MUST TO KNOW IN MYCOLOGY Flashcards by Arriane Cyrel (MTI)

Fluoresces on Wood’s lamp

M. canis

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Agent of dandruff

M. canis

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Medium for dermatophytes

Mycosel

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: inhibit bacteria
: inhibit fungal contaminants

Chloramphenicol

Cycloheximide

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Czapek’s medium

Aspergillus

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Forms at 35’C (yeasts)

Germ tube

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Gram (+), oval cells w/ or w/o buds and pseudomycelia

Candida

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General culture media

SDA

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Basic structure of fungi

Hyphae
Spore

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Chlamydospore

Cornmeal agar

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Media w/ olive oil

M. furfur

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Hair baiting test

T. mentagrophytes

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Infects hair, skin, and nails

Trichophyton

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Red color

T. rubrum

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Teardrop conidia

T. rubrum

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Favic chandelier (antler)

T. schoenleinii

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Phycomycetes/Zygomycetes

Coenocytic (aseptate)

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Ascospores

S. cerevisiae

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(+) Nigerseed/Birdseed
(+) Urease
(+) India ink
(+) Phenoloxidase

C. neoformans

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Arthrospores “TACGeo”

Trichosporon beigelii
Aureobasidium
Coccidioides
Geotrichum candidum

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Blastospores

Yeast

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Direct microscopic
Clearing

10% KOH

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Commonly used stain (fungal culture)

LPCB

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: preservative
: killing agent
: stains the chitin (cell wall)

Lactic acid

Phenol

Cotton blue

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Unsuitable for fungal culture Suitable for bacteria and viruses only

Swab

Wright’s/Giemsa stain

H. capsulatum

Dichotomous

Aspergillus

Thrush

C. albicans

Diaper rash

C. albicans

Moniliasis

C. albicans

In tissue: mold/yeast form

C. albicans

Macroconidia and microconidia

Dermatophytes

Specimen for systemic mycoses

Sputum

ID of yeast Free of CHO

CHO assimilation

Dimorphic fungi

BHIA

High risk to lab personnel

C. immitis

Filamentous Room temperature

Molds

Round/tissue form 37’C

Yeasts

Fungal cell wall

Chitin

Fungal cell membrane

Ergosterol

Aerial/vegetative/substrate Reproductive

Hyphae Spores

Aseptate hyphae Septate hyphae

Sporangiophore Conidiophore

Sexual spores

Ascospores Basidiospores Oospores

Asexual spores : rounding terminal hyphae or intercalary (within) : budding off (yeasts) : dermatophytes : barrel-shaped, produced from fragmentation of mycelium

Chlamydospores Blastospores Conidia (macro/micro) Arthrospores

Skin and hair

10% KOH preparation

Hyphae, yeasts, spores

10% KOH preparation

Dissolves tissue

10% KOH preparation

Fluorescent dye

Calcofluor white

Stains the chitin (cell wall)

Calcofluor white

Green

Calcofluor white

H. capsulatum: the only fungi seen in blood smear

Giemsa/Wright’s stain

Periodic Acid Schiff Glycogen: No glycogen:

purple pink

Black

Gomori silver

SDA + phenol red SDA + cycloheximide + gentamicin

Dermatophyte test medium Inhibitory mold agar

Pigment

Potato dextrose agar

Chlamydospore of C. albicans

Cornmeal Tween 80 agar

(+) M. canis (-) M. audouinii

Rice medium

C. neoformans (black colonies)

Staib’s nigerseed agar

(+) T. mentagrophytes (+) C. neoformans

Urea agar

Cottonseed agar

B. dermatitidis

Biochemical tests

API20C, ID32C

CHROMagar

Candida

For unknown yeast

Germ tube test

Dalmau-Plate

Cornmeal agar

Candida

Mannan antigen

Galactomannan

Aspergillus

Fungicidal agents Target:

Ergosterol -Amphotericin B: systemic fungi (most common) -Nystatin -Azole (Fluconazole): fungistatic -Griseofulvin (IV): dermatophytes

AST methods

Broth microdilution method E-test method

Microscopic exam

Genus-spp ID (molds)

Non invasive

Superficial Mycoses

No immune response from the host

Superficial Mycoses

MOT: person-to-person (garment)

Superficial Mycoses

Superficial Mycoses:

-M. furfur -P. hortai -T. beigelii -P. werneckii

Ptyriasis versicolor (An-An)

M. furfur

Hypo/Hyperpigmentation of skin

M. furfur

KOH: budding yeast cells and hyphae

M. furfur

PAS: “Spaghetti and meatballs”

M. furfur

Lipophilic: require lipids for growth (SDA + olive oil)

M. furfur

Hair

Piedra agents

Black piedra

P. hortai

Dematiaceous

P. hortai

Ascospores (sexual)

P. hortai

White piedra

T. beigelii

Cream colored

T. beigelii

Arthrospores (asexual)

T. beigelii

Tinea nigra

Phaeoannelomyces (Exophiala) werneckii

Brownish spot (dark pigmentation)

Phaeoannelomyces (Exophiala) werneckii

Dematiaeous: moist, shiny-black and yeast-like colonies

Phaeoannelomyces (Exophiala) werneckii

Cutaneous mycoses = skin, hair, nails = skin, hair = skin, nails

Trichophyton Microsporum Epidermophyton

Keratinophilic

Dermatophytes

Tinea or ringworm

Dermatophytes

Dermatophytes Endothrix: Ectothrix:

-T. tonsurans: dandruff (Tinea capitis) -T. violaceum -M. gypseum -M. canis -T. verrucosum

Beard Scalp Body Groin Foot (Athlete’s foot) Nails

Tinea barbae Tinea capitis Tinea corporis Tinea cruris Tinea pedis Tinea unguium

Red pigment

T. rubrum

Microconidia: tear-drop shaped (side)

Macroconidia: pencil-shaped

T. rubrum

Colony: Fluffy white w/ red color reverse

T. rubrum

Microconidia: grape-like (cluster)

T. mentagrophytes

(+) Hair penetration test: V-shaped (+) Urease

T. mentagrophytes

Thiamine-requiring

T. tonsurans

Black dot tinea capitis

T. tonsurans

Microconidia: Balloon-shaped

T. tonsurans

Colony: Creamy white w/ crater-like depression

T. tonsurans

Require thiamine and inositol

T. schoenleinii

Cause Favus: chronic hair infection

T. schoenleinii

Favic chandelier hyphae

T. schoenleinii

Microconidia: clavate/pyriform

T. verrucosum

Macroconidia: rat-tail/string-bean-shaped

T. verrucosum

Macroconidia: spindle-shaped, echinulate

M. canis

(+) Wood’s lamp

M. canis

Geophilic

M. gypseum

Macroconidia: oblong (ellipsoidal) w/ blunt/rounded ends

M. gypseum

(-) Wood’s lamp

M. gypseum

Anthropophilic

M. audouinii

Tinea capitis (old)

M. audouinii

(NG) Rice medium

M. audouinii

Macroconidia: club shaped in pairs

E. floccosum

Dutch pants fuseaux

E. floccosum

Tx: Dermatophytes Local antifungal creams: Oral

-Miconazole -Tolnaltate -Griseofulvin -Ketoconazole

MOT: skin trauma/prick Habitat: soil

Subcutaneous Mycoses

Biopsy, granules (PAS, H&E)

Subcutaneous Mycoses

Subcutaneous Mycoses:

-Sporothrix schenckii -Madura foot agents -Chromoblastomycosis agents -Rhinosporidium seeberi -Loboa loboi

Mold form: flowerette/daisy-like conidia

S. schenckii

Yeast form: Cigar-shaped (asteroid) bodies

S. schenckii

Rose Gardener’s disease

S. schenckii

Cord-like multiple subcutaneous nodules

S. schenckii

Colony: White to black

S. schenckii

Maduromycosis:

mycetoma

Madura foot agents:

P. boydii = most common cause Madurella Leptosphaeria Actinomycetes

Tissue form: Granules

Madura foot agents

Lesion: granulomatous lesions on foot w/ multiple draining sinus tracts

Madura foot agents

Cleistothecia

P. boydii

:Teleomorph/sexual form : anamorph/asexual form

P. boydii Scedosporium apiospermum

Dematiaceous fungi

Chromoblastomycosis agents

Type of sporulation: ID of genus and species

Chromoblastomycosis agents

Chromoblastomycosis agents:

1. Phialophora verrucosa 2. Fonsecaea pedrosoi 3. Cladosporium carrionii

-Vase-like -TOS: phialophore

1. Phialophora verrucosa

-Short chain -w/ 1’, 2’, 3’ conidia -TOS: Acrotheca

2. Fonsecaea pedrosoi

-Long chain (branching) -TOS: Cladosporium

3. Cladosporium carrionii

Infected tissue: Brown sclerotic bodies (Medlar bodies)

Chromoblastomycosis agents

Lesion: Cauliflower-like

Chromoblastomycosis agents

Colonies: Dark w/ jet black reverse

Chromoblastomycosis agents

Size: 300 μm

R. seeberi

Rhinosporidiosis

R. seeberi

MOT: swimming

R. seeberi

Polypoid masses in nose and pharynx

R. seeberi

Tissue form: Sporangium (sac-like structures filled w/ endospores)

R. seeberi

Lobomycosis

L. loboi

Lesion: Keloid-like subcutaneous nodule involving the extremities

L. loboi

Tissue form: Multiple budding cells in chain (Sampaloc-like/Longganisa-like)

L. loboi

Dimorphic

Systemic Mycoses

MOT: inhalation of spores (mold) – infectious

Systemic Mycoses

Specimen: Sputum

Systemic Mycoses

BSL III = require BSC class II

Systemic Mycoses

No person-to-person contact

Systemic Mycoses

Colonies: cottony white to brown

Systemic Mycoses

Systemic Mycoses Exoantigen test:

-B. dermatitidis -P. brasiliensis -H. capsulatum -C. immitis

Agent of: -North American blastomycosis -Chicago disease -Gilchrist disease

B. dermatitidis

Pneumonia and skin infections

B. dermatitidis

Mold form: Lollipop in appearance

B. dermatitidis P. brasiliensis

Yeast form: Single-budding yeast w/ broad based (double centered) – diagnostic

B. dermatitidis

Mickey mouse cap

B. dermatitidis

Culture: Medium w/ cycloheximide Tissue: KOH

B. dermatitidis

Tx: Amphotericin B

B. dermatitidis

Agent of: -South American Blastomycosis -Lutz Splendore-Almeida disease

P. brasiliensis

Infects spleen, liver, lymph node, skin, lungs

P. brasiliensis

Yeast form: Multiple budding yeast resembling Mariner’s wheel

P. brasiliensis

RES parasite

H. capsulatum

Agent of Darling’s disease

H. capsulatum

“Arise of Tutankhamen’s tomb”

H. capsulatum

USA = endemic dimorphic ffungus

H. capsulatum

MOT: inhalation of spores from guanos (droppings of birds, starlings/chickens, and bats)

H. capsulatum

Mold form: Tuberculate macroconidia

H. capsulatum

Yeast form: Yeast cells intracellular in macrophages

H. capsulatum

Culture: Medium w/ cycloheximide (BM, sputum, tissue)

H. capsulatum

Double cell

H. duboisii

Figure of 8

H. duboisii

Causes African histoplasmosis

H. duboisii

Grows in ↑ N2

H. duboisii

Tuberculate macroconidia (resembles H. capsulatum)

Sepedonium

To differentiate: monomorphic

Sepedonium

Agent of: -Desert fever -San Joaquin Valley fever

C. immitis

Alkaline soil

C. immitis

High humidity

C. immitis

Lab. acquired infection

C. immitis

Mold form: Barrel-shaped arthroconidia (200μm)

C. immitis

Yeast form: spherule w/ endospores

C. immitis

Culture: medium w/ cycloheximide (30’C)

C. immitis

Not processed ---(send)---> Reference lab

C. immitis

Lab. Diagnosis (Systemic mycoses) 1. Direct examination a. Histoplasma: b. Blastomyces, Paracoccidioides, Coccidioides: 2. Cultures = RT’ (mold) = 37’C (yeast) 3. Immunological tests a. Coccidioidin b. Histoplasmin

Wright’s/Giemsa - KOH: tissue form - PAS - H&E a. SDA b. BHIA + blood

Exoantigen Test Principle: = B. dermatitidis = C. immitis = H. capsulatum = P. brasiliensis

Double Diffusion A HS, HL, F H & M 1, 2, 3

Normal flora (Ex. Candida) At risk: immunocompromised person

Opportunistic Mycoses

Opportunistic Mycoses:

-Candida -Cryptococcus -Aspergillus

Normal flora: skin, mucous membrane, GIT

C. albicans

Produce yeast and hyphae in vivo

C. albicans

-Germ tube -Chlamydospore -Blastoconidia -Pseudohyphae -True hyphae

C. albicans

(+) Sucrose

C. albicans

EMB: feathering colonies

C. albicans

Diseases: -Thrush -Diaper rash -Candidiasis (Old: Moniliasis): cutaneous, oropharyngeal (white patches), esophagitis, cheilitis, onchomycosis, vulvovaginitis, invasive infection (CNS, blood)

C. albicans

Predisposing factors: -Prolong antibiotic use especially broad spectrum = disturb pH -Pregnancy -DM -Malnutrition

C. albicans

Lab. Diagnosis: 1. (G) SDA 2. Screening test: Germ tube test = Organisms + Serum ---(35’C for 2-3 hours)---> (+) Germ tube 3. Confirmatory test: Chlamydospore Cornmeal agar = C. albicans → Inoculate: CMA ---(RT’ for 48-72 hours)---> (+) Chlamydospore

C. albicans

Other Candida spp = infection on immunocompromised host = 2nd common spp = hematological disorders = endocarditis

C. tropicalis C. glabrata C. krusei C. parapsilosis

Lab. Diagnosis (Candida) Vaginal pH: Rule out vaginosis/Trichomoniasis = Vaginal discharge = Fungal culture

4.5 alkaline pH 10% KOH (Whiff’s test)

(+) Germ tube test = (+) Chlamydospores, (+) Sucrose, (G) 42’C = (-) Chlamydospores, (-) Sucrose, (NG) 42’C = (+) Arthrospores = (NG) 42’C, double conidia

C. albicans C. stellatoidea G. candidum C. dubliniensis

Found in pigeon droppings and soil