The objective of this study was to demonstrate the bactericidal, fungicidal, and sporicidal efficacy of the in-use concentration of ozone in solution generated continuously by the JLA Limited OTEX validated ozone laundering system against a range of typical microorganisms associated with laundering applications. The study used the JLA HC 100 laundering machine as the containment vessel. Test microbial suspensions were introduced directly into the drum of the laundering machine containing a known level of water in the presence of soluble ozone. Using standard microbiological techniques, the numbers of surviving bacteria, mold spores or bacterial spores were determined, and reductions in viable counts were calculated. The following test organisms were employed: Pseudomonas aeruginosa, Methicillin-Resistant Staphylococcus aureus (MRSA), Enterococcus faecalis, E. coli, Acinetobacter baumannii, Aspergillus niger (spores), and Clostridium difficile (spores). All tests used multiple contact times of 0, 1, 3, 5, 7, 11 and 15 minutes at 20 °C±2 °C. Levels of ≤0.25 mg/L of dissolved ozone had minimal effect on the bacteria. Levels of >0.45 mg/L ozone showed good biocidal activity, except against Acinetobacter baumannii, which showed strong resistance to ozone. Against fungal spores, dissolved ozone levels of >1.2 mg/L were required to obtain a 2.8 log reduction. For Clostridium difficile spores, a dissolved ozone level between 0.90 and 1.20 mg/L gave >1.6 log reduction with no discernable spore recovery. In every instance, the control tests (no ozone) recovered levels of microorganisms that proved that the action of the laundering machine has no antimicrobial activity, and that the log reductions obtained are solely due to the effects of dissolved ozone.
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