Ozone: Science & Engineering

ISSN: 0191-9512 (Print) 1547-6545 (Online) Journal homepage: http://www.tandfonline.com/loi/bose20

Inactivation of Giardia muris Using Ozone and

Ozone-Hydrogen Peroxide

Charles W. Labatiuk , M. Belosevic & Gordon R. Finch

To cite this article: Charles W. Labatiuk , M. Belosevic & Gordon R. Finch (1994) Inactivation of
Giardia muris Using Ozone and Ozone-Hydrogen Peroxide, Ozone: Science & Engineering, 16:1,
67-78, DOI: 10.1080/01919519408552381

To link to this article: http://dx.doi.org/10.1080/01919519408552381

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 OZONE SCIENCE & ENGINEERING 0191-9512/94 $3.00 + .00
Vol. 16, pp. 67-78 International Ozone Association
Printed in the U.S.A. Copyright © 1994

Inactivation Of Giardia muris Using Ozone And

Ozone-Hydrogen Peroxide

Charles W. Labatiuk1', M. Belosevic2 and Gordon R. Finch1

1. Environmental Engineering and Science Program

Department of Civil Engineering
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2
Departments of Zoology and Immunology
University of Alberta
Edmonton, Alberta, CANADA T6G 2G7

Received for Review: 2 1 June 1993

Accepted for Publication : 7 December 1993

Abstract

The disinfection effects of the ozone molecule alone and that of ozone
decomposition products when inactivating Giardia muris cysts were
investigated at bench-scale using two different ozone demand-free laboratory
buffer systems. The first water was a 0.05 M phosphate buffer with hydrogen
peroxide added at a 10:1 weight ratio. The second water was a 0.05 M
phosphate - 0.01 M bicarbonate buffer which quickly scavenged radical
species from ozone decomposition. The C3H/HeN mouse model was used to
assess the infectivity of ozone treated cysts.

The phosphate-bicarbonate buffer system had significantly greater ( P < 0.05)

inactivation of G. muris cysts than that observed in the phosphate buffer -
peroxide system where ozone was completely decomposed in less than 120 s.
Consequently, the design of ozone disinfection processes should maintain
ozone residual for disinfection prior to the addition of hydrogen peroxide for
the oxidation of other compounds.

Introduction
Two primary oxidation reaction pathways for ozone in water have been proposed:
direct oxidation which is a selective oxidation of compounds by the ozone molecule
and oxidation by radical intermediates [Hoigne and Bader, 1975). When ozone reacts

At the time of the study, C. Labatiuk was a Ph.D. candidate at the University
of Alberta. He is now an Environmental Engineer with the City of Edmonton,
9803-102A Avenue, Edmonton, Alberta, T5J 3A3.
67
 68 C.W. Labatiuk et al.

with substances in natural waters, it is very likely reacting in a combination of the

direct oxidation and oxidation by radical intermediate pathways (Glaze et al., 1987).
It is not fully understood whether direct oxidation or oxidation by radical intermediates
predominate in disinfection reactions. Hoigne and Bader (1975] stated that
disinfection was the result of direct ozone oxidation. They noted that the reactions
of hydroxyl radicals with molecularly dissolved substrates often occurred in micro-
seconds and that the radicals were consumed before they encountered a microorgan-
ism. Others have suggested an hydroxyl radical mediated mechanism in disinfection
reactions (Bancroft et al., 1984].
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Hydrogen peroxide accelerates the rate of ozone decomposition, increasing the

quantity of high-energy, short-lived hydroxyl radicals (Wolfe et al., 1989a). A pilot-
scale investigation of the disinfection efficacy of ozone-hydrogen peroxide and ozone
in two untreated surface waters was conducted by the Metropolitan Water District of
Southern California (MWDSC) (Wolfe et al., 1989a; Wolfe et al., 1989b; Ferguson et
al., 1990; Scott et al., 1992). The test organisms used included G. muris, MS2 and
f2 coliphages, Escherichia coll and R2A-heterotrophic plate count bacteria. It was
found that ozone-hydrogen peroxide was less efficient at higher hydrogen peroxide to
ozone ratios, and this was postulated to be due to a concomitant lower ozone residual.
This result suggests that the more active agent in the ozone-hydrogen peroxide
process is molecular ozone. Researchers with the MWDSC have coined the term
PEROXONE for the ozone-hydrogen peroxide process. This process is but one of a
number of advanced oxidation processes in which one oxidant is catalyzed by another
or ultraviolet radiation to take advantage of radical reaction pathways (Glaze et al.,
1987). Ozone-hydrogen peroxide has been used in the control of trihalomethane
formation potential, in the control of compounds associated with taste and odors and
for the oxidation of organic compounds. The controlling factors for the oxidative
destruction of different types of organic substrates appear to be pH and the hydrogen
peroxide to ozone weight ratio (Ferguson et al., 1990).

Little is known regarding the effectiveness of ozone-hydrogen peroxide in disinfection.

Duguet et al. (1989) studied Bacillus cereus inactivation using hydrogen peroxide to
ozone weight ratios of 0.4 to 1.0. They concluded that ozone-hydrogen peroxide was
applicable for disinfection but was approximately 1.5-logless efficient than ozonation
alone. Hydrogen peroxide alone has been reported to be an ineffective disinfectant
at the relatively low concentrations appropriate for use in municipal water disinfection.
The U.S. National Research Council (1980) reported that 90 percent inactivation of
E. coll required contact times in excess of 300 min and hydrogen peroxide concentra-
tions of 30-90 mg/L.

The objective of the research reported here was to distinguish between the disinfection
effects of the ozone molecule alone and the effect of ozone decomposition products
when inactivating G. muris cysts under controlled conditions. The results reported
are from laboratory experiments conducted using G. muris, laboratory water, and the
C3H/HeN mouse-6. muris model.
 G. MURIS INACTIVATION WITH 0 3 AND (yH 2 O 2 69

Experimental Protocol
OZONE APPARATUS AND METHODS

Test Waters. Two different ozone demand-free laboratory buffer systems were used.
The first water was 0.05 M phosphate buffer [pH 5.7) with a final hydrogen peroxide
(Perhydrol®; E. Merck] to ozone weight ratio of 10 to 1. It was found that this ratio
completely decomposed ozone in 120 s or less, ensuring a large supply of decomposi-
tion radical species. Lower ratios were found to leave significant concentrations of
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residual ozone which would have confused the interpretation of the results. The
second water was 0.05 M phosphate-0.01 M bicarbonate buffer {pH 6.4) selected to
scavenge radical species from ozone decomposition quickly.

The 0.05 M phosphate buffer was prepared using potassium dihydrogen orthophos-
phate and disodium hydrogen orthophosphate (AnalaR grade; BDH). The 0.05 M
phosphate-0.01 M bicarbonate buffer was prepared by a 1 in 20 dilution of a stock
solution consisting of 127.92 g KH2PO4,8.43 g Na,HPO4 and 16.80 g NaHC03 (AnalaR
grade; BDH) dissolved in 1,000 mL of Milli-Q® water. Deionized laboratory water was
obtained from a Milli-Q9 system (OM-140; Miltipore Corp.) operated at a resistivity of
at least 18 MQ/cm. Deionized water was used for preparing all analytical and test
solutions including the stock ozone solution.

Ozonation. A concentrated stock solution of ozone was prepared using the same
procedure described previously (Labatiuk et al., 1992a). The stock solution ozone
concentration typically was 18 to 20 mg/L at 22°C. Ozone concentration in the
aqueous phase was determined continuously using ultraviolet absorbance at 260 nm
and using a molar absorption coefficient of 3,300 M ' cm'1 [Gordon et al., 1988). Test
solution absorbance was integrated for 0.5 s with a 1 s cycle time using a diode array
spectrophotometer (8452A; Hewlett-Packard). Ozonation was performed at bench-
scale using side-stream injection of the concentrated ozone solution. A 250 mL
Erlenmeyer flask containing approximately 200 mL of liquid was used as the reaction
vessel. The reactor and test water were made ozone demand-free prior to use as
described elsewhere (Labatiuk, 1992; Labatiuk et al., 1992a). Mixing was accom-
plished using a Teflon®-coated magnetic stir bar. Residual ozone was neutralized in
the reactor at the end of the contact period using 50 mL of 1.0 M sodium formate.
The concentration of hydrogen peroxide in aqueous solution was determined using the
cobalt method (Masschelein et al., 1977). Residual hydrogen peroxide was neutralized
using 0.600 mL of 20,000 enzyme units/mL catalase [from bovine liver in a solution
containing glycerol and ethanol; BDH).

GIARDIA METHODS

Parasite. The strain of G. muris used in this study was isolated originally by
Roberts-Thomson et al. (1976) and was obtained from B.J. Underdown, McMaster
University. Specific-pathogen-free 6 to 8 week-old, male, inbred C3H/HeN mice were
used and were obtained from Charles River Breeding Laboratories [St. Constant,
Quebec). The procedures for parasite maintenance, animal care, and the isolation of
the required quantity of highly purified G. muris cysts were described in detail
previously [Labatiuk et al., 1991; Finch et al., 1992a). After isolation cysts were
 70 C.W. Labatiuk et al.

stored at 4°C for 18 h prior to use and were used within 48 h of preparation. Hie
absorbance at 260 nm of a cyst suspension of 104 cysts/mL was used as a quality
control measure. For use in disinfection experiments, cysts at that concentration
could have no more than 0.03/cm absorbance.

C3H/HeN Mouse-G. muris Model. The C3H/HeN mouse-G. muris model [Roberts-
Thomson et al., 1976) was used to assess G. muris cyst infectivKy. The C3H/HeN
mouse exhibits high susceptibility to infection (Belosevic et al., 1984). The basis of
the model is the correlation between the size of the inoculum (cysts/mouse) and the
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time when all mice in a group were releasing cysts in the feces (days). All mice
inoculated with 10s, 104, 10J,10*, and 10 cysts were positive on days 3, 4, 5, 6 and
8, respectively. If all the mice in an ozonated trial did not become positive, the last
day which showed an increase in the number of positive mice was used to determine
the infective dose. The relation used for estimating inactivation (N/No) from mouse
model data was (Labatiuk et al., 1992b):
n x
JL = I m
No nQx l0

where n is the number of positive mice in an ozonated trial; I is the infective dose
after ozonation, estimated using the latent period; n,, is the number of positive mice
in the control; and !<, is the inoculum size in the control, estimated by hemocytometer
count.

PROCEDURE

Buffer was added to a 250 mL Erlenmeyer flask reactor. In the first buffer system,
hydrogen peroxide was added to the reactor to provide a final concentration of about
17 mg/L. The total initial volume in both buffer systems was approximately 200 mL.
The test solution was seeded with G. muris cysts to provide a concentration that was
typically 104 cysts/mL. The concentration of stock ozone solution was measured twice
immediately before a calculated volume of ozone solution was added to the buffer
using a mass-calibrated pipette. This was the applied ozone dose. Over the duration
of the experiment, the test solution was pumped through a thermostatically controlled
35 |iL, 1-cm light path length flow cell at approximately 8 mL/min. At the end of the
contact time, a slight excess of 1.0 M sodium formate was added to the reactor. The
drop in absorbance was assumed to be caused by formate reacting with ozone.
Residual hydrogen peroxide was neutralized using a slight excess of catalase.
Hydrogen peroxide, sodium formate and catalase were used in the controls.

Following ozonation, but prior to concentration for the infectivity testing procedure,
four replicate hemocytometer counts of cyst density were made. This was used to test
the homogeneity of the cyst suspension. If the variation amongst replicates exceeded
that from chance alone [P < 0.05) the data were discarded. It was not necessary to
discard any data due to this criterion.

All disinfected liquid was centrifuged in 175 mL plastic conical centrifuge tubes at
800 x g for 10 min. The supernatant was removed, leaving a 1.5 mL centrifugate, of
 G. MURIS INACTIVATION WITH O3 AND O ^ J O J 71
3

which 1.0 mL was used for infecting mice. A minimum of four hemocytometer counts
were made on the 0.5 mL portion of the sample in order to determine the total
number of cysts. Each one of a group of 5 C3H/HeN mice was orally inoculated with
0.2 mL. The size of the inoculum typically was 104 cysts/mouse. During experiments,
mice were housed individually and the feces were checked daily for G. muris cysts
from day 3 or 4 until day 8, or when they became positive. On day 8, necropsy was
performed on all mice which were negative for G. muris cysts, and the small intestine
was examined for the presence of trophozoites (Labatiuk, 1992).
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Experimental Design

This experiment was designed as a series of trials using two different ozone demand-
free laboratory buffer systems. The first water was a 0.05 M phosphate buffer with
hydrogen peroxide added at a 10:1 weight ratio. The second water was the 0.05 M
phosphate-0.01 M bicarbonate buffer designed to scavenge radical species from ozone
decomposition quickly. Each trial in each buffer was an independent observation of
G. muris cyst inactivation.

The hypothesis being tested was that if the large quantity of radical intermediates
produced in the ozone-hydrogen peroxide process was more effective than ozone
alone, then the peroxide system would result in more inactivation than ozone alone.
The mean applied ozone dose was 1.63 ± 0.07 mg/L. All trials were conducted in
triplicate at room temperature (22°C) and with a 2-min contact time. Data was
analyzed using a t-test. Data Desk Professional® v.3.0 [Odesta Corporation,
Northbrook, IL, USA) was used to perform routine statistical analyses.

Results and Discussion

Details on animal responses and inactivation results for the three trials using each
laboratory buffer system are presented in Table 1.

All mice in control groups invariably became positive: on day 3 for the 10s cysts per
mouse inoculum and on day 4 for the 104 cysts per mouse inoculum. The control
group results also indicated that sodium formate, hydrogen peroxide and catalase had
no effect on G. muris.

Figures 1 and 2 show ozone concentration versus time for the three trials using each
laboratory buffer system. The concentration factor used for ultraviolet absorbance is
14.55 cm-mg L-1 when 3,300 M-'cm"1 is used as the molar absorption coefficient.
Figure 2 indicates the beneficial effects of bicarbonate in quenching the ozone
decomposition reaction. The half-life of ozone in the 0.05 M phosphate-0.01 M
bicarbonate buffer after immediate demand was satisfied was approximately 10 min,
while the half-life of ozone in the phosphate buffer hydrogen peroxide system was
approximately 25 s after immediate demand.

The mean G. muris cyst inactivation in the phosphate-bicarbonate buffer system was
3.9 + 0.9 log inactivation (mean ± one standard deviation] compared with 2.3 ± 0.2
log inactivation in the hydrogen peroxide system. The phosphate-bicarbonate buffer
 72 C.W. Labatiuk et a!.

system, which had little decomposition of ozone over the duration of the experiment
[mean residual ozone of 1.5 ± 0 . 1 mg/L at 2 min which was 92 ± 2 percent of the
applied ozone dose), had significantly greater [P < 0.05) inactivation of G. muris
cysts than that observed in the phosphate buffer hydrogen peroxide system where the
ozone was completely decomposed in less than 120 s. Thus, the disinfection effect
of the stabilized system is superior to that of the free radical system. By inference,
the presence of large quantities of short-lived radical intermediates is not as effective
in the inactivation of G. muris cysts as the ozone molecule alone for a sufficient
contact time.
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Table 1

Ozone and ozone hydrogen-peroxide experiment: Number of C3H/HeN mice

infected with G. muris as a function of days after infection and inoculum size

Inocu- Number Days After Infection Inacti-

Trial Buffer lum of mice (No. of mice positive for cysts) vation
(cysts per per 3 4 5 6 7 8 81* (log
mouse) trial N/NQ)

1 PBP 2.0E+04 5 . 0 1 2 2 2 2 2.5

2 PBP 5.0E+04 5 0 0 5 . . . 2.2
3 PBP 6.5E+04 5 1 1 5 . . . 2.2
4 PCB 1.1E+03 5 . 0 1 1 1 2 2 3.4
5 PCB 5.0E+04 5 0 1 3 4 4 4 4 3.3
6 PCB 4.2E+04 5 0 0 0 0 1 1 1 4.9
Cl* PBP 1.4E+04 5 - 5 - 0.0
C2 PBP 1.5E+05 5 5 . . . . 0.0
C3 PCB 1.1E+04 5 . 5 0.0
C4 PCB 1.6E+05 4 4 - - _ 0.0

*C1, control l;
#
8T, number of mice positive for cysts or trophozoites on day 8;
PBP, 0.05 M phosphate buffer with 10 to 1 peroxide to ozone weight ratio;
PCB, 0.05 M phosphate-0.01 M bicarbonate buffer.

In each of the peroxide system trials the average ozone residual present during the
time it took the ozone to decompose was calculated by numerical integration. The
integrated ozone residuals were found to be 0.37 mg/L, 0.22 mg/L, and 0.46 mg/L in
trials 1,2, and 3, respectively. These concentrations and the corresponding G. muris
cyst inactivations were plotted with other available data (Table 2] to determine if the
inactivation in the peroxide system could be attributed to the ozone residual that was
present during decomposition. The 2.3-log inactivation in the peroxide system can be
attributed to the average ozone residual (0.35 ± 0.12 mg/L) that was present over the
120 s it took the ozone to decompose. This amount of ozone in demand-free 0.05 M
phosphate buffer is sufficient to inactivate at least 99.9 percent of the G. muris cysts
in 120 s (Figure 3).
 G. MURIS INACTIVATION WITH O3 AND Oj/H2O2 73

i
•3 Trial 1

c
o
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U
o
C

Figure 1. Ozone decomposition in 0.05 M phosphate buffer with a 10 to 1 hydrogen

peroxide to ozone weight ratio.

Trial 6
to
6
c
o
•a
£
u
G
O
U
o

§
o
Time, s

Figure 2. Ozone decomposition in 0.05 M phosphate-0.01 M bicarbonate buffer.

Researchers working with B. cereus and G. muris concluded that ozone-hydrogen

peroxide was applicable for disinfection but was less efficient than ozone alone,
particularly at higher hydrogen peroxide to ozone weight ratios (Duguet et al., 1989;
Wolfe et al., 1989b). Observations made with E. coli and two buffer systems similar
to those reported here could be explained either in terms of ozone decomposition
products or the ozone molecule; however, inactivation continued in the presence of
an ozone residual and not in the peroxide system where the ozone residual was below
the detection limit by the end of 60 s (Finch et al., 1992b). These findings in
conjunction with the work reported here suggest that ozone alone is a more effective
disinfectant.
 74 C.W. Labatiuk et al.

o-,
O peroxide system -120 s contact time
A phosphate-bicarbonate system -120 s contact time
D phosphate buffer-120 s contact time
to
o • phosphate buffer-300 s contact time
-2-
O O

-3-
o
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>
-4-
• •
-5-

3 4 5 6 7 10
Integrated Ct (mg»min/L)
Figure 3. G. muris cyst inactivation in various laboratory buffer systems as a
function of the integrated (ozone residual x contact time] product (22°C;
pH 5.7-6.7).

Table 2

Ozonation conditions for G. muris in laboratory buffer systems (22'C; pH 5.7 - 6.7)

Contxt Applied nitial ozone Final ozone Integrated Inactivation

Trial Buffer lime ozone dose residual residual ozone residual (log N/N o ) Reference
(min) (ma/L) img/L) (mg/L) (ms>/L)
1 PBP 2 1.55 0.85 0.06 0.37 2.5 This study
2 PBP 2 • 1.62 0.74 0.00 0.22 2.2 This study
3 PBP 2 1.68 0.94 0.00 0.46 2.2 This study
4 PCB 2 1.58 1.44 1.42 1.43 3.4 This study
5 PCB 2 1.60 1.58 1.50 1.54 3.3 This study
6 PCB 2 1.73 1.71 1.59 1.65 4.9 This study
7 PB 5 0.62 0.47 0.26 0.35 4.2 Labatiuk et al., 1991
8 PB 5 0.64 0.52 0.37 0.44 4.2 Labatiuk et al., 1991
9 PB 5 0.26 0.18 0.08 0.12 3.1 Labatiuk et al., 1991
10 PB 5 0.30 0.22 0.11 0.15 3.1 Labatiuk et al., 1991
11 PB 5 2.55 2.14 1.29 1.66 4.4 Labatiuk et al., 1991
12 PB 5 0.59 0.51 0.35 0.42 >5.1 Labatiuk et al., 1992b
13 PB 2 0.53 0.48 0.41 0.44 3.2 Labatiuk etal., 1992b

t. based on initial and final ozone residual over the contxt time, numerical integration used for trials 1- 3;
>, detection limit of C3H/HeN mouse model;
PBP, 0.05 M phosphate buffer with 10 to 1 peroxide to ozone weight ratio;
PCB 0.05 M phosphate-0.01 M bicarbonate buffer.
PB, 0.05 M phosphate buffer.
 G. MURIS INACTIVATION WITH O3 AND CyH 2 O 2 75

The Surface Water Treatment Rule requires that 99.9 percent of Giardia lamblia
cysts and 99.99 percent of enteric viruses be inactivated or removed for surface water
supplies (U.S. EPA, 1989). The disinfectant concentration x contact time (Ct) product
is used to establish disinfection credits. Appendix 0 in the Guidance Manual [U.S.
EPA, 1991) gives recommendations on the definitions of C and t in ozone disinfection.
The bench-scale ozonation protocol used in this work may be regarded as a very short,
continuously stirred-tank reactor segment during ozone addition followed by a reactive
flow segment for the specified contact time. The Ct product in the reactive flow
segment can be estimated by integrating the ozone residual over the contact time.
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For example, in the phosphate-bicarbonate buffer system trials, ozone decomposition

was a first-order reaction with respect to ozone concentration. Therefore the
integrated ozone residual, C, in each trial was:

(\nC. + \nC,)
C = exp '-\

where C; is the initial ozone residual and C, is the final ozone residual. The initial
ozone residual is the dissolved ozone concentration observed in the reactor at the
beginning of the contact period. The initial ozone residual would be analogous to the
transferred ozone dose if ozone was added as a gas rather than in dissolved form.
The final ozone residual is the ozone concentration at the end of the contact time.
In many full-scale ozone contactors, it is not practical to measure ozone profiles
directly. In this case, the exiting ozone residual for the chamber is recommended as
a conservative measure of C (U.S. EPA, 1991). In a batch reactor the actual contact
time can be used as t. However, in full-scale systems t should be interpreted as the
t10 requirement of flow-through reactors in the EPA Guidance Manual to compensate
for nonidealities such as short-circuiting.

Figure 4 indicates that the apparent Ct for 3 log-units of G. muris cyst inactivation
by ozone is about 0.45 mg x min/L, and for 4 log-units about 0.90 mg x min/L. The
regulatory Ct value for 3 log-units of G. lamblia cyst inactivation by ozone is 0.62 mg
x min/L and for 4 log-units, 1.22 mg x min/L (22°C; extrapolation from Table 0-2; U.S.
EPA, 1991). Differences in the experimental protocol used here and that used in the
research on which Table 0-2 in the Guidance Manual was based are discussed
elsewhere (Labatiuk et al., 1992a). The regulatory Ct values contain a safety factor
of two. Without this factor, they are smaller than the Ct values required to inactivate
the G. muris surrogate. Wolfe et al. (1989b) determined that the Ct for 2 log-units
of G. muris cyst inactivation by ozone in untreated surface water (14°C, pH 8] also
was higher than the regulatory value. These results indicate that there still may be
uncertainty regarding the inactivation of Giardia spp. by chemical disinfection.
Furthermore, should any part of a chemical disinfection system fail, or strict
operational criteria of contact time, disinfectant residual, and influent water quality
not be closely monitored; the single barrier provided by chemical disinfection could
be compromised.
 76 C.W. Labatiuk et al.
o -,
O peroxide system - 120 s contact time
A phosphate-bicarbonate system -120 s contact time
• phosphate buffer-120 s contact time
60 • phosphate buffer - 300 s contact time
O
•a -3-

I
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-4-
in
-5- < -5.il
0 1 2 3 4 5 6 7 8 9 10
Ct(mg»min/L)
Figure 4. G. muris cyst inactivation in various laboratory buffer systems as a
function of the final ozone residual x contact time product (22°C; pH 5.7-
6.7).

Summary And Conclusions

It may be inferred from these results that the direct action of the ozone molecule is
the most important pathway for inactivation of G. muris cysts. Significantly greater
inactivations were obtained in the presence of ozone residual (P < 0.05]. The
G. muris cyst inactivation that occurred in the peroxide system could be attributed
to the ozone residual that was present during decomposition. Little additional
inactivation appeared to occur due to the presence of large quantities of radical
intermediates. While the efficacy of ozone - peroxide has been demonstrated for
oxidation processes, it is apparent from these results that ozone alone is a better
disinfectant. Consequently, the design of ozone disinfection processes should maintain
a stable ozone residual for disinfection prior to the addition of hydrogen peroxide for
the oxidation of other compounds.

A ckn o wledgemen t

The American Water Works Association Research Foundation provided funding for this
work.

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 78 C.W. Labatiuk et al.

Key Words

Ozone; Ozone-Hydrogen Peroxide; Disinfection; Inactivation; Giardia muris cysts;

Infectivity; Animal Model;

Resume

L'action desinfectante de la molecule d'ozone seule ou des produits de reaction de

l;ozone lors de I'inactivation des cysts de Giardia muris a ete etudiee en utilisant
Downloaded by [Nanyang Technological University] at 08:25 11 June 2016

deux milieux tampons n'ayant aucune demande chimique en Ozone. Le premier milieu
etait un tampon phosphate 0,05M additionne de peroxyde d'hydrogene dans un
rapport massique 10=1. Le second milieu etait un tampon phosphate 0,05M +
hydrogeno carbonate 0,01M qui est capable de pieger rapidement les radicaux issus
de la decomposition de I'ozone. Le modele souris C3H/HeN a ete utilise pour etablir
le pouvoir infectant des cystes traites a I'ozone.

Le tampon phosphate hydrogeno-carbonate a permis une inactivation significativement

plus importante (P < 0,05) des cystes de Giardia muris que le tampon phosphate-
peroxyde dans lequel I'ozone etait entierement decompose en moins de 120 s. En
consequence, la conception du procede de disinfection par I'ozone doit maintenir un
residuel djozone en vue de la disinfection avant injection du peroxyde d'hydrogene
utilise pour I'oxydation d'autres composes.

Zusammenfassung

Die Desinfektionswirkung des Ozonmolekiils allein und diejenige der Zersetzungs-

produkte wurden im Hinblick auf die Inaktivierung von Giardia muris untersucht.
Dabei wurden im Labormassstab zwei verschiedene Puffersysteme verwendet, die
selbst keinen Ozonbedarf aufwiesen. Das eine Wasser war ein 0.05 m Phosphatpuffer
mit einem im Gewichtsverhaltnis 10:1 zugefiigten H202-Anteil. Das zweite Wasser war
ein 0,05 m Phosphat-0,01 m Bicarbonatpuffermit schnell abgefangenen radikalen aus
der Ozonzersetzung. Das C3H/HeN Maus Model wurde eingesetzt, um die Inaktivierung
der mit Ozon behandelten Giardia muris Zellen zu iiberpriifen.

Das Phosphat'/Bicarbonatpuffersystem zeigte eine signifikant grossere (P < 0,05)

Inaktivierugsleistungvon Giardia muris als das Phosphatpuffer-H^-System, in dem
Ozon innerhalb von 120 sec. vollstandig zersetzt war. Daraus folgt, dass eine
Ozonresidual zur Desinfektion vorhanden sein sollte, bevor zur besseren Oxidation von
organischen Verbindungen H202 zugesetzt wird.