Laboratory of Microbiology

Faculty of Medicine
Universitas Brawijaya
Learning Objectives
 After this learning session, you are expected to
1. Be able to explain fungi caused diseases cutaneus
and subcutan mycosis in human
2. Be able to explain laboratory diagnosis for detection
fungi caused diseases cutaneus and subcutan
mycosis in human
 Dermatophytes

Dermatomycoses
 Infection on superficial tissues rich in keratin: skin, hair,
and nail
 The disease named: tinea …../ringworm, based on
the location of lesion
 Caused by Dermatophytes, classified as
Deuteromycetes
 There are 3 genera of Dermatophytes:
1. Epidermophyton
2. Microsporum
3. Trichophyton
 Dermatophytes

Epidermophyton Microsporum Trichophyton

• Epidermophyton • M.audouinii, • Species:
floccosum  the M.canis, T.mentagrophytes,
only species of M.gypseum T.rubrum,
Epidermophyton • Infect skin and T.tonsurans,
• Infects skin and hair T.schonleini,
nail T.concentricum,
T.violaceum,
T.ferrugineum,
T.verrucosum,
T.megnini, T.gallinei
• Infect skin, nails,
and hair

Colony Morphology and
Microscopic Appearance
Epidermophyton floccosum on
SDA
Colonies grow slowly at 25 o C and
appear white to tan . The surface buffy
and powdery. The reverse side may
have on orange –brown pigmentation
Dermatophytes:
Epidermophyton
macroconidia
Epidermophyton floccosum stained
LPCP(x500)have club –shaped smooth thin
walled macroconidia that usually grow in cluster
directly from the septa hypha. It has no
microconidia.
 Dermatophytes:
Colony Morphology and Microsporum
Microscopic Appearance macroconidia

Microsporum audoinii colony on SDA Microsporum audoinii stained with

.The colony white to tan ,flat and have LPCB .The hypha are septate with
suedelike surface .The underside is intercalary and terminal spindle
frequently yellow –red-brown shaped chlamydospore
Colony Morphology and Dermatophytes:
Microsporum
Microscopic Appearance
macroconidia

Microsporum canis on SDA . The colony white to

tan , whith yellow – green lemon color at the periphery.
The surface of colony often has aradiate wooly
appearance .The underside is frequently golden yellow
or brown
Microsporum canis stained with LPCB . The
macroconidia are spindle or fusiform in
shape with a thick irregular rough wall
containing between 5-15 cells. The
characteristically have knoblike end
Colony Morphology and Dermatophytes:
Microsporum
Microscopic Appearance
macroconidia

Microsporum gypseum colony on Microsporum gypseum stained

SDA. The colony white to tan surface,a with LPCB .The macroconidia have an
white, starburstlike border, and ellipsoidal to fusiform shape with
suedelike appearance. The underside is a thin irregular wall
frequently red-brown
 Dermatophytes:
Coiled Trichophyton
Colony Morphology and hyphae

Microscopic Appearance macroconidia

microconidia

Trychophyton mentagrophytes Trychophyton mentagrophytes

on SDA . The colony usually cream –
tan in color, although they may darker
with age. The surface of colony
appears flufy and powdery.The
underside of colony may have a color
ranging from tan to brown and dark
red
stained with LPCB . Thiis fungus
typically displays sphericall or
pyriform microconidia growing
singly or in cluster. There are
characteristic septate spiral or
coiled hyphae.

Colony Morphology and
Microscopic Appearance
Genus Trichophyton
Trychophyton rubrum on SDA .
The colony usually white, velvety, or
fluffy. Characteristically will turn
red after longer incubation. The
wine red pigment produces this.
The underside of colony may have a
color red
Dermatophytes:
Trichophyton
microconidia
Trichophyton rubrum
Trychophyton rubrum stained
with LPCB . Thiis fungus typically has
septate hyphae with lateral, tear –
shaped microconidia.The
microconidia may form directly form
macroconidia

Colony Morphology and
Microscopic Appearance
Genus Trichophyton
Trychophyton schoenleinii on SDA .
The colony usually became irregular with
folded surfaces with age.
Dermatophytes:
Trichophyton
Favic
chandelier
Trichophyton schoenleinii
Trychophyton schoenleinii stained with
LPCB . This hyphal ends are swollen, giving a
nailhead morphology and a result of
branching. Often give the appearance
of favic chandeliers
 Dermatophytes:
Trichophyton
Colony Morphology and
Microscopic Appearance microconidia

Trichophyton tonsurans on SDA.

The colony can display a wide variety
of color white, creamy, yellow, tan and Trychophyton tonsurans
pink. although they may darker with stained with LPCB . The
age. The surface of colony appears septate hyphae have
flat or raised with velvety or powdery microconidia that mary
appearance.The underside of colony significantly in size and
may have a color ranging from yellow- shaped. Macroconidia are
brown to red-brown smooth,thin walled, irregular,
and clavate
macroconidia
 Dermatophytes:
Trichophyton
Colony Morphology and
Microscopic Appearance
macroconidia
Genus Trichophyton Trichophyton verrucosum

Trychophyton verrucosum on
Trychophyton verrucosum stained
SDA. The colony usually white,
with LPCB . Thiis fungus produces many
creamy color, although they may
chlamydospores that are sometimes
darker with age. The surface of colony
referred to as chains of pearls.
appears velvety and the center may
be raised
 Dermatophytes:
Trichophyton
Colony Morphology and
Microscopic Appearance

Genus Trichophyton Trichophyton violaceum

Trychophyton violaceum on SDA. Trychophyton violaceum stained with

The colony produces a waxy, LPCB . These figure demonstrate the large,
irregular shaped, raised colony with branched, irregular shaped hyphae
areas that have a violet color aand containing numerous intercalated
others that are white. chlamydoconidia.
Epidemiology

 Anthropophilic - native to human, the major group of

disease causing in humans
 Zoophilic - native to animals
 Geophilic - native to soil
 Dermatophytes:

Epidemiology

 Anthropophilic (native to human)

Are groups that often cause human diseases.
Example:

M. audonii T. schoenleinii
T. soudanese T. mentagrophytes var
mentagrophytes
T. violaceum T. rubrum
T. tonsurans
 Dermatophytes:

 Zoophilic

M. canis M. gallinae

M. equinum T. verrucosum
M. manum T mentagrophytes
var. interdigitale

 Geophilic

M. gypseum T. ajelloi

M. fulvum T. terretre
 Dermatophytes:

Tinea pedis (athlete’s foot)

 Chronic infection in interdigital webs & soles
 Lesion: vesicular, ulcerative, scaly, fissures,
erythema, hyperkeratosis
 May have an acute onset with inflammation, vesicles,
and pustules
 Secondary infection by bacteria may occur

Tinea manus
 Ringworm in palms
 Dermatophytes:
Dermatophytosis:
Clinical Manifestation

Tinea Pedis: is transmitted

via the feet by
desquamated skin scales in
substrates like carpet and
matting
 Dermatophytes:

Tinea Unguium, Onychomycosis

 Fungal infection in nail and nailbed
 Caused by: T.mentagrophytes, T.rubrum, E.floccosum
 Starts from the distal part of nails
 Lesion: subungual debris, nails become yellow, brittle
and thickened
 In white onychomycosis  white patch on the surface of
the nail
 Dermatophytes:

Tinea unguium:
Clinical
Manifestation
 Dermatophytes:

Tinea Corporis, Tinea Circinata

 Fungal infection in human body

 Caused by: T.mentagrophytes,
T.rubrum, M.canis

 Lesion: annular, sharply

marginated with central
healing, raised border,
single, multiple, or confluent

 Mild type: erythema, scaly

 Severe type: vesicles,
pustules, and erythema
 Dermatophytes:

Tinea Imbricata
 Is an unusual form of tinea corporis
 Caused by T. concentricum
 Characteristic lesion: scaly, concentric rings of scaling
spread out peripherally over many years
 Dermatophytes:

Tinea Capitis
 Ringworm in scalp and hair
 Hyphae  scalp  keratin in hair follicles  alopecia,
scales (+), itch

 Ectothrix infection  hyphae produce chain of

arthrospores surround hair; caused by Microsporum spp.
 Endothrix infection  spores are produced within hair
shaft  black dots; caused by T.tonsurans

 T.schonleini  favus: infection on hair shaft with crust

production surround hair follicles
 Dermatophytes:

A
C

Tinea capitis and tinea favosa.

A.Tinea capitis (gray patch type ) caused by Microsporum sp.
B. Tinea capitis caused by M. canis showing large kerion consisting of crusts,
matted hair, exudate, and scalp debris.
C, Tinea favosa caused by Trichophyton sp. showing crusts and scutula
covering the scalp.
 Dermatophytes:

Tinea Barbae, Barber’s itch

 Ringworm on bearded area

of men
 Caused by:
T.mentagrophytes,
T.rubrum, T.violaceum,
M.canis
 Cause severe inflammation,
resembles pyogenic
infection
 Erythema on face and neck,
scaly, often folliculitis
 Dermatophytes:

Tinea Cruris, Jockey’s itch

 Dermatophytosis in glabrous skin (skin which lack of
hairs)
 Caused by: T.rubrum, T.mentagrophytes, E.floccosum

 Lesion: annular, elevated margin, central healing,

vesicles

 The fungi grow on keratinized, dead tissue, produce

antigen that diffuse to epidermis  erythema, vesicles.
Lesion spreads centrifugally.

 Symptoms: pruritus, pain

 Dermatophytes:

 Geophilic and zoophilic mycoses  severe clinical

infection
 Tinea cruris  usually in men, involving perineum,
scrotum and perianal area; may spread to buttock and
thigh
 The appearance of tinea cruris can be seen in other
intertriginous area, such as under the breasts, axilla, and
around umbilicus in obese patients
Dermatophytes:
 Dermatophytes:

Similarities between clinical manifestation of

superficial and cutaneous fungal infection:

 Lesion spread from center to margin (centrifugal

widening)
 Center part heals first than margin (central healing)
 Margin part is active part where fungus grows, usually
appears as slight elevation
 Lesion is usually donned with scales
 Lesion is itchy maculae with or without inflammation
 Dermatophytes:

 Diagnosis
 Direct examination: specimen (skin & nail scrapings,
hair stumps) + KOH/NaOH 10%
 Culture: Sabouraud Dextrose Agar, incubation at
room temperature for ± 30 days  always examine
macroscopic and microscopic feature to determine
the pathogen
For macroscopic (colony) examination), always
examine both upper side and base side of colony
since traits of both parts are very helpful in diagnosis
 For tinea capitis  Wood’s ultraviolet lamp 
shows fluorescence
 Dermatophytes:

 Therapy
 Local: keratolytic agent (salicylic acid), azoles,
selenium sulfide, tolnaftate
 Oral: griseofulvin, ketoconazole, terbinafine
 Dermatophytes:
What is your working diagnosis?
 SUBCUTANEOUS MYCOSES

• Etiologic fungi are ubiquitous in nature

• Infection by skin or subcutaneous
trauma (iatrogenic inoculation)
• Usually granulomatous lesion
• Spread slowly, via lymphatics drainings
• Usually confined to subcutan tissue,
rarely cause systemic involvement
 Subcutan mycosis:
Sporothrix
Sporotrichosis
 Caused by: Sporothrix schenckii
 Is a thermal dimorphic fungus
 Saprophytic in plants
 Morphology:
 at room temperature  branching septate hyphae
and conidia,
 at 370C  cigar-shaped blastoconidia/spores
 Pigment: white – brown – black

Colony Morphology and
Microscopic Appearance
Colony of Sporothrix schenckii
on SDA . The colonies are moist
and have wrinkled
surface.The color of the
surface is white –tan and turns
dark brown or black with age.
Subcutan mycosis:
Sporothrix
microconidia
Sporothrix schenckii stained with LPCB .
These figure demonstrate the round oval
and pear shaped conidia. The septate
hyphae short
 Subcutan mycosis:
Sporothrix
Clinical manifestations:
 Conidia/hyphal fragments enter
the body via injured skin 
nodules  granulomatous 
ulcerative & necrotic lesion;
multiple subcutaneous nodules
and abscess along lymphatic
drainage

 Usually in extremities
 Rarely becomes systemic
infection
 Subcutan mycosis:
Sporothrix
 Diagnosis:
 Specimens: exudate, tissue biopsy
 Direct examination: KOH, FAT, histologic using HE:
cigar-shaped asteroid bodies
 Culture on Sabouraud Dextrose agar
 Serology: agglutination

 Therapy:
 Itraconazole or other azoles
 Amphotericin B  systemic disease
 Subcutan mycosis:
Chromomycosis
Chromoblastomycosis
(chromomycosis, verrucous
dermatitis)
 Caused by dematiaceous fungi:
 Phialophora verrucosa
 Fonsecaea pedrosoi
 Fonsecaea compacta
 Rhinocladiella aquaspersa
 Cladophialospora carrionii
 Traumatic inoculation, chronic infection, wart-like
lesion, granulomatous, ulceration
 Subcutan mycosis:
Chromomycosis
Colony Morphology and
Microscopic Appearance
phialides

Phialophora
verrucosa stained
with LPCB. These
figure
demonstrate the
flask shaped /
vase shaped or
elliptical
phialides or
conidiophores
with wide flared
pigmented
collaretes.
 Subcutan mycosis:
 Spread: by lymphatics draining, may Chromomycosis
cause elephantiasis
 Colonies: compact, velvet-like, brown to
black in color
 On tissues: spherical brown cells 
muriform / sclerotic bodies
 Subcutan mycosis:
Chromomycosis

 Diagnosis
 Specimens: tissue scraping or biopsy
 Direct examination + KOH: sclerotic bodies
 Culture, DD with saprophytic dematiaceous fungi (no
growth at 370C, no gelatin hydrolysis)

 Therapy
 Drugs: systemic anti-fungal drugs (flucytosine or
itraconazole)
 Surgical excision
 Subcutan mycosis:
Phaeohypomycosis

Phaeohypomycosis
 Caused by dematiaceous fungi:
 Exophiala jeanselmei
 Phialospora richardsiae
 Bipolaris spicifera
 Wangiella dermatitidis
 Alternaria spp. & Curvularia
spp.
 Infect health and
immunocompromised person
 Clinical manifestations:
subcutaneous cysts, brain
abscess
 Subcutan mycosis:
Phaeohypomycosis
Colony Morphology and
Microscopic Appearance

Colony of Exophiala
jeanselmei on SDA. The
colonies are brown or green –
Exophiala jeanselmei stained with LPCB .
These figure demonstrate the tubular
black, moist and
elongated conidiophore
glistening. With age they
become covered with velvety
grayss hyphae.The reverse is
black.
 Subcutan mycosis:
Mycetoma

Mycetoma
 Chronic subcutaneous infection induced by traumatic
inoculation with any of fungi or actinomycetous bacteria
normally found in soil.

 Two types:
 Actinomycetoma  caused by actinomycete i.e.
Nocardia brasiliensis, Streptomyces somaliensis,
Actinomadura madurae
This form is more invasive than the other one.
 Eumycetoma  caused by fungus, i.e.
maduromycosis / Madura foot.
 Subcutan mycosis:
Mycetoma

 Causative fungi:
 Pseudallescheria boydii
 Madurella mycetomatis
 Madurella grisea
 Exophiala jeanselmei
 Acremonium falciforme
 Subcutan mycosis:
Mycetoma
Clinical findings:
 Both types of mycetoma
cause granulomatous
lesions in subcutaneous
tissue, which may lead to
suppuration and abscess

 Usually in extremities

 Sinus formation, contains

granules (microcolonies),
can spread to bone and
tissue
 Subcutan mycosis:
Mycetoma
Diagnosis
 Specimen: tissue biopsy, pus
 Culture
 Based on granules:
 White granules  P.boydii & A.falciforme
 Black granules  M.grisea & E.jeanselmei
 Dark red – black  M.mycetomatis

In maduramycoses / eumycetoma: granules are hard,

consists of intertwined hyphae;

in actinomycetoma: tissue is filled by Gram positive

bacteria
 Subcutan mycosis:
Mycetoma

Therapy
 Surgical excision/debridement and systemic therapy

 Actinomycetoma: streptomycin, cotrimoxazole, dapsone

 Maduromycosis: systemic itraconazole and other azole,

flucytosine, amphotericin B
THANK YOU !!