OZONE EFFECTS ON SPECIFIC BACTERIA AND VIRUSES (

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Bacterial infections and the spread of viruses relating to athletic gear are becoming a bigger concern each year. Numerous articles are being written and public awareness is growing rapidly to the health risks. This study and independent laboratory testing was performed to analyze the effectiveness of the Sports-O-Zone system to help control this growing problem. There are numerous ways to kill bacteria. Each way entails direct contact of the virus or bacteria with an oxidant or disinfectant. To effectively claim a 99.99% kill rate of these microorganisms, the disinfectant would have to be in contact with 99.99% of the microorganisms. One of the problems to overcome in direct contact with all of the microorganisms is the clumping of bacterial cells. As noted in the Pennsylvania State University study, (Bactericidal Effects of High Airborne Ozone Concentrations, 1998) there is a tendency of micron-sized particles to clump naturally. The clumping of bacterial cells protects a small percentage of bacteria and causes them to behave as if they had a much higher resistance There are numerous claims of 99.99% kill rate by systems offered in the market place. The problem with athletic equipment is finding an effective way to get the disinfectant into all the crevices of the equipment for direct contact with the bacteria and breaking down the clumping effect of the microorganisms. The Sports-O-Zone machine s answer is the use of OZONE gas to get into all those crevices and effectively work on attacking and eliminating all the bacteria and breaking down the clumping effect. Ozone (O 3 ) is one of the strongest oxidants and disinfectants available, ranking ahead of hydrogen peroxide, bleach, and iodine, without any residue or odor. Ozone is nature s way of purifying air and water. It is colorless and occurs naturally in the air we breathe everyday. Ozone is also one of the food sanitizing ingredients and processes being used in food safety. The U.S. Environmental Protection Agency, in conjunction with the Safe Drinking Water Act of 1991, confirmed that ozone was effective in ridding water of hazardous pathogens, including chlorine-resistant Cryptosporidium (USDA Gov/publications/agoutlook/June1998). Gaseous ozone is also listed as an alternative disinfectant for water-sensitive produce (USDA Guide to Minimizing Microbial Food Safety Hazards for Fresh Fruits and Vegetables). The flexibility of Ozone spans various industries and is well documented as a very effective disinfectant that leaves no residue. OZONE EFFECTS ON SPECIFIC BACTERIA AND VIRUSES (www.ozoneapplications.com) Bacteria are microscopically small, single-cell creatures having a primitive structure. The bacteria body is sealed by a relatively solid-cell membrane. Ozone interferes with the metabolism of bacterium-cells A sufficient amount of ozone breaks through the cell membrane, and this leads to the destruction of the bacteria. Viruses are small, independent particles, built of crystals and macromolecules. Unlike bacteria, they multiply only within the host cell. They transform protein of the host cell into proteins of their own. Ozone destroys viruses by diffusing through the protein coat resulting in damage of the viral RNA.

CASE STUDIES There were several independent laboratory studies done on the effectiveness of the Sports-O-Zone process. The independent laboratory tested the machine s kill rate effectiveness on Methicillin-Resistant Staphylococcus aureus (MRSA), Staphylococcus aureus (Staph), and Candida. It is well documented that ozone produced 99.99% death rates of these bacteria when exposed for 10-480 seconds to ozone concentrations between 300 and 1500 ppm (Pennsylvania State University study, Bactericidal Effects of High Airborne Ozone Concentrations, 1998). The Laboratory test performed on our machine analyzed the kill rate at 3 minute, 5 minute, and 11 minute intervals. Staphylococcus aureus (Staph) Staphylococci (staph) are Gram-positive spherical bacteria that occur in microscopic clusters resembling grapes. Bacteriological culture of the nose and skin of normal humans invariably yields staph. Human staph infections are frequent, but usually remain localized at the portal of entry. The portal may be a hair follicle, but usually it is a break in the skin which may be a minute needle-stick. Staph infection is characterized by inflammation, an elevated temperature at the site, swelling, and the accumulation of pus (www.textbookof bacteriology.net). Serious consequences of staph infections occur when the bacteria invade the blood stream. With staph readily available on the skin, it can easily be transferable from person-to-person, or from person to equipment to person. The warm moist heat of athletic competitions is the ideal environment for these bacteria to grow.

The bacterial cultures of staph were deposited on solid media in petri dishes. The control dish below is on the left, with treated petri dishes containing bacteria ozonated by the Sports-O-Zone machine at 3 minute, 5 minute and 11 minute intervals. Dishes ozonated at Control Dish 3 minutes 5 minutes 11 minutes Conclusion: The independent laboratory study showed the Sports-O-Zone system killed more than 99.98% of Staphylococcus aureus after 5 minutes. concentrations bacteria Staphylococcus aureus kill rate 8.00E+05 6.00E+05 4.00E+05 2.00E+05 0.00E+00 0 3 5 M inute s 11

Methicillin-Resistant Staphylococcus Aureus (MRSA) This is the staphylococcus aureus that are usually resistant to a variety of different antibiotics. A few strains are resistant to all clinically useful antibiotics except vancomycin, but vancomycin-resistant strains are increasingly being reported. It is becoming more of a need to kill these bacteria before it finds a portal of entry into the body. One important defense of these bacteria is eliminating it off the athletic equipment. The bacterial cultures of MRSA were deposited on a solid media in petri dishes. The control dish below is on the left, with treated petri dishes containing bacteria ozonated by the Sports-O-Zone machine at 3 minute, 5 minute and 11 minute intervals. Dishes ozonated at Control Dish 3 minutes 5 minutes 11 minutes Conclusion: The independent laboratory study showed the Sports-O-Zone system killed more than 99.96% of Staphylococcus aureus MRSA after 5 minutes. concentrations bacteria Staphylococcus aureus MRSA kill rate 3.00E+04 2.00E+04 1.00E+04 0.00E+00 0 3 5 M inute s 11

Candida Candida is a parasitic imperfect fungus that resembles yeast and is commonly found in the mouth, intestines, and vagina as a part of the human body s flora. It can cause candidiasis of the mucous membranes of the mouth (also called thrush) and of the vagina (yeast infection). The bacterial cultures of candida were deposited on solid media in petri dishes. The control dish below is on the left, with treated petri dishes containing bacteria ozonated by the Sports-O-Zone machine at 3 minute, 5 minute and 11 minute intervals. Dishes ozonated at Control Dish 3 minutes 5 minutes 11 minutes Conclusion: The independent laboratory study showed the Sports-O-Zone system killed more than 99.76% of Candida albicans after 11 minutes. concentrations bacteria Candida albicans 8.00E+04 6.00E+04 4.00E+04 2.00E+04 0.00E+00 0 3 5 M inute s 11

Independent Laboratory Testing Local Hockey Player s Gear The second study was to examine the effectiveness of the machine when loaded with a set of hockey gear. All systems cannot anticipate the bacteria load and the amount of disinfectant needed to attack the amount of bacteria within the system. The Sports-O- Zone system was designed to overcompensate for the bacteria load to achieve the maximum bacteria kill rate, and then convert the excess ozone back into simple oxygen. Bacteria from various areas of the player s equipment were collected prior to it being treated in the machine. Then the equipment was treated for 5 minutes in our Sports-O- Zone machine. The equipment was then swabbed again to collect the bacteria that still remained. The two samples were analyzed and concluded that we achieved up to a 97% reduction in bacteria. Bacteria colony counts - April 2005 80,000 60,000 40,000 20,000 0 Helmet Equip. Bag Chest Protector Glove Before After