Journal of Hospital Infection (2006) 64, 386e390
Efficacy of super-oxidized water foggingin environmental decontamination
J. Clark , S.P. Barrett M. Rogers , R. Stapleton
a Department of Microbiology, Charing Cross Hospital, London, UKb Sterilox Technologies International ltd, Beaconside, Stafford, UK
Received 5 September 2005; accepted 2 July 2006Available online 14 October 2006
The efficacy of decontamination using Sterilox fog was assessed
against meticillin-resistant Staphylococcus aureus (MRSA) and Acinetobacter
baumannii. Ceramic tiles were inoculated with the test organisms and, once
dried, were subjected to Sterilox fogging using a stationary vaporizing ma-
chine sited at a distance of 3 m for 10 min and then left for a further hour. In a second experiment using the same organisms, the first 10-min foggingperiod was followed by a directed fogging period of 30 s at a distance of1 m. Organisms were cultured from the tiles, plated on to tryptone soya agarand incubated for 48 h. Initial counts of approximately 109 colony-formingunits/mL for both organisms were reduced approximately 104 fold for MRSAand 105.8 fold for A. baumannii when using a single fogging. The second fog-ging resulted in 106.8-fold reductions for both organisms. Sterilox fog is safeand simple to use, and can reduce levels of nosocomial pathogens by a factorof almost 107. It is worthy of clinical evaluation in clinical settings to deter-mine whether it maintains its microbicidal effects against a variety of organ-isms on different surfaces. ª 2006 The Hospital Infection Society. Published by Elsevier Ltd. All rightsreserved.
The use of disinfectants to decontaminate hospi-tals has mixed success in eliminating organisms
Meticillin-resistant Staphylococcus aureus (MRSA),
from the environment, and novel methods of
and the means of controlling it, continue to be of
cleaning have been explored previously.Various
major interest to the healthcare community
methods of transmission of these organisms havebeen identified, and many infection control mea-sures have been tried with different degrees of
* Corresponding author. Address: Department of Microbiology,
West Middlesex University Hospital NHS Trust, Isleworth TW7
Decontaminating the clinical environment after
a patient has been infected with MRSA, or with
0195-6701/$ - see front matter ª 2006 The Hospital Infection Society. Published by Elsevier Ltd. All rights reserved. doi:10.1016/j.jhin.2006.07.019
a multi-resistant Gram-negative bacterium, is
made in maximum recovery diluent [MRD, (peptone
thought to be a sensible precaution in stopping
nosocomial transmission of these organisms. French
dilutions were plated on to TSA, incubated for
et al. reported the use of a vaporized disinfectant
48 h, colonies were counted and the initial bacterial
system that reduced environmental MRSA contami-
concentration was calculated. Ceramic tiles measur-
nation, although further work is required to deter-
ing 10 cm  10 cm were cleaned using detergent fol-
mine the effect of environmental decontamination
lowed by 70% isopropyl alcohol, wrapped in
on MRSA infection rates.However, work performed
aluminium foil and autoclaved at 121 C for 15 min.
by Wilcox et al. indicated that using specific dis-
Ten drops (100 mL/drop) of bacterial suspension
infectants when decontaminating hospital wards
[109 colony-forming units (CFU)/mL] were evenly
reduced the incidence of Clostridium difficile in-
distributed on to 13 tiles (‘positive tiles’), and 10
fection.Whilst the interest of the popular press
drops (100 mL/drop) of sterile MRD were evenly dis-
has focused on MRSA, multi-resistant Gram-negative
tributed on to two tiles (‘negative tiles’) as controls.
bacilli have attracted far less interest. Acineto-
They were left to dry at room temperature for 2 h.
bacter baumannii is a Gram-negative coccobacillus
A Dyna-Fogâ Model 2739 Hurricane ‘Cold Fog’
that is frequently resistant to virtually all anti-
ULV/Mister (Dynafog, Indianapolis, USA) fogging
biotics.It has been found resident on intensive
machine was used for this experiment (capacity
care units, with reservoirs such as curtains being
3.8 L, maximum output 19 L/h). The Sterilox solu-
identified, and can be a particular problem to treat
tion contained 180 parts per million of available
due to its broad antibiotic resistance profile.The
free chlorine at pH 5.2. Five positive tiles were po-
financial impact of healthcare-associated infec-
sitioned horizontally on a laboratory workbench
tions is well recognized with an increase of inpatient
and five tiles were positioned vertically. Addition-
stay and morbidity and mortality, and an estimated
ally, three positive tiles and the two negative tiles
cost to the UK National Health Service in the region
were sealed inside a laminar flow cabinet in the
laboratory, which was not in use, to create positive
Steriloxâ is an established disinfectant for heat-
and negative controls inside a sealed environment.
labile flexible endoscopes, and has a broad
The fogging machine was positioned 3 m away with
spectrum of activity against mycobacteria, fungi,
an unobstructed path to the tiles, and was run for
viruses, bacterial endospores, and Gram-positive
10 min on maximum output. Afterwards, the labo-
and Gram-negative bacteriSterilox is some-
ratory was left for 1 h to allow time for the fog to
times termed ‘super-oxidized water’ and its princi-
settle and act upon the exposed tiles. In the
pal ingredient is hypochlorous acid, which is safe
second modified procedure, tile preparation and
to use and not harmful to the environment. The
fogging were performed as above, but at the end
present study examined the decontamination effi-
of the 10-min initial fogging, the fogging machine
cacy of Sterilox fog against MRSA and acineto-
was held approximately 1 m from the tiles and
bacter dried on to environmental surfaces.
a further 30-s fogging was performed. The tilesreceived no physical cleaning action, i.e. the tilesurface was not wiped in any way during the
fogging process. After this stage, it was necessaryto dry the laboratory using a mop due to the accu-
This study was carried out using two strains of MRSA
mulation of liquid caused by the fogging process.
and two strains of A. baumannii. The MRSA strains
Each tile was placed in a plastic bag containing
comprised a clinical isolate (sensitive to fusidic
100 mL MRD with 1% sodium thiosulphate for Steri-
acid, vancomycin, teicoplanin, linezolid, rifampicin
lox neutralization. The tiles were then agitated
and mupirocin, and resistant to erythromycin, tri-
manually within the bag for 1 min, and serial
methoprim and tetracycline) and a type strain (Na-
10-fold dilutions were made in MRD. Since each
tional Collection of Industrial and Marine Bacteria,
tile was eluted into 100 mL MRD, and 1 mL (i.e.
NCIMB 50143). The acinetobacter strains comprised
2 Â 0.5 mL aliquots) of this was plated neat on to
a clinical isolate (resistant to all commonly used an-
TSA, the limit of detection was 100 organisms/
tibiotics except colistin) and a type strain (NCIMB
tile, i.e. 1 CFU/mL neat eluate. This process was re-
12457). Two days prior to the study, a pure culture
peated for the positive controls, and the negative
of bacteria was plated on to tryptone soya agar
controls were plated using 0.5 mL of neat dilution
aliquots alone. The plates were then incubated at
at 30e35 C. A bacterial suspension to 5 McFarland
35 C for two days and colonies were counted
units (equivalent to 109 organisms/mL) was pre-
(with no growth being equivalent to <100 colonies
pared and a serial 10-fold dilution to 10À7 was
on the tile; the limit of sensitivity). A neutralization
validation was performed to determine the ability
type strain of acinetobacter (NCIMB 12457) gave
of sodium thiosulphate to neutralize Sterilox resi-
a mean colony count of 4.6 Â 102 CFU/tile from
due. This involved adding either 10 mL fogging solu-
the horizontal tiles and 1.1 Â 104 CFU/tile from
tion (test) or 10 mL MRD (control) to 100 mL MRD
the vertical tiles on the first fogging run. The sec-
containing 1% sodium thiosulphate, and inoculating
ond fogging run gave counts of 1.0 Â 102 CFU/tile
both solutions with 1 mL of the 104 dilution of each
for both horizontal and vertical tiles. The clinical
test organism, plating after 1 min and incubating
isolate of acinetobacter showed a mean count of
for 48 h to ensure complete microbial recovery.
3.4 Â 102 CFU/tile for the horizontal tiles and1.7 Â 103 CFU/tile for the vertical tiles on the firstrun. The second fogging run yielded mean counts
of 3.8 Â 102 CFU/tile from the vertical tiles and3.2 Â 102 CFU/tile from the horizontal tiles. The
The experiment was conducted twice, with the
standard method used on the first run and the
Pooling the results for horizontal and vertical
modified method involving the second hand-held
tiles, the type strain of MRSA gave log10 reduction
fogging on the second run. After each run, the tile
factors of 4.05 and 6.64 for the first and second
elutes were plated on to TSA. Recovery of the
fogging experiments, respectively. For the clinical
organisms from the positive controls demonstrated
isolate of MRSA, the corresponding figures were
that the organism remained viable once dried on to
3.75 and 6.99. The acinetobacter type strain
the ceramic tiles. Mean recovery from the MRSA
gave log10 reduction factors of 5.65 and 6.99 for
controls was 1.0 Â 109 CFU/tile on the first fogging
the first and second fogging runs, respectively,
run and 1.6 Â 109 CFU/tile on the second run for
and for the acinetobacter clinical isolate, the cor-
the type strain, and 2.1 Â 109 CFU/tile on the first
responding figures were 5.85 and 6.57.
run and 1.37 Â 109 CFU/tile on the second run forthe clinical isolate. Mean recovery from the acine-tobacter
8.5 Â 108 CFU/tile for the first and second runs,and the type strain yielded 4.8 Â 108 CFU/tile and
The use of Sterilox in the fogging studies resulted
1.4 Â 109 CFU/tile for the clinical isolate on its re-
in a 104-fold decrease of the MRSA type strain and
spective fogging runs. The negative controls for all
a 103.75-fold reduction of the MRSA clinical isolate
after a single treatment, and a 106.64-fold de-
On the first fogging run, the MRSA type strain
crease and a 106.69-fold decrease of the type strain
(NCIMB 50143) yielded a mean colony count of
and the clinical isolate after the two-stage treat-
3.8 Â 104 CFU/tile from the horizontal tiles and
ment. Acinetobacter strains showed greater reduc-
1.8 Â 105 CFU/tile from the vertical tiles. The
tions after one fogging compared with MRSA
second fogging run showed horizontal and vertical
(106.65-fold and 105.85-fold reductions for the
mean yields of 5.2 Â 102 CFU/tile and 2.6 Â
type and clinical strain, respectively). After a sec-
102 CFU/tile, respectively. The clinical isolate of
ond fogging, reductions similar to the MRSA results
MRSA gave mean counts of 2.52 Â 105 CFU/tile for
the horizontal tiles and 5.64 Â 105 CFU/tile for
ded two-stage fogging treatment, whilst improving
the vertical tiles on the first fogging run, and
efficacy, does involve more user interaction in
mean counts of 1.4 Â 102 CFU/tile for both hori-
a clinical setting, which may restrict its clinical
zontal and vertical tiles on the second run. The
Log10 mean colony counts of the four fogging experiments
Note: 100 colony-forming units/mL was the limit of sensitivity and a result of 2 signifies that no organisms were recovered. Figuresin parentheses are log10 reduction factors achieved by fogging. MRSA, meticillin-resistant Staphylococcus aureus.
Exner et al. showed that spread of S. aureus
diminished.As yet, there has been no work exam-
within the environment could result from inade-
ining the impact of organic matter contamination
quate cleanThese authors used a suspension
on a fogging system. The manufacturers recom-
of S. aureus of 0.05 mL (3 Â 107 CFU/mL) inocu-
mend that the system should only be utilized at
lated on to a 5 cm  5 cm square of floor and then
the end of a cleaning process, and in such circum-
mopped in a U-shape using a variety of cleaning
stances, any organic contamination should have
agents. After drying, swabs were taken from the ini-
been removed. However, further work needs to be
tial square of floor and three adjacent squares of
carried out to investigate any role of organic con-
identical size, 7 cm apart, and plated to determine
tamination of surfaces on the efficacy of Sterilox
the presence of S. aureus. Mops soaked in water,
in these situations. Work is also needed on the use
quaternary ammonium compounds or alkylamines
of Sterilox fog in the clinical setting to discover
showed incomplete killing of the S. aureus and
any potential problems of using the fog in a func-
caused dissemination throughout the non-inocu-
tioning clinical area with respect to the technical
lated tiles. Only aldehydes and peroxides showed
aspect of using the system, as well as the resultant
complete killing of the S. aureus with no dissemina-
liquid residue interfering with clinical appliances.
tion. Environmental contamination with A. bau-
With the UK Government publishing ‘Winning
mannii in an intensive care setting suggested that
ways’, it is clear that infection control is a major
poor cleaning was associated with increased pa-
public concern, and the cleaning and cleanliness of
tient coloniIn view of the environment be-
hospitals remains high on the political agenda.
ing a potential source of patient contamination,
The reductions observed in this study compare
and since the conventional ‘mop and bucket’ tech-
favourably with the use of alkylamine compounds,
nique appears to risk leaving residual contamina-
and the safety profile of Sterilox means that it is
tion of surfaces, the use of a fogging treatment
a good candidate for decontaminating the hospital
that may be able to permeate into the various re-
cesses that can be found within most clinical set-tings (such as behind drawers, within the bed
This study found that Sterilox is able to reduce
The authors would like to thank Sterilox UK for the
the burden of MRSA and acinetobacter on environ-
kind provision of consumables used in this study.
mental surfaces when fogged. It would clearly be of
Laboratory work was performed at Charing Cross
interest to investigate the activity of Sterilox
against other nosocomial pathogens that can per-sist on surfaces in the clinical environment such asclostridia and enterococci. Organic contamination
of the environment is an important consideration ofany decontamination process, and the manufac-
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