Trichophyton Microsporum Epidermophyton. dermatomycosis. Dematiaceous(pigmented fungi ) Dimorphic fungi Yeast and yeast like saprophyte

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1 Cutaneous candidiasis

2 dermatophytosis Trichophyton Microsporum Epidermophyton dermatomycosis Dematiaceous(pigmented fungi ) Dimorphic fungi Yeast and yeast like saprophyte

3 dermatomycosis Yeast & yeast like phaeohyphomycete Hyphomycete Candida albicans Cryptococcus neoformans Nattrassia mangiferae ( scytalidium dimitiatum) Exophiala dermatididis Exophiala spinifera Curvularia lunata Alternaria alternata Pyrenochaeta romeroi Oh Ochroconisgallapavum Paraconiothyrium cyclothyrioides Aphanoascus fulvescens( Anixiopsis stercoraria ) Onychocola canadensis Aspergillus glucus group Aspergillus candidus link Fusarium solani Fusarium oxysporum Acremonium strictum Scopulariopsis brevicaulis

4 Candidiasis Causative agents Candida albicans, Candida dubliniensis, Candida famata, Candida glabrata, Candida guilliermondii, Candida haemulonii, Candida kefyr, Candida krusei, Candida lusitaniae, Candida norvegensis, Candida parapsilosis, Candida tropicalis, Candida viswanathii Non alb bicans Cand dida ( Cand ida sp)

5 Who is at increased risk? Babies with a nappy rash People with a metabolic disorder ( diabet.) Overweight people Pregnant woman, high dose contraceptive People who work in wet conditions Immunodeficiency ( AIDS )

6 Ask some question! Occupation ( work in wet condition ) Age Underlying disease ( metabolic disorder d, immunodificiency, vitamin dificiency, High dose broad spectrum antibiotics, corticosteroides, chemotherapy.

7 Attention to the symptom purple patches on the skin like eczema Itchy, the itching in these patches is highest near the edge A small degree of scalling Blister resembling spots are often seen around the itchy patches

8 Cutaneous candidiasis intertrigo diaper candidiasis paronychia and onychomycosis Cheilitis

9 paronychia and onychomycosis

10 Cheilitis

11 cutaneous candidiasis Satellite lesions of Satellite lesions of cutaneous candidiasis showing typical collars of scale

12 intertrigo candidiasis

13 bone curette Sampling device

14 Patient prepration 3 days no washing prior sampling No bacterial or fungal treatment during 7 days any ointments or other local applications present should first be removed with an alcohol wipe

15 sampling If multiple lesions are present choose the most recent for scrapings as old loose scale is often not satisfactory. The tops of any fresh vesicles should be removed as the fungus is often plentiful in the roof of the vesicle. scrap the lesions with a bone curette or blunt scalpel as for tinea scaly border should ldbe scraped NOTE: Following the collection of skin scales all scraped lesions should be firmly rubbed with a swab moistened in BHI broth.

16 Laboratory diagnosis of cutaneous candidiasis (direct exam & culture ) 1 direct exam A. Skin scrapings: Make a wet mount preparation in KOH 10% for direct microscopy. Note a Calcofluor stained mount may also be necessary. B. Skin swabs: Smear swab onto heat sterilized glass slide for Gram stain 2 Culture Sabouraud's dextrose agar slopes containing chloramphenicol and gentamicin, but NO cycloheximide Candida chrome agar 3. incubate at 35C. Maintain cultures for 4 weeks

17 Candidiasis (KOH 10% mount )

18 Gram staining

19

20 Candida chrome agar

21 Germ tube test

22

23 Corn meal agar with 1% tween 80

24 C.albicans/ c. dubliniensis

25 C.albicans/ c. dubliniensis

26 api20c

27 PCR RFLP assay ITS S ITS2 regions of fungal rrna genes were amplified with universal primers ITS1, 5 -TCC GTA GGT GAA CCT GCG G-3 ITS4, 5 -TCC TCC GCT TAT TGA TAT GC-3. Digestion of the PCR products with one restriction enzyme, MspI, allowed discrimination of medically important Candida species, including C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, C. krusei, and C. guilliermondii

28 a) Agarose gel electrophoresis of ITS-PCR products of some of Candida species isolated from different cutaneous samples. (b) Restriction digestion of PCR products of Candida strain with the Msp1enzyme. Line 6: C. albicans, Line 5: C. tropicalis, Line 4: C. guilliermondii, Line 3: C. krusei, Line 2: C. parapsilosis and Line 1: C. famata. Line M: 100 bp molecular size marker.

29 PCR primers ITS3, 5 -GCA TCG ATG AAG AACGCA GC-3 ITS4 5 -TCC TCC GCT TAT TGA TATGC-3 ITS1, 5 -TCC GTA GGT GAA CCT GCG G-3

30 Antibiogram CLSI protocol M 44 A Disk diffusion M 27 A3 macrobroth & microtiter

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