R E G U L AT O R Y G R I D L O C K

Water quality
and disinfection kinetics
Water quality significantly affects
predictions of microbial inactivation.

A
Charles N. Haas, Josh Joffe,
Uma Anmangandla,
Joseph G. Jacangelo,
mendments to the Safe Drinking
and Mark Heath Water Act required that surface water suppliers in
the United States filter, disinfect, or both to protect
customer health. The Surface Water Treatment Rule
(SWTR) specified filtration and disinfection treat-
ment requirements for public water systems using
surface water sources or groundwater under the
direct influence of surface water. The US Environ-
mental Protection Agency (USEPA) has issued a guid-
ance manual1 to assist utilities in implementing SWTR
requirements.
Among the organisms capable of transmitting
waterborne diseases, Giardia lamblia has been classi-
fied as a significant pathogen, often causing severe
gastric difficulty. The USEPA Office of Drinking Water
has adopted the contact-time concept to quantify the
inactivation of G. lamblia
cysts by disinfection. The
The contact-time tables of the US Environmental Protection concept was applied to
Agency are based on survival data obtained in buffered demand- both filtered and nonfil-
free water. However, disinfection data from experiments tered systems to ensure
conducted at Drexel University showed that the choice of source adequate disinfection.
water influences the degree of inactivation. Sophisticated The SWTR required
disinfection kinetic models and statistical tests are used to that all surface water
analyze the sources of these discrepancies. This article treatment facilities provide
demonstrates that the quality of a particular source water is a adequate filtration and
significant factor in predicting disinfection effectiveness. disinfection in order to
achieve (1) a 99.9 percent

Copyright (C) 1996 American Water Works Association

MARCH 1 9 9 6 95
FIGURE 1 Basic kinetic models
chloramines, and ozone were used to disinfect E. coli
(bacteria), Giardia muris (protozoa), and bacterio-
phage MS2 (virus). In Table 1, experimental runs
performed for the batch analysis are categorized by
Chick–Watson model Hom model water, disinfectant, concentration, and pH. Table 2
summarizes the water quality characteristics of the
dN n dN n m–1
= –kC N = –kmC t N experimental waters.
dt dt

N = microbial concentration Batch experiment procedure

C = disinfectant residual Three reactor vessels consisting of 1 L of liquid in
t = time a 2-L beaker were run in parallel (Table 3). The first
k,n,m—kinetic parameters
reactor contained microorganisms and the water or
buffer without disinfectant. The second and third
reactors contained microorganisms, disinfectant, and
the water or buffer. The first reactor served as a con-
trol and was used to determine the microbial decay
removal–inactivation of G. lamblia cysts and (2) a 99.99 in the absence of disinfectant. In the second reactor,
percent removal–inactivation of enteric viruses. It was disinfectant decay was measured to obtain a resid-
assumed that for effective filtration, a conventional ual disinfection measurement. In the third reactor,
treatment plant was achieving a 2.5-log removal of microbial inactivation with disinfectant was per-
Giardia and a 2-log removal of viruses. Disinfection formed to obtain a microorganism survival mea-
was required for the remainder of the removal–inac- surement. To initiate the experiment, a suspension of
tivation. Disinfection credit was awarded where C X T microorganisms at the desired density in demand-
was defined as the residual disinfectant concentration free buffer was added to each vessel and then mixed.
(C, in mg/L) multiplied by the contact time (T, in min) At zero time (start of the experiment), a solution
between the point of disinfectant application and the of disinfectant was added to reactors 2 and 3. The
point of residual measurement. The SWTR Guidance volume of the solution was as close to 5 percent of the
Manual provided C X T tables for several disinfectants microbial suspension volume as possible; e.g., if a 1-
and organisms; these tables indicated the specific dis- L working volume was chosen and a 1-mg/L initial
infection or C X T credit awarded for a particular inac- disinfectant concentration was desired, then 50 mL of
tivation. A safety factor of 1.5 was incorporated into the a 21-mg/L disinfectant solution was added. The first
C X T values included in the tables. reactor received the same volume of a disinfectant-
demand-free buffer solution with no disinfectant.
Background At predetermined times, samples were taken from
The C X T tables of the SWTR Guidance Manual the control and survival reactors, and disinfectant
were based on a number of simplifying assumptions. residuals were immediately quenched with excess
First, many of the data were derived from studies con- sterile sodium thiosulfate. In a 120-mL sterile sample
ducted in buffered demand-free water, resulting in container, 0.1 mL of a 10 percent solution of sodium
findings that may be difficult to extrapolate to actual thiosulfate was used to neutralize a sample contain-
waters. Second, a simple Chick-Watson relationship for ing about 15 mg/L of residual disinfectant, following
the microbial inactivation was assumed. This article Standard Methods procedures.2 The neutralized samples
argues for use of more accu-
rate disinfection kinetic mod-
els, including decay of the dis-
FIGURE 2 Kinetic models incorporating first-order residual decay
infectant residual. The kinetic
parameters of these models for
a particular water can then be n
Ni – kC 0
determined. Chick–Watson with k *: In(Si*) = ln ( ) = [1 – exp (–nk*t)]
No nk*
At Drexel University
(Philadelphia, Pa.), batch dis- Ni m m m
Hom with k*: ln(Si*) = ln ( k C 0 [1 – exp (–nk*t )]
No )
n
= –(
infection experiments were nk* ) m
performed on various combi-
nations of water, disinfectants, Model of choice
and microorganisms. The au- Si* = predicted survival ratio
thors studied four waters: N = microbial concentration
buffered demand-free (BDF) N 0 = initial disinfectant concentration
water, Bull Run Reservoir C 0 = initial disinfectant concentration
t = time
water and Willamette River k* = first-order disinfectant residual decay rate
Water (both from Portland, k,n,m—kinetic parameters
Ore.), and Palm Beach (Fla.)
groundwater. Free chlorine,

Copyright (C) 1996 American Water Works Association

96 JOURNAL AWWA
were then subjected to enu- TABLE 1 Summary of batch experimental runs*
meration of viable organisms
using techniques described Number of
later in this article. Water Disinfectant Dose—mg/L pH Experiments

BDF Fre e c hlo rine 0 .5 6 .9 1

Disinfectant stock 1 6 .9 2
oxidant solution 2 6 .9 1
Mo no c hlo ramine 0 .5 6 .9 1
preparation 1 6 .9 2
Stock chlorine solution. 2 6 .9 1
Ozo ne 0 .2 5 6 .9 2
The working stock chlorine 0 .7 5 6 .9 1
solution was prepared to a con- Bull Run Fre e c hlo rine 1 6 .5 2 –6 .5 4 2
centration $150 ± 10 mg/L by 2 6 .9 3 1
Mo no c hlo ramine 1 6 .3 7 –6 .5 5 2
bubbling chlorine gas into a 2 6 .7 0 1
weak alkaline solution, pre- Ozo ne 0 .2 5 6 .5 3 1
pared by adding sufficient 0 .4 6 .5 3 –6 .9 3 2
Willame tte Rive r Fre e c hlo rine 2 7 .3 0 –7 .3 4 2
NaOH to distilled water to 3 7 .1 7 1
achieve a final pH of at least Mo no c hlo ramine 1 7 .4 1 1
2 7 .1 5 –7 .5 4 2
8.0. The stock chlorine solu- Ozo ne 0 .5 7 .3 7 –7 .5 4 2
tion was stored in dark refrig- 0 .7 5 7 .4 4 1
erated conditions and dis- Palm Be ac h Fre e c hlo rine 4 .5 7 .5 2 1
Ozo ne 2 .5 7 .6 3 1
carded when the concentration
decreased to <150 ± 10 mg/L. * Te mpe rature = 1 8 o C fo r all e xpe rime nts

Stock monochloramine
solution. Preformed mono-
chloramine was prepared TABLE 2 Water quality characteristics of experimental waters
daily as needed by mixing
equal volumes of chlorine Water Quality Parameter Bull Run Willamette Palm Beach
and ammonium chloride
solutions at a 3:1 (Cl 2 :N) To tal o rganic c arbo n— mg/L 1 .0 –1 .7 0 .8 –7 .1 1 0 –1 2
True c o lo r— units <5 NA* 2 5 –4 3
weight ratio, yielding a 150 Ammo nia— mg/L as N <0 .0 2 NA 1 .8 –2 .4
± 10 mg/L as Cl 2 solution. pH 7 .0 –7 .2 5 .0 –8 .5 6 .9 –7 .1
To tal hardne s s — mg/L as CaCO3 7 –1 5 NA 2 5 4 –3 3 2
Each solution was prepared To tal alkalinity— mg/L as CaCO3 5 –1 1 1 4 –3 6 2 3 0 –2 6 6
in a pH 8 phosphate buffer. Turbidity— ntu 0 .2 6 –1 .4 8 0 .7 –5 .0 0 .2 8 –0 .8 5
Stock ozone solution.
* NA— no t available
Oxygen carrier gas contain-
ing approximately 5 percent
ozone was bubbled through
an ozone generator* for a TABLE 3 Three disinfection reactors run in parallel for batch experiments
minimum of 20 min at 20oC
Reactor Purpose Description
(40 min at 40oC) through 400
mL of buffered reagent-grade 1 Co ntro l Wate r + o rganis ms
water in a 500-mL gas 2 Re s idual me as ure me nt Wate r + o rganis ms + dis infe c tant
3 Survival me as ure me nt Wate r + o rganis ms + dis infe c tant
absorption flask. Ozone con-
centrations in the stock solu-
tion ranged from 20 to 30
mg/L. Effluent gas was neu- TABLE 4 Comparison of k* values*
tralized by passage through a
Number of Correlation
solution containing 132 g/L Disinfectant Water k*—1/min Experiments Coefficient
of sodium thiosulfate and 3
g/L of potassium iodide3 to Fre e c hlo rine BDF 0 .0 0 8 6 0 .9 9 6 9
Bull Run 0 .0 3 3 4 0 .9 6 5 8
eliminate excess ozone. The Willame tte Rive r 0 .0 4 8 3 0 .8 6 8 9
stock solution was refriger- Palm Be ac h 0 .1 7 6 2 0 .7 2 9 5
ated (4oC) in dark conditions Mo no c hlo ramine BDF 0 .0 0 0 3 6 4 0 .9 9 8 8
Bull Run 0 .0 0 1 5 0 .9 9 5 4
for 15 to 20 min until use. Willame tte Rive r 0 .0 0 1 5 4 0 .9 8 1 2
Ozo ne BDF 0 .0 7 3 3 0 .9 7 5 5
Bull Run 2 .2 0 5 3 0 .9 3 5 9
Microbial preparation Willame tte Rive r 9 .6 9 1 3 0 .9 8 4 9
E. coli. Sterile nutrient Palm Be ac h 3 6 .4 5 1 0 .9 9 9 9
broth preparation. Nutrient
* Re s idual analys is c o mpute d fro m C = C0 e xp (– k* t)

*Model-T408, Polymetrics Inc., Col-

orado Springs, Colo.

Copyright (C) 1996 American Water Works Association

MARCH 1 9 9 6 97
FIGURE 3 Survival plot for experiments with Giardia, 1 mg/L
of monochloramine, and Bull Run River water
Si (observed) Si* (Chick–Watson model)
1
10
0
10
Survival—Si
–1
10
–2
10
10
–3
0 50
Time—min
broth* was prepared according to manufacturer’s
specifications for use in these experiments.
Working stock culture preparation. The working
stock culture was prepared according to the proce-
dures of Kouame and Haas.4 Under sterile conditions,
an aliquot of permanent stock E. coli culture† was
transferred to capped assay tubes containing sterile
nutrient agar slants. Tubes were placed in an incu-
bator at 37oC overnight for growth. Capped tubes
were then sealed with laboratory film‡ and stored at
4oC. This working stock culture was used for a three-
week period. The contents of two assay tubes were
used to prepare a new set of working stock cultures.
The purity of the strain was checked periodically
using a multitest microbial classification system§ or
similar test.
Experimental culture preparation. To prepare the
microorganisms for the inactivation experiments,
assay tubes containing sterile nutrient broth were
inoculated with microorganisms from the working
stock culture. The assay tubes were incubated at 37oC
for 24 h. Then, the microorganisms were harvested
using 10 mL of BDF water and centrifuged at 1,000˘
g for 10 min, and the assay tubes containing the
microorganisms were washed twice with 10 mL of
BDF water or buffer. They were mixed at all stages in
a vortex mixer. By preparing a standard curve of opti-
cal density (660 nm) versus cell counts, the authors
arrived at a known initial concentration of cells. The
microorganisms were held no longer than 2 h at room
temperature or no longer than 24 h at 4oC.
G. muris. Working stock preparation. All G. muris
cysts were obtained from the Oregon Health Sciences
Copyright (C) 1996 American Water Works Association
98 JOURNAL AWWA
University, Portland, Ore. This
strain of G. muris was ob-
tained from specific path-
Si (Hom model)
ogen-free mice.** For the
production of highly purified
cysts, feces were collected
from the host, and cysts were
isolated from the fecal mate-
rial by a sucrose gradient
technique.
Prior to use, the cyst sus-
pension was washed two to
four times by centrifugation
using disinfectant demand-
free buffer. The final cyst
preparation was resuspended
to the desired concentration
(determined by hemocy-
tometer count) using disin-
fectant demand-free water,
placed in a cooled container,
and shipped overnight to
100 150 200 Drexel University. The cysts
were used within a 14-day
period.
Before use, the cyst prep-
aration was monitored for
optical density at 260 nm and
an A260 of less than 0.03 cm–1 was used to ensure
low disinfectant demand exerted by the cysts.5,6
Experimental stock preparation. Cysts were
washed three times in disinfectant demand-free buffer
immediately prior to experimental use. Following
centrifugation at 500X g for 2 min, cysts were resus-
pended in 10 mL of disinfectant demand-free buffer
for inoculation into reactor vessels.
Bacteriophage MS2. Working stock prepara-
tion. MS2 bacteriophage†† was propagated by inoc-
ulating a 500-mL flask containing 100 mL of tryptone
yeast extract‡‡ with the viral host bacteria E. coli,§§
to which 1 mL of 0.1 M sterile calcium chloride
(CaCl2) was added. The bacteria-containing flask was
placed in a shaking water bath maintained at 37oC.
Bacterial growth in the flask was monitored over
time by measuring optical density.
A standard curve of bacterial density versus pre-
viously generated absorbance was used to predict bac-
terial concentrations. When the bacterial density
reached approximately 1X108 colony-forming units/mL
(cfu/mL), an aliquot of the virus stock (approximately
1012 plaque-forming units/mL [pfu/mL]) was added
to provide a multiplicity of infection (MOI) of 0.1.
This MOI was employed to assure two rounds of virus
replication. Incubation of the bacterial culture con-
*#0003-01-6 Difco, Detroit, Mich.
†American Type Culture Collection 13706, Rockville, Md.
‡Parafilm M, American Can Co., Greenwich, Conn.
§api 20ETM, Analytab Products, Plainview, N.Y.
**CF-1
††15597-B1, American Type Culture Collection, Rockville, Md.
‡‡SP medium 1,0769-01 GB, Difco, Detroit, Mich.
§§15597, American Type Culture Collection, Rockville, Md.
tinued until the host cells FIGURE 4 Water comparison plots with the Hom k* fixed model
lysed. After cellular lysis, 0.01
g of crystallized lysozyme and
3 mL of sterile 0.2 M EDTA
were added, followed by addi-
tional incubation for 1 h in a BDF water (observed) Williamette River water (observed)
shaking water bath at 37oC. 1 Bull Run water (observed) Palm Beach water (observed)
10
The propagated virus and cel-
E. coli /Monochloramine
lular debris were then cen-
–1
trifuged at 3,000X g for 20 min. 10
The supernatant was filter-

Si—observed
sterilized using a membrane 10
–3

filter (porosity = 0.08–0.10

µm) that had been pretreated –5
10
and double rinsed with a 10
percent solution of distilled
–7
water.* The supernatant was 10
then extracted using chloro-
form (CHCl3). The aqueous 10
–9
phase was refrigerated at 4oC –9 –7 –5 –3 –1
10 10 10 10 10
until needed. A permanent Si *
stock of MS2 was maintained
frozen. 10

Microbial enumeration Giardia /Monochloramine

At predetermined times, 1
samples were withdrawn from
the test reactors, dechlorinated
Si—observed

with sterile sodium thiosul- 0.1

fate, and assayed for viable
microorganisms. The follow-
ing procedures were used to
0.01
determine microbial viability.
E. coli. Viability of disin-
fected E. coli was determined
0.001
using membrane filtration and
0.001 0.01 0.1 1
the total coliform recovery
Si *
enhancement procedure em-
ploying m-T7 agar.2 Decimal
1
dilutions were taken to obtain 10
accepted numbers of colonies. –0
10 MS2 virus/Free chlorine
Triplicate plates of each vol-
ume were prepared. 10
–1

Incubation. After filtration,

Si—observed

–2
each filter was placed on m-T7 10

agar and incubated at 35oC for 10

–3

22 to 24 h. Yellow coliforms
–4
were counted. None of the 10
experimental waters exhibited –5
10
detectable coliform counts.
G. muris. Enumeration of 10
–6
Giardia cysts was done by the –7
10
procedures outlined by Hoff –7 –6 –5 –4 –3 –2 –1 –0
et al.7 The cysts were concen- 10 10 10 10 10 10 10 10

trated by centrifugation at Si *
500˘ g for 5 min and resus- Observed versus predicted survival ratio plotted for all four waters; 95 percent confidence
pension of the pellet in phos- limits for error bars are included

phate-buffered dilution water.

*TWEEN 80, J.T. Baker Chemical Co.,

Philadelphia, Pa.

Copyright (C) 1996 American Water Works Association

MARCH 1 9 9 6 99

FIGURE 5 Observed survival plots for MS2 virus in three of the
source waters
BDF water
Bull Run water
Willamette River water
1
0.1
Si—observed
0.01
0.001
0.0001
0 0.2 0.4 0.6 0.8
Time—min
2 mg/L of free chlorine used to disinfect MS2 virus for all observations
The cyst densities in the reconcentrated suspensions
were determined by hemacytometer count and
divided into 0.5-mL portions for in vitro excystation.
The survival ratio of cysts was taken as the ratio of the
densities of viable organisms (i.e, excysted and par-
tially excysted trophozoites), using the volumetric
procedure.8
Bacteriophage MS2. The analysis of surviving
phage was performed by plaque assay2 except for the
use of the E. coli host.
Procedure for disinfectant residual
measurement
At least twice during the course of the experi-
ment (at the beginning and the end), samples were
withdrawn from the residual measurement reactor for
the analysis of residuals. These samples were ana-
lyzed immediately after collection to minimize the
possibility of residual loss. The resulting data were
used for analysis of the disinfectant residual.
Free chlorine and monochloramine. Chlorine
residuals were measured using the methods
described2 and at times corresponding to initial, mid-
point, and final experimental conditions as well as
intermediate times as required.
Stock chlorine solution. Chlorine concentrations
were determined by forward amperometric titration
for free chlorine.2
Stock monochloramine solution. After the com-
bined solutions were stirred for 15 min, the resul-
tant solution was checked for free chlorine and mono-
chloramine using forward amperometric titration.2
Ozone. Ozone residuals were measured using the
methods described previously2 and at times corre-
Copyright (C) 1996 American Water Works Association
100 JOURNAL AWWA
sponding to initial, midpoint, and final
times during the experiment and at
intermediate times as required. Ozone
residuals were determined using the
indigo colorimetric procedure.2
Modeling disinfectant residual
decay
Analysis was based on residual mea-
surements taken from the second reac-
tor. Disinfectant residuals were measured
throughout the duration of the experi-
ment. As expected, residuals declined
minimally in BDF water, at rates of <0.25
mg/L for free chlorine, <0.2 mg/L for
monochloramine, and <0.22 mg/L for
ozone. Decreases in residual were most
likely attributable to the demand caused
by organisms in all cases, organic mate-
rial in natural water, volatilization in the
case of ozone and free chlorine, and
1.0 1.2 1.4 1.6 autodecomposition and reaction with
water in the case of ozone.
Residuals decayed according to a first-
order relationship.9 Disinfectant resid-
ual observations were analyzed accord-
ing to the equation
C = C0 exp(–k *t) (1)
in which k* = first-order decay rate, C = observed dis-
infectant residual (mg/L), C0 = initial disinfectant
dose (mg/L), and t = time from start of experiment to
time of sample. The values of k* were determined by
the method of nonlinear regression. In this approach,
the best-fit value of k* was chosen to minimize the
error sum of the squares:
2
3
S Cpredicted – Cobserved 4 (2)
This was achieved using a software application
program.* The results from calculations for all waters
and disinfectants are shown in Table 4. Correlation
coefficients close to 1 demonstrate the validity of the
first-order hypothesis.
A direct relationship relative to demand for each
disinfectant was observed (i.e., k* increases in order of
chlorine + preammoniation < monochloramine < free
chlorine < ozone). The value of k* also varied by water.
Disinfectant demand was observed from greatest to
least for Palm Beach, Willamette River, Bull Run, and
BDF water, respectively. The low value for the BDF
water was expected because dC/dt = 0 is a condition for
demand-free water. In summary, residual demand
was a function of both disinfectant and water.
Models for microbial inactivation
The data analysis of this project 10 used more
sophisticated and statistically accurate kinetic mod-
*Microsoft Excel 4.0 Goal Seek, Microsoft Corp., Redwood, Wash.
els than those used in the
TABLE 5 Two-tailed t test for water comparison using the Hom k* fixed model
SWTR guidance manual.
(p values given)
This analysis incorporated
the basic rate equations given Disinfectant Water E. coli Giardia MS2
in Figure 1, which includes
the Chick–Watson11,12 and Fre e c hlo rine Bull Run <.0 1 (–)* 0 .2 4 † <.0 1 (–)*
Willame tte <.0 1 (–)* 0 .6 3 † <.0 1 (–)*
Hom 13 models. The C X T Palm Be ac h <.0 1 (–)* 0 .0 6 † 0 .0 6 †
tables of the guidance man- Mo no c hlo ramine Bull Run 0 .0 4 (+)‡ 0 .0 2 (+)‡ 0 .6 4 †
Willame tte 0 .6 6 0 .0 6 † 0 .6 7 †
ual only used the Chick– Ozo ne Bull Run <.0 1 (–)* <.0 1 (–)* 0 .1 5 †
Watson model with the dilu- Willame tte <.0 1 (–)* 0 .7 7 † 0 .1 7 †
tion coefficient n = 1. Palm Be ac h 0 .0 2 (–)* 0 .2 1 † §

The first-order decay rate * Kill was le s s than kill in BDF wate r.
for the different water and † The re was no s ys te matic diffe re nc e .
‡ Kill was mo re than kill in BDF wate r.
disinfectant combinations § Only two time data po ints we re available .
had been estimated with a
high degree of correlation
(Table 4); therefore, it was
important to incorporate this term (k*) into the dis- SSE (Uz)/(N – z) z
z
infection kinetic models (Figure 1). These equations
were derived by substituting the first-order residual
}} # 1 + F 1 – a X }}
S SE 0/ ( N – z ) 1 N – z N 2–z
(4)

decay equation (Eq 1) into the original rate expres- in which Uz = the vector of kinetic parameters for
sions for the three kinetic models (Figure 1) and then the bound of the confidence region, N = the number
integrating them.11 The resulting log-survival equa- of observations, z = the number of parameters, a is
tions for both the Chick–Watson and Hom models taken at 5 percent for a 95 percent confidence inter-
with first-order decay are given in Figure 2. The val, and F = the cumulative Fisher F distribution with
approximate solution to the Hom model with decay z and N – z degrees of freedom at the upper 1-a per-
was employed in this analysis.14 centile. This equation was rearranged for the confi-
The Hom and Chick–Watson equations in Figure dence interval:
2 were regressed against the survival data, and the z z
first-order decay rate was fixed according to the
appropriate value given in Table 4. Previous work10
1
SSE (Uz) = SSE0 X 1 + F 1 – a X }}
N – z N2 –z
(5)

has demonstrated that the Hom model provided a A software program* was used to solve the pre-
statistically significant improvement in fit to the inac- vious equation for the error sum of squares SSE(Uz)
tivation process, relative to the Chick–Watson model by varying only one of the three kinetic parameters.
in particular. Therefore, the Hom approach rather An initial high guess for the parameter was used for
than the Chick–Watson approach embodied in the C upper-bound convergence, and an initial low guess
X T tables was deemed the model of choice. The bet- was used for lower-bound convergence. These ini-
ter fit of the Hom model is illustrated in Figure 3 for tial guesses were based on the optimal parameter
the case of Giardia with monochloramine. The Hom values determined using nonlinear regression.
model better accounts for the shoulder in the sur-
vival data. Water comparison analysis
The optimal kinetic model parameters (i.e., k, n, In the water comparison analysis, inactivations
and m) were calculated using nonlinear regression achieved in BDF water were compared with those of
according to the same procedure used for the resid- the Bull Run, Willamette River, and Palm Beach
uals. The SSE0 was minimized for the survival data waters. The authors wanted to test whether disinfec-
according to the equation tion experiments performed with BDF water could be
used to predict performance in other waters. The pre-
SSE0 = S(lnSi – lnSi*)2 (3) dicted parameters k, n, and m from the Hom model
fit to BDF water were substituted into the predicted
in which Si = the observed survival ratio and Si* = the survival equations for the other waters, using the Hom
predicted survival ratio as given in Figure 2. Soft- with k* model of Figure 2. Only the first-order resid-
ware* was used to iterate the values of the model ual decay rate k* was retained because it was unique
parameters to a small tolerance level until the objec- for each of the waters. This led to a new set of predicted
tive function given by Eq 3 was minimized. survival values (Si* in Figure 2) for each of the waters
To calculate a confidence region for the kinetic that deviated slightly from the predicted survival for
parameters and the resulting predicted survival, the BDF water (according to the differences in k*).
model prediction error was computed by propagating The predicted survival values (Si*) were plotted
the errors associated with the determination of the against the actual observed values (Siobserved) for each
kinetic parameters. Upper and lower bounds for each of the waters (Figure 4). A perfect agreement between
of the parameters k, n, and m were determined using
the F ratio test:15 *Microsoft Excel 4.0 Solver, Microsoft Corp., Redwood, Wash.

Copyright (C) 1996 American Water Works Association

MARCH 1 9 9 6 101
 FIGURE 6 Observed survival plots for Giardia in three of the source
nificant difference between prediction
waters and observation—was refuted; in other
words, there was a statistically signifi-
BDF water cant difference between predicted and
Bull Run water observed inactivation values. In a num-
Willamette River water ber of cases, the levels of inactivation
predicted by the use of kinetic parame-
ters obtained in demand-free water dif-
1 fered significantly from those obtained
in one of the natural waters.
The differences between the predicted
and observed inactivations were greater
for E. coli and the MS2 virus than for
Si—observed

Giardia. It is possible that in the case of E.

0.1 coli and the MS2 virus, the failure to
achieve high levels of inactivation (e.g.,
99.9 percent +) can be attributed to some
protection afforded by the turbidity pre-
sent in the natural waters but not in BDF
water. This explanation is consistent with
0.01
the fact that protection by solids may
result in the diminution of microbial
inactivation.16,17
0 50 100 150 200 Survival values for the various waters
Time—min deviated from predictions because water
1 mg/L monochlorine used to disinfect Giardia for all observations quality influences predicted inactivation.
The kinetics observed in demand-free
water did not provide a good indication
of the inactivation process in actual
observed survival and prediction would result in a waters, even when corrected for disinfection demand.
45o line. Observed points for BDF water were ex- Although the reason for this deviation remains
pected to approximate the 45o line because the kinetic unclear, it does indicate that disinfection performance
parameters were predicted using nonlinear regres- should be tested using the water of interest.
sion for BDF water. The predicted survivals for many The plots of Figures 5 and 6 are based on survival
of the water and disinfectant combinations deviated data for three of the four experimental waters. All
from the 45o line. Points plotted to the right of the 45o of these experiments were performed with the same
line indicated more kill than was found in BDF water. organism and dosage of disinfectant. Systematic dif-
Conversely, points to the left of the 45o line indicated ferences among the inactivations of the source waters
less kill. were apparent. For the MS2 virus and free chlorine
The error bars on these figures depict the 95 per- experiments of Figure 5, the inactivation rate was
cent confidence limits for the predicted inactivation greater for BDF water than for other waters. However,
values. Error bars were calculated using the upper in the Giardia and monochloramine experiments of
and lower limits of each of the kinetic parameters k, Figure 6, the kill in the Bull Run water was faster
n, and m. The resulting absolute maximums and min- than the kill in other waters. Although the inactiva-
imums for the predicted survival values were calcu- tions differed systematically for each of the source
lated using the 95 percent confidence space of para- waters, this difference was usually less than 1 log.
meter limits. In some cases, the error bars did not The SWTR guidance manual C X T tables make
intersect the 45o line. Some of the waters deviated adjustments for pH, but the expected pH effects from
systematically from the 45o line, indicating greater the disinfection kinetics are not always demonstrated
or lesser inactivation than that found in BDF water. in the field. In Table 1, for example, the pH of Bull
A more rigorous statistical test was used to compare Run water is less than that of BDF water, and the pH
the inactivations in the different waters. of Willamette water is greater than that of BDF water.
A paired t-test was conducted to compare the pre- From the disinfection kinetics, a lower pH would
dicted inactivations in the Bull Run, Willamette, and increase the hypochlorous acid to hypochlorite ratio
Palm Beach waters with inactivations in BDF water. and increase inactivation.18 This trend is also fol-
For each water and microorganism combination, the lowed for the guidance manual C X T values given for
paired t statistic between the observed log-survival Giardia cysts and viruses for the whole range of tem-
ratio and the predicted log-survival ratio was com- peratures. Therefore, free chlorine is expected to be
puted. The significance level (p value) for the two- more biocidal at low pH. However, Figure 5 indicates
sided test was computed (Table 5). A low p value on that the inactivation in Bull Run water is slower than
this test indicates that the null hypothesis—i.e., no sig- that of BDF water even though the pH is lower. In

Copyright (C) 1996 American Water Works Association

102 JOURNAL AWWA
Figure 6, the kill rate in the Bull Run water is faster
than the other waters, but the C X T tables for chlo-
ramines make no distinction for different pH values.
Only one C X T table is given for chloramines at a pH
range of 6–9. In short, even though the pH is to some
extent accounted for in the C X T tables, it is hard to
imagine that the synergistic effects of all of the water
quality parameters can be adequately modeled.
Furthermore, the disinfectant demand is a func-
tion of water quality but not necessarily a predictor
of inactivation, as a comparison of the residual decay
rates given in Table 4 with the inactivations in Figure
6 makes clear. The Bull Run water shows a higher
first-order decay rate (k*) than BDF water, yet inac-
tivation is faster in BDF water (Figure 6).
Conclusions
• The effect of water type is significant in pre-
dicting microbial inactivation.
• Inactivation data obtained in BDF water do not
appear to be adequate predictors of microbial inacti-
vation in actual waters to be treated. Disinfection
testing should be performed in the water of concern.
• Even after disinfectant demand is taken into
account, other factors appear to influence inactivation
of organisms in various waters. The particular aspects
of water quality that could account for this finding
include turbidity or inorganic chemical composition.
The effect of different water qualities on sublethal
injury and repair processes may also be important.
These variables are worthy of further investigation.
• The use of pH in adjusting the tabulated C X T
values is insufficient to characterize the effect of water
quality on disinfection performance.
Acknowledgment
The authors thank the AWWA Research Founda-
tion for funding this project and personnel at the
Portland (Ore.) Water Bureau for assisting with exper-
imental work. Joel Hornberger assisted with experi-
mental analyses.
References
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About the authors: Charles D.
Haas is L.D. Betz Professor of Envi-
ronmental Engineering, Drexel Uni-
versity, Philadelphia, PA 19104. A
member of AWWA, WEF, and
IAWQ, Haas has previously pub-
lished his research in J OURNAL
AWWA and several other profes-
sional journals. He is past chairman
of AWWA’s Disinfection Committee. Haas has BS and MS
degrees from the Illinois Institute of Technology in Chicago
and a PhD degree from the University of Illinois in Urbana.
Joseph G. Jacangelo is a vice-president of Montgomery Wat-
son, 560 Herndon Pkwy., Herndon, VA 22070. Josh Joffe
is an environmental engineer with Malcolm Pirnie, Inter-
national Blvd., Mahwah, NJ 07495. Uma Anmangandla
is an environmental engineer with PRC Environmental
Management, 1800 JFK Blvd., Philadelphia, PA 19103,
and Mark Heath is an environmental engineer with Mont-
gomery Watson, 700 3rd Ave., Seattle, WA 98104.
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