• Topic: Clinical and laboratory diagnosis of fungal diseases affecting the skin and its appendages
• Student info: Course: Medical Immunology

1. Keep exactly what you gave: Medical Immunology
2. Use requirement-compliant course title: Microbiology, Virology and Immunology

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📋 PRESENTATION DETAILS (for every slide header/footer)

• Title: Clinical and Laboratory Diagnosis of Fungal Diseases Affecting the Skin and Its Appendages
• Course: Medical Immunology
• Student: Zeel Kathiriya
• Group: 11 | Semester: 4
• Instructor: Duyshenkulova Marina

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🖼️ SLIDE 1 — TITLE SLIDE

Clinical and Laboratory Diagnosis of Fungal Diseases Affecting the Skin and Its Appendages

Course: Medical Immunology Presented by: Zeel Kathiriya Group: 11 | Semester: 4 Instructor: Duyshenkulova Marina

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🖼️ SLIDE 2 — TABLE OF CONTENTS

1. Aim & Objectives
2. Overview of Fungal Diseases of Skin & Appendages
3. Dermatophytosis (Tinea Infections)
4. Candidiasis of Skin & Nails
5. Clinical Diagnosis Methods
6. Laboratory Diagnosis — Direct Microscopy (KOH Mount)
7. Laboratory Diagnosis — Culture & Identification
8. Molecular & Serological Methods
9. Antifungal Treatment Overview
10. Conclusion
11. References

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🖼️ SLIDE 3 — AIM & OBJECTIVES

To understand the clinical features and laboratory methods used for the diagnosis of superficial and cutaneous fungal diseases affecting the skin and its appendages.

• Classify fungal diseases by affected structure (epidermis, hair, nails)
• Describe clinical presentations of tinea and candidiasis
• Explain direct microscopy (KOH mount) and its interpretation
• Describe fungal culture on Sabouraud Dextrose Agar (SDA)
• Outline molecular and serological diagnostic methods
• Identify Wood lamp and dermoscopy as clinical adjuncts
• Summarise antifungal treatment principles

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🖼️ SLIDE 4 — OVERVIEW: CLASSIFICATION

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🖼️ SLIDE 5 — DERMATOPHYTOSIS (TINEA INFECTIONS)

• Trichophyton spp. — most common; T. rubrum, T. tonsurans, T. violaceum
• Microsporum spp. — M. canis (zoophilic), M. gypseum (geophilic)
• Epidermophyton floccosum — affects skin and nails only (not hair)

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🖼️ SLIDE 6 — CANDIDIASIS OF SKIN & NAILS

• Skin folds (intertriginous areas): groins, axillae, submammary
• Erythematous macerated patches + satellite pustules at margins
• Risk factors: obesity, diabetes, excessive moisture, broad-spectrum antibiotics

• Nail fold inflammation (paronychia) → nail plate invasion
• White/yellow discolouration, proximal subungual pattern
• Chronic mucocutaneous candidiasis (CMC) in immunodeficiency

• Thick white pseudomembranous plaques on tongue/buccal mucosa
• Underlying erythematous mucosa
• Seen in infants, immunocompromised, antibiotic users, denture wearers

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🖼️ SLIDE 7 — CLINICAL DIAGNOSIS METHODS

• Performed in a darkened room
• Microsporum spp. → apple-green / blue-green fluorescence of infected hair shafts
• Trichophyton spp. — usually non-fluorescent
• Quick, non-invasive, bedside screening tool
• Also useful for Pityriasis versicolor (yellow-green), Erythrasma (coral-red)

• Comma / corkscrew hairs → tinea capitis
• Spiked pattern + longitudinal striae → onychomycosis
• Dermatoscopic "black-dot" pattern in endothrix tinea capitis

• Annular plaque with central clearing → Tinea corporis
• Satellite pustules at lesion edges → Cutaneous candidiasis
• Nail dystrophy + subungual debris → Onychomycosis
• Fluorescence guides targeted sample collection for KOH & culture

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🖼️ SLIDE 8 — LAB DIAGNOSIS: DIRECT MICROSCOPY (KOH MOUNT)

• Skin scrapings (from active border of lesion)
• Nail clippings + subungual debris
• Plucked hairs (include root/bulb)

1. Place specimen on glass slide
2. Add 10–20% KOH solution (± DMSO or Calcofluor white)
3. Gently heat or wait 15–30 min to digest keratin
4. Apply cover-slip; examine at 10× then 40×

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🖼️ SLIDE 9 — LAB DIAGNOSIS: CULTURE & IDENTIFICATION

• Acidic pH 5.6 — suppresses bacteria; selective for fungi
• Low nutrients — mimics keratin environment
• Incubation: 25–28°C for 2–4 weeks
• Add Cycloheximide + Chloramphenicol to inhibit saprophytes and bacteria
• Dermatophyte Test Medium (DTM): colour change red = dermatophyte

• Trichophyton — thin-walled pencil-shaped macroconidia; teardrop microconidia
• Microsporum — large, thick-walled, rough spindle macroconidia (4–15 cells)
• Epidermophyton — club-shaped, smooth, 2–4 cell macroconidia; NO microconidia
• Candida — Germ tube test in serum at 37°C: positive = C. albicans (2 h)

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🖼️ SLIDE 10 — MOLECULAR & SEROLOGICAL METHODS

• Pan-dermatophyte PCR directly from nail/hair specimens
• ITS (Internal Transcribed Spacer) region sequencing — gold standard for species ID
• Results in 24–48 hours (vs. 2–4 weeks for culture)
• Detects mixed infections and antifungal-resistant strains (e.g. T. indotineae)
• High sensitivity ~95%

• Protein fingerprint identification from cultured colonies
• Rapid (minutes per sample), high accuracy for Candida & dermatophytes
• Cost-effective for high-throughput laboratories
• Replaces traditional biochemical tests

• PAS (Periodic Acid–Schiff) stain on nail biopsy → hyphae stain magenta/red
• GMS (Grocott-Gomori Methenamine Silver) → fungal elements stain black
• Indicated when KOH and culture are negative/inconclusive
• Anti-Candida antibodies (serology) — useful in chronic mucocutaneous candidiasis

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🖼️ SLIDE 11 — ANTIFUNGAL TREATMENT OVERVIEW

• Clotrimazole, Miconazole (azoles) — tinea corporis, cruris, pedis, candidiasis
• Terbinafine cream — tinea pedis, corporis (allylamine)
• Ciclopirox nail lacquer — mild onychomycosis
• Nystatin cream/powder — cutaneous candidiasis

• Terbinafine (oral) — drug of choice for dermatophyte onychomycosis & tinea capitis
• Itraconazole — pulse therapy for onychomycosis; Candida; non-dermatophyte molds
• Fluconazole — candidiasis, tinea capitis (alternative)
• Griseofulvin — first-line tinea capitis in children (older, less used now)

⚠️ Emerging resistance: T. indotineae (terbinafine-resistant) — increasing globally. Always confirm diagnosis before starting systemic therapy.

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🖼️ SLIDE 12 — CONCLUSION

1. Fungal diseases of skin and appendages range from superficial to subcutaneous, classified by depth of infection.
2. Dermatophytes (Trichophyton, Microsporum, Epidermophyton) are the principal agents of tinea infections affecting skin, hair and nails.
3. Candida albicans causes cutaneous candidiasis, paronychia, and oral thrush — especially in immunocompromised patients.
4. KOH direct microscopy is the fastest, most accessible first-line laboratory test, demonstrating hyphae or pseudohyphae.
5. Culture on SDA remains the gold standard — enables species identification and antifungal susceptibility testing.
6. Molecular tools (PCR, ITS sequencing, MALDI-TOF) provide rapid, species-level diagnosis in complex or resistant cases.
7. Wood lamp and dermoscopy are valuable non-invasive clinical adjuncts to guide sample collection.
8. Treatment ranges from topical azoles/allylamines for limited disease to prolonged systemic antifungals for nail and scalp involvement.

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🖼️ SLIDE 13 — REFERENCES

1. Murray, P.R., Rosenthal, K.S., Pfaller, M.A. (2021). Medical Microbiology (9th ed.). Elsevier.
2. Brooks, G.F., Carroll, K.C., Butel, J.S. et al. (2022). Jawetz, Melnick & Adelberg's Medical Microbiology (28th ed.). McGraw-Hill.
3. Havlickova, B., Czaika, V.A., Friedrich, M. (2008). Epidemiological trends in skin mycoses worldwide. Mycoses, 51(Suppl 4): 2–15.
4. Aly, R. (1994). Ecology and epidemiology of dermatophyte infections. J Am Acad Dermatol, 31(3): S21–S25.
5. Ameen, M. (2010). Epidemiology of superficial fungal infections. Clin Dermatol, 28(2): 197–201.
6. Nenoff, P. et al. (2014). Mycology — an update Part 1: Dermatomycoses. J Dtsch Dermatol Ges, 12(3): 188–210.
7. Gupta, A.K., Cooper, E.A. (2008). Update in antifungal therapy of dermatophytosis. Mycopathologia, 166(5–6): 353–367.
8. Rezusta, A. et al. (2011). MALDI-TOF MS identification of dermatophytes. Med Mycol, 50(5): 541–545.

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📌 QUICK SUMMARY: Images per slide

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SLIDE 1 — TITLE SLIDE

Clinical and Laboratory Diagnosis of Fungal Diseases Affecting the Skin and Its Appendages

Course: Medical Immunology Presented by: Zeel Kathiriya Group: 11 | Semester: 4 Instructor: Duyshenkulova Marina

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SLIDE 2 — TABLE OF CONTENTS

1. Aim & Objectives
2. Overview & Classification
3. Dermatophytosis — Tinea Infections
4. Candidiasis of Skin & Nails
5. Clinical Diagnosis Methods
6. Laboratory Diagnosis — KOH Mount
7. Laboratory Diagnosis — Culture
8. Molecular & Serological Methods
9. Antifungal Treatment
10. Conclusion
11. References

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SLIDE 3 — AIM & OBJECTIVES

To understand the clinical features and laboratory methods for diagnosing fungal diseases of the skin and its appendages.

• Classify fungal diseases by affected structure
• Describe clinical presentations of tinea & candidiasis
• Explain KOH microscopy and culture methods
• Outline molecular diagnostic tools
• Summarise antifungal treatment principles

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SLIDE 4 — OVERVIEW & CLASSIFICATION

• Affect outermost keratin only
• e.g. Pityriasis versicolor, Tinea nigra

• Invade skin, hair and nails
• Caused by dermatophytes (Trichophyton, Microsporum, Epidermophyton)

• Enter via trauma
• e.g. Sporotrichosis, Mycetoma

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SLIDE 5 — DERMATOPHYTOSIS

• Trichophyton spp. — T. rubrum, T. tonsurans (most common)
• Microsporum spp. — M. canis (zoophilic)
• Epidermophyton floccosum — skin & nails only

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SLIDE 6 — CANDIDIASIS

• Skin folds — erythema + satellite pustules
• Risk: diabetes, obesity, moisture

• Nail fold swelling, nail discolouration
• Chronic mucocutaneous candidiasis (CMC) in immunodeficiency

• White curd-like plaques on tongue/mucosa
• Common in infants & immunocompromised patients

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SLIDE 7 — CLINICAL DIAGNOSIS METHODS

• Microsporum spp. → apple-green fluorescence of hair shafts
• Trichophyton → usually non-fluorescent
• Quick, non-invasive, bedside screening

• Comma/corkscrew hairs → tinea capitis
• Spiked pattern → onychomycosis

• Annular lesion + central clearing → tinea corporis
• Satellite pustules → candidiasis
• Nail dystrophy + debris → onychomycosis

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SLIDE 8 — KOH DIRECT MICROSCOPY

1. Add 10–20% KOH to specimen on slide
2. Heat gently / wait 15–30 min
3. Examine at 10× then 40×

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SLIDE 9 — CULTURE & IDENTIFICATION

• Acidic pH 5.6 — selective for fungi
• Incubate 25–28°C for 2–4 weeks
• Add cycloheximide + chloramphenicol to suppress contaminants

• Microsporum — large spindle-shaped rough macroconidia
• Trichophyton — thin pencil-shaped macroconidia
• Candida — germ tube test (+) in serum = C. albicans

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SLIDE 10 — MOLECULAR & SEROLOGICAL METHODS

• Gold standard for species identification
• Results in 24–48 h vs. 2–4 weeks for culture
• Detects mixed infections & resistant strains

• Rapid protein fingerprint from cultured colonies
• High accuracy for Candida & dermatophytes

• PAS stain → hyphae stain magenta/red
• GMS stain → fungal elements stain black
• Used when KOH and culture are negative

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SLIDE 11 — ANTIFUNGAL TREATMENT

• Clotrimazole / Miconazole — tinea corporis, cruris, pedis
• Terbinafine cream — tinea pedis, corporis
• Nystatin — cutaneous candidiasis
• Ciclopirox lacquer — mild onychomycosis

• Terbinafine oral — onychomycosis, tinea capitis
• Itraconazole — Candida, non-dermatophyte molds
• Fluconazole — candidiasis, tinea capitis
• Griseofulvin — tinea capitis in children

• Skin (tinea) — 2–6 weeks
• Tinea capitis — 6–8 weeks
• Onychomycosis — 3–6 months

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SLIDE 12 — CONCLUSION

• Fungal skin diseases range from superficial (keratin only) to subcutaneous (deep tissue).
• Dermatophytes cause tinea infections of skin, hair and nails; Candida causes candidiasis.
• KOH mount — fastest, cheapest first-line test; identifies hyphae/pseudohyphae.
• SDA culture — gold standard; enables species ID and susceptibility testing.
• Molecular methods (PCR, MALDI-TOF) give rapid species-level results.
• Wood lamp and dermoscopy are useful non-invasive clinical adjuncts.
• Treatment ranges from topical antifungals to prolonged systemic therapy depending on site and severity.

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SLIDE 13 — REFERENCES

1. Murray, Rosenthal, Pfaller. Medical Microbiology, 9th ed. Elsevier, 2021.
2. Brooks et al. Jawetz, Melnick & Adelberg's Medical Microbiology, 28th ed. McGraw-Hill, 2022.
3. Havlickova et al. Epidemiological trends in skin mycoses. Mycoses, 2008; 51(Suppl 4): 2–15.
4. Aly R. Ecology and epidemiology of dermatophyte infections. J Am Acad Dermatol, 1994; 31(3): S21–S25.
5. Ameen M. Epidemiology of superficial fungal infections. Clin Dermatol, 2010; 28(2): 197–201.
6. Nenoff et al. Mycology — an update Part 1. J Dtsch Dermatol Ges, 2014; 12(3): 188–210.
7. Gupta AK, Cooper EA. Antifungal therapy of dermatophytosis. Mycopathologia, 2008; 166: 353–367.
8. Rezusta et al. MALDI-TOF MS for dermatophytes. Med Mycol, 2011; 50(5): 541–545.

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