Assessment of Nonlinear Leastsquares Estimation of Monod Kinetic Parameters From Batch Reactor Experiments

1
ASSESSMENT .OF NONLINEAR LEAST-SQUARES

ESTIMATION OF MONOD KINETIC PARAMETERS
FROM BATCH REACTOR EXPERIMENTS
by
MARCO ANTONIO VILLARREAL RODRIGUEZ
Department of Civil Engineering and Applied Mechanics

McGill University
Montreal, Canada
June 1990
A thesis submitted to the Faculty of Graduate Studies

and Research in partial fulfillment of the requirements
for the degree of Master of Engineering
Marco A. Villarreal. 1990

("
( To my parents ....
(
l
"
l'
An academic career compels young man to scientific pro-

i
~1
- duction, and only strong characteres can resist the temp-
tation of superficial analysis.
Albert Einstein.
11
Abstract
1t is known that the use of batch experiments for the estimation of kinetic coefficients
is a better alternative than continuous flow reactors, in terms of the time needed to per-
form t.he experiments.
ln this study a new metl'iodology to e:;t;mate the four Monod-kinetic parameters from
batch reactor data set is presented. This method fits biomass and substrate plots simul-
taneously via nonlinear least-squares analysis. The nonlinear equations are solved via the
secant method.
The methodology performed satisfactorily with two synthetic data sets. It was also
applied to seven batch reactor data sets available in the literature and to ten data sets
from batch experiments using wastewater and sludge from the municipality wastewater
trcatment plant in Granby, Quebec. The constants estimated vary considerably among
thernselves, and sorne were not within the range of values commonly found in the litera-
turc. The methodology proposed yields parameter estimates which produce the minimum
sum of squares residuals.
ft is bclieved that further stuJies are needed if one is to attempt estimating the four
~Iollod-kjnetics constants using batch reactors experiments.
111
Rsum
L'utilisation d'expriences en mode cuve pour l'estimation des constantes de C1n~tiqll('
est reconnue comme tant une meilleure alternative que le racteur coulement continu,
en terme de temps requis pour raliser les expriences.
Dans la prsente tude, ne nouvelle mthodologie pour estimer les <Illettre !>c\l'al1li-t 1 cs
de la cintique de Monod, d'aprs les donnes de racteurs cuve, sera discute. Cctte
mthode lisse simultanment des donnes de biomasse et -le substrat via une analyse
des moindre carrs non-linaires. Les quations non-linaires sont rsolues utilisant la
mthode de la scante.
La mthodologie agit de faon satisfaisante avec deux ensembles de donne:-. .,illluli>(:-,
Elle a aussi t applique sept ensembles de racteurs cuve disponible dan:> la litt{~1 ature
et dix ensembles de donnes provenant d'expriences en mode cuve utilisant des t'(tlIX
uses et des boues de la station d'puration des eaux uses Granby, Qubec Lps (O!,-
stantes estimes varient considrablement entre elles-mmes, et certaines Ile se tl'OllVCllt
pas dans l'tendu des valeurs normalement trouves dans la littrature. Le~ e<;tilllatioll">
de paramtres qui rsultent de la mthodologie propose sont celles qui produisent 1(
minimum des sommes des carrs des rsidus.
Nous croyons que des tudes supplmentaires sont ncessaires si l'on veut tentcr
d'estimer les quatre constantes de la cintique de Monod d'aprs les exprienccs danf>
des racteurs en mode cuve.
IV
Acknowledgements
The author wishes to express his gratitude to Professor R. Leduc, research supervisor,
for his invaluable assistance throughout the research and preparation of this report.
He is also grateful to Professor V.T.V. Nguyen, for his important assistance and con-
structive critlcism.
Special acknowledgemen't is given to Salvador Flores Guerrero (M. Eng.) for his en-
couragement and invaluable advice which made the complet ion of this study possible.
He is particulary indebted to E. Waldron and 1. Buchanan for their important assis-
tance in the environmentallaboratory.
Thanks also go to the authorities of the wastewater treatment plant in Granby, Que-
bec, Andr Charbonneau and Marc Brodeur. for their complete cooperation throughout
the study.
The author was partially supported by the Instituto Tecno16gico y de Estudios Supe-
1 iores de Monterrey, Monterrey N. L. Mxico.
\'
J
Contents
Abstract III
Rsum IV
Acknowledgements V
Contents VI
List of Tables IX
List of Figures x
List of Symbols and Acronyms XIII
1 Introduction 1
2 Literature Review 4
2.1 Wastewater Treatment
2.2 Biomass Growth and Substrate Utilization Cl
2.2.1 Biomass Growth Rate 7
2.2.2 Yield Coefficient . 8
-,
2.2.3 Endogenous Decay 9
2.3 Estimation of Monod Kinetic Parameters . 10
vi
2.3.1 Continuous-Flow Experiments . 10
l 2.3.2 Batch Reactor Expcriments 12
3 Methods and Materials 22
3.1 Model development . . 22
3.1.1 Mathematical analysis 22
3.1.2 Nonlinear least-squares analysis 28
3.1.3 Aigorithm for estimation of the parameters . 30
3.1.4 Estimation of L 30
3.2 Initial Estimates ... 33
3.3 Laboratory Experiments 36
3.3.1 Sampling 36
3.3.2 Ratch reactor 36

.., 3.3.3 Experiments . 37
."
~
4 Results and Discussions 38
4.1 Synthetic Data ...... . .. .. .. . .. 38
1.2 Braha and Hafner (1987) Data Sets 40
4.3 Data from the Granby Plant .. .51
4.3.1 Experiments .. ........ .52
5 Summary & Conclusions 69
A Sensitivity Analysis 72
A.l Intl'Oduction . . . 72
A.2 l'.lodci and Application 73
A.3 Commcnts . . . . . . . 74
l B Main Computer Program 76
Vil
C Non-biodegradable substrate 80
1 ay vs y
D SSRS and ill 83
E SSRX and ffi

aA vs A 85
1
l, F Analysis for the roots 88
~
Vlll
List of Tables
2.1 Typical values oi kinetic coefficients at 20C 10
2.2 Models used by Simkins and Alexander (1984) 18
3.1 Comparison cf methodologies . . . . . . . . . . . . . . . . . . . . . . . .. 32
4 1 Synthetic data . . . . . . . . . 39
4.2 Original and estimated parameters for the synthetic data analysis 39
4.3 Braha and Hafner (1987) Data Sets . . . . . . . . . . . . . . . 42

4.4 Estimated parameters with Braha and Hafner (1987) data sets 43
4.5 Parameters estimated by Braha and Hafner (1987) . . . . . 51
... 6 Characteristics of the Granby wastewater treatment plant. 52
4.7 Characteristics of sludge and substrate for each experirnent . 54
4.8 Estimated parameters for the Granby Plant data sets ... 55
4.9 Ratio of soluble ta total substrate concentration (COD) at Granby. . 55
., 10 Substrate and biomass data for runs 1 to 5 . . . 56
1.11 SSR 's obtained with the Granby Plant data sets 67
A.1 Parameters studied .. 73

A.~ Thirteen smallest values of the SSRs for the first analysis 74
A.3 SSRSs values for the second analysis . . . . . . . .. . 75
IX
'1
List of Figures
2.1 Fl,~W schematic of the suspended-growth activated sludge 5

2.2 Baeterial growth ............ . . . . . 6
2.3 Diomass growth rate vs Substrate concentration 8
2.4 Complete-mixing continuous flow reactor without recycle 11

2.5 Batch Reactor . . . . . . . . . . . . . . .......... 13
2.6 Biomass growth and substrate degradation vs time in a batch reactor 14

''i''l'
.",
3.1 Substrate as the independent variable . 24
3.2 Time as the independent variable 21
3.3 Substrate vs time for different /(IJ 25
3.4 Aigorithm . . . . . . . . 31
3.5 Wand SSRS versus Y . 34

3.6 Wand SSRX versus A 3.5
ay versus y and !l.a

4.1 Synthetic 1: ffi 8A versus A . 40
4.2 8Y versus y and lli
Synthet ie 2: ffi 8A versus A . 41
4.3 SSR ratio, biomass and substrate graphs for LBI 44
4.4 SSR ratio, biomass and substrate graphs for LB2 4.)
4.5 SSR ratio, biomass and substrate graphs for LB3 46

.Jffi;
".1- 4.6 SSR ratio, biomass and substrate graphs for LB4 47
x
4.9 SSR ratio, biomass and substrate graphs for LB7 . .... 50
4.10 SSR ratio, biomass and substrate graphs for run 1 (COD) 57
4.11 SSR ratio, biomass and substrate graphs for run 1 (TOC) . 58
4.12 SSR ratio, biomass. and substrate graphs for run 2 (COD) 59
4.16 SSR l'atio, biomass a:d ",ubstrate graphs for run 4 (COD) 63
4.17 SSR ratio, biomass a'1d substrate graphs for run 4 (TOC) . 64
4"
'i
F.l -W- vs Y, SSRS vs Y, ~7 vs A and SSRX vs A for LBI . 89
F.2 -W- vs Y, SSRS vs Y, ~7 vs A and SSRX vs A for LB2 . 90
F.4 -W- vs Y, SSRS vs Y, a:: vs A and SSRX vs A for LB4 . 92
F.6 ~ vs l', SSRS vs Y, a:: vs A and SSRX vs A for LB6. 94
F.7 * vs Y, SSRS vs Y, a:: v!: A and SSRX vs A for LB7 . 95

F.8 * vs Y, SSRS vs Y, ~7 vs A and SSRX vs A for Run 1 (COD) 96
F.9 * vs Y, SSRS vs Y, ~7 vs A and SSRX vs A for Run 1 (TOC) 97
F 10 ~ vs Y, SSRS vs Y, ~7 vs A and SSRX vs A for Run 2 (COD) 98
F.ll * vs Y, SSRS vs Y, ~ vs A and SSRX vs A for Run 2 (TOC) 99

f F.l2 W- vs Y. SSRS vs Y, ~ vs A and SSRX vs A for Run 3 (COD) 100
Xl
.j
F.13 ~~ vs Y, SSRS vs Y, ~~ vs A and SSRX vs A for Run 3 (TOC) 101
F.14 ~~ vs Y, SSRS vs Y, fi vs A and SSRX vs A for Run 4 (COD) 102
F.15 ~~ vs Y, SSRS vs Y, fi vs A and SSRX vs A for Run 4 (TOC) 103
F.16 ~~ vs Y, SSRS vs Y, fi vs A and SSRX vs A for Run 5 (COD) 104

F.17 ~~ vs Y, SSRS vs Y, fi vs A and SSRX vs A for Run 5 (TOC) 105
xii
List of Symbols and Acronyms
A constant
B constant
COD chemical oxygen demand
F/M food-biomass ratio
k maximum substrate utilization rate per unit of biomass
kd endogenous decay coefficient
c d estimate of endogenous decay coefficient
kt constant
k2 constant
k3 constant
k4 constant
1<1 first-order rate constant
L constant
L fanal upper limit of L
Lo lower limit of L
Lp value of L when the plateau is reached
mznSSR minimum sum of squared residuals
.,. MLVSS mixed liquors volatile suspendid solids
,.
:;t
q inverse yield
Q influent flow rate
QR recycle line flow rate
Qw sludge wasted flow rate
rd endogenous decay rate
rg biomass growth rate
.
rll
rg
substrate utilization rate
biomass growth net rate
R by-products from substrate consumption
S growth-limiting substrate concentration
Se biodegradable substrate concentration at the end of the experiment;
effluent biodegradable substrate concentration
S, biodegradable substrate at time tj
So biodegradable substrate concentration at the begining of the experiment;
influent biodegradable substrate concentration
St pl'edicted substrate concentration at time t
{
xiii
l
SSR sum of squared residuals

SSRS sum of squared residuals of biodegradable substrate concentration
SSRX sum of squared residuals of the logarithm of biomass concentration
SVl sludge volume index
S average biodegradable substrate in the time interval 6t
t time
ti time at the i th observation
TOC total organic carbon
V volume
Vm maximum substrate consumption
X biomass concentration
Xa average biomass concentration
Xo biomass concentration at the begining of the experimentj
influent biomass concentration
XR biomass concentration recycle tine
X average biomass in the time interval D.t
."..,..
X* substrate concentration required to produce a biomass concentration X
J...
X* 0 substrate concentration required to produce a biomass concentration X o
y total yield coefficient
'Y constant
D.[(s correction term for half-velocity constant
D.S biodegradable substrate depleted in the experiment
D.t time interval
D.X biomass growth in the experiment
D.Y correction term for the total yield coefficient
D.p.m correction term for the maximum specifie growth rate
() hydraulic retention time
J.l specifie growth rate
J.lm maximum specifie growth rate
e constant
4> constant
w constant
XIV
Chapter 1
Introduction
One of the major princip les limiting the formation of human communities has been the
preoccupation with securing and maintaining a suitahle supply of clean water. Unfortu-
nately, water has played an important role as a carrier of wastes throughout the history
of civilizations.
It is known that over 100 different types of viruses and pathogenic bacteria may
he present in untreated domestic wastewater. (Tchobanoglous and Schroeder, 1985 [29]).
The first documented relationship between human wastes, drinking water, and disease
was reported in the mid-nineteenth century. Nevertheless waterborne disease was not
properly accepted un tH more advanced studies in the germ theory of disease were done
by Pasteur and other scientists at the end of the nineteenth century and early twr>ntieth
cent ury (Montgomery, 1985 [14]). It is now known since many decades that microorgan-
isms can be responsible for specifie public health prohlems mcluding bacterial illnesses
such as cholera and gastroenteritis, viral diseases such as hepatitis, amoebic dysentery or
diarrhca which are generated by protozoa, and parasitic helminth (worm) infections such
{ as tapeworms or roundworms (Montgomery, 1985 [14]).
1
1 The first goal for the water treatment professionals in the nineteenth cent ury was
to eradicate waterborne diseases. This was accompli shed successfully in the developed
countries by the virtual elimination of the mos t deadly waterborne diseases such as ty-
phoid, cholera and amebiases. However, since the beginning of the twentieth ccntury,
public health concerns have switched from acute disease to the chronic health effects of
trace quantities of organic, inorganic and microbiological pollutants.
On the other hand, wastewater carries wastes of different types from different sources
including residential, commercial, industrial wastes, ground water, surface water, stormwa-
ter, etc. These \Vastes need to be removed for pollution control before the watel" is dis-
charged into the receptor body (river, lake, etc.). The discipline that deals \Vith \Vastew-
ater treatment is called wastewater engineering.
Wastewater engineering is a branch of environmntal engineering which applies the
basic principles of science and engineering to the problems of water pollution control.
The ultimate goal of this engineering branch is to develop wastewater management tcch-
niques in order to help protecting the environment. Physical, chemical and biological
pro cesses are generally involved in a municipal wastewater treatment plant. Disinfection
and coagulation are examples of chemical processes, while sedimentation, filtration and
screening are examples of physical processes.
Biological treatment processes are a significant and indispensable segment of domestic
wastewater treatment plants. In particular the activated sludge process is extcnsively
used, mainly due to its efficiency in degrading organic matter and suspended solids. The
activated sludge process was deve!oped in England in the carly 1900s (Joint Committee of
2
----------------~----------------------------------- -
the WPCF and the ASCE, 1977 [23]) and was brought into North America in the 1920s.
1 By the 1940s the process has been used throughout the world.
AlI the biological pro cesses present in a wastewater treatment plant take place in
a defined volume. Sueh a volume is eommonly named a reactor. Important changes in

the composition and eonce~tration of organic eompounds occur while the wastewater is
retained in the reactor. These changes are caused by hydraulic transport of materials into
and out of the reactor as well as by reaetions that oeeur within the reaetor. To completely
define a reactor system and design similar ones it is necessary to know the rate at which
the change~ of composition and concentration of biodegradable substance occur and the
extent of the changes.
To design any biological pro cess for a wastewater treatment faci li ty, it is required
to know the rates at whieh various components (such as organic compounds) are de-
graded from the wastewater and the rate at which biomass is produeed in the reactor.
Sueh rates of change are important because they will determine the size of the reactor
required for a specifie degree of water quality.
The most weil known mathematical mode! which describes the rates for the degra-
dation of organic material and the biomass produced in a bateh reactor is the Monod
kinetics model (Monod, 1949 [20]). This model includes four parameters ~hat need to
be estimated to properly design biological units for wastewater treatment. In this study
a new methodology is developed to estimate the parameters involved in Monod kinetics
using various sets of data from bateh reactors.
3
.J
Chapter ~
Literature Review
2.1 Wastewater Treatment
Wastewater is classified into industrial and municipal. The characteristics of inctust.l'ial
wastewater vary with respect to the type of industry, therefore its treatment will be (liffer-
ent for each case. However the characteristics of municipal wastewater are bettel' kllowlI
because they are affected mainly by domestic uses, relathely small hydraulic and organic
loads from industries (with respect to domestic loads), stormwater, infiltrations, among
others. Domestic wastewater contains mainly biodegradable organic matter, suspended
solids, pathogenous microorganisms, nutrients, and a wide variety of compounds in lllllCh
smaller concentrations_
Biological treatment pro cesses are generally involved in domestic wastewatcr treat-
ment plants. In biological processes, microorganisms (mainly bacteria), present in the
wastewater, convert easily biodegradable organic pollutants into degraded products Thf!
biological process can be classified, according to the type of baderial growth, iIlto at-
tached or suspended-growth.
In the attached-growth process a medium is utilized to support the microorganism~.
4
------------------------
This growth process is carried out usiog trickliog filters, rotatiog biological contactors,
activated biofilters, among others (Benefield & Randa1l1980 [3]).
One of the most common suspended-growth processes in wastewater treatment plants
is the activated sludge .. The system comprises three components, which are the aerated re-
actor, the clarifier and a sludge recycle line (see Fig. 2.1). In the reactor, the biodegradable
Clarifier
Reactor
(
Recycle line
Figure 2.1: Flow schematic of the suspeoded-growth adivated sludge
organic matter is degraded by the biomass under appropiate conditions (pH, temperature,
oxygen, etc.). The biomass grows and forms ftock that oormally settle in the clarifier.
The sludge (i.e. biologicai solids, water and other dissolved constituents) is withdrawn
fl'om the bot tom of the clarifier. A portion of that sludge is returned to the aeration tank
and the rernailling is wastcd. As an alternative, sludge can also be withdrawn from the
aeration basin.
i
5 1
j
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2.2 Biomass Growth and Substrate Utilization
Effective environmental control in biological wastewater treatment is based on an lIllder-
standing of the basic principles govern',ng the growth of microorganisms. The gt'Ilcl"al
growth pattern of bacteria in a batch culture is shown in Fig. 2.2. In aCl'obic batch cul-
ture experiments, bacteria are provided with enough soluble food (substrate) and suitabk
.
environmental conditions (pH, temperature, redox, and others).
The growth of microorganisms under aerobic conditions in batch reactors can be
Cl)
II) en Q)
CI) Cl !Il >-
.... CI)
<Il C s::. <Il co
s::. c a. c Cl>
s::.
a. 0
<il
C.
>-
-cin Cl)
CI)
<Il
II)
'0
-
s::.
~ <Il
<Il
s::.
C.
CI)
::l
!Il
s::.
Q) 0
.... <il 0
c
OJ
Q)
"C
a. c Q)
0 Cl
c a. 0
<Il
.... (1) -0 x V
'\
"C
C
/
CI) Q)
!Il
II)
<Il ....
~
s::.
.... -
C,) Cl)
<Il c. (l)
8 Cl (.)
Cl ;:: (.)
ttI
~
/
Tlme t
Figure 2.2: Bacterial growth
divided into six phases (Benefield & Randall 1980 [3]) (see Fig. 2.2).
1. The lag phase is characterized by the adaptation of microorganisms to thcil'
newenvironment. There occurs log generation time and null growth rate, and
cell size and rate of metabolic activity are at maximum.
2. During the acceleration phase there is a decrease in generation time and an
increase in growth rate.
3. The exponential phase is characterized by maximal and constant specific gl'Owth
rate, maximum rate of substrate conversion and constant gcneration time.
6
4. The declining growth phase exhibits an increase in generation time and a de-
crease in specifie growth rate due to the graduaI decrease in substrate concen-
l tration and the incased accumulation of toxic metabolites.
5. In the stationary phase the nutrients are exhausted, leaving behind a high
concentration of toxie metabolites, and maximum physical crowding .

6. In the endogenous phase there is endogenous metabolism, high death rate, and
cell lysis takes place.
2.2.1 Biomass Growth Rate
The rate of growth of biomass during exponential growth (phase 3), can be described by
the following equation:
(2.1)
where
Tg = biomass growth rate, mass vo/ume- 1 time- 1
Jl = specifie growth rate, time- 1
x = concentration of microorganisms, mass volume- 1
In the case when the biodegradable soluble substrate is growth-limiting, the specifie
growth rate can be described using Monod kinetics (Monod, 1949 [20]):
(2.2)
where
ILm = maximum specifie growth rate, time- 1
S = concentration of growth-limiting substrate, mass volume- 1

/(5 = half-velocity constant, mass vo/ume- 1
7
The relationship between the specifie biomass growth rate and the limiting substrate
concentration is presented in Fig. 2.3.

1 lntroducing Eq. ( 2.2) in ( 2.1) gives:
IlmXS
rg = [(6 +S (2.3)
fJ-m ........... _.................... _- ............. __ ................ _...... -- -. _ ............... _-_ ... -......... -_ ........ .
fJ-m
2
Substrate
Figure 2.3: Biomass growth rate vs Substrate concentration
2.2.2 Yield Coefficient
In aerohic culture systems, a portion of the substrate is converted to new cells and the
other portion is oxidized to end products. The quantity of new cells produccd has bccn
observed to be a function of the substrate removed. The following relationship has bccn
developed between the rate of substrate utilization and the rate of biomass growth:
(2.4 )
where
y = total yield coefficient, mass mass- 1

r6 = substrate utilization rate, mass vo/ume- 1 time- 1
8
The total yield coefficient Y refers to any fini te period of exponential growth, and is
defined as the ratio of t.he mass of cells formed to the mass of substrate consumed.
l Introducing Eq. ( 2.3) in ( 2.4), we have:
J.lmXS
(2.5)
r" = Y(K. + S)
Setting k = ~ gives:

kXS
(2.6)
r" = [(" + S
where k represents the maximum substrate utilization ra.te per unit of biomass, (time- 1 ).
2.2.3 Endogenous Decay
In bacterial systems used for wastewater treatment, the distribution of cell ages is su ch
that not aIl the cells in the system are in the same phase. The microorganisms which are
in the endogenous phase do not normally generate new cells, therefore inducing a decrease
in the growth rate called endogenous decay.
Endogenous decay not only takes into account the cells age but also the energy of
maintenance and predation (Herbert, 1958 [13]). The rate of endogenous decay (rd) has
been found to be proportional to the concentration of microorganisms in the system and,
hence, it follows the relationship:
(2.7)
where kd represents the endogenous decay coefficient, (time- 1 )
The net rate of bacterial growth (r~) is written:
(2.8)
Introducing Eqs. ( 2.3) and ( 2.7) in ( 2.8) produces:
(2.9)
9
Table 2.1: Typical values of kinetic coefficients at 20C
Coefficient Units Range
Benefield & Randall (1980) Metcalf & Eddy (199)
day-l 6-S 2 - 10
mg/l COD 25 - 100 15 - 70
mg MLVSS
mg ('OD 0.35 -0.45 0.25 - OA
day-l 0.05 - 0.1 0.04 - 0.075
where r~ is the net rate of b~cterial glOwth, (mass volume- 1 ttme- 1 ). Thc range of values
commonly found for Y, ](a, k, and kd for municipal wastewater are presented in Table 2.1.
2.3 Estimation of Monod Kinetic Parameters
The general characteristics of domestic or municipal wastewaters are weil known. Ncvcl'-
theless, for each case an estimation of the parameters (i.e. the biokinetic coefficlellt) i~
.'
strongly recommended for the design and operation of a wastewater treatment plant is
planned (Vasicek 1982 [34]).
Values for the parameters Y, p'm, Ka and kd must be found for a particulal' case and
used in the design for most effective results. Two methods can be consideree! to detcrrnnc
these coefficients: continuous-flow systems or batch reactors.
2.3.1 Cont inuous- Flow Experiments
To estimate the parameters, the usual procedure with continuous-flow reactrs is t op er-
ate the units over a wide range of retention times (i.e. flowrates), and influent cO!1ceIlLll
tions (5 different retention times ranging from 1 to 10 days are recommcnded by BCllcfidd
and Randall1980 [3] and Met.calf & Eddy 1979 [19]). When steady-statc l~ Icac!l('u fol' t
specifie retention time, the v.ilues of discharge (Q), substrate concentration in thc illnucllt
.'
(Sa), substrate concentration in the effluent (Se), and microorganism!> conccnt.ration in
10
----------~----------------------_.----
l Se , Q , X
- ---
v, x, Se
Figure 2.4: Complete-mixing continuous flow reactor without recycle
the tank (X) are measured.
Applying subc;trate balance to the system and assuming steady-state, the following
equation is obtained:
(2.10)
realTanging ylelds:
(2.11)
\Vriting Sr = So - Se, and () = ~ where () is the retention time (time), this last equation
becomes:
....'
rs = -"r (2.12)
o
Equating Eqs. ( 2.12) and ( 2.6), and linearizing, the following relationship is obtained:
OX = (1(,) ~ + ! (2.13)
Sr k Se k
Dy plotting ~~ vs i. a straight \ine with slope If' and intercept i is obtained.
FUl'thermore, a stcady-state mass balance on the system can be written assuming that
active biomass in the influent is negligible, as:
11
(2.14)
1 Introducing ( 2.9) and ( 2.12) in this last equation, and rearranging, gives the following
linear relationship:
1 Sr
- = y - - kd (2.15 )
() ()X
""
Plotting ~ vs fff gives etimates of Y and kd'
2.3.2 Batch Reactor Experiments
The use of continuous-flow experiments for parameter estimation, involves a considerable
amount of time. It is known that to achieve steady-state in the system, apPl'oximn.tely
two to three weeks are needed after the initiation of the test. Consequently, if diffcl'cnt
retention times are required [Benefield and Randal (1980) [3], and Metcalf & Eddy (199)
[19]] for processing the data, either two to three rnonths will be necessary to complete
the study, or sever al reactors should be run in parallel which implies significant amount
of qualified personnel and equipment.
On the other hand, batch reactors are closed systems in which a specifie concentration
of adapted biomass is allowed to grow in the presence of substrate under a suit,ablc Cllvi-
ronment (pH, oxygen, temperature, etc ... ) as shown in Fig. 2.5. The reaction is genpI'ally
allowed to proceed until the biodegradable substrate is almost completcly dcpldcd by
the biomass. During the process, substrate and biomass concentrations arc mcasUI'cd at
different times. Generally, the experirnent will not last for more than 24 hr when adaptcd
biomass is available. If no adapted biomass is available, there is a nccd t obtaill ~()me
using a batch, or continuous flow reactor.
Fig. 2.6 shows biomass growth, and substrate disappearance as a functioll of time
where So, X Ol Se and Xe indicate the substrate and biomass conccntration at time zero,
12
VI XI S
Figure 2.5: Batch Reactor
and substrate and biomass ccn~entration at the end of the process, respectively.
An essential element in any comparison between batch and continuous-flow experi-
ments, is that batch reactors represent the best alternative for biokinetics estilnation in
( ter ms of the time needed to obtain the parameter estimates. Few methodologies has been
developed to estimate the Monod biokinetics parameters using batch reactor experiments.
Most of them apply numerical methods while others employ either linearization, graphi-
cal, chemical analysis or respirometric measurements. These methodologies are reviewed
hereafter.
(a) Reynolds and Yang (1966) Method [25]
This is an introductory methodology in which the authors used five batch reactors
simultaneously containing different substrate and biomass concentrations. The authors
measured substrate and biomass concentration at time zero and at time 24 hr. in each
flask l'eactor. In this study the equations were rearranged by introducing Eqs. ( 2.4) and
( 2.7) in ( 2.8), yielding:

(
(2.16)
13
kd = 0
............. ........ X
. '
.' . .. '

s.
Time t
Figure 2.6: Biomass growth and substrate degradation vs time in a batch reactor
Discretizing Eq. (2.16) gives:
(2.17)
where 6.X is the biomass growth (Le. Xe - X o ) mass volume-l, b.S is the substrate
depleted (i.e. So - Se) mass volume-l, and b.t = duration of the experiment.
Setting b.t = 1day and X = X a where X a is the average biomass during the experi
ment, [i.e. X a = !(Xo + Xe)] and rearranging yields:
(2.18)
Plotting ~: vs ~: provides estimates of Y and kd'
(b) Robinson and Tiedje (1983) [27]
In this study a nonlinear regression analysis was proposed to estimate Monod kinetics.
Robinson and Tiedje used the Monod integrated form for substrate removal [Eq.( 2 ..5)],
in which biomass was eliminated in Eq. ( ~.5) by using the following relationship:
....
(2.19)
14
Introducing Eq. ( 2.19) in ( 2.5) yields:
( dS Jl.mS [Xo + Y(So - S)]

(2.20)
dt Y(I(3 + S)
Performing the integration they established:
(2.21 )
Robinson and Tiedje discerned that high correlation existed among the variables, and
argued that a nonlinear least-squares technique would not be the appropriate routine to
use, although from the statistic point of view it is considered the best alternative [Bach
(1982) [2]].
The authors proposed instead a nonlinear regression analysis using the foIlowing model:
dS dS dS
St = ~Pm dPm + tlI<3 dK + tlY dY
3
(2.22)
where the derivatives were solved by using implicit differentiation in Eq. ( 2.21).
{
The analysis was carried out using a computer program (MONODCRV) that estimates
the theoretical predicted substrate (St) at time t with Eq. ( 2.22) (given initial estimates
of the parameters) and calculate the residual errors (S - St) for aIl the points, where S
is the observed substrate at time t. tlpm, tlI<s and ~Y were solved via multiple linear
regression and added (either positive or negative) to the initial values. The process con-
tinued until the correction terms were less than 0.01. The initial guesses of the parameters
were evaluated with the linearized forms of Eqs. (2.3) and ( 2.6) which are discussed
hereafter. The model was tested with success using different sets of simulated data and
one set of H 2 -1imited batch growth of Desulfovibrio spp.
The authors defined their method as a nonlinear regression analysis with Gaussian
mcthod. Nevertheless, it is considered that this methodology is rather a combination of
the Newton Raphson and the steepest descent methods.
15
1
.J
(c) Robinson and Characklis (1984) Method [26]

In this study the authors estimated the parameters from the integrated form of the
Michaelis-Menten equation. This equation was used to describe substrate depletion ac-
cording ta the foIIowing relationship:
(2.23)
where
Vm = maximum substrate of consumption, mass volume- 1 time- 1

R = by-products from the substrate consumption, mass volume- 1 time- 1
The model did not consider biomass growth.
The authors estimated the parameters by using Eq. (2.23) in four different fonus:
one nonlinear and three different linearized forms. In the nonlinear approach the authors
used the same nonlinear regression analysis applied by Robinson and Tiedje (1983) [27J,
while they used linear least-squares analysis with the linearized forms.
Robinson and Characklis (1984) [26J found that applying nonlinear regression analysis
ta the integrated form and considering So ta be not free of error, yielded bcttcr results
than any of the other three Iinearized forms by using the !inear least-squarcs technique.
(d) Simkins and Alexander (1984) [28]
The authors introduced a new mass-balance equation as follows:
So + qXo = S + qX (2.24)
where q = is the ceU quota or inverse yield (~) and X o is the initial biomass concentration.
Setting qX equal ta X*, which corresponds to the amount of substrate required ta produce
a biomass concentration equal to X. In the same manner X; is defincd as bcing equal ta
....... qXo. Rearranged, Eq. ( 2.24) can be written as:
(2.25 )
16
Solving for X* from Eq. (2.25) and its derivative with respect to time, and introducing
1 both terms into the differential form of Eq. ( 2.3) yields:
_ dS = /-lmS (S X* - S) (2.26)
dt Ka +S 0 + 0
The authors employed Eq. (2.26), rearranged for a particular set of assumptions. The
assumptions and the respective models are presented in Table 2.2. Data were collected
during an experiment on the mineralization of benzoate byan induction of Pseudomonas
spp. The non!inear regression analysis was carried out using MARQ-FIT, which performs
a minimization of least-squares of data by the method of Marquardt (1963) [18].
In this study, all the models produce positive results, excluding the model where no
assumptions were stated\ i.e. the Monod model with growth. For this set of data the
Jogistic mode! yielded a better fit than the Monod model with growth, eventhough the
value of So was close to that of /(3' The authors offered no possible explanation for that.
If we examine the high correlation among the parameters, as pointed out by Robinson and
Tiedje (1983) [27], and Robinson and Characklis (1984) [26], the parameter correlation
rises as a possible explanation.
(e) Braha and Hafner (1987) Method [7]
The authors presented a new methodology to evaluate Y, K31 /-lm, and kd , while
the other studies (presented above) have only estimated two or three out of the four
parameters. In this study, the authfs utilized the methodology proposed by Rey 10lds
and Yang (1966) [25]. They plotted the data from a batch reactor using Eq. ( 2.18), to
obtain the values of Y and kd through !inear least-squares. The two other parameters (1<3
and /lm) were found through nonlinear least-squares analysis of Eq. ( 2.21) (by using a
computer program the BP7IB application of the curve-fit module).
The use of Reynolds and Yang (1966) [25] methodology in a single batch reactor
l makes this technique a questionable method, mainly due to the units of kd' In their
17
.1
,
1
Table 2.2: Models used by Simkins and Alexander (1984)
ModeZ and characteristics Equations and inequalities
1. Zero Order
Differentiai form -dS - k
dt - 1
Integrated form S = Sa - kIt
Derived parame\er kt = JlmX;
Necessary conditions X; ~ So and So ~ J(,
II. Monod, no growth

Differentiai form -dS _ .J!J.L
dt - K.+S
Itegrated form [(,ln 10 + S - So = -kIt
Derived !>arameter kI = JlmX~
Necessary condition X; ~ So
III. First order J

Differentiai form -dS -
dt -
k3 S 1/,
Itegrated form S = Soe- k3t ~
Derived parameter k - ~"'x~ il

3 - K.
..- Necessary conditIOns X; ~ So and So J(, 1
il'
IV. Logistic
DifferentiaI form -fts = k4 S(So + X; - S)
Integrated forrn S - so+x;
- I+4e"4(50+X~)1
50
Derived parameter k4 -~
- K.
Necessary condition So !(,
V. Monod with growth

-dS _ IJmS(So+X~-S)
Differen tial form dt - K.+S
Integrated form [(,In la = (Sa + X; + K,) ln f~ - (So + X;)llm t
Derived parameter None
Necessary condition None
VI. Loga7'ithmic
DifferentiaI form -fts = J1.m(So + X; - S)
Integrated form S = So + X;{l- elJmt )
Derived pararneter None
Necessary condition So ~ !(,
l
18
original study [Eq. ( 2.18)] the units of kd are day-I because the measurements were done
24 hours after initiation of the test, while Braha and Hafner measured the substrate and
1 bioma&-; concentration throughout the trial (and contemplated aIl the points). If time is
considered in Eq. ( 2.17) i.e.
I:J.X =y t:J.S _ k
X a t:J.t X a t:J.t d
and the methodology is applied, the estimated values of Y and kd are negative.
(f) Van Niekerk et al, (1987) [33]
These authors model the soluble COD uptake in batch reactor using the foIlowing equation
(2.27)
where [(1 is the first-order rate constant, volume mass- 1 time-t, estimated via linear
regression analysis.
The authors pointed out that first-order substrate uptake can be regarded as the sum-
mation of a number of zero-order removals [Tischler and Eckenfelder (1969) [31]].
(g) Templeton and Grady (1988) [30]
In another study, Templeton and Grady (1988) used Eq. ( 2.21) to estimate Ks, JLm
and Y. The parameters were obtained with Marquardt's B-Solve algorithm [Marquardt
(1963) [18], Kuester and Mize (1973) [15]]. The authors used no other model or technique
to do a comparative analysis.
(h) Discretization and linearization techn~ques
The discretization and Iinearization of equations for parameters estimations is a corn-
mon technique used by chemical and environ mental engineers. Renee, by discretizing and
!inea! izing Eqs. ( 2.3) and ( 2.6) the following relationships are obtained:
19
~tX J(.Y 1 Y
---=--=+- (2.28)
~S J-lm S /lm
!ltX J(. 1 1
-=--=+- (2.29)
!lX /lma.x S J.lma.x
where 6..X, IlS, X and S are the biomass growth, the substrate depleted, and biomass
and substrate averages respectively in the time interval 6..t. If 6..t is rclativcly small.

plotting -~: and ~t: ver~us ~,estimates of [(., J.lm and Y are obtained (Robinson and
Tiedje (1983) [27]). Therefore, with a set of data of substrate and biomass versus lime
from a batch reactor, it would be possible to find the constants f(., JLm and lI" (but not kd ).
However several authors have found vulnerable points concerning the lincarization of
equations. Robinson and Tiedje (1983) [27], and Robinson and Characklis (1984) [26]
found that estimating parameters by nonlinear regression analysis generates constants
which fit the data points better than by discretization and linearization techniques. 8raha
and Hafner (1987) [7] pointed out that by-products were generated by the microorganisms
which would affect bacteria growth in a closed system, and that these cffects WCI C Ilot
considered in Eqs. ( 2.28) and ( 2.29).
In summary among the methodologies commented above, only the method by Araha
and Hafner (1987) [7] yields estimates for al! the constants. Yet, this method is qlles-
tionable technique (as explained above and proved in Appendix A). Consequently, if ail
estimation of the four Monod parameters is required, a new methodology is desirablc On
the other hand, it was noticed in the study by Sirnkins and Alexander (1984) [28], that
\Vhen no assumption was made in Eq. (2.26), the pararnetcrs estimated did Hot yicld the
minimum sum of squares residuals (SSR), [the logzstic modcl (k<1 = y.) yielded smallcr
SSR]. It is unknown as to why the computer packages converge to wrong estimates iu
20
the two studies commented above, but from this analysis it is strongly recommended to
perform a sensitivity analysis similar to the one carried on in Appendix A if a computer
package is applied.
Finally it is not th scope of this thesis to coyer either graphical, chemical nor respiro-
metric techniques for parameters estimation sinee this study focusses on numerical ap-
proaches only. However it is important to mention herein that only the methodology
proposed by Blok (1974 [5] and 1976 [4])(respirometric measurements) estimates the four
parameters. On the other hand, Camara and Randall (1984) [9] (chemical) and Gates
and Marlar (1968) [10] (graphical), estimate one and two parameters, respectively.
21
1
Chapter 3
Methods and Materials
In Chapter 2 it is pointed out, that the approaches for estimating the biokinetics yielded
only two or three out of the four Monod parameters. The only rnethodology which yiclds
the four constants was proposed by Braha and Rafner (1987) [7], but it was showll in
Chapter 2 that it is a questionable methodology. Therefore, a new model should he
developed if one attempts to estimate the four Monod parameters from batch reactor
experiments.
In this study a new methodology is proposed and assessed to estimate the f01l1' pa-
rameters of the Monod kinetics expression using various sets of data from batch reactors.
The methodology presented herein fits simultaneously biomass and substrate data sctl>
through nonlinear least-squares analysis.
3.1 Model development
3.1.1 Mathematical analysis
(a) Substrate Utilization

The rate of substrate utilization is defined by Eq. ( 2.20) as follows:
22
dS J1.mS [Xo + Y(So - S)]
-=- (3.1 )
dt Y(I<.. + S)
l Rearranging and integrating Eq. ( 3.1) yields:
J1.
1
m 0
t
dT - -
-
1SSo
K 6 Yd
[Xo + Y(So - )]
-
i
50
5
Xo
Yd
+ Y(So - )
(3.2)
Performing the integr~tion of Eq. ( 3.2) gives:
(3.3)
Substituting the upper and lower limits for substrate and rearranging, yields the following
equation:
(3.4)
Eq. ( 34) is the same as the one used by Robinson and Tiedje (1983) [27], Braha and
Hafner (1987) [7], Braun and Berthouex (1970) [8] and Moser et al. (1973) [21] in their
studies [i.e. Eq. ( 2.21), but arranged differently].
The least-squares techniqae considers the independent variable as being error-free, and
minim.ize1> the sum of square residuals (SSR) with respect to the dependent variable. In
Eq. ( 3.4) least-squares assumes that the measured substrate is considered to be error-free
and the time variable is assumed to contain the source of error (see Fig. 3.1). Bence,
to properly apply the least-squares technique, time must be considered as error-free (i.e.
the independent variable) and substrate the source of error (see Fig. 3.2).
Analyzing the plots of ex peri ment al data presented in the studies by Simkins and
Alexander (1984) [28] and Braha and Hafner (1987) [7], the relat.ionship between l\~ and
80 cau be Jtablished from a plot of substrate degradation versus time for Ks > So,
[{~ ~ So and !(! < So. These relationships are shown graphically in Fig. 3.3.
Uloth and Mavinic (1977) [3:!] and Lawrence and McCarty (1970) [17], pointed out
23
s
1 0 model
0 observation
Cf) er ror
....co
<J)
....
CI) -----0
.0
::J
Cf)
0----
Time t
Figure 3.1: Substrate as the independen t variable
so
model
o observation
(J) er ror
CD
.......
co
"'-
CI) o
.0
::J
(J) o
o
Tlme t
Figure 3.2: Time as the independent variable
l
24
(f)
<D
co
~
(f)
.0
:J
(f)
K8 ) S 0
Time t
Figure 3.3: Substrate vs time for different /(~
that as the complexity of the waste increases, the value of /(5 increases. Hence, consid-
ering that the samples used in our experiments faIl into this group (since two industries
discharge their wastes into the sewage of Granby), and looking at the shape of each of the
( plots of our lahoratory experiments (which are similar to the shape for Je > So , Chapter
4), the value of l<S is considered to be larger than So, and the second term of the right
hand side of Eq. ( 3.4) [i.e. _1 ln Y(So1 l+Xo ] hecomes negligible and therefore it can he
/.lm
eliminated, allowing time to be the independent variable.
From the above assumption Eq. ( 3.4) C:]T) he simplified to the following form:
t = y l(, ln So[Y(So - S) + Xo]

(3.5)
JLm(YSo + X o) XoS
or
(3.6)
where L is a constant defined as
(
25
Furthermore, if the sarnple tested has /<, close to So, this model could still be applied. In
this case, an inapropriate fit would occur when time is small (i.e. when filting the first
points). Therefore sorne accuracy would be lost in that portion of the curve, but the l'est
of the points would have an appropriate fit .

(h) Biomass Growth
In Chapter 2 the cell growth was established by Eq. (2.9), which considcl's endogenolls
decay. Herein this equation is used in its differential form, Le.
dX _ l'm SX _ k X (3. )
dt K, +S d
If substrate is elirninated in Eq. ( 3.7) by using the following relationship S' = So + .\\-:- \"
[see Knowles et al (1965) [16], Pirt (1975) [24] and Robinson and Tiedje (1983) [27]}, a
new equation is established:
_dX _ l'm X (YSo + X o - X) _ kdX (3.8)

dt y (I<, + So) + X o - X
Performing the integral for values of X o at t = 0 and X at t = i, the following cquation

is obtained:
(:3.9 )
where w and 1 are constants defined as:
(:l.1O )
(:3.11)
In Eq. ( 3.9) the variable time is expressed as a dependent variable and it cannot he ar-
ranged to be independent. Therefore, to establish time as an indepcndent variable sorne
26
assumptions may be considered in the analysis.
l If /(6 is considered to be much larger than So, then Scan be eliminated from the
denominator in Eq. ( 3.7), thus creating the following relationship:
dX LX
-dt= - (yY So.+X0 -X)-kdX (3.12)
Effecting the integration and rearranging generates the following equation:
e4>t
X - ----'-,.----- (3.13)
- 1 + ..L
X
- e tPt
o
where ~ and <p are constants defined as:
(3.14)
(3.15)
By using Eqs. ( 3.6) and ( 3.13), it is not possible to estimate the four parameters due to
the fact that the respective values of /lm and /(6 cannot be estimated separately, but only
the value of the ratio L = 71;, [this was acheived in the ~tudy of Simkins and Alexander
(1984) [28]].
An alternative to the above approach consists in substituting the substrate expression
Eq. ( 3.6) in Eq. ( 3.7). This yields the following relationship
(3.16)
where A and 0' are constants defined as:
(3.17)
27
1
Performing the integration of Eq. (3.16), and considering X o at t = 0 and X at t = t, givcs
ot
ln (~) = (B _ kd) t _ YS oin (A+xoe ) (3.18)
Xo A A A+Xo
It was ohserved by inspection in Eq. (3.18), that when t = 0, X = X o and when
t -+ 00, X -+ o.
3.1.2 Nonlinear least-squares analysis
Nonlinear least-squares analysis was performed using Eqs. (3.6) and ( 3.18) for substratc
removal and ceU growth respectively. For the purpose of simplicity, X 0 and So were
considered in this study to he error-free (i.e. the readings for X o and So wele considered
as one hundred percent accurate). This assumption probahly affects in somc llIallllCr the
values estimated, but it is believed that this simplification is appropriate at tlli" ~tagc in
the development of this new methodology.
Rearranging expression ( 3.6) for least-squares analysis yields:
(3.19)
where n is the number of substrate-time observations.
Performing the derivative of Eq. ( 3.19) with respect to Y gives:
(YSo + Xo)2(So _7P~L(So+~)t.)] + SoS,(l'So + Xo(So+~Jt.)2

{ (Y So + XoeL(So+~Jt')3
_ (l'So + Xo)[S; + S,(So - L;~t. eL(So+~Jt.)] }
..:..-.----.:..:..-=---~----tx------'-'- (:3.20)
(YSo + x o(So+-v Jt ')2
For biomass Eq.( 3.18) was rearranged for least-squares analysis as follows:
28
(3.21)
(
where m is the number of biomass-time observations.
The derivatives of F2 with respect to A and kd give:
a:; = 2t. r11~ (A;:~:t') [(! -kd) - Y1 ln (A;:t) + ln (~)]

YSoXo (1- eat .) [(~ - kd) - qaln (A~!~:") + ln (t)]
(A + X o) (A + Xoe at .)
- ~' [C! -kd) - 1 y :t) +

ln ( A; ln ( ;- ) ]} (3.22)
F2=2~{(B_k)t2_YSotll (A+xoeat')+tl o)} (X (3.23)

kd f;; A dIA n A +X o 1 n X,
Equating thf' right-hand-side of Eq. ( 3.23) to zero and solving yields the estimate of
kd , denoted kd :
m m m
~t ln 1&1
+L-
Q1
2
ll.'\"t
A~ 1
-
A ~ 1 ln IAxQC
r.&'\"t AX0 , 1
X,1
kd = 1=1 1=1 m 1=1 (3.24 )
Lt~
1=1
Eqs. ( 3.22) and ( 3.24) do not correspond to the SSR of biomass concentration [i.e.
L:(X, -X)2 where X is the biomass concentration from Eq. ( 3.18)], but rather to the SSR
of the logarithms of the biomass concentrations [i.e. 2:(ln(X,) -ln(X))2]. By minimizing
the sum of square residuals of the logarithms (denoted SSRX), it is possible to simplify
the allalysis due to the fact that ka car be expressed as a function of A. By attempting
using 2:(X, - X)2 to estimate SSR, the values of kd and A would have to be estimated
sillluitancously by a numerical method as such as the secant method. This would make
l"
the equations and the numerical analysis more complex.
' '"
29
l.
3.1.3 Aigorithm for estimation of the parameters

1 Fig. 3.4 represents the algorithm developed in this study. The value of L* is varicd from
an initial (Lo) to a final (LJanaI) value with a given increment. To establish the f(,;l! iblc
region for L* (Lo < L* < L lanal ) it was observed that for values of L < Lo, le and ILm
were negativej therefqre, values of L lower than Lo were not considcred. The valut' of
LImai was established when' both SSR's tend to increase when L* increases. 1
The algorithm was cornputerized in a program, written in Turbo PASCAL v. 4.0 [6].
The prograrn creates an ASCII file, whe"eby for each value of L* the estimate of }l' is
found with Eq. ( 3.20). With the same value for L* and the estirnated value of r, the
estirnates of A and kd are calculated using Eqs. ( 3.22), and ( 3.24) (sec Fig. 3.4). Once
the pararneters are estimated, the surn of square residuals of substrate (dcnoted SSRS)
and SSRX are evaluated.
The secant method is formulated as follows:
(3.25 )
where f() is a function of x, Xn and Xn-l are initial estimates, and Xn+l is tht' ne\\'
estirnate.
3.1.4 Estimation of L
Finding an overall minimum by adding both sums of square residuals and plottilJg the
resulting surns vs L, was considered as being not an adequate technique. Thi5 i~ due tu
the faet that Eqs. ( 3.22) and ( 3.24) rninirnize the logarithm of the rnicroorganisfll~ LOB-
centration while Eq. (3.20) minimizes the soluble substrate concentratiOll. ConseqlJ('ntly,
rnostly the sum of square residuals of the substrate would contribute to d('temlll)(~ the
value of L.
1 Except for Run 4 COD, Run 5 COD and Run 5 TOC. See page 55
30
.J
1Begin 1
l
l IL* = Lo 1
Estimate Y by the secant method using Eq. ( 3.20) for

a given value of L*.
Once the value of Y is estimated, the value of A is found

by the secant mthod using Eq. ( 3.22). kd is evaluated
for each iteration of A using Eq. ( 3.24)
With the estimate of A, kd is estimated using Eq. ( 3.24)
After finding the values for all the parameters for a given
value of L*, SSRS and SSRX are calculated and saved in
the output file
[L* = L*+~LI
No
IIfL*=L jmal l
Yes
End
Figure 3.4: Algorithm
As an alternative, the SSR's divided by their respective minimum SSR were plotted
over the total range of values of L. In the following step, the minimum mtersection point
is locatedj this point is defined by the intersection of the two curves that corresponds to
the smallest m~~~R' This point represents the minimum percent that both curves are
"'off" from their optimum fit.
A comparison among the techniques available in the literature and the methodology
l proposcd hercin 15 shown in Table 3.1.
31
1
Table 3.1: Com2arison of methodologies

Reference Mathmatical Comments Paramctel's
technique esfzmated
Reynolds and Linearization Five or more batch reactors }r, kd

~
Yang (1966) are required.
: ~
;
~
Xo, Sa error-free
<
i-
Robinson and Gaussian Minimization of S, - St ILm, !\!, y

Tiedje (1983) X o, So error-free
Time as independent variable
Robinson and Gaussian Michaelis- Menten model Yrnax, !\'m

Characklis (1984) Time as independent variable So, R
-,
Simkins and Marquard Substrate as independent variable X;, So
Alexander (MARQ-FIT) Better fit with logistic model I!.m.
K.
( 1984) eventhough J(! was close to So
Braha and Linearization, Substrate as independent variable JLm, }/

Hafner Marquard X o, So error-free f{!, kd
(1987) Questionable methodology
This study Least-squares Time as independent variable

X o , So error-free
Assumption: f{5 > So
Non biodegradable
substrate
.i
f 32
1
3.2 Initial Estimates
To perform the secant method in Eqs.( 3.20) and ( 3.22) initiaIs estimates of Y and A are
required. Renee, to evaluate these initial estimates and to ensure that the roots obtained
were the optimum values, the following analyses were performed.
(a) Y from Substrate Plots
For a given value of L, ~~ [Eq. ( 3.20)] and SSRS were evaluated as a function of Y.
The results of these calculations are plotted in Fig. 3.5.
3.5 the initial estimate of Y ean be found precisely
*.
By looking at the plots of Fig.
either by finding at the graph the minimum value of SSRS, or the root of Also it can
be seen that only two roots are feasible for ~, one which minimizes the SSRS and the
other that maximizes it (when Y = 00).
(b) A from Biomass Plots
Once the value of Y has been estimated by the secant method through Eq. ( 3.20) for
the fixed L, Eq. ( 3.22) and SSRX were evaluated for different values of A. The plots are
shown in Fig. 3.6.
Looking at Fig. 3.6, it can be seen that only one value of A minimizes the SSRX
and two maximize it (A = a and A = 00, for both of which SSXR -+ 0 ). It is noted also
that. Eqs. ( 3.18) and ( 3.22) are discontinuous at A = O.
In the algorithm the value of L* was varied from Lo to L'anal. Therefore, initial
<'stimates of Y and A for each value of L* were required. Hence, the initial values for Y
{ and A for the Ilcxt L" were the roots obtained for the previous L*. Jt was found that when
the incremcIlt of L* was small (0.000001) these new estimates performed satisfactorily.
33
1
LI..
"0 o
y
CI}
cr
CI}
CI} 0
Figure 3.5: * and SSRS versus Y
34
Discontlnulty at A = 0
Dlscontlnulty
at A = 0
X
Il:
II)
II)
o A
Figure 3.6: !!fi and SSRX versus A
35
3.3 Laboratory Experiments
3.3.1 Sampling
The sample of sewage used in the analysis was a daily composite sample obtained from the
Granby (Quebec) wastewater treatment plant, which is an extended acration activated
sludge system.
A daily composite sample consisted in small samples taken every 15 minutes during
24 hrs in the influent of the basins. The daily samples were refrigerated at 4 oC un ti! the
last of the consecutive sampling days was reached.
The mixed liquor (called sludge hereafter) tested consisted in a grab sam pie frolll the
aeration basins. The composite sewage and the sludge sample were transported to the
Environmental Engineering Laboratory at McGill University immediately after the sludge ~
sample was collected. To ensure that the microorganisms had enough food. the ~Iudgc
(four to five liters) was fed with one liter of the composite sample of sewage, and acrat.ed
overnight at 4 oC in the laboratory. Batch reactor experiments stated the next 1ll0l1liIlg.
3.3.2 Batch reactor
For the initial population of microorganisms, approximately thrce to four liters of the
above sludge were left to settle for approxima.tely 30 minutes, thcn the supcrnatant was
removed. The settled sludge (0.5 to 1 L) was used in the batch rcactor expel illlcnt.
Four to five liters of influent were poured into the batch reactor togcther with the set-
tled sludge, and placed in an incubator (10 or 20 OC). Aeration and mixing were provided
to achieve complete mixing.
The analyses carried out at different time intervals during the expcriments (sec Table
4.10) were:
36
Mixed liquor volatile suspended solids (MLVSS) (triplicates),
Soluble chemical oxygen demand (COD) (triplicates),
Soluble total organic carbon (TOC) (triplicates),
Dissolved oxygen (DO).
MLVSS and COD (closed reflux) were performed following the outlines presented in
the Standard Methods (1976) [1]. TOC was measured using the Dorhman DC-80 auto-
mated laboratory analyzer (Dorhman, Santa Clara, California). DO measurements were
obtained using a Y.S.I. Model 54 Oxygen meter (Yellow Spri.lgs, Ohio).
3.3.3 Experiments
As commented in Chapter 2, two industries irregularly discharge their wastes into the
.\
sewage. Therefore, three preliminary runs were performed to determine the time required
by the microorganisms to degrade the biodegradable substrate.
For the preliminary runs the sewage used was a dail} composite sample of the previous
24 days. These runs lasted for 3, 24 and 30 hrs. From the last two runs, it was concluded
that ni ne hours was the approximate time required for the microoganisms to deplete the
biodegradable substrate.
37
Chapter 4
Results and Discussions
Estimation of the constants was carried out using three different data sets: (1) synthctic
data, (2) data from the study of Braha and Hafner (1987) [7], (3) data from five ex-
periments using wastewater and sludge from the wastewater treatment plant of Granby,
Quebec.
4.1 Synthetic Data
Initially the model used two sets of synthetic data for both substrate and biomas~. 1'wo
sugnificant figuref were used because this represents the accuracy of the me<tslIl'{'mcnt'i
performed in oU'. laboratory. The synthetic data were obtained by fixing thp value" of
the four pararl1eters in Eqs. (3.6) and ( 3.18) and assuming values of 50 and X Theil
the sub~trate and biomass concentrations were evaluated at diffel'ent times. Both the
synthetic data and the estimated constants are enclosed in Tables 4.1 and 4 2.
The estimated values of 1(, and Pm are 7.2 and 0.36 percent "off" of the original COIl-
stants, for SI and S2 respectively. Nonetheless, these new estimates yicld smallcr SSIt'~,
for both biomass and substrate, than the original parameters.
38
Table 4..
l' Synthetic data
Time (hr) SI S2
COD MLVSS COD MLVSS
0 100.00 1000.00 100.00 1500.00
0.25 94.57 1005.37 86.83 1505.33
0.5 89.40 1010.34 75.30 1509.00
0.75 84.48 1014.94 65.24 1511.21
1.00 79.80 1019.17 56.47 1512.13

1.25 75.34 1023.05 48.85 1511.93
1.50
. 71.11 1026.59 42.22 1510.74
1.75 67.10 1029.81 36.48 1508.70
2.00 63.29 1032.71 31.49 1505.91
2.25 59.67 1035.31 27.18 1502.49
2.50 56.25 1037.63 23.45 1448.52
2.75 53.01 1039.68 20.23 1494.08
3.00 49.94 1041.46 17.44 1489.24
3.25 47.04 1043.00 15.03 1484.06
3.50 44.30 1044.31 12.96 1478.58
3.75 41.71 1045.38 11.16 1472.86
4.00 39.26 1046.25 9.26 1466.94
Table 4.2: Original and estimated parameters for the synthetic data analysis
J(~ /lm y kd SSRS SSRX
( ~,) (hr-I) (mg biomas~) (hr-t)
m ~ub~trate
SI 3000.0 1.00 1.500 0.010 1.53 xl0- 4 1.44 X 10- 10
Estimated 1 3226.5 1.07 1.495 0.010 1.52 xlQ-4 1.06 X 10- 10
S2 4000.0 1.50 1.000 0.020 1.55 xlQ-4 5.16 X 10- 11
Estimated 2 4014.3 1.51 1.000 0.020 1.48 xl0- 4 4.08 X 10- 11
Figs. 4.1 and 4.2 show the feasible roots for Y and A. Looking at these plots the
same shape can be seen as those presented in Chapter 3, Figs. 3.5 and 3.6 (W vs Y
and ~ vs A). During the execution of the computer program, the values of Y and A
COIlvcl'ged, and SSRS and SSRX were minimized (Y, A, SSRS and SSRX were displayed
on the screen during the execution of the program).
39
S1
20
1 0
-20
-40
-so
-10
-100
-120
~ -140
...
~
-110
-110
-200
-220
-240
-260
-210
-lDO
0 2 1 1
Y
-1
-2
~
~,l,
.....
-3
,2-
~
.- E -.

-5
-6
-7
-1
-0.2 0 0.2 04 06 0.11
(Th .......... )
A
ay versus Y and ffi

Figure 4.1: Synthetic 1: .El aA versus A
4.2 Braha and Hafner (1987) Data Sets
In the study carried out by the above authors, seven experiments in batC'h reactors wer<'
performed to assess their methodology. Braha and Hafner l'an in parallel a complete
mixing, continuous-flow reactor for a period of approximately six months to providc ac
climated sludge for the batch experiments .

.... In the continuous-flow reactol', microorganisms were Ced with industrial W.l.stcwatcr
40
--------------- ------
52
10
'0
40
lO
20
10
-10
-20
~...
-lO
-40
-'0
" -10
-70
-80
-90
-100
-110
-120
-130
-140
2 4 1
Y
0.5
-05
-1
;n -1 :1
.!.
~2
....
..
f "
e
-2
-2.5
-l
-l.5
-4
-45
-200 0 200 400
,.
Figure 4.2: Synthetic 2: ffi

Y
versus y and BF2
BA
versus A
containing organic pollutants at different hydraulic retention times (4 - 17.5 h). For each
hydraulic retention time, the process was operated between 15 to 22 days to achieve
quasi steady-state (transitional periods 7-10 days not included). For each continuous-flow
experiment, samples were collected and a batch experiment was l'un on each of these
samples. The results of the seven batch reactors are shown in Table 4.3.
The methodology was applied to the seven data sets. The resulting graphs of the SSR
41
f; $.1 " ~ ...
Table 4.3: B!aha and Hafner (1987) Data Sets - ----
Expenment Analysis Time (hr)

0 0.25 0.5 1 2 3 4 5 6 7.5 10 12.5 15 24
LBI TOC !!!!l.
1 277 1 270 266 249 222 198 172 158 144 119 75 65 48 19
MLVSS T 5880 - - 570 - - - 5918 - - 6042 6075
- 6105
LB2 TOC !!!S.
1 281 272 264 249 221 197 174 154 141 119 79 61 46 20
MLVSS 7 4310 2 - - 4265 - - -4390 - 4481 - . - 4540 4596
LB3 TOC~
1 319 308 298 281 248 222 195 173 155 126 96 72 51 36
MLVSS 7 4190 - - 4148 - - -4319 - - 4412 - 4458 4488 .
LB4 TOC !!11l
1 296 286 278 261 233 207 184 161 142 121 88 60 49 33 1
AILVSS T 4020 - - 3970 - - -4128 - - 4246 - 4279 42931

LB5 TOC !!!S.
1
281 272 263 250 223 199 183 172 - - 92 58 5] 27 1
MLVSS~ 3680 - - 3641 - - -3756 - - 3861 - 3929 3948
LB6 TOC !!!S.
1
273 - 255 244 219 203 186 - 131 -118 84 50 42 19
~
l'\j
MLVSS~ 5020 - - 4929 - - - - - - 5150 - - 5237 5260
LB7 TOC~
1
228 220 217 197 178 158 119 97 85 77 63 55 38 -
AfLVSS 0/ 3880 - - 3821 - - - 3997 3 - - 4030
------ -----
4084
- - -
- -
Il 7 in the paper.
z430t3 ln the paper
34097 in the paper, beheved t be 3997.
ratio, and of the fitted substrate degradation and biomass growth curves (both obtained
with the parameter estimates) appear in Figs. 4.3 to 4.9. Table 4.4 shows the feasible
region of L and the estimates obtained.
In the computer program, the value of kd was constrained to be non-negative in any
iteratiol1. To constrain the value of kd , a value of zero is assigned to it if it is lower than
zero from Eq. ( 3.24) 'during the search procedure for A (see algorithm Fig. 3.4).
Table 4.4: Estimated parameters with Braha Lnd Hafner (1987) data sets
Expe7'iment Lmm Lmax K5 J.tm y kd L
1
mQ hr
1
mq hr
!!:.S.
1
hr- J m!lMLVSS
m~Q TOC
hr- 1 1
m~Q hr
LBi 0.000015 0.00020 297.53 0.0068 1.192 o+ 0.000023

LB2 0.000026 0.00010 4868.41 0.1266 0.958 ot 0.000026 t
LB3 0.000027 0.00030 216127.90 5.836 0.961 ot 0.00002i t
LB4 0.000029 0.00016 12777.22 0.3705 0.993 ot 0.000029 t
LBS 0.000029 0.00038 3215.28 0.0965 1.017 ot 0.000030
LB6 0.000020 0.00100 36.61 0.0037 4.805 0.0003 0.000100
LB7 0.000033 0.00073 12041.24 0.3974 0.922 ot 0.000033 t
t L = Lman t Constraint WOlS active
The constants obtained are shown in Table 4.4; the results vary greatly. Eventhough
the sludp:c age varies among the experiments, this could not account for the differences
in the parameters estimated. On the other hand, the shape of substrate degradation
and biomass growth curves are similar for the seven batch experiments and should yield
similar values for each of the parameters.
It IS notcd in six (6) out of the seven (7) SSR ratio graphs (Figs. 4.3, and 4.5 to 4.9)
that the m~:;Y:U plots reach a plateau with shallow slope at certain value of L, denoted
here as Lp. For values of L < Lp, the constraint on kIJ was active (i.e. kd < 0 -+ kd = 0),
while for L > Lpl kd was > O. For LB2 the value of k i remained constrained throughout
the fClslblc Icgioll of L.
Silllil,\! to the s)"nthetic data analysis, the partial derivatives of FI and F2 VCI sus }~
43
lB1
1.1:1
~.
1.14
;+
;"
.+
1.13
'r
i
1.12
c
1.11

~ .+
1.1
~ 101

III
4
1.011
107
~ +
tl 1.01S
f 1.D!5
+
1.04
R
1.03
1.02
+
1.01
1
000001 O.OOOOl 0.0000' 000007 000009 000011
L 810 ...... _
Subatnlt. +
Biomass
&.12
&.1
&.08
&.05
&.04
&02
li
'.98
~~
"'"
EC:
~::
U5
'.94
en"
~
%
592
'9
SB8
SB6
'.84
, 82
'8
'.78
0
'.75
0 12 16 24
trno (hr)
Substrate
280
250
240
220
200
180
Q
160
~
! 140
8... 120
100
80
50
40
20
0
0 , 12 \6 20 24
trno (hr)
Figure 4.3: SSR ratio, biomass and substrate graphs for LBl
44
'J
LB2
1.5
l ..,
i
c:
~
1.4
~
t!
4 I.l
~
la
i 1.2
~
1 1.1
0.00004 0.00006 O.OOOOB 00001

0.00002
L
- Substrat. + 1........
Biomass
4.5
4.'8
4."
4'4
4.'2
4.5
4.41
., ."
~-;'
ec:
'"'l:
4.46
444
~l
442
.... 4.4
%
411
4.l6
434
412
4.l
4.28
4.26
0 12 \6 24
tme (tir)
$ubstrate
lOO
280
260
240
220
200
110
~
.s 160
140
~ 120
100
la
60
40
20
0
,
'1 0 1 12 \6 24
~ t.rn. (tir)
Figure 4.4: SSR ratio, biomass and substrate graphs for LB2
45
'J
LB6
1.5....-----------------l
1.1
0.0003 000034
00001 000014 0 DOOn 0.00022 0.00026
000002 000006
- - Substrat.
Biomoss
5.l-r-----------------,
525
52
4.95
D
'9J---,---,----r--~a~-,--~12---.---:.,6~-.---:~-.~~2.
o
lino (h.)
Substrote
2ao~----------------------------------------~
25C
240
220
200
IBO
~ 160
.g 140
~ 120
100
aD
&0
40
20
a 12 16 20 24
o 4
'J. lino (IIr)
Figure 4.8: SSR ratio, biornass and substrate graphs for LB6
49
.j
1.7
LB7
l
\.5
ic 1.5
~
~ \.4
t!
4
I! \.3
~
l 1.2
:.:r
i
\.\
1
0.00002 000006 0.0001 000014 000018 000022 000026 00003 000034
L
- Su~tnrte + 8iG....
Biomass
4.1
4.08
4.01
4.04
4.02
~';'
..... 3.98
e c
"'l: 3.96
111"
~~
::lE
J.94
3.92
3.9
J 88
J.86
J.84
3.B2
a 2 4 10 12 14
lft-.. (h')
Substrate
210
220
210
200
190
IBO
170
160
150
~ 140
5 IJO
8
~
120 c
110
100
c
90
Ba o
--
70
60 c
______ c
50
40
la
a 5 la 12
lft-.. (h')
Figure 4.9: SSR ratio, biomass and substrate graphs for LJ37
50
and A respectively (see Figs. 3.5 and 3.6) were evaluated at the minimum intersection
( point (i.e. lowest intersection point in the SSR ratio graph)j the shapes of the partial
derivat.ive.'> show a single root. The plots are presented in Appendix F.
Table 4.5 shows the original constants estimated by Braha and Hafner (1987) [7].
The table show the SSRS with the parameters found in the original study, and the SSRS
obtained by the methodology used herein. It can he seen that the sum, of squares ohtained
in this stl:dy are smaller than those given by the original study.
Table 4.5: Parameters estimated hy Braha and Hafner (1987)

Experiment Y J(~ Pm 4 SSRS SSRS SSRX
ms. MLVSS !!!S. hr- 1
Braha and This study 10- 3
mg TOC 1
Hafner (1987) [7]
LBl 1045 14000 00400 223.2 220.7 2.50
LB2 1.35 13000 00482 103.4 93.8 5.17
LB3 1.37 13000 0.500 509.4 442.7 7.11
LB4 1.38 14000 0.568 384.1 352.7 7.13
LBS 1.56 15000 0.776 1096.4 341.1 7.26
LB6 1.42 16000 0.548 725.3 413.8 3.66
LB7 0.78 14800 0.383 1490.1 1125.9 3.41
4.3 Data from the Granby Plant
The Granby wastewater treatment plant treats the sewage from the municipality of
Gran by, Quebec. It is known that two industries (cheese processing and textile) dis-
charge their waste irregularly into the city sewage. The organic load from the cheese
processing industry is approximately one third of the total organic load of the sewage.
The load from that industry is mostly in the form of organic suspended solids.
The wastpwater treatment plant consists of an activated sludge pro cess that is op-
t'rated in the extended aeration phase. Theoretically, th,~ extended aeration process is
4The Ulllts 10 the orrgmai study are d- 1 , but upon our calculations the units of Jl.m are hr- 1
51
designed such that aU substrate removed is converted into energy metabolism and oxi-
1 dized, so no ex cess biomass is produced and sludge handling is eliminated. Hencc, the
rate of growth can be written
(".1 )
.
In other words, aIl the biomass produced is theoretically oxidized and metabolizcd by
microorganisms. This assumption was proved by Gaudy et al. (1970 [12], 1971 [lI}, 1974
[35]), and Obrayashi and G \.udy (1973) [22].

The plant consists of six stages as indicated in Table 4.6.
Table 4.6: Characteristics of the Granby wastewater treatment plant

Stages , Pumping statwn
, Sand removal
, Screening
, Aeration basms
, Secondary clanfier."
Ch/orinatwn
Equivalent population 126000
3
Inflow average 50000 dBy
m
Aeration basins (4) 25693 m3 total

Average hydraulic retention time 12.3 hr
Sludge age 40 days
4.3.1 Experiments
Initially, three preliminary experiments were performed at 20C in order to monitor the
behaviour of biomass growth and substrate degl'adation. From thc!>e exrcriment~ th('
presence of non-biodegradable substrate in the sewage was obscrvcd. lt is bc]ieved that
most of the concentration of non-biodegradable substrate cmanated from the textile ill-
dustry. Also, it was noticed that thp. degradation of the substrate was achicved within
nine hours or less. Nevertheless, substrate measurements wcre obtaincd in Runs l, 2 and
52
.5 for 2.5, 24 and 48 hours after initiation of the tests, respectively, but the concentration
levels of substrate did not decrease after the tenth hour.
The (ive experiments were performed in the following dates: June 28, 1989, August
23, 1989, December l, 1989, February 3, 1990 and February 10, 1990. Unfortunately, the
conditions of the biomass in the plant changed considerably from experiment to another.
In the last week ~f July 1989 and first three weeks of August, major repairs \Vere
perfOl med in the apration system of the basins. During that period, once a week, . dif-
fcrent tank was emptled for such repairs. This forced the biomass to adapt to the new
condItions, which were different than those in the first experiment.
During the second week of November 1989, sludge bulking occured in the plant.
Bulking is the result of preferential growth of filamentous baeteria among the various
(non-filamentous) speeies of microorganisms normally present in the aeration tanks. The
filamcnlolls bacteria usually hinder the settling of flocs in the seeondary clarifier and, con-
..
sequently, large amounts of biomass is lost in the effluent. To extinguish the filamentous
bactel iJ, the basins were chlorinated during the fourth week of November. As indicated
previously, the third experiment was started in December 1, 1989 using samples (i.e. chlo-
1 inatco) from the fourth week of November.
The last two experiments were similar between them, but climatologie conditions (i.e.
much lower operating temperature) were considerably different than those in the first
th rel' batch 1 li n!>
The ,>ewag<. tcsted presented a certain amount of non biodegradable substrate, and the
mode! llscd ;n this study (Monod kinetics) consid~rs only the biodegradable substrate.
lIenec a I1Icthodology is developed to assess the amount of non-biodegradable substrate
ill tlll' sdll1ples.
The ITldhodology consits in giving different concentrations of non-biodegradable sub-
..,trelte ,\Ild thpll subtraeting this amount from the total substrate (measuremcnts). Then
53
the sum of squared residuals is obtained following a negative exponential trcnd mode!.
The model proposed is given by the following equation:
where S, is the biod~gradable substrate at time ti (i.e. total substrate mlJlus nOIl-
biodegradable) and bis a constant. Rearranging for least-square analysis it yiclds:
n
Fo(b) = ~)Soe-bt, - S,)2 (<1 :1)
1=1
Performing the derivatives with respect to band rearranging gives:
F
_0
b = 2S 0
{n0
~t
S L- ' e- 2bt , - n S e- }
~t
L..J 1 e
bt ,
(1.4 )
1=1 1=1
where bis estimated by the secant method when the derivative in Eq. ( 4 4) is set to be
equal to zero.
The Ilon-biodegradable substrate estimate is varied over a wide range. The mo(l!1
yields one minimum SSRS for each substrate-time data set, which was con"'ld(>ICU tu 1)('
the non-biodegradable substrate, although this concentration did not corr('.,polld to II\('
last substrate concentration measured. The computer program is glvcn in Appcndix ('
The sludge age of the biomass on the last day of sampling, the temperature at which
the experilY'~nts were carried, and the nonsoluble substrate ale pre!:>cntcd III Tdb!c .,
Results of the five experiments are presented in Table 4.10.
Table 4 ..
7' Characteristics of sludge and substrate for each experiment
Run Sludge Age AfLVSS SVI F/M Temperature N on-bzod cgmdablc.
days !!13.

!E.'l
J
day-l oC COD (T) TOC (7)
1 35 - - - 10 44.31 :n O
2 46 3007 57 - 20 63 ..58 2808
3 48 2300 59 0.10 20 98.00 ~2.20
4 43 2316 71 0.11 10 123 87 19.24
.5 40 2316 71 0.11 10 132.07 .56.DG
54
Contrary to the analysis with the data sets from Braha and Hafner (1987) [71 paper,
the value of kd was always positive, therefore no constraint was needed.
On the other hand, L fmal was constrained to values su ch that the corresponding l(~
were larger than 12. This constraint was considered due to the fact that large values
fol' A's <tIC cxpf'cted (Simkins and Alexander (1984) [28], Braha and Hafner (1987) [7],
Uloth alld Manavic (1977) [32] and Lawrence and McCarty (1970) [17]). The constants
cstillliLLcd and the plots for each data set are shown in Table 4.8 and Figs. 4.10 to 4.19,
respectively.
Table 4.8: Estimated parameters for the Granby Plant data sets
Run Lmm Lmar J(~ J1.m y kd L
1
1
mq hr
1
mq hr
!!!S.
1
hr- 1 m2\fLVSS
ma Substrate
hr- l
mq .'Ir
1 COD 0.000149 0000395 637.0 0.103 0.268 0.002 0.000161
TOC 0.000423 0.001930 1313.6 0.552 1.092 0.002 0.000423 t
2 COD 0.000141 0.000540 2783.0 0.392 0.416 0.00.5 0.000141 t
TOC 0.001410 0.006310 3464.1 ~.884 1.284 0.006 0.001410 t
-
:3 COD 0000:312 0.000761 883491.0 275.650 0.606 0.009 0.000312 t
TOC 0002490 0009490 2757.8 6.867 4.577 0.011 0.002490 t
.,.
4 COD 0.000082 0.000422 338.3 0.032 0.081 0.002 0.000094 +
TOC 0000280 , 009380 1802.4 0.505 0.704 0.00:3 G 000280 t
5 COD 0.00011 0 0.003240 14.1 0.006 0.395 0.002 0.000420 ~
+
TOC 0000800 J.004100 155.1 0.132 0.583 0.001 0.000850 +
t L = Lmm + Constraint was active
Tahle 4.9: Ratio of soluble to total substrate concentration (COD) at Granby

m~an standard devzatzon number of analysls
Influent 0.3?14 0.0646 10
Eillucnt 0.7439 0.1284 1.5
Analyzing the ploU, of runs 4 and 5 (COD), it can be seen that the phenomenon
of contact stabilization may have taken place in our batch experiments considering that
f Illost of the mcasurcmcnts after the 6 th hour are above the substrate predicted and the
1
'1
1
.55 1
j
- ----- 0 _ _ i Ji , uza'
s-";
....
Tahle 4.10: Snb.,t rate and biomass data for runs 1 to 5

Run 1 Run 2 Run 3 Run ..( Run 5
Time COD TOC MLVSS COD TOC MLVSS COD TOC MLVSS COD TOC MLVSS COD TOC MLVSS
/zr ~ ~ ~ ~ ~ ~ !!.!9. !!.!9. !!.!9. ~ !!.!9. !!!1l. !!!1l. !!!1l. !!!1l.
T T T T T T T T T T T T T T T
0 121.0 46.4 1830 160.0 62.27 1421 212.5 39.93 651 271.0 73.74 770 242.3 75.13 909
0.5 73.0 37.Cl 1818 120.8 - 1156 180.0 4?.l 702 237.0 - 771 209.4 76.3 918
1 68.0 34.1 1857 120.0 33.97 1510 171.0 30.2 685 205.0 69.8 781 - - -
1.5 69.0 30.9 18n 125.0 31.18 1406 151.5 32.0 671 146.3 54.44 776 165.2 50.01 926
2 63.0 30.7 1862 100.0 31.78 1429 156.5 29.l 670 133.5 55.83 780 135.2 50.48 910
3 - - - 80.0 30.55 1468 145.0 28.8 6SS 126.5 61.24 772 128.4 58.36 908
-1 - - - 69.0 27.67 1427 133.0 25.7 &65 115.1 70.1 766 121.3 50.63 912
CJ1
Cl 4.5 42.0 24.3 1812 - .- - - - - - - - - - -
.5 - - .- 83.0 29.5 140tl 130.0 29.3 666 - - - - - -

5.5 - -- - - - - - - 110.4 53.94 772 135.8 61.12 918
6 42.0 23 ..5 1839 - - - - - - - - - - -
7 - - - 70.9 28.9 1:388 104.0 21.9 660 134.0 48.53 768 145.0 58.34 907
S - - - - - 970 23.7 679 - - - - - -
$5 - - .. .. - 132.1 49.04 2 126.6 53.22 904
9 - - - 9q ~ 21..1 666 - - - - - -
10 10.0 22.2 lK12 58.0 2.3.12 lt 10 - - 112.0 51.00 no 139.2 60.67 916
11.5 . - - . - - 128.8 49 10 765 137.9 .59.23 904 :
RUN1
1 DOS
10011
l i
C
1.007
~~
1006
1 DO!!
4
&1 1004
t1
!l 100J
E
~ 1002
1.001
000014 o OOOIS 000022 0.00026 o OOCl 000034 000038
Subatnrte
L
.. !hon.o
Biomoss
1.9
189
1 Sil
I.S7
186 C
0
1 S5
~~ 1 S4
"'''
EC 1.83
v~
1/1" I.S2
c
~~
C
::E I.SI
'l'
1S
'. 179
17S
177
1 76
1 75
0 2 4 ID
t..... (hr)
Substrote
130
120
110
100
90
~
BD
.s
0
70
0
0 60
50
0 0
40
3D
20
ID
0 2 4 ID
~ liN (hr)
" Figure 4.10: SSR ratio, biomass and substl'ate graphe;; for l'un 1 (COD)
57
Figure 4.11: SSR ratio, biomass and substrate graphs for run 1 (TOC)
58
RUN 2
108
COD
1 07
ic
1 06
~ 1.05
~
~
~ 1.04
1/1
~
~
1 Dl
~
;;;. 1 02
~
101
1
00001 00002 o OOOl 00004 00005
L
Subolnll. + 810"",.
Biomass
1 ~6
1 ~4
1.~2
15
148
,,...,........ 1.46 0
0
e c:
~::
1"
"'~ 1 42
~. ~
::t 0 0
1.4
0
1 lB
1 l6
134
1 l2
13
8 la
!me Chr)
Substrate
180
170
150
.,0
140
IJO
120
~
.s
Q
110
100
0
V 90
80
70 0 0
60
~o
40
JO
6 la
tme (hr)
Figure 4.12: SSR ratio, biomass and substrate graphs for run 2 (COD)
59
RUN J
1.22
-1
121
1.2
1.19
)(
1.la
~
1 17
1 16
c 1.15
{ 1.14
~ 1.13
~
. 1 12
1.11
Il
X1
1.09
tl 1 oa
~ 1 07
;;;. 1 06
~ 1 05
1 04
1 03
1 02
1 DI
1
00006 o ODOS
00003 00004 0.0005
L
- Subtltnrte + 810""'.
Biomoss
740
730
no
710
700
<:>
"-
co
690 0
0
4' t 6S0
0
1 ~
~
670
0 C
0
660
650
640
630
620
0
tme (hr)
Substrote
240
2JO
220
210
200
190
ISO
,.
<> 170 0
g 160
150 0
Q
0
0 140
130
120
110
0
100 0
90
SO
70
t 0
trne
4
(hr)
6 a
Figure 4.14: SSR ratio, biomass and substrate gl'aphs for l'un 3 (COD)
61
RUN 4
COD
I.J'
I.J2
( I.J
1.28
i 1 25
12'
"
~ 1.22
~ 1~
t'! 1.18
4
fil Ils
0:
t'! 1.1'
1 12
"
;,. Il
~
1 08
1 06
la'
1.02
1
000005 000015 000025 0.000J5 0000"
L 81a .... _
- Substrole +
Biomoss
790
785
a 0
780
C
<> 775
"- c c
~
.s'"
fil
770

il ~ c
:1
765
760
755
750
5 la 12
trM Chr)
Substrote
300
2BO
260
2~
220
~ 200
.s
Cl
0
IBO
U
160
1~ a
c c c
C
120
0
C
100
f
80
a , 5 8 10 12
" \me Chr)
Figure .t.16: SSR ratio, biomass and substrate graphs fol' l'Un 4 (COD)
63
RUN 4
TOC
1.22
1 12
I.IB
i
c
1.15
~ 1.14
~
..tI 1.12
~ Il
tI
lOB
~
~ 106
~ 1.04
1 02
0 0002 0004 0006 o OOB 001

L
- Subslnlte + 810""'_
Biomoss
790
7B5
C
780
775
~
... C
!
... 770
~
:E C ~~
765
760
755
750
0 2 4 B 10 12
lrne Ch')
$ubstrote
100
90
BD
70
~
! 60
g
1-
50 C C
40
JO
~ 20
0 2 4 6 B 10 12
...t trr. Ch')
FigUl'e 4.17: SSR ratio, biomass and substrate graphs fol' l'un <1 (TOC)
64
RUN 5
COD
122
l 1.2
1.111
1
c
1.11

1.14
itI 1.12
~ Il
t'I
c 1.011

~
1.06
1.04
1.02
0 0.0004 00008 00012 0.0016 0.002 0.0024 000211 o 00J2
L
Substnlte + liII""'.
Biomoss
~Jo
c
925
920
C C
C
915
~
!'"
C
-d
910
~
::E
a
905
D D
900
1195
1190
0 2 4 5 ID 12
ti11e (hr)
Substrate
2So
270
260
250
240
2JO
220
210
~ 200
! 190
Q
0 1110
u
170
160
150
a
140 a a
a a
IJO a 0
120 0
110
100
'", 0 2 4
ti11e (hr)
5 1 10 12
Figure 1.18: SSR ratio, biomass and substrate graphs for run 5 (COD)
65
RUN 5
TOC
1.17
1
1.111
1.115
1.14
1 <:
1.13
1.12
~~
1.11
1.1
..
I!
1.01
1.01
~ 1.07
<: 1.06
E
~ 1.015
i~
1.04
1.03
1.02
1.01
1
0.00011 00012 00016 0002 o OOH 000211 00032 00036 000'
L
- - Substrnle + 1110 ......
Biomoss
930
D
925
120
D D
..
~
E
'-'
Sl5
910 a
~
%
905
0 0
900
B95
B90
0 4 10 12
tme Ch')
Substrote
100
90
110
a
70
Q
~
! 60
D
C
0
C
0
8
1- D
50 o D 0
40
JO
20
2 10 12
tme (h,)
Figure 4.19: SSR ratio, biomass and substrate graphs for l'un 5 ('fOC)
66
Table 4.11: SSR's obtained with the Gr anby Plant data sets
l Run SSRS SS RX
10- 4
"
1 COD 483.81 5 .93
TOC 8.54 5 .97
2 COD 839.53 20.1
TOC 37.90 3 9.2
. 3 COD 624.34 24.9
. TOC 72.52 6 4.9
4 COD 2251.76 3.0
TOC 366.99 3.7
5 COD 839.19 5.2
TOC 335.14 4.4
fc\\' Illcsurements prior to the 6th hour are below the substrate predicted. Ratios of sol-
uble t total sllbstrate concentration (COD) were obtained from tests performed by the
persone! of the Gran by wastewater treatment plant during the months of November 1989
clnd ,Jcl/luary 1990. The results are presented on Table 49 which indicate that contact
''1.
slctbilization was probably taking place in the Granby plant.
l\Clthcr the differences of the characteristics of sludge and substrate in each experi-
l11Cllts liaI the contact ~tabilization phenomena could account for the values of the pa-
ranll'ters c5tlfnated in this study. The parameters are far from being simlilar, except kd
(Ind } (in SOITlC cases). It is known also that the differences in tempe rature during the
pel formancc of the experiments affect the parameters estimated; however it is Bot possible
ta e\'lluate prccisely such effects.
~~.! vs } - and ~/~; vs A \Vere evaluated for each run and only one foot was found in
each UISC (the plots are shown in Appendix F). As in the previous analyses, Y, A, SS' R.','
and SS'RX werc di5played on the screen of the monitor and it was observed that while V
(Illd A l'llll\Clged, SS RS' and SSRX \Vere minimized. Substrate degradation and biomass
gJl)\\,th were weil fitted in ail the plots, Table 4.11 shows the SSR's obtained for these
dat a sets.
67
In summary, the analyses carried out using data sets from Braha and Hafner (198i) [il
1 and the Granby Plant have yielderl values for 1<3 and !Lm that are far flOm bt'inp; simil.1I
betwecn them. Nevertheless, the estimated values for Y and kd were snnilal lnd \\'Ithlll
the values commonly found in the literature. The main concern rcmaillS in 't.he l'!>timdtioll
of /{s andwhich seems to be a very difficult task due to the high H!e!atiun bl'l\\"('('ll
Jlm

these two constants [see Robinson and Tiedje (1983) [27] and :\ppcndlx A].
On the other hand it is known that the minimum sun! of square resldllals is t 1)('
criterion which has to be satisfied in numerical approaches for kinetic est; rllation III t I\('
chemical and environmental fields. This study has shown the missesttnmtioll ln pn'\'IOIIS
studies of the Monod kinetic parameters which did not mect thi.., niterIOll [BI(lha alld
Haller (198/) [7] and Simkins and Alexander (1984) [28] (~lonod \\ith g\O\\th)] The ".,-
sessment developed herein has shown (Appendix F) that tlw 1lIl1limum SSH 's wa" Illet ill
every data ~,et analyzed. Unfortunatcly, the estimates of A. and !lm WCH' Hot (,Oll!->Istant.
and several times out of the values commonly found in the literature.
I
68
Chapter 5
Summary & Conclusions
To properly design a wastewater treatment unit, it is recommended to knO\v the
l'ales of su bslratc: uptake and biomass growth. The best known model to Pledict
these rdles IS the Monod kinetic model (which contains four parameters). The
estimation of Monod kinetics can be carried out either by continuous flow 01 batch
reactors. It is known that batch reactors is the best alternative in terms of time
needed to pel form the experiments .
It was lemarked herein that the parameters estimated in the studies by Braha and
Hafner (1987) [7J and Simkins and Alexander (1984) [28J (Monod with growth) did
not conespond to the minImum SSRS. Therefore there IS a need to develop a ne\\'
approach, to estimate the four parameters from batch reactor data sets, that meets
th", minimum SSR cnterion. Optimum estimates of parameters are obtained whell
this critcrion is met .
The Ilonlinear least-squares methodology presented herein, fits substrate utilization
and biomass growth curves simultaneously. The secant method was used to solve
the nonlinear Icast-squares equations.
69
The methodology developed in this study \Vas applied to three diffclent dat.a sets:
(1) synthetic data, (2) data from the study of 8raha and Hafner (19S7) [7], (3) data
1
from five experiments performed with wastewater and sludgc fro111 t.he Wd"t('\\',tI ('1
treatment plant of Granby, Quebec. The values found of f{$ dIld JI", wlth delta
sets (2) and (3) were dissimilar among them, and sorne of thcm are Ilot wit.hiIl tl\('
values cornmonly found in tht literature. lt is believed that the 1l1l(OIlCl'it.l'IHy of
the estirnated values for these two parameters is due to the high COI relat iOll bl'l.\\'('('11
them.
This assessment appears to be the first to estimate the four Monod-klllctic p,t!,lIn-
eters from a single nonlinear least-squares formulation. by fittmg silllltlL-IIl('oll"ly
substrate uptake and biomass growth.
Based on the criterion that species of mieroorganisms depletmg slrllllar ~lIb . . tl(ttl'
under similar envl;onmental conditions (pH, oxygen, tempclature, Lee. ) ~h()\Ild
yield about the same values for the parameters, it appears to he qucst.ionabk t.u
use this ne\\' methodology for the estimation of the four MOIlod-kinctlc!'> plI illlH't('I"
before more studies are performcd.
The minimum SSR is a eriterion tnat needs to be fulfilled to obtain the optlllllllll
estimates, which has been achleved with this new methodology
However, obtainmg accu rate estimates of the parameter5 seelll~ t he ct VCly cldli( 1Iit
task given that the response surface of the SSICs functioll scems to be quite fiat III
the vicinity of the minimum SSR point, as shown in Appendix A.
The parameters obtained in this research work cannot be cornpalf'd directly with
the values estimated in previous studies [e.g. Braha and IIafner (1087) [7JJ <'IIIU' \V('
have shown that these values do not fulfill the critcrJon of minimum SSR
70
Further studies need to be done in this topie, for example by running in parallel
1 continuous-flow and batch experiments with synthetic waste and comparing the
constants estimated in both systems. Also, different numerical approaches need to
bc attempted that could possible handle the high correlation among the constants.
71
1
Appendix A
Sensitivity Analysis
The analysis presented herein shows the existence of several sets of parlllleters whl( h
yielded smaller SSR than the parameters estimated by Braha and Hafllel (19R7) [] ill
thelr study.
A.l Introduction
This study consists in a sensitivity analysis which gives different values to each cou..,t,LIIt
and then in calculating the corresponding sum of squares residuals (SSR) wlth resp('ct to
the substrate, which is defined as:
n
S'S'R = 2)St - SI)2 (A 1)
1=1
where
s, = measured substrate concentration,
St = calculated substrate concentration, as obtained with Eq. ( 221)
72
1
,-
A.2 Model and Application
SSRS were evaluated by giving an initial and a final value for fJ-m, Y and J(! with their
respective increments. The computer program calculated the SSRS for each possible
cOlllbination of variables .
.
Duc to the [act that a value of St is reguired in Eg. (A.1) and since Eg. (2 ~1)
canllot be written in ter ms of substrate as a function of time, the St value was found via
the ~cCtnt mcthod. The InItiaI guess value was the corresponding S,.
Braha and Hafner (1987) [7] proposed a methodology in which the yalues of Y and
kd were round USIng Eg. (2.18) and the values of /{! and fJ-m were estimated through
Ilollltllear lea!>t-squdfes analysis with Eg (2.21).
l1egardlcfls of wbat these authors stated, and analyzing Table 2 in their paper, it seems
that the value of Y wa~ also estimated simultaneously with Ks and fJ-,n vIa nonlinear least-
'>quarf'S lnalysls, alld not usmg Eq,( 2 18) to estimate Y a~ It was established.
Due to the fact tbat we could not assure what methodology was applied, two analyses
\\"CI e pcrformed The first consisted In varying the value Y and the second used the value
obtailleu \Vith Eq. ( 2.18).
Inltially a SSRS \Vas computed using the parameters estimated by Braha and Hafner
III their study \Vith the first ex periment (f{s = 14000 mq~OD, fJ-m = 0.4 hr- I and Y =
1 15 Hl7~,'~~~~~~/I) The SSRS [('und \Vas 223.188; the proposed inItial and final values for
crlch of the pal amctcrs and their increments are presented In Table A.l. The outcome
Tahle A.1: Parameters studied

parameter initial value final value increment
/(3 100 20000 100
}" 0.5 4.1 0.1
Ilm 0.1 2.1 0.1
of this lllalysis gave 855 different combinations that yield sm aller SSRS than the one
73
proposed by Braha and Hafner (1987) [7]. The SSRs vary from 223.187 to 207.585. The
l values of the parameters that yield the thirteen smallest SSRSs are presentl-d III '1'<11>1(,
A.2.
Table A :~: Thirteen smallest values of the SSRs for tlH' first analysis
y [{~ J.lm SSR li.....
J'rn
2.8 9300 05 207.748 1~600
2.8 11200 0.6 207.791 1S667
2.9 10800 OC 207.707 18000
2.9 16300 0.9 207.702 1S111
3.0 14000 O.S 207.621 17500
3.0 19300 1 1 207 795 17515
3.1 15300 0.9 207.620 17000
3.1 17000 1.0 207 .5~5 17000
3.1 18700 1.1 207 721 17000
3.2 16500 1.0 2075SB 16.500
32 19800 1.2 207 :?6 16,500
3.3 14400 09 207.727 16000
3.3 16000 1 0 207.702 16000
3.4 18700 1 2 207.684 1,1.183
In the second analysis the value of}" was set to 1.'1.5 (since this was thc vetlue c~tlll\at('rI
for Y by the authors) and J{~ and J.lm \Vere varied from the same inltlLl to filla l val W'""
with the same increment as in the first analysis. Fourtecn differcnt combill,iolls )'ll'ld
lower SSRS values. The output is shown in Table A 3.
A.3 Comm~nts
The results obtained from this analysis show that estimates that ylcld SSRS lowcl, thall
those obtained by the mdhod of Braha and Hafner (1 9S) [7J.
These authors used the computer prograrn BP7 1 B with the dpplication module CIII v(!
fit They suge~ted that a nonlinear least-squales analysis wotlld be a reasollable tcchniqlle
to use. The algorithm in this computer program is Ilot known, but it scerns that the fitting
74
Table A..3' SSRSs values for the second analysis
l y J(~ J1.m SSR lia.
u_
1.45 3400 0.1 208.260 34000

1.45 6900 0.2 222.197 34500
1.45 7000 0.2 211.222 35000
1.45 10500 0.3 216.358 35000
1.45 10600 03 213.263 35333
1.45 10700 0.3 222.701 35667
1:45 14100 0.4 215 240 35250
1 45 U200 04 214 477 35500
1.45 14300 04 220.744 35750
1.45 17600 0.5 219.575 35200
1.45 17700 0.5 215.135 35400
1.45 17800 0.5 215 266 35600
1 45 17900 0.5 219.888 35800
technique employed nonlinear least-squares analysis.
After performing the SdIllC analysls with different initial and final values \Vith different
illClcments, it is observed that there is no pattern to be followed (between the variables
and the SSR) if one desires to estimate the parameters with the tnal-and-enol technique
It I~ unknowll if the computer Pdckage used in the original study estimates the pa-
1 dllletCI ~ by considenng eithcr the time or the substrate as the independent \'aI iable,
thcrefore, the same analysis was carried on but obtaining the SSR \VIth respect to time
(i.e substrate as independent variable). Varying Y, ](3 and /lm with the same initial and
filial values and inclement'i (similar ta the first analysis), 3369 sets of vanables yielded a
"'I\l,dler SSR.
75
Appendix, B
Main Computer Program
The algorithrn presented in Chapter 3, Fig. 3.4, is cornputirized in the program showlI
in thlS Appendix.
76
pragram Algorithm;
var infile,outfile : text;
tx,tt,ss,xx: array[O .. 20) of real;
srs,5rx,suml,sum2,5um3,sum4,sum5,sum6,sum7,sum8,5um9,nosol : real;
A,B,C,Xo,An,llinc,llfinal,lln,al,Yn,Y,So,Ks,mu,kd,t,s,x : real;
i,n,m : integer;
procedure captura;
begin
assign(infile,'c:\input.txt');
assign(outfile,'c:\output.txt');
reset (infile) ;
n:=O;
m:=Q;
readln(infile,n,m);
for i := 0 ta n do
begin
readln(infile,t,s);
tt[i):=t;
55 [ i) : =s;
end;
for i := 0 to m do
begin
readln(infile,t,x) ;
tx [i) : =t;
xx [i) : =x;
end;
close (infile) ;
rewrite(outfile);
end;
function ssrs(Y:real):real;
begin
suml := 0;
for i:= 1 to n do
suml := suml + 5qr(y*so+Xo)*So)/(Y*So+Xo*exp(11n*tt(i)*(So+Xo/Y)-ss[i);
ssrs . - suml;
end;
function ssrx(kd,A:real):real;
begin
suml : = 0;
for i:=l to n do
suml := suml + sqr(ln(Xo*expB/A-kd)*tx[i]-C/A*lnA+Xo*exp(al*tx[i]/(A+Xo
ssrx .- suml;
end;
function real) :real;

var den : rL _,
begin
suml:=O; sum2:=O; sum3:=Q; sum4:=0;
for i := 1 to n do
begin
den := Y*So+Xo*exp(lln*tt[i]*(So+Xo/Y;
suml .- suml + sqr(So) * (Y*So+xo)/sqr(den) ;
sum2 := sum2 + So*sqr(Y*So+Xo) * (So-11n*Xo*Xo*tt(i]/Y/Y*exp(11n*tt[i]* (So+
Xo/Y)/exp(3*ln(den;
sum3 .- sum3 + ss[i]*So/den;
sum4 := sum4 + ss[i]*(Y*So+Xo)*(Sa-lln*Xo*Xo*tt[iJ/Y/Y*exp(lln*tt[i]*(So+
Xo/Y)/sqr(den) ;
77
end;
fYy := suml - sum2 - sum3 + surn4;
end;
l
j procedure secY;
var Y_l,fY_l,fYn,Yl real;
begin
Y_l:=0.95*Yn;
fY_l:=fYY(Y_l) ;
repeat
fYn:=fYY(Yn);
Yl:=(Y l*fYn - Yn*fY_l)/(fYn - fY_l}:
y l:=Yn;
fY_l:=fYn:
Yn:=Yl:
srs := ssrs(Yn}:
writeln(Yn:8:6,' , ,11n:8:7,' , ,srs:10:6):
until (abs(fYY(Yl < 0.00000001):

end;
function fkdkd(A:real):real:
var gar : real;
begin
suml:=O: sum2:=0: surn3:=O; surn4:=O:
for i := 1 to m do
begin
sum1 :=surn1 + B/A*sqr(tx[i]}:
sum2 :=sum2 + C/A*tx[i]*lnA+Xo*exp(al*tx[i]})/(A+Xo:
sum3 :=sum3 ~ tx[i]*ln(Xo/xx[i]):
sum4 :=sum4 + sqr(tx[i]);
end;
gar := (surn1 - sum2 + su~3)/surn4:
if gar < 0 then fkdkd .- 0
else fkdkd := g~r;
end;
function fAA(A:real) :real:

begin
kd : = fkdkd (A) ;
sum1:=O; sum2:=0: surn3:=O: sum4:=0: sum5:=Oi sum6:=Oi sum7:=O; sum8:=Oi sum9:=
for i:=1 to m do
begin
sum1 := sum1 + B/A*(B/A-kd)*sqr(tx[i);
sum2 : = sIlm2 + C*B/A/A,"tx[ il *ln( (A+Xo*exp (al*tx[ i]}) / (A+Xo) ) ;
sum3 := sum3 + B/A*tx[i]*ln(Xo/xx[i]);
sum4 := sum4 + (B/A-kd)*tx[i]*C*Xo*(l-exp(al*tx[i]/(A+Xo)/(A+Xo*exp(al*tt[i
sum5 := sum5 + c*C*Xo*(l-exp(al*tx[i]/A/(A+Xo)/(A+Xo*exp(al*tX[l] *
ln( (A+"{o*exp(al*tx[i]) )/(A+Xo:
surn6 := sum6 + C*Xo*(l-exp(al*tx[i]/(A+Xo)/(A+Xo*exp(al*tX(l]*
ln (Xo/xx [ i J) ;
sum7 := sum7 + (B/A-kd)*tx[i]*C/A*lnA+Xo*exp(al*tx(i]/(A+Xo i
surnS := sum8 + sqr(C/A*lnA+Xo*exp(al*tx(i)/(A+Xo))):
sum9 := sum9 + C/A*lnA+Xo*exp(al*tx[i])/(A+Xo*ln(Xo/xx[i]) i
end;
fAA:= (suml - sum2 + sum3 + surn4 - sum5 + sum6 - sum7 + sum8 - sum9) i
end;
78
,
'1
procedure s~cA:
var A l,fA l,fAn,Al real;
begin- -
t A 1:=0.98*An;
fA- l:=fAA(A 1);
-
repeat
fAn:=fAA(An) ;
A1:=(A_l*fAn - An*fA_l}/(fAn - fA_l)i
A l:=An;
fA_l:=fAn;
An:=Al;
if An < yn*so then An := yn*so + 0.0001;
Ks:= (Yn*So+Xo)*So/(An-Yn*So);
mu := Iln*Ks;
srx := ssrx(kd,An);
WRITELN(An:8:5,' ',Ks:10:3,' ',mu:lO:4,' ',srx:lO:6};
until (abs (fAA (Al < O. 000000001} ;
kd := fkdkd(An);
end;
begin
captura;
J\osol :=0;
Yn := initial gess ;
lln := Lmin ;
llinc := Linc
lIn := lIn - llinc;
llfinal := Lmax;
An := initial gess;
repeat
lIn := lIn + llinc;
secY;
B := 11n*So*(Yn*So+Xo);
C := Yn*So;
al := Iln*(So+Xo/Yn);
secA;
Ks:= (Yn*So+Xo)*So/(An-Yn*So);
mu:=lln*Ks;
writeln(outfile,lln:18:1l,' ',Ks:10:4,' ',mu:lO:4,' ',Yn:10:4,' ',kd:10:4,' "
writeln(lln:8:6,' ',Ks: 10:4,' ',mu:10:4,' ',Yn:lO:4,' ',kd: 10:4,' ',srs: 12:9,'
close(outfile);
end.
79
J
1
Appendix C
N on- biodegradable substrate
Herein the computer program used to estimate the non-biodegradablc substrale is pre-
sented. The computer program was writtcn in Turbo PASCAl v. 4.0. [6J.
80
.J
proqram substrate;
var infile,outfile : text:
ss,tt : array[O 15] of real;
bn,s,t,b : real:
1 i,n : integer:
procedure captura;
begin
assign(infile,'c:\inputfile');
assign(outfile,'c:\outputfile'):
reset(infile);
n :- -1;
repeat
n := n + 1:
readln(infile,t,s):
ss[n] :- s;
tt[n] := t:
until eof(infile):
close(infile);
end:
function bb(b:real):reali
var suml,sum2 : real:
begin
suml := 0; sum2:=O:
for i := 1 to n do
begin
suml := suml + tt[i]*sqr(ss[0])*exp{-2*b*tt(i]):
sum2 := sum2 + tt(i]*ss(i]*ss[O)*exp(-b*tt(i]):
end:
bb := suml - sum2;
end;
function ssr(b:real):real:
var surol : real;
begin
surol := 0:
for i :=l. to n do
suml := suml + sqr(ss(i) - ss(O)*exp(-b*tt(i));
ssr := suml;
end;
procedure secant;
var b l,bl,fb l,fbn real;
begin- -
b l := 0.9*bn:
fb_l := bb Cb_l) ;
repeat
fbn := bb (bn) ;
bl := (b_l*fbn - bn*fb_l)/(fbn - fb_l):
b l := "n:
fb l := fbn;
bn-:= bl:
until absb1 - b_l)*10000) < 0.00001;
end;
procedure Mean:
var as,ssrr,nosolin,nosolinc,nosolmax real;
begin
rewrite (outfile) i
81
noso11n := Non-soluble initial;
noso11nc :=Non-soluble increment;
nosolmax := Non-soluble final:
bn ;= 0.05:
for i :: 0 to n do
ss[i] := ss[1] - noso11n;
repeat
for 1 := 0 ta n do
ss[1] := ss[i] - noso1inc;
noso11n := noso11n + nosolinc;
secant;
ssrr := ssr(bn):
wr1teln(noso11n:8:3,' ',bn:15:12,' ',ssrr:20:10);
wr1teln(outfile,nosol1n:8:3,' ',bn:15:12,' ',ssrr:20:10);
until nosolin >= nosolmax;
close (outfile) :
end;
begin
captura:
Mean;
end.
(
82
,
f
Appendix D
SSRS and ~f) vs Y
The computer program presented herein was used to estimate the initial guesscs for Y and
to verify the feasible roots of FI' The computer program was written in Turbo PASCAl
v. 4.0 [6].
83
program Fl;
var infile,outfile : text~
tt,ss,xx : array[O 20] of real;
srs,srx,suml,sum2,sum3,sum4,sum5,sum6,sum7,sum8,sum9,nos01 real;
al,Ks,m,Xo,lln,Yn,Y,Yinc,Yfinal,So,t,s,x : real:
i,n : integer;
procedure captura;
begin
assign(outfile,'c:\output.txt');
reset(infile);
n:=O;
readln(infile,n);
for i := 0 to n do 4
begin
readln(infile,t,s) ;
tt[i] :=t;
ss( i] : =s;
end;
close(infile);
rewrite(outfile);
end;
function ssrs(Y:real):real~
begin
suml := 0;
for i:= l to n do
suml := suml + sqr(y*so+Xo)*So)/(y*so+Xo*exp(lln*tt[i]*(So+Xo/Y)-ss[i});
ssrs := suml;
end:
function fYY(Y:real):real;
var den: real;
begin
suml:=O; sum2:=0; sum3:=Oi sum4:=O;
for i := 1 to n do
cegin
den := Y*So+Xo*exp{lln*tt(i]*(So+Xo/Y~
suml := suml + sqr(So)*(Y*So+Xo)/sqr(den);
sum2 := sum2 + so*sqr(y*so+xo)*(So-11n*Xo*Xo*tt(i)/Y/Y*exp(11n*tt(i]* (So+
Xo/Y)/exp(3*ln(den ;
sum3 := sum3 + ss(i]*So/den;
sum4 := sum4 + sS(i]*(Y*So+Xo)*(So-lln*Xo*Xo*tt(i]/Y/Y*exp(lln*tt[i]*(So+
Xo/Y)/sqr(den);
end;
fyy := suml - sum2 - sum3 + sum4;
end;
begin
captura;
lIn := given value of L;
Yn := Initial value of Yi
Yinc := Increment of y;
Yn := Yn - Yinc;
Yfinal := Final value of y;
repeat
Yn := Yn + Yinc;
writeIn(outfile,Yn:l8:ll,' ',fYY(Yn):2l:11,' ',ssrs(Yn):12:9);
writeIn(Yn:18:11,' ',fYY(Yn):21:11,' ',ssrs(Yn):12:9);
until Yn >= Yfinal;
f close(outfile);
end.
84
Appendix E
SSRX and ~FJ vs A
The computer program presented herp!n was uscd to estimate the initial gucsses for A
and to verify the feasible roots of F2 - The program Wa'5 written in Turbo PASCAl v. -1.0
[6] _
85
program F2:{definitive}
var infile,outfile : text:
tx,tt,ss,xx : array(0 . 20] of real;
srs,srx,sumJ,sum2,sum3,sum4,sum5,sum6,sum7,~um8,sum9,nos01 : real;
1 A,B,C,Xo,An,llinc,llfinal,lln,al,Yn,Y,So,Ks,mu,kd,t,s,x : real;
i,n,m : integer;
procedure captura:
begin
assign(outfile,'c:\output.txt') ;
reset (infiIe) ;
n:=O:
m:=O;
readln(infile,n,m);
for i := 0 to n do
begin
readln(infile,t,s);
tt (il: =t;
ss [il :=s;
end:
for i := 0 to m do
begin
readln(infile,t,x);
. tx [il :=t;
xx [i] :=x;
end;
close(infile);
rewrite(outfile) :
end;
function ssrx(kd,A:real):real;
begin
suml : = 0;
for i:=1 to n do
suml := suml + sqr(ln(Xo*expBjA-kd)*tx(i]-CjA*lnA+Xo*exp(al*tx(i)/(A+Xo
ssrx := suml;
end;
function fkdkd(A:real):real;
var gar : real;
begin
sum1:=0: sum2:=0: sum3:=O: sum4:=0:
for i := 1 to m do
begin
suml :=s'lml + BjA*sqr(tx[i));
sum2 :=sum2 + CjA*tx(i]*lnA+Xo*exp(al*tx[i]/(A+Xo:
sum3 :=5um3 + tx[i]*ln(Xojxx[i]):
sum4 :=sum4 + sqr(tx(i) ;
end:
gar := (sumi - sum2 + sum3)jsum4;
if gar < 0 then fkdkd := 0
eise fkdkd := gar;
end;
function fAA(A:real) :real;
begin
kd : = fkdkd (A) ;
suml:=O: sum2:=O: sum3:=0; sum4:=0; sum5:=O; sum6:=O: sum7:=0: sum8:=O; sum9:=
r for i:=1 to m do
...
86
begin
suml := sum1 + B/A*CB/A-kd)*sqr(tx[i]):
sum2 .-sum2 + C*B/A/A*tx[i]*lnA+Xo*exp(al*tx[i)/(A+Xo));
sum3 := sum3 + B/A*tx[i)*lnCXo/xx[i);
), .-
sum4 .- sum4 + (B/A-kd)*tx[ij*C*Xo*(l-exp(alwtx(i]/(A+Xo}/(A+Xo*exp(al*tt[i
sum5 := sum5 + C*C*Xo*C1-exp(al*tx(i]/A/(A+Xo)/(A+Xo*exp(al*tx[i])} *
lnA+Xo*expCal*tx[i)/(A+Xo)) ;
sum6 := sum6 + C*Xo*(l-exp(al*tx(i]/(A+Xo)/(A+xo*eXp(al*tx[i))*
ln(Xo/xx[iJ) i
sum7 := sum7 + CB/A-kd)*tx[iJ*C/A*lnA+Xo*exp(al*tx(i)/(A+Xo));
sumS := sumS + sqr(C/A*lnA+Xo*exp(al*tx[i]j(A+Xo);
sum9 := sum9 + C/A*lnA+Xo*exp(al*tx[i)j(A+Xo))*ln(Xojxx[i]);
end:
fAA:= (sum1 - sum2 + sum3 + sum4 - sumS + sum6 - sum7 + sum8 - sum9) ;
end:
beqin
captura;
Yn := qiven value for Yi
lln := qiven value for L:
B := Iln*So*(Yn*So+Xo):
C := Yn*Soi
al := lln*(So+Xo/Yn);
An := Initial value for A;
Aninc := Increment;
An := An - Aninc;
Anfinal := Final value for A;
repeat
An := An + Aninc:
writeln(outfile,An:1S:11,' ',fAA(An):21:11,' ',ssrx(fkdkd(An),An):21:11)i
wri teln (An: 18: 11,' " fAA (An) : 21: 11,' ',ssrx (fkdkd (An) , An) : 21: 11) ;
," until An >= Anfinal;
close (outfile) :
end.
87
1
Appendix F
Analysis for the roots
In the [ollowing figures the SSR's and the roots that correspond to the minimum SSR's are
shown al the mlTllmUm znlersectioTl poznt for the Braha and Hafner (1987) [i] data sets,
and for the data sets from the experiments perforrned in this study. It can he ohserved
that in sorne graphs (Figs. F.I and F.3 to F.6) that a more or less sud den s!ope change
tales place ln the *" vs A plots. During the computations it was obs'r ved (on the screen
of tlw llIorlltor) that the slope change occurs when the non-negative constraint 011 kd
be(omcs active.
88
1
...
ili S
a il CO
III
...J nt:.
s:<
-
... f. ....J
...
/
-
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,
'"
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ijJ
rr,
:::
~
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N ... .. .,1
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.,.... ..
"
on
'"
al
"0 ~
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"' ......

-<:
'" ""c>
1 1 1 0
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0 0 <> 0 C>
(-JOI .. wu)
YI>/l:JP
0
0
<>
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XIISS
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C>
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-
t..rJ
,...,
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t f..
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if;
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fi cf,
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o
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e ...
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al

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7
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(.puo'"O~J)
s~s
89
~
LB2
~".~
L82
-
100
50
j /
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~
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E
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00 OS 12 16 -10
24 2B
-06 -04 -0 2 0 02 00 06 OS
Y 12 10
(ThauSOndS)
...
ao
000059
(0 000059
0 70 -1 / 000057
000056
60 -1 ./
000055
000054
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~:
"'il
.
~
t
:j
JO / )(
!li
III
000053
000052
000051
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20 -1 \ /
000047
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ID -1 \ /
0000'5
000044
o 1 1'-- i ----= 000043
04 OB 12 16 2 20 2B
-06 -0 -02 0 02 0' 06 OS 12 1.
Y (ThauSOndS)
...
Figure F.2: lli

J)' vs y ,','
':;':;Re.,'
vs }' , f2f2.
,)" vs A an d ........
C,'C,'RX vs A Cor LB2
-~----- -- 2llI
. li ......
LB3
LB3
100 ]
~
000001
50
a
a -0 00001
-0 00002
-50
-000003
-100 -0 00004
"-
;;: ~-o 00005
... -150 ~
~-o 00006
-200 -000007
-0 oooos
-250 JI -0 00009
-lOO
-0 0001 j \
-350 1 -0000' 1
05 15 25 J5 -0 00012
y -0 6 -0 4 -0 2 a 02 04 06 08
(Thouao_>
'"
140 OOOOU
:::~ ::::::~
~ IJO
- / 1 \
100 000041
90
- 0000.
: 80
~c x
!if ; 70 ~ a 00039
~o ~
. 60 a 0003!
50
40 OOOOJ7
:: J\ 1
/ 000036
000035
'D
a 1 ? i~ 0~~4
05 15 25 35 -0 6 -0 -0 2 a 02 C4 C6 ce
(lhou>CndS)
y '"
Figurt' F :3: ':!.fi

(Il
vs} .~SRS VS } 1t VS A and ssn.\: \'5 A for LB3
~
LB4
JI.. '"
..
LB4
000001
'::j ~ -0 00001
-50 / -0 00002
-0 OOOOl ~
>- -0 00004 /
/
-100]
"
"- ~
~ ;:;-000005
" -150 ....
j " -OOOOO!
-200
-250 1 -000007
-000008
-lOO J1 -000009
-0 0001
-350
04 -000011
08 12 16 2 24 28 J2 -06 -04 -02 a 02 04 06 08
y (Thau"'_l
A
100
(,0 a 000~8
t-..:)
90
0000"
80
000056
70
.
000055
~
60
000054
"
Ille:
21::
IIIg 50
g 0.00053
t 40 '"
0.00052
JO
000051
20
00005
la
000049
a
000048
04 08 12 16 2 24 28 J2 -0 6 -04 -0 2 a 02 04 06 08
y (Thauoa_l
A
Figure F.4: ~ vs l", S'SRS vs Y, 9ft vs A and SSRX vs A for LB4

<;; ~ ". ~
...
LB5 LB5
l
=i
20
~ 2
0
-20 0
-40 -1
-60 -2
~
...
)-
-BO ~.!.
"-
;;:
... -100
-\20
~
V"
.
"-~
E
-3
-4
-\40 -5
-150 -5
-IBO -7
-200 -8
-220 -9
-240
-\0
0.4 08 \2 16 2 24 28 32
-06 -0 4 -0 2 a 02 04 06 03
y (Tl\......fIds)
Jo.
::j
0.0007
(.0
~ 000068
50 / 000066
000064
000062
~
:] \ /
en" 0.0006
~::
III~ ~
III 0 000~8
o OOO~6
20
000054
1 000052
I~ L
la
00005
o !
04 08 12 16
000048
24 28 32
-06 -0 4 -02 0 02 0.4 06 08
y
..
(Tl\OUlGndI)
Fi~lIre F.5: ~ VS }", SS'RS' VS Y, ~I~ VS A and S'SRX vs A for LB5

... ". . "\ .fi..-:A
L86 L86
OoO.J------T-----------------------____________________,
"1 _1 00012
'0 a DO"
0001
5 00009
o oooa
00001
00006
~ 0005
-5 00004
~ ;S a 0003
~
-=v -la
!;;
00002
00001
Q
==-- ,
-l' -0 0001
-Q 00Q2
-Q 0003
-20
-0 0004
-00005
-25 -00006
-00M1
-Jo;I----~--~----._--_r----._--_.--~r_--,_--~
3
-O.OOOIS
-0.0009 -1 I-----l----.------.----r----r----.--__r-_--.___-I
5 9 \1 -\Il la JO 50 10
(Th ...... ndI)
'f
A
OOllr--~r-------------------------- ------------,
~
,;:..
=j
35
0009
D.DDI
JO 0.007
';'
v 25 0006
Ole
2f~
.,"
6
o
20 i 0.005
0004
.5
0.003
10
0.002
5
0001
01 1~i
3 5 9 0;1----;---~----~---r----.---_r--~r_--~--_4
y
" -;10 la JO 50 10
(Th......ndll
A
Figure F.6: ~ vs Y, SSRS vs Y, Wf vs A and SSRX vs A for LB6

......................~------------~----------~~~~~'~-~ ,~
'~ .........r, -~ ...-~~ ~~ .. ~ ..... - - . : '....... Tf" ... '~- "'i"'~yr:;r -.~t,,,"~,,,
- "'..>r;I""'" . . ~.....~.-
! .....
LB7
80
LB7
&0 04
02
40
0
2Q
-0 2
a
-0 ,
-20
-06
-40
~ -08
>-
-50 1
~
... -80
~ ...
,~
-1
v -12
-100 ~"
v"
E -1 ,
-120 e -1 &
-140 -18
-150: -2
-180 -22
-200 -2 ,
-220 -25
04 08 12 16 2 24 28 -28
-&00 -400 -200 Q lOG 40G
Y 600 800
A
4~
0.00059
40
/ 1
c.o a 0005S
0- J~ J /'
30
/ 000057
";
v 25
<ne:
5~
~
X
fil ~
&;
C
20 / !li
'"
00005&
15
000055
"]
5
0
\ / 000054
04 OS 12 000053
16 24 28
-600 -400 -200 a 200 400 &00 SOO
y
A
Figlllt:' F.i: ~ VS L .-;SRS VS y', ~~tt VS A and SSRX vs A for LB7

"'~ ....,.i:<\
..
Run 1
Run 1
:J ---
COD
01
0
COD
------
-01
lO -i /
-02
20 -0 J
-04
10 -0 ,
~
,"
~
0
~.!.
,2
-0.6
-07
-=" -10 ~. -0 Il
v -09
-20
-1
-lO -II
-1 2
~:11 -1 3
-1.4
-1.5
-60 1
-10
0.1 03 0' 07 09 Il 13 15 17 19 21 -06 -04 -02 02 04 06 011
y (ThOU .._ )
A
10 DDDD61,--------------------,------______________________________- ,
c:.o
(j)
o 00D609
9
00006011
0000607
l
Il
o OD0606
7 o DDD6D5
~.~~~:~~
';' 6
v
Ille:
5:: x
5
III~ ~O 000601
00006
4
0000599 /
0.000598
J
0000597
2 o 00D596
0000595
0000594
0.00059l
0+1~-r.-,-~._r-. .~_._r_.,_~._._. .~ 00005924---r-~--,__,r_~--;__,r__r--~_,--_r--~_,--~--~~--~
0.1 03 05 07 09 Il 13 15 17 19 21 -06 -04 -02 02 04 06 011
(Thou .. _ )
y
A
Figure F.S: *'" vs Y, SSRS vs 'V, ~ vs A and SSRX vs A for Run 1 (COD)
i .~ ~ ..
Run 1 Run 1
TOC
4 TOC
01
l
a
2
-0.1
0 -0.2
-1
-0 l
-2
-3
"<II -0 ~
~ o!.
~:
-0.5
"-
;:: -~
v -06
-5
"
-5 -07
-7
-0 B
-1
-09
-9
-la -1
-11 -1.1
-12
-12
Q~ 15 25 J5 45 55 -IBD -ua -100 -60 -20 2Q 60 100 140 180
Y A
00005989
600
CO 00005988
-.J 00005987
/
00005986
500 -l
00005985
a 00059!l~
400 -l 00005983
00005982
i 0005981
~ /
VI
.,~ JOO '" 0000598
00005979
00005978
200 -1 / 00005977
00005976
00005975
100 -1\ / 00005974
00005973
00005972
o 1 -IBO -140 -100 -60 -20 2Q 60 100 ..0 IBO
05 15 25 35 45 55
y
...
Figure F.9: ~ vs },., S'SRS' vs }", 7ft vs A and SSRX vs A for Run 1 (TOC)
.. ~ ~~ ..
Run 2 Run 2
COD
~~
COD
1 1
:so
40
~ 0
:~j
-1
,.v
"-
.:v /
J
~.1
,~
~
v"
~
.
E
E
-2
-3
-4
-"V
-20
-30
-40
-~
-6
-50
-7
0.3 0' 07 09 Il 13 1~ 17 19 21 -1111 -140 -100 -60 -20 20 60 100 140 180
Y
A
I~
14
1 00020111 ------
~ 13..1 /' 0002017
00
0002016
/
12 ]
Il
10 0002015
-;-
:~
0002014
Ole
5f~
" x
.,~ gfo 002013
t. (Il
0002012
0002011
J
0.00201
/'
1 1 1
0002009
03 05 07 09 1.1
-1 0002008
1J 15 17 19 21 -160 -140 -100 -60 -20 20 60 100 140 1110
Y A
Figure F .10: Wvs Y, SSIlS vs }", i?ft vs A and SSRX vs A for Run 2 (COD)
;. " .....
Run 2
, i
TOC Run 2
TOC
01
-01
Ji / -02
-03
'1 /
-0 ~
>- ~
"
"-
;;: ~ ...
1 -05
... ,~
f! ....E
-0.6
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-08
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-II
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-1110 -140 -100 -60 -20 20 60 100 140 180
Y
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0 2 10 -1110 -140 -100 -60 -20 20 60 100 140 IBO
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Fi gUI (' F.l/. ~ VS Y, SSHS \'5 }', 4f VS A and SSRX vs A for Run 2 (TOC)
"'",. " il>- '"
...
Run 3 Run 3
COD
60 COD
a oooos
50
40
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./ -0
la
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~:
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l 000255
2
00025
o , 0002"
-IBO -140 -100 -50 -20 2D 50 100 140 IBO
01 OS 07 09 Il 1J 15 17 19 2.
Y A
Figure F.12: ~ vs Y, SSRS vs F, fJff vs A and SSRX vs A for Run 3 (COD)

____________________________ ... ..... .. ..... .. .. ~... 'I.... .. ~I~~~.,... --~ .. i:~'l''''3l'--'#l1 ..,(J1't"'
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" <ri ....

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TOC Run 3
'.5 ~OC
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Flg\lle F.13: *" VS }'. SSRS vs }'. ~ vs A and SSRX vs A for Run 3 (TOC)
.. ~ ..
Run 4
05
COD Run 4
05
04
03
02
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.
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t~
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(Thou ...,.,.)
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y ...
Figure F.14: ~ vs V. SSRS vs Y, ~ vs A and SSRX vs A for Run <1 (COD)
t l; '!I ...
Run 4 Run 4
TOC
12 TOC
0.5
10
:~
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2
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...,. ,2
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O' OS 12 16 2 28 -110 -90 -70 -50 -30 -'fJ ICI JO ~ 70 90 lI0
y ..
Fig\llc F.15: *" VS } ' , SSRS VS }'", 9ft VS A and SSRX vs A for Run 4 (TOC)
~ ... "iIL- _ _ . " ' _

~
.,,,~
.-'~<\
Run 5 Run 5
COD COD
60 0001
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02 04 06 oa 12 14 16 1 a 2 -1 3 $ 7 11 13 1$
y
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Y (Thau ... _> 15
Figure F.16: * vs Y, S'SliS vs Y, 2ft vs A a.nd SSRX vs A for Run 5 (COD)

">
-
Run 5 Run 5
TOC 01
roc
1
6 0
5 -01
4 -02
3
-0.3
2 ~
1 -04
~"'
,2
~
..... 0 /:!.
v
-05
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E
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-7
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-200 -100 o 100 200
24 2B 32
'(
...
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0000454
0000453
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., 0008
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400-l /' 0000443
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JOO 200
-200 -100 100
o. 08 1l 16 2 2B J 2
...
Flgtlrt~ FA: ~ vs }', SSH8 vs Y, W1 vs A and SSRX vs A for Run .5 (TOC)

Bibliography
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109

Assessment of Nonlinear Leastsquares Estimation of Monod Kinetic Parameters From Batch Reactor Experiments

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1

ASSESSMENT .OF NONLINEAR LEAST-SQUARES

MARCO ANTONIO VILLARREAL RODRIGUEZ

Department of Civil Engineering and Applied Mechanics

A thesis submitted to the Faculty of Graduate Studies

Marco A. Villarreal. 1990

An academic career compels young man to scientific pro-

- duction, and only strong characteres can resist the temp-

tation of superficial analysis.

sum of squares residuals.

~Iollod-kjnetics constants using batch reactors experiments.

en terme de temps requis pour raliser les expriences.

La mthodologie agit de faon satisfaisante avec deux ensembles de donne:-. .,illluli>(:-,

stantes estimes varient considrablement entre elles-mmes, et certaines Ile se tl'OllVCllt

de paramtres qui rsultent de la mthodologie propose sont celles qui produisent 1(

minimum des sommes des carrs des rsidus.

des racteurs en mode cuve.

He is particulary indebted to E. Waldron and 1. Buchanan for their important assis-

tance in the environmentallaboratory.

1 iores de Monterrey, Monterrey N. L. Mxico.

List of Symbols and Acronyms XIII

2.1 Wastewater Treatment

2.2 Biomass Growth and Substrate Utilization Cl

2.2.1 Biomass Growth Rate 7

2.2.2 Yield Coefficient . 8

2.3 Estimation of Monod Kinetic Parameters . 10

l 2.3.2 Batch Reactor Expcriments 12

3 Methods and Materials 22

3.1 Model development . . 22

3.1.1 Mathematical analysis 22

3.1.2 Nonlinear least-squares analysis 28

3.1.3 Aigorithm for estimation of the parameters . 30

3.2 Initial Estimates ... 33

3.3 Laboratory Experiments 36

3.3.2 Ratch reactor 36

4 Results and Discussions 38

4.1 Synthetic Data ...... . .. .. .. . .. 38

1.2 Braha and Hafner (1987) Data Sets 40

4.3 Data from the Granby Plant .. .51

4.3.1 Experiments .. ........ .52

5 Summary & Conclusions 69

A.2 l'.lodci and Application 73

l B Main Computer Program 76

E SSRX and ffi

2.1 Typical values oi kinetic coefficients at 20C 10

2.2 Models used by Simkins and Alexander (1984) 18

3.1 Comparison cf methodologies . . . . . . . . . . . . . . . . . . . . . . . .. 32

4.3 Braha and Hafner (1987) Data Sets . . . . . . . . . . . . . . . 42

... 6 Characteristics of the Granby wastewater treatment plant. 52

4.7 Characteristics of sludge and substrate for each experirnent . 54

4.9 Ratio of soluble ta total substrate concentration (COD) at Granby. . 55

., 10 Substrate and biomass data for runs 1 to 5 . . . 56

A.1 Parameters studied .. 73

A.3 SSRSs values for the second analysis . . . . . . . .. . 75

2.1 Fl,~W schematic of the suspended-growth activated sludge 5

2.4 Complete-mixing continuous flow reactor without recycle 11

2.6 Biomass growth and substrate degradation vs time in a batch reactor 14

3.5 Wand SSRS versus Y . 34

ay versus y and !l.a

4.5 SSR ratio, biomass and substrate graphs for LB3 46

4.8 SSR ratio, biomass and substrate graphs for LB6 49

F.3 -W- vs Y, SSRS vs Y, ~7 vs A and SSRX vs A for LB3 . 91

F.4 -W- vs Y, SSRS vs Y, a:: vs A and SSRX vs A for LB4 . 92

F.5 -W- vs Y, SSRS vs Y, ~7 vs A and SSRX vs A for LB5 . 93

F.13 vs Y, SSRS vs Y, vs A and SSRX vs A for Run 3 (TOC) 101