Mycological Study of Dermatomycosis in Population of Cairo, Egypt

Salama A. Ouf1, Tarek A. Moussa1, Samar M. R. Eltahlawi2, Alshimaa S.M. Abd- elmegeed1

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 immune-depressed 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 anti-microbial 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, Egypt2- 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

TotalTotal male

/Total female

Age groups (yr)Clinical types of tinea

61 - 70 51 - 60 41 - 50 31 - 40 21 - 30 11 - 20 0 - 10

183120 - - - - - 22 98 male

Tinea capitis63 - - - - - - 63 female

9544 - - 6 - 2 8 28 male

Tinea corporis51 - 4 6 14 4 3 20 female

4421 1 2 2 2 8 2 4 male

Tinea cruris23 - - 8 5 5 2 3 female

13545 5 5 8 10 8 5 4 male

Tinea pedis90 1 16 32 29 12 - - female

4913 - 1 4 6 2 - - male

Tinea unguium36 - 2 16 14 2 1 1 female

7441 - - - 5 16 20 - male

Tinea versicolor33 - - - 11 7 9 6 female

6026 - - - - - - 26 male

Candidiosis34 - - - - - 1 33 female

640 310/330 7 30 82 96 66 73 286 Total

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

Table 2. Survey of patients suffering from dermatophytic and non-dermatophytic infections in three hospitals, classified according to clinical types and etiological agents

Culture results of different ringworms

Positive cultures

Negative

cultures

Total

DermatophytesNon - dermatophytes

Total

Non-filamentous Filamentous

Microsporum gypseum

Trichophyton interdigitale

Microsporum canis

Trichophyt

on tonsurans

 

Trichophyt

on mentagrophyte

s

Trichophyton rubrum

Malassezia

furfur

Unicellular ( Yeast

cells)

Aspergillus

spp

Fusarium spp

Scopulariopsis spp

Male 9 7 43 69 22 13 27 80 6 0 4 28047Female

5 17 42 50 27 26 26 101 11 2 6 313

Total14 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

00.5

11.5

22.5

33.5

4

Tinea capities Tinea corporis Tinea cruris Tinea pedis Tinea unguium Tineaversicolor

MaleFemale

Infected cases (%)

Figure 2. Occurrence of different clinical types of tinea in 82 from total 280 patients suffering from cancer

Infected cases (%)

0

1

2

3

4

5

6

Tinea capities Tinea corporis Tinea cruris Tinea pedis Tinea unguium Tineaversicolor

MaleFemale

Figure 3. Occurrence of different clinical types of tinea in 30 from total 210 patients suffering from heart diseases

0

0.5

1

1.5

2

2.5

3

Tinea capities Tinea corporis Tinea cruris Tinea pedis Tinea unguium Tinea versicolor

MaleFemale

Infected cases (%)

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)

Dermatophyte fungiOzonized oil b Gaseous ozone

Spore germination growth Spore germination growth

0.25 0.5 4 4 Microsporum gypseum

0.25 0.5 4 4 M. canis

1.0 2.0 8 16 Trichophyton interdigitale

1.0 2.0 8 16 T. mentagrophytes

0.25 1.0 8 8 T. rubrum

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

Dermatophyte fungi

Sporulation (log CFU/ml)

% 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 72.06a 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. 

Dermatophyte fungi

Conductivity after 8 hrs(µmohs/g fresh weight)

Leakage as% of total

conductanceControl 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.

Dermatophyte fungiKeratinase activity (U/ml)

% reductionControl 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.

Dermatophyte fungi

Recovery (%)b

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.

THANK YOU