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MYCV 311
LABORATORY / SECOND SEMESTER
Mr. Joseph Casinsinan

WEEK 4: SUPERFICIAL, CUTANEOUS AND SUBCUTANEOUS MYCOSES

FUNGAL IDENTIFICATION ▪ Solitary, irregular, pigmented (brown to black) macules

SPECIFIC MYCOSES
▪ no scaling or invasion of hair follicles, and the infection is not contagious
SUPERFICIAL MYCOSES
▪ Causative agent:
Confined to the outermost layer of skin and hair (epidermis only) - affect only the
cornified layers (stratum corneum) of the epidermis HORTAEA WERNECKII (Exophialla werneckii)

Elicit little or no host immune response and are nondestructive – asymptomatic - colony: shiny, black, and mucoid

Do not elicit severe infections thus, Usually of cosmetic concern only and are easy to - microscopic: thick, septate, branching hyphae that contain a dark pigment in their
diagnose and treat walls (dematiaceous)

Includes Non-Dermatophytic Tinea: Likely contracted by traumatic inoculation of the fungus into the superficial layers of
- Tinea versicolor (pityriasis versicolor) the epidermis.
- Tinea nigra
- White Piedra
- Black piedra

WHITE PIEDRA

Clinical Manifestations
TINEA VERSICOLOR (Pityriasis Versicolor)

▪ Characterized by the appearance of firm, irregular nodules along the hair shaft (soft,
Clinical Manifestations mycelia mat surrounding the hair shaft)

▪ Characterized by discrete or confluent macular areas of discoloration or depigmentation ▪ axillary, pubic, beard, and scalp hair may be infected
of the skin

▪ Causative agent: TRICHOSPORON SPP.

▪ Most prevalent site of infection: face, chest, trunk and abdomen

TRICHOSPORON BEIGELLI
▪ Causative agent: Malassezia

- colony: straw to cream yeast-like, smooth/wrinkled, dry/moist, velvety

MALASSEZIA FURFUR (Pityrosporum orbiculare)

- microscopic: produce arthroconidia, hyphae, and blastoconidia

- colony: cream to yellowish, typically smooth to slightly wrinkled

Other species: Trichosporon ovoides, T. asteroides, T. cutaneum, T. inkin, and T. asahaii

- Microscopic: budding yeasts (approx. 4-8 μm) along with septate, sometimes branched,
hyphal elements (“spaghetti with meatballs appearance”)
PIEDRA - Occurs in tropical and subtropical regions - related to poor hygiene
- Wood’s lamp: yellow fluorescence of infected body sites
Affects the hairs of the groin and axillae (mustaches and beards, on eyelashes and
eyebrows, and in armpit and pubic hair)
Pale patches – darkly-pigmented skin

Surrounds the hair shaft and forms a white to brown swelling along the hair strand.
Fawn-colored liver spots – individuals with fair complexion

Infection does not damage the hair shaft

MALASSEZIA – lypophilic yeast-like fungus

T. Beigelli - grow rapidly on primary fungal media

- considered part of the cutaneous mycobiota

T. asahaii, - which is implicated in severe and frequently fatal disease in

Growth in media overlayed with olive oil - reveal yeast cells with daughter cells being immunocompromised hosts
produced from collarettes also known as bud scars

Require FATTY ACIDS to grow

BLACK PIEDRA

Clinical Manifestations

▪ Presence of hard, dark brown to black gritty nodules that are firmly attached to the
TINEA NIGRA
hair shaft

Clinical Manifestations ▪ endemic in tropical areas of Africa, Asia, and Latin America

▪ A localized infection of the stratum corneum, particularly of the palms (tinea nigra ▪ Causative agent:
palmaris)

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PIEDRAIA HORTAE Microscopic: MICROCONIDIA - hyaline, smooth-walled, spherical to subspherical

- colony: black-greenish, limited and pointed colonies MACROCONIDIA – smooth, thin-walled, clavate-shaped, multi-celled

- microscopic: Thick-walled rhomboid cells containing ascospores are seen *occasional – spiral hyphae

Nodules consist of ASCI (sack-like structures) containing eight (8) Ascospores Trichophyton SHOENLEINII

Colony: ss: waxy or suede-like with a deeply folded honey-comb-like thallus

CUTANEOUS MYCOSES rs: no coloration

RINGWORM (Tinea) Microscopic: MICROCONIDIA – non-existent routinely

Clinical Manifestations MACROCONIDIA – non-existent routinely

▪ Various forms of dermatophytosis are referred to as “tineas” or ringworm * ”favic chandeliers” hyphae

▪ Group of closely related filamentous fungi that infect ONLY SUPERFICIAL MICROSPORUM AUDOUINII
KERATINIZED TISSUES—the skin, hair and nails
Colony: ss: flat, greyish to light tan-white, dense suede-like to downy surface (mouse
▪ classified according to the anatomic site or structure affected fur in texture)

▪ Causative agents: Trichophyton, Epidermophyton, Microsporum rs: yellow-brown to reddish-brown

Trichopyton rubrum, Trichophyton mentagrophytes, Trichophyton shoenleinii, Microscopic: MICROCONIDIA – non-existent routinely
Microsporum audouinii, Microsporum canis, Microsporum gypseum, Epidermophyton
floccosum
MACROCONIDIA – non-existent routinely

*PECTINATE HYPHAE - look like little combs

*sterile hyphae with terminal chlamydospores

Wood lamp – hair fluoresce

MICROSPORUM CANIS

Colony: ss: flat, greyish to light tan-white, dense suede-like to downy surface (mouse
fur in texture)

rs: yellow-brown to reddish-brown

Microscopic: microconidia – A few pyriform to clavate

macroconidia – spindle-shaped with 5-15 cells, VERRUCOSE, thick-

walled and often have a terminal knob

*pectinate hyphae - look like little combs

*sterile hyphae with terminal chlamydospores

MICROSPORUM GYPSEUM

Trichophyton RUBRUM
Colony: ss: flat, cinnamon-colored and powdery

Colony: ss: downy to suede-like, white to cream to deep red

rs: light tan to lavender color

rs: yellow-brown to wine red

Microscopic: MICROCONIDIA – rare

Microscopic: MICROCONIDIA - slender clavate to pyriform, sessile

MACROCONIDIA – thick-walled, rough, elliptical, and multiseptate
(walls are a little thinner and the tips are more rounded compared to canis)
MACROCONIDIA - smooth, thin-walled multi-septate
*pectinate hyphae - look like little combs
slender and cylindrical to cigar-shaped
*sterile hyphae with terminal chlamydospores
Trichophyton MENTAGROPHYTES
Wood’s lamp – hair do not fluoresce
Colony: ss: flat, white to cream, powdery to granular surface
Causes Ectothrix Tinea capitis
rs: yellow-brown to reddish-brown
EPIDERMOPHYTON FLOCCOSUM

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Colony: ss: greenish-brown or khaki-coloured with a suede-like surface, raised and CHROMOBLASTOMYCOSIS (Verrucous dermatitidis)
folded in the centre, submerged fringe of growth
Clinical Manifestations
rs: light tan to lavender color
▪ Characterized by the slow development of progressive granulomatous lesions that in
Microscopic: microconidia – NO microconidia! time induce HYPERPLASIA of the epidermal tissue

macroconidia – smooth, large, thin-walled multi septate; borne singly or in ▪ Infections appear as multiple, large, warty, “cauliflowerlike” growths that are usually
clusters of two or three directly from the hyphae clustered within the same region

* occasional raquet hyphae ▪ SCLEROTIC BODIES – seen in lesions as Copper-Colored Cells

▪ Causative agents:

DERMATOPHYTES – Classification depending on habitat: Fonsecaea pedrosoi, Cladosporium carionii, Phialophora verrucosa

1. Anthropophilic species: Human beings are the main or only hosts, may be Also known as CHROMOMYCOSIS - chronic, localized disease of the skin and
transmitted directly or indirectly from person to person subcutaneous tissues

 T. rubrum, M. audouinii and E. floccosum All are dematiaceous fungi which are darkly pigmented fungi

2. Zoophilic species: parasitize the hair and skin of animals but can be transmitted to Fonsecaea pedrosoi
humans
colony: flat to heaped and folded, suede-like to downy, olivaceous to black
• T. verrucosum in cattle and M. canis in dogs and cats
rs: olivaceous to black
3. Geophilic species: live in the soil and are occasional pathogens of both animals and
humans, less pathogenic for human beings
- microscopic: dark brown hyphae and suberect conidiophores loosely branched

• M. gypseum and T. ajelloi

Cladosporium carionii

HAIR PERFORATION TEST colony: ss: mostly olivaceous-brown to blackish-brown but also sometimes grey, buff or
brown, suede-like to floccose, often becoming powdery due to the production
of abundant conidia
T. rubrum - surface erosion of hair shafts
rs: olivaceous-black
T. mentagrophytes – perpendicular penetration pegs in the hair shafts
- microscopic: Dematiaceous hyphomycete forming branched acropetal chains of
 Positive penetration: M. canis conidia, each with a distinct hilum
 Negative penetration: M. equinum
Phialophora verrucosa

colony: initially dome-shaped, later becoming flat, suede-like and olivaceous to black in
colour

TYPES OF HAIR INFECTION Microscopic: Phialides are flask-shaped or elliptical with distinctive funnel-shaped,
darkly pigmented collarettes
1. ECTOTHRIX: arthroconidia are formed on the outside of the hair - M. audouinii, M.
canis and T. mentagrophytes Colarettes - a frill or collar (often cup-shaped) of outer wall material remaining at the
apex of a phialidic conidiogenous cell
2. ENDOTHRIX: arthroconidia are formed inside the hair - T. tonsurans and T.
violaceum

3. FAVUS (Favic): hyphae, arthroconidia, and empty spaces resembling air bubbles MYCETOMA
(“honeycomb” pattern) are formed inside the hair - T. schoenleinii

Clinical Manifestations

▪ a chronic granulomatous infection that usually involves the lower extremities but may
SUBCUTANEOUS MYCOSES occur in any part of the body

Introduced traumatically through the skin - deeper layers of the dermis, subcutaneous ▪ gradually progresses to involve the bone, muscle, or other contiguous tissue and
tissue, and bone - rarely spread to distant organs ultimately requires amputation in most progressive cases

Chronic and insidious; once established, the infections are stubborn to most antifungal ▪ seen among people living in tropical and subtropical regions
therapy

▪ Causative agents:
Exposure is largely occupational or related to hobbies (e.g., Gardening, wood gathering)

Scedosporium apiospermum, Pseudallescheria boydii, Actinomyces spp., Nocardia spp.

sporotrichosis, chromoblastomycosis, eumycotic mycetoma, phaeohyphomycosis,
subcutaneous zygomycosis
The infection is characterized by swelling, purplish discoloration, tumorlike deformities
of the subcutaneous tissue, and multiple sinus tracts that drain purulent material
containing yellow, white, red, or black granules

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seen among people living in tropical and subtropical regions of the world whose outdoor ▪ a general term used to describe any infection caused by a dematiaceous organism
occupations and failure to wear protective clothing predispose them to trauma
▪ clinical forms vary from solitary encapsulated cysts in the subcutaneous tissue to
2 TYPES OF MYCETOMA sinusitis to brain abscesses
- Eumycotic
- Actinomycotic
▪ Causative agents:

Exophiala jeanselmei
EUMYCOTIC MYCETOMA

Colony: ss: initially smooth, greenish-grey to black, mucoid and yeast-like; with age,
Clinical Manifestations
often becoming dome-shaped and suede-like in texture

▪ Also called MADUROMYCOSIS, Madura foot

rs: olivaceous-black

▪ caused by a heterogeneous group of fungi that have septate hyphae

Microscopic: Conidiogenous cells are predominantly annellidic and erect

▪ Subcategorized as white grain mycetoma or black grain mycetoma, a distinction

Wangiella dermatitidis (Exophiala dermatitidis)
determined by the pigmentation of the infecting agent’s hyphae

▪ Causative agents: Colony: ss: initially yeast-like and black, becoming suede-like, olivaceous-grey with the
development of aerial mycelium with age

Scedosporium apiospermum
brown pigment is often produced in the agar

Colony: ss: greyish-white, suede-like to downy

Microscopic: The yeast-like cells are hyaline and thin-walled when young becoming
darkly pigmented (thick walled when matured)
rs: greyish-black reverse
With the development of mycelium, flask-shaped to cylindrical annellides
Microscopic: single-celled, pale-brown, broadly clavate to ovoid conidia, 4-9 x 6-10 µm, are produced
rounded above with truncate bases are observed
Cladosporium trichoides (Caldophialophora bantiana)
- larger cleistothecia (compared to boydii)
Colony: ss: moderately fast growing, olivaceous-grey, suede-like; grow at temperatures
Cleistothecia - a globose, completely closed fruit body with no special opening to the up to 42-40C
outside (released only by decay or disintegration)
Microscopic: Conidia are formed in long, sparsely branched, flexuose, acropetal chains
*heterothallic - require two compatible partners for mating to occur from undifferentiated conidiophores

Scedosporium boydii (Pseudallescheria boydii)

Colony: ss: greyish-white, suede-like to downy SPOROTRICHOSIS

rs: greyish-black reverse Clinical Manifestations

Microscopic: Conidia are borne singly or in small groups on elongate, simple or branched ▪ primarily a chronic mycotic infection of the cutaneous or subcutaneous tissues and
conidiophores or laterally on hyphae adjacent lymphatics

- smaller cleistothecia ▪ Also known as “Rose Gardener’s disease”

*homothallic - self fertile with a single individual capable of sexual reproduction even in ▪ Acquired through trauma (thorns, splinters, bites, scratches) usually to the hand, arm or
solo culture feet

▪ Considered an occupational hazard for farmers, gardeners, florists, nursery workers, and
miners
ACTINOMYCOTIC MYCETOMA
▪ Causative agent:
Clinical Manifestations
Sporothrix schenckii complex
▪ caused by a fungus-like bacteria
s. Shenckii complex = set of five dimorphic fungi
▪ Same clinical manifestation as eumycotic mycetomas
S. Shenckii complex includes:
▪ Causative agents:
 S. schenckii sensu strictu
 S. brasiliensis
Actinomyces spp.
 S. globose
 S. Mexicana
Nocardia spp.  S. luriei 

PHAEOHYPHOMYCOSIS SUBCUTANEOUS ENTOMOPHTHOROMYCOSIS

Clinical Manifestations Clinical Manifestations

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▪ Also known as subcutaneous mucormycoses and zygomycoses

▪ Chronic subcutaneous form of mucormycosis that occurs sporadically as a result of

traumatic implantation

▪ Causative agents: Entomophthorales

 Basidiobolus ranarum – infects children; proximal limbs

 Conidiobolus coronatus – infects adults; facial area

Basidiobolus ranarum

Colony: ss: moderately fast growing at 30C, flat, yellowish-grey to creamy-grey,

glabrous, becoming radially folded and covered by a fine, powdery, white surface
mycelium

Microscopic: production of “beaked” zygospores is characteristic of the genus (smooth,

thick-walled zygospores that have two closely appressed beak-like appendages)

Conidiobolus coronatus

Colony: ss: flat, cream-coloured, glabrous becoming radially folded and covered by a
fine, powdery, white surface mycelium and conidiophores

Microscopic: Conidiophores forms terminal conidia which are spherical, single-celled

and have a prominent papilla

- conidia may also produce hair-like appendages, called villae

RHINOSPORIDIOSIS

Clinical Manifestations

▪ Chronic granulomatous disease characterized by the development of large polyps or

wart-like lesions (nose, conjunctiva)

▪ >90% of the cases have been reported from India, Sri lanka and South America.

▪ Causative agent:

RHINOSPORIDIUM SEEBERI

Colony: (culture of fungus unsuccessful)

Microscopic: in tissues - presence of mucosal and submucosal cysts (sporangia) that

contains numerous endospores

Rhinosporidium seeberi✓ If autoclaving is unavailable, then fugal culture can be

decontaminated by soaking the plates and tubes overnight in a fresh 1:10 bleach solution

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