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The Use of Ozonized Oil in the Treatment of Dermatophitosis Caused by Microsporum Canis in Rabbits

The ozone is effective against most microorganisms due to its high oxidant power. Low concentrations and short-term contact are sufficient to inactivate bacteria, mold, yeast, parasites, seaweeds, protozoa and fungi. Microsporum canis is an important agent of dermatophitosis in human and animal. The aim of the current study was to assess the efficacy of ozonized oil over Microsporum canis in rabbits. Eighteen male New Zealand white rabbits, weight ranging from 2 to 3.2 kg were depilated in the cranial dorso-lateral and right caudal, and cranial and left caudal regions. The regions were inoculated with Microsporum canis, excepting the right caudal region, and were denominated TM, O, OM and M, respectively. After seven days, the treatment of lesions in TM began with 0.12g of terbinaphine 1% cream; in OM and O with 0.12g of ozonized oil; all animals were treated once a day for 28 days. Region M was not treated. Material was collected from those regions for cultivation in Sabouraud agar at day 28 of treatment. In the evolution of the treatment with terbinaphine, of 14 contaminated regions with Microsporum canis ten evolved to cure. With the ozonized oil, of 15 contaminations, four were cured. Clinically, that is, the macroscopic evaluation of lesions showed improvement in the TM and OM treated regions. We can conclude that there was statistical evidence of the protection action of the oil against the dermatophyte.


INTRODUCTION
The fact that the ozone is a potent oxidant accounts for its high reactivity. This gas dissolves in the plasma, extracellular fluids, the thin layer of water which covers the skin and in regions coated by mucosa, triggering reactions with a number of molecules present in biological fluids, such as antioxidants, proteins, carbohydrates, and, mainly, with polyunsaturated fatty acids. By these reactions, reactive oxygen species (ROS) and lipid oxidation products (LOPs) are generated, both responsible for successive and multiple biochemical reactions that occur in different body cells (5).
The ozone in its gaseous or aqueous phases is effective against the majority of microorganisms tested by numerous research groups (2,4,5,6,26). Low concentrations and shortterm contact are enough to inactivate bacteria, mold yeast, protozoa algae and fungi (10,11,12,20). The mechanism of antimicrobial action occurs due to the membrane lysis of the agents, after the oxidation process (14). It is believed that the primary attack is against the cellular wall of the microorganism *Corresponding Author. Mailing address: R. Dr. Cesário Motta Junior, 112 -Vila Buarque -CEP 01221-020 São Paulo, Brasil.; Fone: (11)  .; E-mail: fvdaud@terra.com.br Dermatophitosis caused by M. canis and, following, on penetrating the interior of cell, the ozone promotes oxidation of amino and nucleic acids. The cellular lysis depends on the extension of such reactions (25).
Microsporum canis is an important agent of dermatophitosis in human and animal (7,22). This worldly distributed canis variety is the most common zoophilia species in human infections. The lesions caused by this dermatophite are usually located on the skin and pelage and commonly cause tinea capitis and tinea corporis in children and adults who had contact with dogs or cats (mainly reservoirs), or infected children (13). In animals, while the dogs develop tegumentary lesions, the cats, otherwise, remains asymptomatic or shows tiny lesions; such peculiarity makes the cats a serious threat to the control of the microsporum dermatophitosis (7).
The aim of the present study was to assess the efficacy of the ozonized oil against Microsporum canis in rabbits.

MATERIAL AND METHODS
Eighteen male New Zealand white rabbits, weighing

Mycological examination
The mycological examination was performed at days 0 (prior to inoculation) and 7, 14, 21 and 28 post-inoculation. Precision scales were used daily (Denver Instrument Company AA-200) to weigh the medication.

Ozonized oil
The ozonized oil Bioperoxoil®, originally from Barcelona, Spain, was used for the treatment of regions O and OM.

Statistical methodology
To assess the qualitative variables, the oucomes were presented through absolute and relative frequencies. These analyses were performed using the McNemar Test. It was established the significance level of 5%.
Resume measurements were calculated for the quantitative variables. The comparison between the quantitative variables was performed using the Paired t Test. The significance level was established at 5%. Dermatophitosis caused by M. canis

RESULTS
Twenty-eight days after the onset of the treatment, the actions of the terbinaphine 1% cream and the ozonized oil were equivalent ( Table 1).
The individual evolution of each treatment is shown in Tables 2 and 3. The regions treated with the ozonized oil (OM) and the non-treated areas are shown in Table 4, whereas the regions treated (TM) and non-treated with terbinaphine 1% cream are in Table 5.
It is noteworthy that the statistical analysis of Tables 4 and   5 Figure 4 shows the evolution of lesion ( Figure 3A) treated with terbinaphine 1% cream for 21 days. Figure 5 refers to the treatment of lesion of figure 3B with ozonized oil for 21 days. Finally, Figure 6 shows the initial lesion ( Figure 3C) without any treatment, after 21 days.

DISCUSSION
The statistical evaluation of data shows that the action of the ozonized oil and terbinaphine cream was similar (Table 1).
However, the individual evolution of each treatment was different (Tables 2 and 3