Chlorine dioxide effectively reduces infective

Clostridium difficile spores on inoculated glass surfaces

T. Norén
Department of Laboratory Medicine, Microbiology, Örebro University Hospital, Örebro, Sweden

Introduction Results
Clostridium difficile infection (CDI) is the most common Repeated testing in triplicate using ten minutes exposure by 1600 ppm ClO2
cause of antibiotic-associated diarrhea and virulent strains effectively reduces high-concentrated spore-suspensions of C. difficile on
spread effectively and globally. Long-time surviving spores glass-carriers comparable to hypochlorite (Fig 1). Furthermore, exposure
within health-care environment challenge hospital clean- intervals down to 1 minute does not reduce efficacy (Fig2).
ing as an important part of hygiene precautions.
Hypochlorite has been a sporicidal agent used commonly
Conclusion
8 Chlorine dioxide of 1600 ppm
in cleaning but both corrosion and irritant gas emission
have been draw backs. These negative side effects can be 7
concentration is shown to be
reduced by using chlorine dioxide, which is well document- Fig. 1. sporicidal in clostridium difficile
ed disinfectant for drinking water but very few publications Median reduction of
6 Clostridium difficile PCR when tested in clean conditions
address Clostridium difficile spores and surface cleaning. ribotype 027 spore inoculum
of 2.4 x 107when exposed in
on glass-carriers.
We evaluated the efficacy of chlorine dioxide (LifeClean®) 5
triplicate on glass carriers for
on highly concentrated C. difficile spore suspensions (≥85%). 10 minutes by ethanol 70%, A rapid disinfection in one minute
Spore innoculum
4 2.4 x 107
1/10 hypochlorite (NaOCl) and
chlorine dioxide (LifeClean®)
makes this an interesting agent
Free viable count 1600 ppm (mg/L). for everyday hospital cleaning
3 Free viable count was how
where wipes adds mechanical
Methods
Total viable count many spores that could be
2
germinated from filtered
solution and total viable
support.
In the absence of European Clostridium difficile sporicidal count included the sessile
standards we designed a reproducible carrier test modi- 1 viable spores mechanically Almost all ClO2 emits from exter-
fied according to AOAC (association of official agricultural
removed from the glass
surface.
nalized solution in 4 minutes
chemists). 0 (data not shown) giving less risk
NaCl Ethanol 70% NaOCl LC 800 ppm LC 1600 ppm
We prepared a spore suspensions culturing Cd-strains of corrosion and sustained
(PCR ribotype UK 023, NCTC 13366) on FAA media anero- respiratory irritants compared
bically for 48 h followed by a seven day bench rest in air. to hypochlorite.
9 Log 10
A gentle harvest into NaCl followed by alcoholshock 1:1 cfu
and washing x 4 (centrifuging at 3000 rpm for 10 minutes 8
and resuspending the pellet) held on ice +4°C. Finally we Fig. 2.
calculated free spores using phase-contrast microscope 7 Median Clostridium difficile Contact
Free viable count (PCR ribotype UK 027 spore torbjorn.norén@regionorebrolan.se
and Bürker chamber (107-108 cfu/mL) corresponding to inoculum of 4.8-9.6 x 107)
6 Total viable count
viable counts. viable count reduction on
glass carrier in triplicate test
After inoculation of 0.1 mL spore suspension (108cfu/mL) 5 run for 1 to 10 minute expo-
on a sterile glass carrier 0.2 mL disinfectant or negative sure of 1600 ppm chlorine
4 dioxide (LifeClean®).
control (NaCl) was covered on top of the dried inoculate

Posterdesign - Dept. of Medical Photography, Örebro University Hospital

Free viable count was how
inactivated for 1-10 min. Neutralization- was done by 3 many spores that could be
dilution (submerging carrier in 250 mL NaCl) and by germinated from filtered
solution and total viable
chlorine inactivation using 0.7% sodiumtiosulphate. 2 count included the sessile
After shaking for 20 minutes we filtered the whole viable spores mechanically
volume through 0.47µm and culturing both filterpaper 1 removed from the glass
surface.
and filtrate. Finally a standardized dubbel-S cotton-swab
0
was mechanically rubbed over the inoculated surface and 0 min 1 min 2 min 3 min 4 min 5 min 6 min 7 min 8 min 9 min 10 min
adherent residue was cultured for viable counts.