Transcript of ozonized oil

Mycological Study of Dermatomycosis in
Population of Cairo, Egypt
Salama A. Ouf1, Tarek A. Moussa1, Samar M. R. Eltahlawi2, Alshimaa S.M. Abdelmegeed1
1Botany
and Microbiology Department, Faculty of Science, 2Dermatology Department,
Faculty of Medicine2, Cairo University, Giza 12613, Egypt
Introduction
• Dermatophytes are a group of closely related keratinophilic fungi that infect
keratihized tissues such as hair, nails and skin. Virtually, there is no human
population free from these mycotic diseases.
• Dermatophytes have increased over the past few decades particularly in immunedepressed patients. Although the research regarding the synthesis of new
antifungal drugs has intensified, however the treatment with systemic antifungal
chemical agents such as ketoconazole, fluonazole and itraconazole derivatives
has side effects, in particular, when these chemicals are used in long-term
application. This has open the way for finding out an alternative to chemical
drugs.
• One of the possible approach is to use ozone therapy which has powerful antimicrobial properties against viruses, bacteria, parasites and fungi due to
oxidation reaction which occurs upon any collision between an ozone molecule
and a molecule of an oxidizeable cellular components, particularly those
containing double bonds, sulfhydryl groups, and phenolic rings.
Aim of study
1- The aim of this investigation was to study the occurrence and
etiological agents of dermatomycosis and candidiasis among 640
patients attending dermatology clinics at three hospitals in Cairo, Egypt
2- Parallel study was undertaken to determine the prevalence of
dermatophytosis among the patients who were suffering from
diabetes, cancer and heart diseases. This study included 620 patients
attending National Endocrine and Diabetes Institute, 280 at National
Cancer Institute and 210 at National Heart Institute.
2- To investigate the effect of ozone on growth, cellular leakage and
keratinolytic activity of the recovered dermatophytes
Table 1. Survey of patients suffering from skin mycosis due to dermatophytes and other fungi in
three hospitals at Cairo classified according to age group
Age groups (yr)
Clinical types of tinea
Tinea capitis
Tinea corporis
Tinea cruris
Tinea pedis
Tinea unguium
Tinea versicolor
Candidiosis
Total
0 - 10
11 - 20 21 - 30
31 - 40
41 - 50
51 - 60
Total male
/
61 - 70
Total female
male
98
22
-
-
-
-
-
120
female
63
-
-
-
-
-
-
63
male
28
8
2
-
6
-
-
44
female
20
3
4
14
6
4
-
51
male
4
2
8
2
2
2
1
21
female
3
2
5
5
8
-
-
23
male
4
5
8
10
8
5
5
45
female
-
-
12
29
32
16
1
90
male
-
-
2
6
4
1
-
13
female
1
1
2
14
16
2
-
36
male
-
20
16
5
-
-
-
41
female
6
9
7
11
-
-
-
33
male
26
-
-
-
-
-
-
26
female
33
1
-
-
-
-
-
34
286
73
66
96
82
30
7
310/330
Total
183
95
44
135
49
74
60
640
Table 2. Survey of patients suffering from dermatophytic and non-dermatophytic infections in
three hospitals, classified according to clinical types and etiological agents
Positive cultures
Aspergillus spp
Fusarium spp
Scopulariopsis spp
22
13
27
80
6
0
4
Total
280
47
Female
Total
Total
Unicellular ( Yeast cells)
69
Malassezia furfur
43
Filamentous
Trichophyton rubrum
7
Nonfilamentous
Trichophyton
mentagrophytes
9
Trichophyton tonsurans
Microsporum canis
Male
Trichophyton interdigitale
Culture results of
different ringworms
Microsporum gypseum
Dermatophytes
Negative cultures
Non - dermatophytes
5
17
42
50
27
26
26
101 11
2
6
313
14
24
85
119
49
39
53
181 17
2
10
593
47
640
Figure 1. Occurrence of different clinical types of tinea in 81 from total 620 patients suffering
from diabetes
4
3.5
3
Infected
cases
(%)
2.5
2
Male
Female
1.5
1
0.5
0
Tinea capities Tinea corporis Tinea cruris
Tinea pedis Tinea unguium
Tinea
versicolor
Figure 2. Occurrence of different clinical types of tinea in 82 from total 280 patients suffering
from cancer
6
5
Infected
cases
(%)
4
Male
Female
3
2
1
0
Tinea capities Tinea corporis Tinea cruris
Tinea pedis Tinea unguium
Tinea
versicolor
Figure 3. Occurrence of different clinical types of tinea in 30 from total 210 patients suffering
from heart diseases
3
2.5
Infected
cases
(%)
2
Male
Female
1.5
1
0.5
0
Tinea capities
Tinea corporis
Tinea cruris
Tinea pedis
Tinea unguium Tinea versicolor
Table 3. Minimum inhibitory concentration (MIC) of ozone applied as a gas for 2 hours or as ozonized oil
for 2 minutes against growth and spore germination of the test dermatophyte fungi.
MIC a (µg/ml)
Ozonized oil b
Gaseous ozone
Dermatophyte fungi
growth
Spore
germination
growth
Spore germination
Microsporum gypseum
4
4
0.5
0.25
M. canis
4
4
0.5
0.25
Trichophyton interdigitale
16
8
2.0
1.0
T. mentagrophytes
16
8
2.0
1.0
T. rubrum
8
8
1.0
0.25
aThe minimum inhibitory concentration (MIC) is defined as the lowest ozone concentration that
reduced growth or spore germination by 80% in comparison to the ozone-free controls.
bOil was dissolved in dimethyl sulfoxide (DEMSO).
Table 4. Effect of the minimum inhibitory concentration (MIC) of ozonized oil (for growth)
on sporulation (log CFU/ml) of the test dermatophyte fungi
Sporulation
(log CFU/ml)
Dermatophyte fungi
% reduction a
Control
Ozonized oil **
Microsporum gypseum
6.79 ± 0.32
4.90 ± 0.27
98.71
M. canis
6.79 ± 0.21
5.26 ± 0.32
97.05
Trichophyton interdigitale
5.65 ± 0.12
4.92 ± 0.26
81.33
T. mentagrophytes
5.61 ± 0.17
4.75 ± 0.32
86.34
T. rubrum
5.83 ± 0.08
5.28 ± 0.11
72.06
a Reduction in sporulation calculated as percentage from the control value
Table 5. Change in conductance of the bathing solutions containing mycelia of some
dermatophyte fungi previously treated with the minimum inhibitory concentration (MIC) of
ozonized oil.
Conductivity after 8 hrs
(µmohs/g fresh weight)
Dermatophyte fungi
Leakage as
% of total
conductance
Control
Ozonized oil
Total
conductancea
Microsporum gypseum
24.00 ± 2.41
44.03 ± 3.82
49.44 ± 2.98
40.51
M. canis
24.64 ± 3.98
41.90 ± 4.87
49.50 ± 6.32
34.87
Trichophyton interdigitale
22.98 ± 3.02
32.51 ± 4.01
46.22 ± 4.93
20.62
T. mentagrophytes
21.51 ± 2.21
31.03 ± 3.18
44.97 ± 5.32
21.17
T. rubrum
22.92 ± 3.88
28.02 ± 2.92
44.29 ± 3.98
11.52
a Total
conductance was measured by adding 1ml chloroform.
Table10. Sugar amount in bathing solutions after 8 hours incubation of mycelia of some dermatophyte
fungi previously treated with the minimum inhibitory concentration (MIC) of ozonized oil.
Dermatophyte fungi
Sugar amount in bathing solution
(µg/ml)
% increase in sugar
permeability
Control
Ozonized oil
Microsporum gypseum
154 ± 18
312 ± 14
102.6
M. canis
133 ± 09
243 ± 15
82.7
Trichophyton interdigitale
108 ± 11
159 ± 11
47.2
T. mentagrophytes
110± 10
171 ± 10
55.5
T. rubrum
95 ± 08
131 ± 10
37.9
Table 11. Effect of the minimum inhibitory concentration (MIC) of ozonized oil on the activity
of keratinase of some selected dermatophyte fungi.
Keratinase activity (U/ml)
Dermatophyte fungi
% reduction
Control
Ozonized oil **
Microsporum gypseum
8.21±3.86
1.05±0.98
87.21
M. canis
26.32±3.86
6.11±3.33
76.79
Trichophyton interdigitale
9.14±2.06
4.43±3.04
51.53
T. mentagrophytes
18.98±4.50
6.17±2.70
67.49
T. rubrum
22.00±4.70
11.08±5.74
49.64
Table 12. Effect of ozonized oil and terbinafine applied at 2 µg/ml on recovery of guinea pigsa
artificially inoculated with some dermatophytes.
Recovery (%)b
Dermatophyte fungi
Control
Ozonized oil
(2 µg/ml)c
Terbinafine as reference
drug
(2 µg/ml)
Microsporum gypseum
0.00
100
100
M. canis
0.00
85.8±6.1
92.9±4.9
Trichophyton interdigitale
0.00
87.5±3.6
83.3±4.8
T. Mentagrophytes
0.00
82.5±2.2
87.5±6.3
T. rubrum
0.00
79.7±5.0
79.7±4.1
aThirty
two guinea pigs were employed for this experiment.
bGuinea pigs were examined after five weeks post inoculation.
COzonized oil was applied once a day.
Conclusion
• Tinea capitis was the most common clinical disease (28.6%) in the clinic attending patients,
followed by tinea pedis (21.1%) and tinea corporis (14.8%). Tinea cruris and tinea unguium
were the least in occurrence (6.9 and 7.7%, respectively). Tinea versicolor was detected in
11.6%.
• The skin diseases due to Candida spp. were reported in 9.4% of skin mycosis.
• The most susceptible persons were children below 10 years (44.7%) followed by those aging
31-40 years (15%).
• Unicellular yeast was the most common etiological agent and Trichophyton tonsurans was
the second most frequent causative agent accounting for 18.6%, followed by Microsporum
canis (13.3%).
• Diabetes looks a risk factor for tinea pedis (6.61% of diabetic patients) and tinea corporis
(3.23%).
• The effect of ozone in the form of ozonized oil on the sporulation, mycelia leakage of sugar
and nutrients as well as activity of keratinase of the pathogens is apparently more efficacious
than gaseous ozone. Generally, M. gypseum and M. canis were the most susceptible fungi
while T. interdigitale and T. mentagrophytes were relatively resistant.
• On a trail to apply a control measure, the percentage recovery of M. gypseum- infected
guinea pigs reached 100% in on application of 2 µg/ml ozonized oil. This result is equivalent
to that obtained by using 2 µg/ml terbinafine as antifungal reference drug.
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