B.

CUTANEOUS FUNGI

 Invade/ destroy keratinized – hair, skin, nails

o Keratinolytic keratinase
o With inflammation  can deep to cutaneous
 Dermatophytes  dermatophytosis

CLASSIFICATION:

Origin

Antropophilic Geophilic Zoophilic

Source: Humans soil Animal
*infected  uninfected
No. of conidia (culture medium) Few Most numerous Moderate
Tissue reaction Mild Severe Moderate
Examples Epidermophyton floccosum Microsporum gypseum Microsporum canis
Tricophyton equinum

Morphology

Epidermphyton Microsporum Tricophyton

Tissue/s infected Skin Hair Hair
Nails Skin Skin
Nails
Presence in tissues NO Some species NO
Teleomorph None/ unknown (Imperfect) Arthroderma/ Nannizia Arthroderma
*Anamorph
*Arthroconidia – able to damage the hair

A. Skin Infection

TINEA:

 Circular patterns, patches *numerous

 Reddish, itchy, scaly
 “ringworm”

SKIN Tinea capitis Tinea barbae Tinea corporis Tinea cruris Tinea manuum Tinea pedis Tinea unguium Tinea imbricate
INFECTIONS
 Characteristics: Ringworm of the Barber’s itch Body ringworm Ringworm of the *peeling – Ringworm of the Overlapping ring-
scalp “ringworm” groin interdigital areas nails like lesions
*Alopecia *scaly lesions Jock’s itch *Bullae – fluid- Nails  brittle, *Tricophyton
*circular patterns filled vesicle thickened, concentricum
discolored
Affected site: Head  scalp Bearded areas of Trunk Groin area Hands Soles Nails
Eyelashes the face Shoulders *Perineum and Palms Feet
Eyebrows Neck *Skin other than perianal areas Fingers
bearded area,
scalp, groin,
hands, or feet
*T. unguium - difficult to treat; oral anti-fungal drugs

B. Hair infection  Brittleness  loss of hair  Inside

 Growth of fungi along the length of  Inner layer
1. Ectothrix
the hair  Fungi – arthroconidia
 Outside
2. Endothrix 3. Favic
 Outer layers
 Worst type  Destroys inner, outer,  Destruction of tissues  almost  Mousy odor  pus formation 
subcutaneous death of the tissues yellow crust
 Inflammation  pus formation

Laboratory Diagnosis  Hyphae  Nonselective SDA

o Small: 2-3 microns o Do not inhibit saprophytic fungi
Specimen:
o Hyaline, septate, branching o 25-30°C for at least 1 week
 Skin scrapings  Presence of arthroconidia  Maximum of 3 weeks
 Hair o Endothrix & ectothrix infection vs. favic  Selective SDA
 Nail clippings type o Saboraud Dextrose Agar with
antimicrobial agents:
Exposure to Wood’s Light  Chloramphenicol
 Yellow-green fluorescence  Calcofluor White (CFW) staining  Cycloheximide
o M. audouinii  Hyphae o Includes:
o M. canis o Branching  Mycosel (BBL)
o Fluoresces  Mycobiotic (Difco)
Direct Microscopy  Dermatophyte test medium
Culture: (DTM)
 KOH Preparation
o Do not distinguish the dermatophytes  Primary isolation  Modified SDA
o 25-30°C for at least 1 week
  Maximum of 3 weeks  Chlortetracycline  Dermatophytes  alkaline
Dermatophyte test medium (DTM)  Gentamicin products  reddish
o Selective for dermatophytes o Differential discoloration of agar
 Cycloheximide  Phenol red

Microsporum Microsporum canis Microsporum Tricophyton mentagrophytes Tricophyton rubrum

audouinii gypseum Type I Type II Type I Type II
Classification Zoophilic Geophilic Zoophilic Anthropophilic Anthropophilic
Wood’s Light Yellow-green Yellow-green (-) (-) (-) (-) (-)
fluorescence fluorescence
Colonial Flat, velvety, thin Cottony and Flat and granular Flat and granular to Flat and powdery to Cottony and velvety Powdery to low
Characteristic granular powdery cottony velvety
Pigmentation
Obverse Light: white to buff Cinnamon-colored yellow cream to buff yellow cream to buff White to reddish Cream to deep red
white, light brown

Reverse Pale salmon to pale Yellow, rarely pale Light tan Pale to red-brown Pale to red-brown Wine-red Wine-red
brownish:
Orange-red
Conidia Uncommon More conidia in Type I than in Type II More conidia in Type II than in Type I
*Ovoid if present
Macroconidia 6 – 15 cells 3-9 celled, broadly Uncommon Pencil-shaped
Asymmetrical spindle- shaped, Cigar/ pencil/club-shaped Cigar-shaped
beaked apex rough-walled Thin, smooth walls Thin walled
Terminal ends may 3-6 cells Broad bases
be rounded 3-8 cells
Microconidia Clavate or pyriform if present, single or Globose & unicellular Clavate or pyriform Tear-shaped
Laterally attached to in small clusters Produced singly along Attached singly in Club-shaped
hyphae (*ovoid) the hyphae on short sleeves en thryses En thryses
pedicles – “en grappe” along the hyphae by
pedicles – “birds on
trees”
Hyphae o Hyaline o Hyaline o Hyaline Hyaline, septate, branched
o Septate o Septate o Septate *Spiral hyphae seen in 30% of isolates
o Branched o Branched o Branched
o Terminal
chlamydospores
o Pectinate
 Tricophyton tonsurans Tricophyton Tricpohyton Tricophyton Tricophyton Epidermophyton
concentricum verrucosum schoenleinii violaceum floccosum
Classification Anthropophilic
Wood’s Light (-) (-) (-) (-) (-) (-)
Colonial Characteristic Flat and velvety Folded and furrowed Commonly small Convoluted to folded Verrucose and Flat, slightly granular at
Convoluted with raised Glabrous to slightly glabrous first
center and flat velvety
periphery with some Often submerged into
submerged growth surrounding medium
Glabrous to slightly “mousy odor”
velvety
Pigmentation *color differ to medium
Obverse Yellow White but becoming White or cream White to tan Cream then becoming White which becomes
honey-brown lavender khaki-green brown
-center is folded
Reverse Yellow-brown to Yellow Colorless or salmon Colorless or light Yellow-brown with
chestnut red-brown yellow observable folds
Conidia Uncommon No conidia Absent/ uncommon
macroconidia and
microconidia
Macroconidia Uncommon Uncommon “Snowshoes”
Absent or rare, 3-5 cells “Paddles”
distorted Thin-walled “Beaver’s tail”
Small pencil-/club- “Rat-tail extension”
/cigar-shaped
Thick walls
3-8 cells
Microconidia Various size and shapes Abundant No microconidia
with flattened base Large
Abundant Clavate
Teardrop-/club-shaped
“Balloon forms”
- Aged pleomorphic
microconidia
- Broad matchstick
Hyphae Terminal Masses of tangled Favic chandeliers, Swollen hyphae
chlamydospore hyphae with antlers, nail head containing cytoplasmic
chlamydospores granules
PHYSIOLOGICAL TESTS  Acquire strands of hair → LPCB → Microscope
 Positive result: wedge-shaped perforation into the hair
Polished Rice Test
 Negative result: no perforeation
 Purpose: M. canis vs. M. audouinii
UIrease test
M. canis Positive growth
 Culture medium
M. audouinii Negative growth
o Christensen’s agar
 Procedure:
o Stuart’s broth
o Medium: Rice grains (8 grams) with distilled water (2.5ml)
 Purpose: T. mentagrophytes vs. T. rubrum
o Sterilize: 15 mins @ 125°C (autoclave)
T. mentagrophytes Positive
o Plate
T. rubrum Negative
o Transfer colonies from primary isolation medium
 Result (Christensen’s medium)
o Incubation:
o Positive: red to purple color in less than 4 days of incubation
 Room temperature
o Negative: No color change
 6-10 days
Growth factor test
 25-30°C
 Casamino acid agar/ Tricophyton agar
 Result: Rice grains → Yellowish = positive
o Nicotinic acid
Hair Perforation test
o L-Histidine
 Purpose: T. mentagrophytes vs. T. rubrum
o Growth factors
T. mentagrophytes Positive
 Thiamine
T. rubrum Negative
 Inositol
 Other dermatophytes with positive results:
 2 Plates:
o M. canis
o Plate 1
o M.gypseum
 Nicotinic acid
o T. tonsurans
 L-Histidine
o T. violaceum
 Inositol
 Culture medium: yeast extract in distilled water
o Plate 2
o 10% yeast extract – 10 grams yeast + 100ml d.water
 Nicotinic acid
o Add strands of hair
 L-Histidine
 Juvenile hair (≤1 cm)
 Inositol
o Sterilize: 15 mins @ 125°C (autoclave)
 Thiamine
o Plate → filter paper → overlay yeast extract (2-5 ml) → 10-12 hair
 T. verrucosum: (+) inositol
strands
 T. violaceum (+) inositol and thiamin
o Incubate up to 4 weeks at room temperature