Hand hygiene skin ATP study: Hand washing with tap water and soap vs. ozonated water and soap vs. antiseptic and ozonated water vs. antiseptic and tap water

Alvaro Liceaga, M.D.; Elvin Mercado, C.S.T.; Mary Narcaroti R.N.MSHA; Lisa Liceaga B.S. -

A Unique Comparison in regard to Adenosine Triphosphate (ATP) Reduction on the Skin, as a Measure Of Biologic Material Reduction.

Hand hygiene is of greatest importance because the most common mode of transmission of pathogens is through the hands, especially in settings such as hospitals, operating rooms, medical offices, specialty clinics, therapy centers and outpatient surgery centers. Clean hygienic hands are the single most important factor in preventing the spread of pathogens and in reducing the incidence of healthcare associated nosocomial infections.

Handwashing physically removes particulates, soil and organic material and reduces microbial contamination acquired by contact with the local environment. Unfortunately, water alone cannot remove all contaminants, such as oils, fats and microorganisms on hands, thus requiring the use of soaps or detergents to dislodge and dissolve those materials. Proper hand hygiene techniques are described by governmental agencies, such as use of soap or detergent to thoroughly rub on all surfaces of both hands followed by rinsing and drying (6). Despite best efforts to clean hands, handwashing with soaps is only partially effective in this regard.

A number of sanitizing chemicals have become commonplace in the medical setting:
Anti-Bacterial Hand Soaps had been in widespread use. Solutions contain soap with or without additives like Triclosan 0.375% or Chloroxylenol 0.95% (PCMX). In 2016, the FDA issued a rule banning the use of Triclosan, Triclocarban and 17 other chemicals in hand and body washes, which are marketed as being more effective than simple soap (2). Manufacturers replaced Triclosan with Chloroxylenol (PCMX) Cl, Benzalkonium Cl, and or Benzethonium.

Sanitizing alcohol based gels containing 70% Ethanol have the downside of gel buildup that reduce effectiveness and do not effectively kill all pathogens.

Betadine (Povidone-iodine) has long demonstrated anti-bacterial effectiveness for procedural applications or surgical prep but some bacteria have developed resistance.

Isopropyl Alcohol 70% solution may be used as a hand sanitizer. It is germicidal and antiseptic are used in many healthcare settings, such as doctor's offices and hospitals.

Chlorhexidine (Hibiclens) is used as a surgical hand scrub and for preoperative skin preparation in surgery. It is active against gram-positives, gram-negatives, anaerobes, aerobes, yeasts but may be ineffective against some viruses.

Hydrogen Peroxide (H2O2) is used for the sterilization of surfaces, such as surgical tools and for wound disinfection. H2O2 has broad-spectrum properties against bacteria, viruses, yeasts, and spores. It can be harmful to skin tissue.

Ozonated water may substantially reduce residual contaminants on the skin. Unfortunately, there are only a few studies regarding hand sanitation with ozonated water. More are needed.

Background:
Ozone has a very characteristic odor and is a powerful oxidant. It has been used in industrial applications, e.g. water treatment. Ozonated water is produced using an ozone generating electrical device. Ozone is created on demand using an ultraviolet ozone generator located proximate to the faucet. The gas is dissolved into the water via a venture-type mixing system so that the ozone concentration at the tap outlet remains constant for a given flow rate. Typical tap ozone concentrations can range from 0.5 to 4 ppm.

In water, ozonated water is produced when gaseous oxygen (O2) molecules are dissociated by an electrical potential into nascent oxygen atoms (O-) which then subsequently collide with oxygen molecules to form the unstable ozone gas molecule (O3) which has extremely high oxidizing capacity.
Ozonated Water (O3-H20) free radical oxygen molecules react with microbes and effectively kill them via oxidation and degenerate organic compounds. Ozone quickly kills all microorganisms non-selectively, including bacteria, viruses, molds and spores. According to the US EPA Office of Water, ozone is one of the most potent and effective germicides when used in water treatment.

The University of Dundee evaluated the anti-microbial efficiency of ozone water from an ozone generating faucet (19). They found that its' strong antimicrobial properties are 100 times stronger and reacts 3000 times faster than chlorine as a disinfectant within 1 min. Ozone leaves no trace of residual products upon oxidative reaction. After 20 minutes, ozonated water reverts into plain water and oxygen gas. They believed that the use of ozone water may reduce the use of toxic chemical disinfectants in use by up to 50%.

Ozonated water has been proposed as an alternative disinfection agent to prevent deformation or other damage during conventional autoclave disinfection and is approved by the US Center for Disease Control for the rapid disinfection of medical surfaces and equipment (CDC, 2008) (72).

Since ozonated water (O3-H20) is known to possess antimicrobial properties, we proposed this study as the first in our series of tests to compare the effectiveness of ozonated water. We evaluated whether the bioload of viral, bacterial and/or other organisms present on the skin, as measured by ATP readings, are lowered by several handwashing techniques.

Materials: Lenova Faucet
The ozone generator devise used in this study is the Aqualogic Faucet by Lenova (Kona) which is commonly used in residential and some commercial applications for surface disinfection. The faucet system consists of a stainless steel faucet connected to a typical water supply lines and a connection to an AC outlet. The ozone production is through closed self-contained system that produces ozonated water by a patented ozone gas generator and valve system, and a Venturi mixing tube, designed to work at public water pressure. Water outflow is at a consistent flow rate of about 1.2 gallons per minute (same as any common faucet) with an ozone concentration between 0.8 and 1.4 ppm that is considered safe to use and is very effective as an anti-microbial that can reportedly kill most pathogens in under 2 minutes of contact time.

Water3Water10

HYGIENA
Hygiena's SystemSURE plus ATP Cleaning Verification System is a tool used to monitor and improve the cleanliness levels of surfaces in healthcare facilities. It uses bioluminescence technology to identify and measure adenosine triphosphate (ATP). ATP is an energy molecule found in all living organisms, including skin, blood, mucus, saliva, viruses, fungi, molds, spores and bacteria. The persistence of ATP on the hand may be an indication of incomplete or improper cleaning technique or a measure of residual ATP from human skin cells.

The Hygiena's SystemSURE Luminometer, is used to measure ATP collected with the swab and results are expressed numerically as Relative Light Units. (RLU).

Methods and Procedures
Control: 10 healthy volunteers were selected. Before initial cleaning, the palms of both hands were swabbed using the UltraSnap ATP detection device and measured with the SystemSURE Plus Luminometer. The control numbers were recorded.

Cleaning Technique was standardized as follows: Before swabbing, volunteers were asked to perform a standard hand-washing technique as recommended by the CDC in healthcare settings. The volunteers were asked to wet their hands and apply the selected agent and rub their hands together vigorously for 20 seconds covering all surfaces of the hands and fingers. Next, they were asked to rinse with running tap water for thirty (30) seconds. Hands were then air dried for five (5) minutes; volunteers were asked not to touch any surface or objects during the drying time. Once dried, both palms were simultaneously swabbed using the UltraSnap ATP device and measured with the SystemSURE Plus Luminometer. The results were then recorded. Skin Subjects were asked to report any skin irritation or skin abnormality following washing.

Studies performed:
1. H2O + Soap Cleaning Technique: Standard soap solution. The results were then recorded.
2. O3-H2O + Soap Cleaning Technique: A week later, Standard soap and O3-water. The results were recorded.
3. Tap H2O + Hibiclens Cleaning Technique: A week later, Hibiclens antiseptic solution and rinse with running tap. The results were then recorded.
4. O3-H2O with Hibiclens Cleaning Technique: A week later, Hibiclens antiseptic solution and O3-water. The results were then recorded.

O3 Concentration Monitoring: We measured ozone conditions during testing: O3 content, pH, water temperature and exposure time. The physical conditions of the ozone contained in the water was tested and monitored to evaluate if they met the appropriate conditions and permissible levels to achieve desired sanitation. Ozonesolutions.com Monitoring Device measured ozone content in the water in parts per million. Testing included the content of ozone in the water, pH, temperature and exposure time at the time of use. Parameters: Ozone content PPM, Water pH, Temperature, and Exposure Time:

Recap: Ozonated water properties Range: Ozone content PPM: 0.7ppm – 1.39ppm; Ave 1.18. Water pH: 7.59 - 8.13; Ave 7.83. Temperature (F): 77.4 - 82.8 F; 79.9. Exposure Time <1 min.

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Control (Black) - skin tested prior to any form of cleaning. H20 (HORIZONTAL GREEN) - skin tested after cleaning with soap and rinse with tap water. Measured Results: An Average of 85.76% reduction in ATP levels during supervised standard handwashing with tap water.

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Control (Black) - tested prior to any cleaning. HIBICLENS (Pink Diagonal Stripe) skin tested after cleaning with Hibiclens soap and tap water rinse. Measured Results: An average of 94.2 % reduction in ATP levels during supervised standard handwashing using Hibiclens antiseptic solution and rinsing with tap water.

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Control (Black) - skin tested prior to any form of cleaning. O3-H20 (Solid Blue) - skin tested after cleaning with soap and ozonated water. Measured Results: An average of 97.3% reduction in ATP levels during supervised standard handwashing with soap and ozonated water rinse.

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Control (Black) skin tested prior to any form of cleaning. O3-H20 (Red Diamond) - skin tested after cleaning with soap and ozonated water. Measured Results: An Average of 97.5% reduction in ATP levels during supervised standard handwashing with Ozonated water using Hibiclens antiseptic solution.

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Discussion: Subjects reported no skin irritation or skin abnormality following washing.

Summary of Observations: The palms of the volunteers showed a wide variation in ATP load.

The hands cleaned with soap and rinsed in tap water resulted in average of 85.76% reduction in ATP levels.

The hands cleaned with Hibiclens antiseptic solution and rinsed in tap water resulted in an average of 94.2 % reduction in ATP levels.

The hands cleaned with soap and rinse with ozonated water resulted in an average of 97.3% reduction in ATP levels.

The hands cleaned with Hibiclens antiseptic solution and rinsed with ozonated water resulted in an average of 97.5% reduction in ATP levels. These hands tested with Hibiclens antiseptic solution and ozone water rinse resulted in only an additional 0.2% reduction in measured ATP.

Authors Note on Naturally Occurring ATP on Host Skin: The skin as a living organ has naturally occurring levels of ATP that should not attributed to exogenous microorganisms, eg. virus, fungi, spores or bacteria. It is believed impossible to eliminate all ATP from skin cells to achieve a zero "0" RLU result.

Clean hygienic hands are the single most important factor in preventing the spread of pathogens and in reducing the incidence of healthcare associated nosocomial infections, especially with the growing risk of infection with resistant pathogens.

Routine handwashing in patient care is valuable to remove organic material and microbial contamination acquired by contact with patients or the environment. Tap water may contain a variety of microorganisms and pathogens and may be a source of nosocomial infections. A plausible route for transmitting these organisms from water to patient could be through the use of contaminated water.

We contend that the addition of 0.5 to 1.5 ppm of ozone dissolved in water, if maintained and delivered at 'standard' physical conditions, will substantially reduce levels of undesirable organisms on the skin. It may further be postulated that the O3 reduces the bio-contaminates in the water itself. Additional testing is recommended to confirm the above hypothesis, perhaps including bacterial, viral and fungal culture testing.

This preliminary study helps confirm that ozonated water possesses excellent antimicrobial properties and should have a place in effective hand hygiene protocols. The addition of ozonated water to the handwashing regimen is recommended by the authors, as it has proven to be of value in reducing ATP levels on the skin following this preliminary study. Further studies are needed to issue additional recommendations.

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