There are many types of equipment of making acidic electrolyzed water from tap water. We utilized one of them : OXILYZER which is made by Miura Electric Co., Akita, Japan. Aqua oxidized water (AOW) is acidic water with a pH under 2.7 and is processed by electrolysis through a small NaCI mediated specific membrane. It has over 1, 100 mV of oxi-redx potential and includes Cl
2, OH·, O
3, ClO.
This acidic water has a strong bactericidal effect to bacteria and virus but there are few reports concerning its bactericidal activity to periodontopathic bacteria. We investigated the bactericidal effect of AOW to periodontopathic bacteria, two strains of
Actinobacillus actinomycetemcomitans (Aa), two strains of
Porphyromonas gingivalis (Pg) and a strain of
Fusobacterium nucleatum (Fn) .
Staphylococcus aureus (Sa) 209 P,
Escherichia coli (Ec) B and
Salmonella enteritidis (Se) SER were used for comparison of the bactericidal activity of AOW to periodontopathic bacteria.
AOW was obtained from an OXILYZER and stored in a polyethilene bottle and kept in a dark cold place totally blocked from light.
Bacterial strain Aa : Y4 and ATCC 29522, Pg : 381 and ATCC 33277, Fn : ATCC 23726 were grown at 37°C in BHI (5% CO
2), BHI contained hemin, menadion and yeast extract (anaerobic condition), and GAM broth (anaerobic condition), respectively. After washing by centrifugation, each bacteria was suspended in PBS and adjusted to the optical density of 0.3 at 540 nm light length with 0.1% gelatin containing Hanks' balanced salt solution (GHBSS) .
Serum and saliva were obtained from volunteers of healthy student. Saliva was sterilized through a millipore filter with a 0.45μm pore size.
Bactericidal activity was performed by colony count seeding and grown on BHI or Anaero columbia blood agar or GAM medium after treatment with AOW.
The following results were obtained:
1. Bactericidal effect and effective concentration of AOW
Periodontopathic bacteria were AOW sensitive and were killed in one minute, from the original 25% AOW, while it needed more time and a higher concentration to kill Ec, Sa and Se.
2. The effect of serum and saliva on bactericidal activity of AOW.
To Aa and Pg, the addition of serum or saliva in a concentration of 1% of AOW had the effect of bactericidal activity. However, the bactericidal activity was completely lost by the addition of 10% serum in AOW. To Fn, the bactericidal effect was lost in only 1% of serum or 10% salivary addition.
3. Stability of bactericidal activity of AOW.
AOW was kept in a polyethilene bottle at 4°C with shading from light. The bactericidal activity was examined by withdrawing AOW from the shielded bottle weekly. The activity was well preserved until 5 months. After 21 weeks, a slight decrease of bactericidal activity was observed in 25% AOW.
4. Cause of decrease of bactericidal activity by addition of 10% serum to AOW.
The addition of serum to AOW caused a decrease of oxi-red potential from 1144.8 to 479.0. pH from 2.52 to 5.60, and HClO from 19.4 ppm to undetectable.
In conclusion, AOW is a useful disinfectant of periodontopathic bacteria
in vitro. The oral environment is very complex. We must pay careful attention not only to the decrease of bactericidal activity of AOW by salivary flow and bleeding, but also the effect of AOW on the mucosa.
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