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MUST To KNOW in Mycology and Virology
MUST To KNOW in Mycology and Virology
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Molds Filamentous
Room temperature
Yeasts Round/tissue form
37’C
Chitin Fungal cell wall
Ergosterol Fungal cell membrane
Hyphae Aerial/vegetative/substrate
Spores Reproductive
Sporangiophore Aseptate hyphae
Conidiophore Septate hyphae
Sexual spores Ascospores
Basidiospores
Oospores
Asexual spores Chlamydospores: rounding terminal hyphae or intercalary (within)
Blastospores: budding off (yeasts)
Conidia (macro/micro): dermatophytes
Arthrospores: barrel-shaped, produced from fragmentation of mycelium
10% KOH preparation Skin and hair
Hyphae, yeasts, spores
Dissolves tissue
Calcofluor white Fluorescent dye
Stains the chitin (cell wall)
Green
Giemsa/Wright’s stain H. capsulatum: the only fungi seen in blood smear
Periodic Acid Schiff Glycogen: purple
No glycogen: pink
Gomori silver Black
Dermatophyte test medium SDA + phenol red
Inhibitory mold agar SDA + cycloheximide + gentamicin
Potato dextrose agar Pigment
Cornmeal Tween 80 agar Chlamydospore of C. albicans
Rice medium (+) M. canis
(-) M. audouinii
Staib’s nigerseed agar C. neoformans (black colonies)
Urea agar (+) T. mentagrophytes
(+) C. neoformans
Cottonseed agar B. dermatitidis
Biochemical tests API20C, ID32C
CHROMagar Candida
Germ tube test For unknown yeast
Cornmeal agar Dalmau-Plate
Mannan antigen Candida
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
Genus-spp ID (molds) Microscopic exam
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Superficial Mycoses Non invasive
No immune response from the host
MOT: person-to-person (garment)
-M. furfur
-P. hortai
-T. beigelii
-P. werneckii
M. furfur Ptyriasis versicolor (An-An)
Hypo/Hyperpigmentation of skin
KOH: budding yeast cells and hyphae
PAS: “Spaghetti and meatballs”
Lipophilic: require lipids for growth (SDA + olive oil)
Piedra agents Hair
P. hortai Black piedra
Dematiaceous
Ascospores (sexual)
T. beigelii White piedra
Cream colored
Arthrospores (asexual)
Phaeoannelomyces Tinea nigra
(Exophiala) werneckii Brownish spot (dark pigmentation)
Dematiaeous: moist, shiny-black and yeast-like colonies
Cutaneous mycoses Trichophyton = skin, hair, nails
Microsporum = skin, hair
Epidermophyton = skin, nails
Dermatophytes Keratinophilic
Tinea or ringworm
Endothrix:
-T. tonsurans: dandruff (Tinea capitis)
-T. violaceum
Ectothrix:
-M. gypseum
-M. canis
-T. verrucosum
Tinea barbae Beard
Tinea capitis Scalp
Tinea corporis Body
Tinea cruris Groin
Tinea pedis Foot (Athlete’s foot)
Tinea unguium Nails
T. rubrum Red pigment
Microconidia: tear-drop shaped (side)
Macroconidia: pencil-shaped
Colony: Fluffy white w/ red color reverse
T. mentagrophytes Microconidia: grape-like (cluster)
(+) Hair penetration test: V-shaped
(+) Urease
T. tonsurans Thiamine-requiring
Black dot tinea capitis
Microconidia: Balloon-shaped
Colony: Creamy white w/ crater-like depression
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T. schoenleinii Require thiamine and inositol
Cause Favus: chronic hair infection
Favic chandelier hyphae
T. verrucosum Microconidia: clavate/pyriform
Macroconidia: rat-tail/string-bean-shaped
M. canis Macroconidia: spindle-shaped, echinulate
(+) Wood’s lamp
M. gypseum Geophilic
Macroconidia: oblong (ellipsoidal) w/ blunt/rounded ends
(-) Wood’s lamp
M. audouinii Anthropophilic
Tinea capitis (old)
(NG) Rice medium
E. floccosum Macroconidia: club shaped in pairs
Dutch pants fuseaux
Tx: Dermatophytes Local antifungal creams:
-Miconazole
-Tolnaltate
Oral
-Griseofulvin
-Ketoconazole
Subcutaneous Mycoses MOT: skin trauma/prick
Habitat: soil
Biopsy, granules (PAS, H&E)
-Sporothrix schenckii
-Madura foot agents
-Chromoblastomycosis agents
-Rhinosporidium seeberi
-Loboa loboi
S. schenckii Mold form: flowerette/daisy-like conidia
Yeast form: Cigar-shaped (asteroid) bodies
Rose Gardener’s disease
-Cord-like multiple subcutaneous nodules
Colony: White to black
Madura foot agents Maduromycosis: mycetoma
P. boydii = most common cause
Madurella
Leptosphaeria
Actinomycetes
Tissue form: Granules
Lesion: granulomatous lesions on foot w/ multiple draining sinus tracts
P. boydii Cleistothecia
Teleomorph/sexual form
Scedosporium apiospermum: anamorph/asexual form
Chromoblastomycosis Dematiaceous fungi
agents Type of sporulation: ID of genus and species
1. Phialophora verrucosa
-Vase-like
-TOS: phialophore
2. Fonsecaea pedrosoi
-Short chain
-w/ 1’, 2’, 3’ conidia
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-TOS: Acrotheca
3. Cladosporium carrionii
-Long chain (branching)
-TOS: Cladosporium
Infected tissue: Brown sclerotic bodies (Medlar bodies)
Lesion: Cauliflower-like
Colonies: Dark w/ jet black reverse
R. seeberi Size: 300 μm
Rhinosporidiosis
MOT: swimming
Polypoid masses in nose and pharynx
Tissue form: Sporangium (sac-like structures filled w/ endospores)
L. loboi Lobomycosis
Lesion: Keloid-like subcutaneous nodule involving the extremities
Tissue form: Multiple budding cells in chain (Sampaloc-like/Longganisa-like)
Systemic Mycoses Dimorphic
MOT: inhalation of spores (mold) – infectious
Specimen: Sputum
BSL III = require BSC class II
No person-to-person contact
Colonies: cottony white to brown
Exoantigen test
-B. dermatitidis
-P. brasiliensis
-H. capsulatum
-C. immitis
B. dermatitidis Agent of:
-North American blastomycosis
-Chicago disease
-Gilchrist disease
Pneumonia and skin infections
Mold form: Lollipop in appearance
Yeast form: Single-budding yeast w/ broad based (double centered) –
diagnostic
Mickey mouse cap
Culture: Medium w/ cycloheximide
Tissue: KOH
Tx: Amphotericin B
P. brasiliensis Agent of:
-South American Blastomycosis
-Lutz Splendore-Almeida disease
Infects spleen, liver, lymph node, skin, lungs
Mold form: Lollipop in appearance
Yeast form: Multiple budding yeast resembling Mariner’s wheel
H. capsulatum RES parasite
Agent of Darling’s disease
“Arise of Tutankhamen’s tomb”
USA = endemic dimorphic ffungus
MOT: inhalation of spores from guanos (droppings of birds, starlings/chickens,
and bats)
Mold form: Tuberculate macroconidia
Yeast form: Yeast cells intracellular in macrophages
Culture: Medium w/ cycloheximide (BM, sputum, tissue)
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H. duboisii Double cell
Figure of 8
Causes African histoplasmosis
Grows in N2
Sepedonium Tuberculate macroconidia (resembles H. capsulatum)
To differentiate: Sepedonium is monomorphic
C. immitis Agent of:
-Desert fever
-San Joaquin Valley fever
Alkaline soil
High humidity
Lab. acquired infection
Mold form: Barrel-shaped arthroconidia (200μm)
Yeast form: spherule w/ endospores
Culture: medium w/ cycloheximide (30’C)
Not processed ---(send)---> Reference lab
Lab. Diagnosis 1. Direct examination
(Systemic mycoses) a. Histoplasma: Wright’s/Giemsa
b. Blastomyces, Paracoccidioides, Coccidioides
- KOH: tissue form
- PAS
- H&E
2. Cultures
a. SDA = RT’ (mold)
b. BHIA + blood = 37’C (yeast)
3. Immunological tests
a. Coccidioidin
b. Histoplasmin
Exoantigen Test Principle: Double Diffusion
A = B. dermatitidis
HS, HL, F = C. immitis
H & M = H. capsulatum
1, 2, 3 = P. brasiliensis
Opportunistic Mycoses Normal flora (Ex. Candida)
At risk: immunocompromised person
-Candida
-Cryptococcus
-Aspergillus
C. albicans Normal flora: skin, mucous membrane, GIT
Produce yeast and hyphae in vivo
-Germ tube
-Chlamydospore
-Blastoconidia
-Pseudohyphae
-True hyphae
(+) Sucrose
EMB: feathering colonies
Diseases:
-Thrush
-Diaper rash
-Candidiasis (Old: Moniliasis): cutaneous, oropharyngeal (white patches),
esophagitis, cheilitis, onchomycosis, vulvovaginitis, invasive infection (CNS,
blood)
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Predisposing factors:
-Prolong antibiotic use especially broad spectrum = disturb pH
-Pregnancy
-DM
-Malnutrition
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
Other Candida spp C. tropicalis = infection on immunocompromised host
C. glabrata = 2nd common spp
C. krusei = hematological disorders
C. parapsilosis = endocarditis
Lab. Diagnosis (Candida) Vaginal pH: 4.5
Rule out vaginosis/Trichomoniasis = alkaline pH
Vaginal discharge = 10% KOH (Whiff’s test)
Fungal culture
(+) Germ tube test C. albicans = (+) Chlamydospores, (+) Sucrose, (G) 42’C
C. stellatoidea = (-) Chlamydospores, (-) Sucrose, (NG) 42’C
G. candidum = (+) Arthrospores
C. dubliniensis = (NG) 42’C, double conidia
Cryptococcus Found in pigeon droppings and soil
(Filobasidiella) neoformans MOT: inhalation of airborne organism
Encapsulated yeast (India ink)
Diseases:
-Meningitis
-Torulosis (Old: Cryptococcosis)
-Pneumonia
Lab. Diagnosis:
-Old: India ink preparation = CSF, pulmonary tissue
-New: Latex agglutination (Ag): capsule
-Biochemical tests:
= (+) Inositol
= (+) Urease
= (-) NO3 assimilation = yellow
= (+) Phenoloxidase = makes the colonies black
= (+) Phospholipase
-(G) Birdseed agar
= Assimilate creatinine
= Colonies: Yeastlike, mucoid, cream to brown
-Culture: SDA w/o cycloheximide
Tx: Amphotericin B, Fluconazole
Rhodotorula Resemble C. neoformans
Encapsulated
To differentiate: Rhodotorula = pink colonies, (+) Urease, (+) NO3 assimilation
Aspergillus Bread mold
Septate hyphae
Found in nature: soil, plant, air
Vesicles: swollen conidiophores
A. flavus Aflatoxin (toxicoses) = HCC