Learning Objectives

• e. General properties and classification of

fungi (morphological classification)
• f. Infections produced by fungi and their
diagnosis
• Eukaryotic
• Two structures are more important
– Chitinous cell wall
• N- Acetylglucosamine
• Beta – glucan
– Ergosterol cell membrane
• Yeast

• Molds

• Dimorphism
• Single cells
• Asexual reproduction by budding
• Long filaments
• Group of hyphae forms mycelium
• Septate or non septate
37oC yeast

25oC mold
• Most of them are obligate aerobes
• Requires preformed organic source of
carbon
• Natural habitat is environment
– Except Candida albicans
• Generally sexual and asexual
• Sexual spores
– Zygospores
– Ascospores
– Basidiospores
• Asexual spores
• Otherwise conidia
• Arise from the sides or the ends of
specialised structures
– Arthrospores
– Chlamydospores
– Blastospores
– Sporangiospores
Arthrospores
Chlamydospores
• Pseudohyphae
– Multiple buds that do not detach

– Candida albicans
 Find out the answer
Asexual spores Example

Arthrospores ?

Chlamydospores ?

Blastospores Cryptococcus
neoformans
Sporangiospores ?
• Formation of granulomas
• Mainly in systemic fungal infection
– Blastomycosis, Histoplasmosis
• Cell mediated immune response Is
involved
• Acute suppuration, charecterized by the
presence of neutrophils in the exudate
– Aspergillosis, sporotrichosis
• May lead to delayed type of
hypersensitivity reaction
Classification
Laboratory diagnosis of fungal
infections
• Direct microscopy
• Culture
• DNA probe test
• Serology
 Specimen
Microscopy and culture
• Superficial and cutaneous mycoses
– Skin scrapings
– Hair
– Nail clippings
• Subcutaneous mycoses
– Exudates from the lesions
• Systemic mycoses
– Sputum, lung biopsy, CSF etc
Serology
• Blood
 Collection
• Hair
– Pluck the hair from follicle or only the shaft
– Hair brush sampling technique
• Nail
– Clippings with sterile instruments
• Skin
– Clean the site with 70% alcohol
– Scrapings with sterile scalpel
• Other specimens by appropriate means
 Transportation
• Specimens for superficial mycoses are
best transported in sterile paper covers
• Other specimens in appropriate containers
 Direct microscopy
• KOH mount
– Add 10-20% of KOH to the specimen
– KOH will digest the keratin
– Mount with a cover slip and observe
– Skin, nail and hair
– Mixing with KOH also help to clear off host
cellular debris in other specimens
• Staining
– Gram stain for yeast infections
– Special stains
• Calcofluor white stain
• Dark ground microscopy
– India ink preparation for Cryptococcus
• Histopathology stains
– For tissue biopsy
• H&E stain
• Periodic acid schiff stain
• Grocott- Gomori’s methanamine silver stain
 Culture
• Sabouraud’s dextrose agar
– Low pH inhibit bacterial contamination
– Addition of chloramphenicol and cycloheximide inhibit
bacterial and saprophytic fungal growth
• Incubation at 370C for most of the fungus
• Dimorphic fungus forms mycelium at 250C
• Grow slowly
• 7 – 10 days for most of the fungus
• Candida and Aspergillus may grow in 24 – 72
hours
 Serology
• Rise in antibody titer is demonstrated in
systemic fungal infection by complement
fixation test
• Capsular polysaccharide antigen of
Cryptococcus neoformans in the CSF is
demonstrated by latex agglutination test