms (OHOs), and
parameters, evaluate toxicity and inhibition, process optimization, phosphorus-accumulating organisms (PAOs) with the help of respi-
and treatment process design (Ellis et al., 1996; Chu et al., 2003). rometric testing is crucial for the biofilm models.
In design/retrofitting theme, the biofilm model is calibrated The third approach is to use biofilm heterogeneous (layered)
using operational data from an existing plant and subsequently properties, i.e. number of layers through biofilm and mass transfer
used to design a new biofilm systems i.e. unaerated (anoxic) and boundary layer thickness (LL) to calibrate the biofilm performance.
aerated (aerobic) bioreactors. Operational data in this case is de- These proposed calibration approaches are used only for the simu-
fined by main and recycle flow rates, reactor volumes, and design lation/optimization and retrofitting themes since they required
hydraulic retention time (HRT). Information on hydrodynamic data operational data for calibration as opposed to the educational them
and axial solid–liquid distribution are estimated from experimen- which based on literature data.
tal data obtained from literature/pilot-scale biofilm studies. It is
recommended to determine the biofilm kinetics and stoichiometry 2.3.5. Stage V: Sensitivity analysis
parameters using respirometric techniques for maximum specific To verify the predictability of the biofilm model and the effi-
growth rate for heterotrophic biomass (lmax), biomass true yield ciency of the proposed calibration protocol, sensitivity analysis
(YH), Monod half saturation coefficient (KS), and specific lysis (de- of the kinetics, hydrodynamic, and biofilm heterogeneous (lay-
cay) rate constant for heterotrophic (bh) in order to meet the re- ered) parameters was undertaken. Sensitivity analysis ranks mod-
quired effluent characteristics. el parameters based on their contribution to overall error in the
model predictions. Sensitivity analysis of the biofilm parameters
2.3.3. Stage III: Data analysis are evaluated using the normalized sensitivity coefficient (Si,j)
Quality assessment of the collected data includes two steps: and the mean square sensitivity measure (djmsqr). The normalized
data evaluation and mass balance. Evaluation of the collected data sensitivity coefficient as shown in Eq. (7) is defined by US EPA as
is used only for simulation and retrofitting purposes. In this step, a ratio of the percentage change in the output variable (yi) to a
the information collected is important to understand the capacity change in the input variable (xi) (US EPA, 2009). For each param-
and behavior of the biofilm system. Therefore, the design, opera- eter, the default values were varied by ±50% to provide upper and
tional, and experimental data should be evaluated and processed lower limits in a sensitivity analysis to assess its impact on the
for understanding of the entire process. Outlier detection and output parameters. The influence of each parameter on the model
interpolation of the data can be used to improve the available data output is defined using the method proposed by Petersen et al.
(Vanrolleghem et al., 2003). Different statistical methods such as t- (2003) into the following 4 categories: (a) no significant influence
tests (comparing the means of different data sets) and f-tests (com- (NI: Si,j h 0.25), (b) influential (I: 0.25 6 Si,j h 1), (c) very influen-
paring the differences between standard deviations of data sets) tial (VI: 1 6 Si,j h 2), and (d) extremely influential (EI: Si,j h 2)
can be also used to estimate the uncertainty in the collected data. (Petersen et al., 2003).
Mass balances of the flow, organic matter, nutrient, phosphorus,
 
and sludge are useful to maintain an acceptable quality of the col- DY i =Y i 
Si;j ¼   ð7Þ
lected data. This stage is recommended to maintain a high quality DX i =X i 
of the verification and validation.
The mean square sensitivity measure (djmsqr) is defined by Brun
et al. (2002) as shown in Eq. (8). This sensitivity measure is de-
2.3.4. Stage IV: Model calibration
signed to assess the individual parameter importance in a least
In this stage, steady-state biofilm model implemented in Bio-
squares parameter estimation context and a high value of djmsqr
WinÒ software is used to fit the experimental data and calibrated
indicates that a parameter has an important influence on the sim-
using three sequential approaches: attachment/detachment ap-
ulation results, whereas the value of zero means that the simula-
proach, kinetics and stoichiometry approach, and biofilm proper-
tion results do not depend on a parameter.
ties approach (such as number of layers in the anoxic/aerobic
zone and mass transfer boundary layers). It interesting to note vffiffiffiffiffiffiffiffiffiffiffiffiffiffi
u
that applying the aforementioned approaches sequentially is very u1 X n
dmsqr ¼t  S2 ð8Þ
important. The biofilm attachment/detachment step has to be ap- j
n i¼1 i;j
plied first before changing the kinetics/stoichiometry and biofilm
 A. Eldyasti et al. / Bioresource Technology 111 (2012) 111–121 117

3. Results and discussion Hydrodynamic configuration and axial solid–liquid distribution

data of the CFBBR system was obtained from the online pressure
In order to evaluate the proposed calibration protocol, a case measurement and calculated using the aforementioned method
study of a pilot-scale CFBBR system for biological treatment of mu- and Eqs. (4)–(6). Lava rock particles with an average diameter of
nicipal wastewater, at empty bed contact times (EBCTs) of 1.5– 600 lm were used as the carrier media (bioparticles) for biofilm
3.0 h and volumetric nutrients loading rates of 1.43–4.29 kg COD/ attachment in the CFBBR. The maximum possible surface area
(m3 d), 0.17–0.48 kg N/(m3 d), and 0.021–0.093 kg P/(m3 d), was (SSAmax) in the anoxic and aerobic bioreactors was calculated con-
used for the biofilm model implemented in BioWinÒ. The results sidering zero void ratio and biofilm thickness of 500 and 120 lm,
determined by applying different biofilm model calibration stages and a bare lava rock particles of diameter 600 lm as 3750 and
in CFBBR’s case study, outlined in Fig. 2, are described in the fol- 7060 m2/m3, respectively. Considering variable bed porosity, sur-
lowing sections. face area, for the anoxic and aerobic reactors, of 2100–3113 m2/
m3 and 3320–6990 m2/m3 were used translating to a total surface
area (TSA) of the carrier media for the entire anoxic and aerobic
3.1. Definition of objectives, layout, and data collection (modeling reactors considering the compact bed was 170 and 1070 m2,
stages I and II) respectively as calculated using Eq. (1).
Since the thin aerobic biofilm thickness of the CFBBR of 100–
The biofilm model of the pilot-scale CFBBR system was used for 190 lm is in the same range of activated sludge ‘‘flocs’’ (70–
simulation of BNR performance and validated the second theme of 125 lm), kinetic coefficients derived from conventional suspended
the proposed calibration protocol (Fig. 2). Four different organic growth respirometry are applicable (Andreadakis, 1993; Jorand
and nutrient loading rates were used. The CFBBR was modeled using et al., 1995). However due to the much thicker biofilm in the anoxic
basic reactors available in BioWinÒ, i.e. influent, unaerated media riser of 500–600 lm, the biofilm kinetics implemented were the
bioreactor, aerated media bioreactor, nitrate recirculation, clarifiers, ones derived from fluidized bed respirometric studies (Chowdhury
effluent, and sludge wastage effluent as shown in Fig. 1d. The riser et al., in press) of an observed maximum specific growth rate (lmax)
was simulated using two unaerated media bioreactors followed of 3.69 ± 0.44 1/d and biomass true yield (YH) of 0.36 ± 0.03 g COD/g
by three aerated media bioreactors as a downer and a solid–liquid COD.
separator to collect the excess biomass from the system. The CFBBR
system was implemented in the aforementioned layout to simulate 3.2. Data analysis (modeling stage III)
the change in the bioparticles distribution from the dense to the di-
lute phase (Fig. 1c) in the same bioreactor and calculated using Eqs. Data collected from the pilot scale CFBBR system operating for
(4)–(6) of the experimental data collected along the reactors. It is more than 255 days was evaluated using the data comparison
noteworthy that the pressure drop measured experimentally in techniques i.e. t-tests (comparing the means of different datasets)
the middle of the riser and at three location in the downer i.e. bot- and f-tests (comparing the differences between standard devia-
tom, middle, and top of fluidized bed, were used to calculate voidage tions of datasets). The paired student t-test and f-test were con-
(e), media fill (eS) in accordance with Eqs. (4)–(6). ducted to determine the statistical significance of the observed
The influent enters the riser with a downer–riser liquid and ni- differences between the experimental data for TCOD, SCOD, TBOD,
trate recirculation collected from the last downer of aerated reac- SBOD, TN, NH4–N, NO3–N, NO2–N, TP, PO4–P, TSS, and VSS at the
tor. The combined fluid flows from riser to the downer. Finally, 95% confidence level.
the effluent from the downer goes to the downer solid–liquid sep- Mass balances for COD, nitrogen, phosphorus, and alkalinity
arator, shown as a clarifier, with the provision for sludge wastage. were evaluated to validate the mass balance around the CFBBR sys-
The cross sectional area of anoxic and aerobic reactors was consid- tem including the anoxic and aerobic bioreactors as shown in Table
ered equal to the actual cross sectional area of the column in the S2. Approximately 40% of the influent COD was utilized in the an-
pilot-scale. In order to simulate the fluidization regime of CFBBR oxic column by denitrification while about 45% of the influent COD
system and the change in biofilm thickness with the change in was oxidized in the aerobic column. Anoxic COD consumption was
the voidage ratio and the filling factor (Fig. 1c), the shear factor 328, 508, 921, and 974 g COD/d in phases I, II, III, and IV respec-
was calibrated separately in each reactor with respect to expanded tively. COD consumption for denitrification (3.5–3.7 mg COD/mg
fluidized bed by a detachment rate coefficient in BioWinÒ model. It NO3–N) was estimated using Eq. (9) (Metcalf and Eddy, 2003), con-
is interesting to note that the properties (i.e. roughness, porosity, sidering the observed biomass yield of 0.12–0.16 g VSS/g COD.
and chemical adsorption) and the weight of carrier media in Bio-
WinÒ model are not explicitly defined but implicitly as SSA and 2:86
COD consumption for denitrification ¼ ð9Þ
% fill of the bioparticles. ð1  1:42  YÞ
Table 1a illustrates the municipal wastewater characteristics for
a duration of 255 days, characterized predominantly by a carbon to COD (%) closure was calculated using influent and effluent COD
nitrogen ratio of 8:1, TCOD/VSS ratio of 2:1 and TBOD/TCOD of concentrations, and COD in the biomass wastage from the CFBBR
0.65. Table 1b shows the detailed operational conditions and de- system with an average percentage closure of 92%. Mass balance
sign parameters of the CFBBR. At this stage the experimental data of the nitrogen uptake in the CFBBR system shows a higher per-
of phase III (highlighted) was used to calibrate the biofilm model centage closure of 98%. Phosphorus balance closed at an average
while the other phases were used for validation. The aforemen- of 92%. Table S2 shows that 95–263 g NO3–N/d were removed in
tioned influent characteristics of the municipal wastewater, simu- the anoxic column, which generate 339–939 g alkalinity as
lated using the influent specifier associated with BioWinÒ, revealed CaCO3/d. In the aerobic column, 106–280 g NH4–N/d were nitrified
the carbonaceous and nutrient fractions according to the biode- and utilized 757–2071 g alkalinity as CaCO3/d. Therefore, alkalinity
gradable and unbiodegradable organic matter in the influent mass balance closures were about 88% in all four phases.
wastewater as shown in Table S1b. Although the influent specifier
associated with BioWinÒ was able to simulate most of the influent 3.3. Model calibration (modeling stage IV)
characteristics, TSS, VSS, and SBOD were always off from the exper-
imental data, revealing the limitations of the influent specifier A benchmark model was running with the collected data from
associated with the model. the previous steps to fit the steady state CFBBR performance during
118 A. Eldyasti et al. / Bioresource Technology 111 (2012) 111–121

255 days of biological nutrient removal from municipal wastewa- The third step was to calibrate the biofilm properties including
ter. At this stage, three sequential steps were used to calibrate the total thickness of mass transfer boundary layers (MTBL) and
the BioWinÒ model starting by adjusting the biofilm thickness fol- the number of biofilm layers. In the anoxic reactor, which has a
lowed by adjustment of the heterotrophic/autotrophic kinetic biofilm thickness of 500–700 lm, the number of biofilm layers
parameters and biofilm properties including the total thickness was set to 3 layers with a total MTBL of 600 lm. In the aerobic
of mass transfer boundary layers (MTBL) and the number of biofilm reactor, which has oxygen diffusion throughout the biofilm, the
layers. number of biofilm layers was set to 2 layers with a total MTBL of
First, the biofilm thickness was adjusted by attachment/detach- 400 lm. It is noteworthy that the boundary layer thickness is cal-
ment approach using the detachment rate (g/m3 d) associated with ibrated only by trial and error in order to fit the experimental efflu-
BioWinÒ followed by kinetics/stoichiometry and biofilm properties ent quality.
approaches. Two different scenarios have been applied in the first Two calibration methods were tested: one where the aforemen-
approach, according to the fluidization regime in liquid/solid and tioned sequence of calibrating biofilm thickness followed by kinet-
gas/liquid/solid fluidization regimes, defined as two and three ics and stoichiometry and then biofilm properties denoted
phase fluidization regimes, respectively. In the anoxic bioreactor, henceforth as method 1, and the other where kinetics and stoichi-
liquid/solid fluidization regime has been applied with a constant ometry were adjusted first followed by biofilm thickness and lastly
detachment rate of 1.4  105–1.5  105 throughout the entire bio- biofilm properties referred to as method 2. It is interesting to note
reactor and different filling factors while a three phase fluidization that applying the attachment/detachment approach first main-
regime applied in the aerobic bioreactor. In the aerobic bioreactor, tained the biofilm thickness within a 5% of the experimental bio-
a higher detachment rate of 1.7  106 was applied at the bottom of film thicknesses of 600 and 190 lm in the anoxic and aerobic
the tank and a smaller detachment rate of 1  106 at the top of the reactors, respectively after applying the second approach. As illus-
reactor representing the direct effects of air diffusers and air bub- trated in Table 2, using method 1, the effluent characteristics and
bles as shown in the bioparticles distribution in Fig. 1c. It is note- the measured biofilm thickness were modeled in 12 iterations
worthy that the biofilm thickness in the aerobic bioreactor is while applying the method 2 calibration took 20 iterations. It is
significantly smaller than the anoxic bioreactor which represents noteworthy that with the current biofilm process configuration,
a higher effect of bioparticles abrasion in the aerobic zones trans- each run lasted for a minimum of 1 h using traditional laptop or
lates to a higher detachment rate by one order of magnitude rela- desktop computers, with some of the runs lasting for more than
tion to the anoxic zone. A significant reduction of the detachment 3 h. Thus, the computational effort using the proposed approach
rate (about 40%) was applied to the third reactor, which represents (method 1) is significantly lower than method 2.
the upper section of the aerobic fluidized bed bioreactor. In other Scrutiny of the data for method 1 presented in Table 2 clearly
biofilm reactors, i.e. moving bed bioreactor (MBBR) with a lower indicates that fitting the biofilm thicknesses i.e. runs 1–4 already
fill factor, the detachment rate can be reduced significantly. Apply- modeled the effluent COD, SCOD, and ammonia within the range
ing this approach in other biofilm simulation packages can be done of observed average experimental values, plus or minus the stan-
using other parameters, e.g. shear factor (0–5) in AQUIFASÒ; dard deviation. From runs 5 to 12, the adjustment of the kinetic
detachment rate (kg/m2 d) and the internal solids exchange rate parameters mainly refined effluent nitrates, TKN, and consequently
(m/d) for GPS-XÒ. TN concentrations. However in method 2, initial calibration (runs
After the calibration of biofilm thickness, adjusting the kinetics 1–10), by fitting all water quality parameters, erroneously overpre-
and stoichiometry parameters has been used to fit the steady state dicted biofilm thicknesses. Subsequent attempts to fit biofilm
CFBBR performance. It is interesting to note that the biofilm kinetic thickness (runs 11–13) completely threw off the effluent nitrate
parameters of ammonia oxidizing bacteria (AOBs) and nitrite oxi- predictions, necessitating another round of kinetic parameters
dizing bacteria (NOBs) in a very thin biofilm are similar to the adjustment (runs 14–19), before final fitting using biofilm proper-
existing ASM values and can be used as a guide for biofilm kinetics ties i.e. aerobic mass transfer layer thickness.
modification for the AOBs and NOBs. Therefore, conducting a respi- The steady-state CFBBR model using BioWinÒ focused on vari-
rometry study is only important for the ordinary heterotrophic ous aspects of process performance i.e. reactor effluent characteris-
organisms (OHOs) when the biofilm thickness is thick and sub- tics, nutrient removal rates, biofilm thickness, total biomass in the
strate utilization for energy in preference to biomass assimilation reactor, and process yield as well as the COD uptake, nitrification,
under substrate limited conditions. and denitrification rates. Table 3a shows a comparison between
The kinetics of the ordinary heterotrophic organisms (OHOs) model prediction and experimental data for all phases. The model
have been measured using a new respirometry technique (devel- was calibrated using the experimental data of phase III and fol-
oped by Nakhla and coworkers (Chowdhury et al., in press)) emu- lowed by validation using the other phases. In phase III, the model
lating the hydrodynamic conditions in the CFBBR. The observed predicted effluent NH4–N of 0.9 mg/L, NO3–N of 5.4 mg/L, and TKN
maximum specific growth rate (lmax) of 3.69 ± 0.44 1/d, biomass of 3.8 mg/L compared well to observed NH4–N of 0.9 ± 0.6 mg/L,
true yield (YH) of 0.36 ± 0.03 g COD/g COD, and endogenous decay NO3–N of 5.4 ± 1.3 mg/L, and TKN of 2.3 ± 0.6 mg/L, in the pilot-
coefficient (b) of 0.26 1/d in the fluidized bed respirometers were scale CFBBR system. The BioWinÒ model predicted the effluent
measured using Eqs. (2) and (3). The kinetic parameters of other NH4–N, NO3–N, and TKN of the other phases in agreement with
types of bacteria such as ammonia oxidizing bacteria (AOBs) and the experimental data. As illustrated in Table 3a, the average
phosphorus-accumulating organisms (PAOs) which were not mea- percentage error (APE) in phase I, calculated as the summation of
sured experimentally have been modified by ±30% of the default the absolute difference between the experimental and predicted
model as shown in Table S3a. It is interesting to note that the sub- values divided by the experimental values, averaged over the num-
strate half saturation concentration of nitrite oxidizing bacteria ber of data points, revealed that the discrepancy between predicted
(NOBs) in BioWinÒ was 24 times smaller than the calibrated value. and measured final effluent TCOD, SCOD, PO4–P, TP, and SBOD was
A comparison between the calibrated and literature values of the 15%. Comparatively, a higher APE of 30% was observed between
selected kinetics parameters shows that all the adjusted values simulated and measured final effluent TCOD, SCOD, PO4–P, TP,
were within a range of ±30% except for the nitrite oxidizing bacte- and SBOD in phases I, II, and IV. In phase II, the BioWinÒ model over-
ria (NOBs) of substrate half saturation coefficient as shown in Table predicted SCOD, TKN, and PO4–P by 20% while the other final efflu-
S3b. ent characteristics were in agreement with the experimental data.
 A. Eldyasti et al. / Bioresource Technology 111 (2012) 111–121 119

Table 2
Impact of biofilm thickness, kinetics, and biofilm properties approaches on effluent characteristics and biofilm thickness.

Trial Iteration Parameter (mg/L) Biofilm thickness

(lm)
COD SCOD NH4–N NO3–N TKN TN Anoxic Aerobic
Method I: Appling the biofilm thickness approach followed by kinetic and biofilm properties approaches
1 Default 36⁄ 25 0.6 11.2 3.9 15.7 870 910
2 DRanx: 1E + 5, DRaer: 1E + 5 37 26 0.5 10.4 3.9 14.8 750 790
3 DRanx: 1.2E + 5, DRaer: 1.4E + 6 49 34 0.5 19.2 3.9 23.6 690 210
4 DRanx: 1.4–1.5E + 5, DRaer: 1.0–1.7E + 6 49 34 0.6 18.9 3.9 23.4 610 190
5 DR done + (OHO: lmax and Y of 3.69 and 0.26) 46 30 0.7 18.0 3.5 22.2 610 190
6 DR done + (OHO: lmax and Y) + (AOB: KS 0.8) 46 30 0.7 17.9 3.6 22.3 610 190
7 DR done + (OHO: lmax and Y) + (AOB: KS 0.9) 46 30 0.7 17.0 3.6 21.3 610 190
8 DR done + (OHO: lmax and Y) + (AOB: KS 1) 46 30 0.7 15.1 3.6 19.5 600 190
9 DR done + (OHO: lmax and Y) + (AOB: KS) + (NOB: KS 1) 46 30 0.7 13.5 3.6 17.9 600 190
10 DR done + (OHO: lmax and Y) + (AOB: KS) + (NOB: KS 1.5) 46 30 0.7 11.8 3.6 16.2 590 190
11 DR done + (OHO: lmax and Y) + (AOB: KS) + (NOB: KS 2) 46 30 0.7 10.4 3.6 14.7 580 190
12 DR done + (OHO: lmax and Y) + (AOB: KS) + (NOB: KS 2.4) + 2 layers of biofilmaerobic 46 30 0.9 5.4 3.8 10.1 580 190
Method II: Appling the kinetic approach followed by biofilm thickness and properties approaches
1 Default 36 25 0.6 11.2 3.9 15.7 870 910
2 OHO: lmax and Y of 3.69 and 0.26) 35 22 0.5 10.7 3.3 14.4 870 950
3 (OHO: lmax and Y) + (AOB: KS 1) 35 22 0.4 10.7 3.2 14.3 900 950
4 (OHO: lmax and Y) + (AOB: KS 1.5) 35 22 0.2 11.1 3.0 14.4 910 950
5 (OHO: lmax and Y) + (AOB: KS 0.7) + (NOB: KS 1) 35 22 0.5 9.3 3.2 13.0 900 950
6 (OHO: lmax and Y) + (NOB: KS 2) 35 22 0.5 8.2 3.2 11.9 870 950
7 (OHO: lmax and Y) + (NOB: KS 2.4) 35 22 0.5 7.8 3.2 11.5 870 950
8 (OHO: lmax and Y) + (NOB: KS 3) 35 22 0.5 7.0 3.3 10.7 870 950
9 (OHO: lmax and Y) + (NOB: KS 3.5) 35 22 0.5 6.7 3.2 10.4 870 950
10 (OHO: lmax and Y) + (NOB: KS 3.9) 35 22 0.5 6.7 3.2 10.4 870 950
11 Kint done + DRanx: 1E + 5, DRaer: 1E + 5 35 23 0.4 6.5 3.2 10.1 760 820
12 Kint done + DRanx: 1.4E + 5, DRaer: 1E + 6 46 31 0.4 10.6 3.2 14.3 620 240
13 Kint done + DRanx: 1.4E + 5, DRaer: 1.0–1.7E + 6 46 30 0.5 2.2 3.4 6.1 580 190
14 DR done + (OHO: lmax and Y) + (NOB: KS 3) 46 30 0.5 8.3 3.4 12.2 590 190
15 DR done + (OHO: lmax and Y) + (NOB: KS 3) + (AOB: KS 1) 46 30 0.7 7.9 3.6 12.3 590 190
16 DR done + (OHO: lmax and Y) + (NOB: KS 2.4) + (AOB: KS 1) 46 30 0.7 10.4 3.6 14.7 600 190
17 DR done + (OHO: lmax and Y) + (NOB: KS 3.5) + (AOB: KS 1) 46 30 0.7 5.1 3.6 9.4 580 190
18 DR done + (OHO: lmax and Y) + (NOB: KS 3.5) + (AOB: KS 1) + 2 layers of biofilmaerobic 46 30 0.9 0.6 3.9 5.4 560 190
19 DR done + (OHO: lmax and Y) + (NOB: KS 3) + (AOB: KS 1) + 2 layers of biofilmaerobic 46 30 0.9 1.5 3.8 6.2 560 190
20 DR done + (OHO: lmax and Y) + (NOB: KS 2.4) + (AOB: KS 1) + 2 layers of biofilmaerobic 46 30 0.9 5.4 3.8 10.1 580 190
⁄
Shaded parameters are the problematic ones that were outside the average ± SD range.

Table 3a
Experimental and simulated effluent quality.

Parameter Phase I Phase II Phase III Phase IV

Simulated Exp.⁄ Simulated Exp.⁄ Simulated Exp.⁄ Simulated Exp.⁄
pH 7 6.9–7.3 7 6.9–7.2 7 7.0–7.4 7 6.8–7.2
Alkalinity⁄⁄ 113 89 ± 23 124 116 ± 12 103 75 ± 38 100 79 ± 24
COD (mg/L) 22 26 ± 3 27 39 ± 8 46 41 ± 14 54 45 ± 7
SCOD (mg/L) 16 13 ± 4 18 15 ± 4 30 20 ± 8 39 23 ± 5
NH4–N (mg/L) 0.4 0.6 ± 0.5 0.6 0.9 ± 0.3 0.9 0.9 ± 0.6 3 3.9 ± 0.9
NO3–N (mg/L) 4 3.6 ± 1.2 4.8 4.7 ± 1.3 5.4 5.4 ± 1.3 3 2.8 ± 0.6
TKN (mg/L) 2.3 2.5 ± 0.4 2.7 3 ± 0.6 3.8 2.3 ± 0.6 6 6.7 ± 1.2
TN (mg/L) 6.6 6.2 ± 1.1 8.3 7.6 ± 1.3 10.1 7.7 ± 0.8 10 9.8 ± 1.2
PO4–P (mg/L) 0.9 0.7 ± 0.1 0.6 0.5 ± 0.1 0.9 0.7 ± 0.4 0.8 0.6 ± 0.2
TP (mg/L) 1.2 1.0 ± 0.1 0.8 1.2 ± 0.2 1.4 1.3 ± 0.2 1.1 1.2 ± 0.4
TSS (mg/L) 7 11 ± 2 8 22 ± 6 15 41 ± 20 14 27 ± 6
VSS (mg/L) 5 9±2 5 16 ± 5 10 22 ± 11 9 21 ± 6
BOD (mg/L) 6 16 ± 5 7 19 ± 3 15 21 ± 8 16 23 ± 3
SBOD (mg/L) 3 3±1 4 4±2 8 7±2 10 7±2
⁄ ⁄⁄
Average ± SD (number of samples, 20) with a frequency of a sample every 3 days; (mg CaCO3/L).

It is interesting to note that the model overpredicted the final by APE of 30–60% due to underprediction of effluent TSS and VSS
effluent alkalinity in all cases while it predicted the nitrogen com- concentrations.
ponents by an APE of 20%. Furthermore, the effluent solids as TSS As illustrated in Table 3b, anoxic COD removal in different
and VSS were underpredicted in all phases within an APE of 50%, phases were close to the experimental data with an APE of 3–5%
reflecting the limitation of the BioWinÒ model to predict the liquid whereas the aerobic COD removal were overpredicted with an
solids separation of the CFBBR system efficiently. Moreover, the APE of 5–14%. Nitrification and denitrification rates of 0.095–
model significantly underpredicted the effluent BOD in all phases 0.27 kg N/d and 0.084–0.25 kg N/d, respectively, predicted by
120 A. Eldyasti et al. / Bioresource Technology 111 (2012) 111–121

Table 3b
Simulated results and measured parameters for nutrient removal rates.

Parameter Phase I Phase II Phase I Phase II

⁄ ⁄ ⁄
Sim. Exp. Sim. Exp. Sim. Exp. Sim. Exp.⁄
Anoxic COD consumption (kg/d) 0.32 0.33 ± 0.05 0.50 0.51 ± 0.06 0.88 0.92 ± 0.1 0.92 0.97 ± 0.1
Aerobic COD consumption (kg/d) 0.44 0.42 ± 0.04 0.60 0.55 ± 0.1 1.0 0.80 ± 0.15 1.0 0.87 ± 0.18
Yield (g VSS/g COD) 0.18 0.12 ± 0.04 0.19 0.13 ± 0.04 0.2 0.13 ± 0.04 0.22 0.16 ± 0.04
Aerobic N removal (kg/d) 0.095 0.11 ± 0.01 0.14 0.16 ± 0.01 0.28 0.30 ± 0.01 0.27 0.28 ± 0.01
Anoixc N removal (kg/d) 0.084 0.095 ± 0.01 0.13 0.15 ± 0.01 0.23 0.26 ± 0.01 0.25 0.26 ± 0.01
⁄
Average ± SD (number of samples, 20) with a frequency of a sample every 3 days.

BioWinÒ were comparable with the observed nitrification and very influential (VI: 1 6 Si,j h 2) for effluent characteristics of TCOD,
denitrification rates, estimated from the amount of nitrogen nitri- SCOD, SPO4–P, and TBOD while the other effluent characteristics
fied and denitrified within APE of 1–5%. were extremely influenced (EI) by the changes of the OHO yield.
Biomass yield in the pilot-scale CFBBR calculated as the sum of Moreover, other kinetics parameters of lmax-OHO, KOHO, and bOHO
the net change in attached biomass, sludge wastage, and effluent were influential (I) for all effluent characteristics except the NO3
solids divided by the total COD consumed in the process was was significantly impacted (VI) by ±50% change in lmax-OHO.
0.12, 0.13, 0.13, and 0.16 g VSS/g COD in phases I, II, III, and IV, Changing the number of anoxic and aerobic biofilm layers (LF)
respectively with overall sludge production ranging from 109 to from 1 to 4 layers, and anoxic and aerobic mass transfer boundary
439 g VSS/d according to the solids in influent and produced from layers thickness (LLanoxic, LLaerobic) which were varying from 100 to
the biomass. BioWinÒ predicted that 15–52 g VSS/d biomass were 500 lm, very significantly affected effluent NO3 concentrations
lost in the effluent of CFBBR system with an overall sludge wastage (category IV). All other effluent parameters impacted by biological
ranging from 175 to 730 g VSS/d in phases I–IV. Considering the nitrogen removal i.e. ammonia, TKN, and TN, as reported in Table
aerobic and anoxic nutrient mass removal rates, the mean cell res- S4a, were influenced by the biofilm properties. The mean square
idence time, decay coefficient, and the simulated COD removal sensitivity measures (djmsqr) were used to rank all the parameters
ranging from 760 to 1920 g COD/d in phases I to IV, the simulated as illustrated in Table S4b. The most important kinetic and stoichi-
biomass yield with BioWinÒ were calculated as 0.18, 0.19, 0.2, and ometric parameters were lmax-NOB followed by bNOB and K NO2 . For
0.22 g VSS/g COD in phases I, II, III, and IV, respectively which are the biofilm parameters, the aerobic mass transfer boundary layer
approximately 50% higher than those observed experimentally. (MTBLAerobic) was the most important parameter followed by fol-
Although the predicted aerobic and anoxic attached biomass lowed by the number of anoxic biofilm layers. Although the Mass
thicknesses of 190 and 580 lm respectively were in close agree- transfer boundary layer (MTBLAerobic) was the most important bio-
ment with the experimental values of 150 and 600 lm in the aer- film parameter but it was the second one in the overall ranking as
obic and anoxic reactors, the total biomass in both models was shown in Table S4b. It is interesting to note that AOB and NOB yield
underpredicted by 20%. The total model predicted biomass in the coefficients had a minimal influence among all other parameters.
anoxic and aerobic reactors ranging from 1530 to 1570 g VSS and Overall evaluation of the model parameters highlights the areas
2576 to 2590 g VSS, compared to the measured biomass which var- that the model is sensitive for max. spec. growth rate (lmax-NOB),
ied from 1828 to 1886 g VSS and 3200 to 3243 g VSS in phases I–IV, mass transfer boundary layer thickness (MTBLAerobic), aerobic de-
respectively. cay rate (bNOB), substrate (NO2) half sat. (K NO2 ), yield (YOHO), num-
ber of anoxic layer, max. spec. growth rate (lmax-AOB), and number
of aerobic layer in addition to the detachment rate.
3.4. Sensitivity analysis (modeling stage V)

In order to verify the predictability of the calibrated biofilm

model using the proposed protocol, sensitivity analysis comprised 4. Conclusions
19 biofilm kinetic and stoichiometric parameters and four of the
biofilm properties parameters were evaluated using the normal- Although the existing calibration protocols of ASM can be ap-
ized sensitivity coefficient (Si,j) and the mean square sensitivity plied partially for the attached growth system, the proposed cali-
measure (djmsqr). Kinetic and stoichiometric parameters associated bration protocol sets a complete strategy to model particulate
with growth and decay of AOB, NOB, OHO, and PAOs were sub- biofilm reactors and proposes a method to collect the data and
jected to sensitivity analysis within a change of ±50% of the cali- translate it to useful information. The detailed calibration proce-
brated values. In this study, all the kinetic and stoichiometric dures presented here will not only help the process engineers de-
parameters (7 parameters) of PAOs had no significant influence sign and retrofit plants but also plan sampling and monitoring
(NI: Si,j h 0.25) and the total number of 12 kinetic and stoichiome- requirements for process optimization. Sensitivity analysis is a
tric parameters were presented in Tables S4a and S4b. very helpful tool to identify the most important parameters of bio-
As illustrated in Table S4a, the value of normalized sensitivity film model and guide experimental measurements. The application
coefficient (Si,j) were calculated with regard to 12 effluent charac- of Si,j and djmsqr rank the kinetics, stoichiometric, and biofilm prop-
teristics. However, for one of them, i.e. TSS, the values of Si,j were al- erties parameters in BioWinÒ. Supplementary materials Carbona-
ways under 0.25 and thus TSS was excluded from Table S4a. For ceous and nutrient fraction estimated for wastewater and
AOB bacteria, the biofilm model was extremely influential (EI: Si,j assumed for municipal wastewater in BioWinÒ are summaries in
h 2) with the change of maximum specific growth rate (lmax-AOB) Table S1a. Table S1b illustrating the simulated characterization
for both nitrification and denitrification processes while it was very compared to the experimental influent characterization for cali-
influential (EI) for the other kinetic parameters of AOB. As shown in brated phase (phase III). Overall mass balances of pilot-scale of cir-
Table S4a, the NOB was extremely influential (EI) with the change of culating fluidized bed bioreactor are shown in Table S2. Calibrated
lmax-AOB, K NO2 , and bNO2 . In case of OHO bacteria, the yield coeffi- kinetics parameters have been used in BioWinÒ compared to
cient (YOHO) was influential (I: 0.25 6 Si,j h 1) for effluent VSS and literature data are summaries in Table S3a and b. Table S4a and
 A. Eldyasti et al. / Bioresource Technology 111 (2012) 111–121 121

b summaries the sensitivity coefficient and parameters of effluent Hulsbeek, J.J.W., Kruit, J., Roeleveld, P.J., van Loosdrecht, M.C.M., 2002. A practical
protocol for dynamic modelling of activated sludge systems. Water Science and
characteristics used in biofilm model.
Technology 45 (6), 127–136.
Jorand, F., Zartarian, F., Thomas, F., Block, J., Bottero, J., Villemin, G., Urbain, V.,
Acknowledgements Manem, J., 1995. Chemical and structural (2D) linkage between bacteria within
activated sludge flocs. Water Research 29 (7), 1639–1647.
Kappeler, J., Gujer, W., 1992. Estimation of kinetics parameters of heterotrophic
The authors gratefully acknowledge Trojan Technologies, Can- biomass under aerobic conditions and characterization of wastewater for
ada, Natural Science and Engineering Research Council of Canada activated sludge modeling. Water Science and Technology 25, 125–140.
(NSERC), Ontario Center of Excellence (OCE), Canada, and City of Langergraber, G., Rieger, L., Winkler, S., Alex, J., Wiese, J., Owerdieck, C., Ahnert, M.,
Simon, J., Maurer, M., 2004. A guideline for simulation studies of wastewater
London, ON, Canada for their endless support and interest at every treatment plants. Water Science and Technology 50 (7), 131–138.
stage of this research project. Metcalf and Eddy, 2003. Wastewater Engineering: Treatment and reuse. 4th ed.,
McGraw-Hill.
Nakhla, G., Zhu, J., Cui, Y. 2004. Liquid–solid circulating fluidized bed wastewater
Appendix A. Supplementary data treatment system for simultaneous removal of carbon, nitrogen, and
phosphorus, US Patent No. 7261,811, Int’l PCT patent awarded (2005).
Nicolella, C., van-Loosdrecht, M.C.M., Heijnen, J.J., 2000. Wastewater treatment with
Supplementary data associated with this article can be found, in
particulate biofilm reactors. Journal of Biotechnology 80, 1–33.
the online version, at doi:10.1016/j.biortech.2012.02.021. Orhon, D., Yildiz, G., Cokgör, E.U., Sözen, S., 1995. Respirometric evaluation of the
biodegradation of confectionary wastewaters. Water Science and Technology
References 32, 11–19.
Petersen, B., Gernaey, K., Henze, M., Vanrolleghem, P.A., 2002. Evaluation of an
ASM1 model calibration procedure on a municipal–industrial wastewater
Andreadakis, A.D., 1993. Physical and chemical properties of activated sludge floc. treatment plant. Journal of Hydroinformatics 4 (1), 15–38.
Water Research 27 (12), 1707–1714. Petersen, B., Gernaey, K., Henze, M., Vanrolleghem, P.A., 2003. Calibration of
Boltz, J.P., Morgenroth, E., Sen, D., 2010. Mathematical modelling of biofilms and activated sludge models: a critical review of experimental designs. In: Agathos,
biofilm reactors for engineering design. Water Science and Technology 62 (8), S.N., Reineke, W. (Eds.), Biotechnology for the Environment: Wastewater
1821–1836. Treatment and Modelling. Waste Gas Handling. Kluwer Academic Publishers,
Brun, R., Kuhni, M., Siegrist, H., Gujer, W., Reichert, P., 2002. Practical identifiability Dordrecht.
of ASM2d parameters – systematic selection and tuning of parameters subsets. Takács, I., Bye, C.M., Chapman, K., Dold, P.L., Fairlamb, P.M., Jones, R.M., 2007. A
Water Research 36 (16), 4113–4127. biofilm model for engineering design. Water Science Technology 55 (8–9), 329–
Chowdhury, N., Nakhla, G., Zhu, J., 2008. Load maximization of a liquid–solid 336.
circulating fluidized bed bioreactor for nitrogen removal from synthetic US EPA, 2009. Guidance on the Development, Evaluation and Application on
municipal wastewater. Chemosphere 71, 807–815. Environmental Models EPA 100/K-09-003. US EPA, Washington, DC, USA.
Chowdhury, N., Nakhla, G., Zhu, J., in press. A novel fluidized bed respirometric Vanrolleghem, P.A., Insel, G., Petersen, B., Sin, G., De Pauw, D., Nopens, I., Weijers, S.,
technique for determination of biofilm kinetics. Environmental Technology. Gernaey, K., 2003. A comprehensive model calibration procedure for activated
Chowdhury, N., Nakhla, G., Zhu, J., Islam, M., 2010. Pilot-scale experience with sludge models. In: Proceedings of WEFTEC 2003, 76th Annual Technical
biological nutrient and biomass yield reduction in a liquid–solid circulating Exhibition and Conference, 11–15 October 2003, Los Angeles, CA, USA.
fluidized bed bioreactor. Water Environmental Research 82 (9), 772–781. Wanner, O., 1986. A multispecies biofilm model. Biotechnology and Bioengineering
Chu, K.H., van-Veldhuizen, H.M., van-Loosdrecht, M.C.M., 2003. Respirometric 28, 314–328.
measurement of kinetics parameters: effect of active sludge floc size. Water Wanner, O., Eberl, H., Morgenroth, E., Noguera, D., Picioreanu, C., Rittmann, B., van
Science and Technology 48, 61–68. Loosdrecht, M.C.M., 2006. Scientific and Technical Report No. 18, Mathematical
Eldyasti, A., Andalib, M., Hafez, H., Nakhla, G., 2011. Comparative modeling of Modeling of Biofilms. IWA Publishing, London.
biological nutrient removal from landfill Leachate using a circulating fluidized Wanner, O., Gujer, W., 1984. Competition in biofilms. Water Science and
bed bioreactor (CFBBR). Journal of Hazardous Materials 187 (1–3), 140–149. Technology 17, 27–44.
Eldyasti, A., Chowdhury, N., Nakhla, G., Zhu, J., 2010. Biological nutrient removal Wanner, O., Reichart, P., 1996. Mathematical modeling of mixed-culture biofilm.
from Leachate using a pilot liquid–solid circulating fluidized bed bioreactor Biotechnology and Bioengineering 49, 172–184.
(LSCFB). Journal of Hazardous Materials 181, 289–297. Water Environment Federation (WEF), 2011. Biofilm reactors. WEF Press, New York,
Ellis, T.G., Barbeau, D.S., Smets, B.F., Grady Jr., C.P.L., 1996. Respirometric technique McGraw-Hill.
for determination of extent kinetics parameters describing biodegradation. Xavier, J.B., Picioreanu, C., Van Loosdrecht, M.C.M., 2005. A framework for
Water Environmental Research 68, 917–926. multidimensional modeling of activity and structure of multispecies biofilms.
Epstein, N., 2003. Liquid–solids fluidization: Handbook of fluidization and fluid- Environmental Microbiology 7, 1085–1103.
particle systems. Marcel Dekker, New York, pp. 707–708.