Mycology, M(ASCP)

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No cross walls

Aseptate hyphae

Cell cross walls very evident

Septate hyphae

Looks like antlers of a buck deer

Favic chandeliers

Trichophyton schoenleinii

Favic chandeliers are diagnotic for which fungus

Knots of twisted hyphae

Nodular organs

Resemble tennis racquets

Racquet hyphae

Molds at room temp (25-30C) and yeast at body temp (35-37C)

Dimorphism

Asexual reproductive structure, only formed in septate fungi

Conidia

Conidium produced by budding (i.e. Candida, Cladosporium)

Blastoconidia

Formed by daughter cell pushing through a minute pore in the parent cell (i.e. Bipolaris)

Poroconidia

Conidia formed on a tube or vase-shaped conidiogenous structure, a phialide (i.e. Penicillum, Aspergillus)

Phialoconidia

Conidia that are grown from inside a vase-shaped conidiogenous structure, an annellide (i.e. Scopulariopsis) BOWLING PIN

Annelloconidia

Multicelled conidia
Only used when Microconidia appear also

Macroconidia

Conidium that arises from a hyphal element, is aseptate
Only used when with Macroconidia

Microconidia

Thick walled survival conidia, formed when unpleasant environment conditions exist

Chlamydoconidia

Conidium produced by fragmentation of hyphal strands through septation points. May be adjacent or have disjunter cells in between (i.e. Coccidioides immitis - disjuncter cells)

Arthroconidia

Only formed in ASEPTATE fungi, formed by internal cleavage of the contents of a sac called a sporangium. Supported on a base, COLUMELLA, on a stalk, SPORANGIOPHORE.

Sporangiospores

Sexual spore, contained in ASCI cells in a protective sac, ASCOCARP

Ascospore

Sexual spore, where protrusions at the tip of the BASIDIUM form (Cryptococcus neoformans sexual stage)

Basidiospore

Sexual spore formed by fusion of 2 hyphal arms (zygophores)

Zygospore

Powdery colonies, flat and crumbly because of dense production of conidia

Granular colonial texture on agar plate

Waxy colonies, smooth surface because they produce no aerial mycelium... ussually yeasts

Glabrous colonial texture on agar plate

Deep furrows irregularly radiating from center of culture (like rays of the sun)

Rugose topography on agar plate

Bottonlike central elevation

Umbonate topography on agar plate

Wrinkled, convoluted surface (like mountains)

Verrucose topography on agar plate

Phenol kills organisms, while lactic acid preserves fungal structures. Cotton blue stains chitin in fungal cell walls - permanent stain.

Lactophenol Cotton Blue (LPCB) wet mount

Potassium hydroxide will dissolve keratin in skin/hair/nail specimens, making fungal elements more visible

Potassium hydroxide (KOH) preparation

Used to stain for Nocardia spp.

Modified acid-fast stain

Used to observe capsules around yeast, esp. Cryptococcus neoformans

India Ink preparation

Calcofuor white is taken up into the chitin of fungal cell walls and produces an apple green fluorescence - can be mixed with KOH prep too

Calcofluor White stain

Choice medium for general initial isolation - supports bacteria, fungi, dermatophytes, and yeasts

SABHI: Sabouraud Brain Heart Infusion Agar

Nutritionally richer than SABHI, should be reserved for normally sterile sites and anaerobic actinomycetes

BHIB: Brain Heart Infusion with Blood Agar

An acid pH 5.6 and nutritionally poor, inhibits growth of bacteria, but still basic media

SDA: Sabouraud Dextrose Agar

Contains gentamicin, which inhibits bacteria

IMA: Inhibitory Mold Agar

Blood is nutritious, and gentamicin and chlorampheicol inhibit bacteria. Nocardia and aerobic actinomycetes will not grow.

Brain Heart Infusion Agar with Blood, Gentamicin, and Chloramphenicol

Best used for fastidious fungal pathogens, Bacteria, aerobic actinomycetes and opportunists will not grow.

Brain Heart Infusion Agar with Blood, Gentamicin, Chloramphenicol, and Cycloheximide

Contains antibiotics plus phenol red indicator. When dermatophytes grow, they change the color from yellow to RED.

DTM: Dermatophyte Test Medium

Media used to subculture yeasts because it has less glucose and a neutral pH compared to regular SDA

Neutral Sabouraud dextrose agar

Media used to subculture molds, it promotes sporulation and pigmentation of colonies

Potato dextrose agar

Forms mature colonies in 5 days or less

Rapid growers

Forms mature colonies in 6 to 10 days

Intermediate growers

Forms mature colonies in 11 to 21 days

Slow growers

Most are rapid growers, saprobic (living on decaying matter in soil), and normally inhaled

3 Common properties of fungal opportunists

SABHI room temp - Woolly gray colony, reverse colorless

Absidia sp. culture

ASEPTATE, with BRANCHING SPORANGIOPHORES BETWEEN RHIZOIDS on the stolons. Sporangia are PEAR-SHAPED

Absidia sp. microscopic

SABHI room temp - Cottony white colonies, becoming off-white to yellow with age. Grows at up to temps of 42C

Apophysomyces sp. culture

ASEPTATE? FUNNEL SHAPED COLUMELLA, FOOT CELLS at base of sporangiophores.

Apophysomyces sp. microscopic

SABHI room temp - White, fluffy mycelium, rapid growth, becoming gray with age

Mucor sp. culture

ASEPTATE, single or BRANCING SPORANGIOPHORES with round sporangia. NO RHIZOIDS OR STOLONS

Mucor sp. microscopic

SABHI room temp - white, dense, cottony aerial hyphae rapid growth, with ag become dotted with brown/black sporangia

Rhizopus sp. culture

ASEPTATE, UNBRANCHED SPORANGIOPHORES arise OPPOSITE RHIZOIDS at the nodes. STOLONS connect the groups of rhizoids

Rhizopus sp. microscopic

SABHI room temp - woolly white, rapid growth

Saksenaea sp. culture

ASEPTATE, FLASK-SHAPED SPORANGIA, RHIZOIDS are produced opposite the sporangiophores

Saksenaea sp. microscopic

SABHI room temp - White cottony rapid growth, turning gray with age. May grow at 42C also.

Cunninghamella sp. culture

ASEPTATE, sporangiophores terminate in VESICLES(swollen cells) on which one celled sporangia form at the tips of DENTICLES (toothpick-like projections)

Cunninghamella sp. microscopic

SABHI room temp - woolly dark rapid growth

Syncephalastrum sp. culture

ASEPTATE, sporangiophores terminate in VESICLES(swollen cells) surrounded by sporangia with rows of sporangiospores, looks like a SUNFLOWER

Syncephalastrum sp. microscopic

SABHI room temp - light gray, woolly rapidly matures to dark green/black or brown with black reverse

Alternaria sp. culture

DARK, CHAINED POROCONIDIA which have HORIZONTAL & VERTICAL SEPTA. (#alternate)

Alternaria sp. microscopic

SABHI room temp - initially shiny white yeastlike -> with age, become shiny black and leathery with white fringe BLACK YEAST

Aureobasidium sp. culture

Light to DARK brown conidiophores are NOT differentiated from hyphae. Short denticles support hyaline conidia. (Looks like Nostoc)

Aureobasidium sp. microscopic

SABHI room temp - at first is velvety grayish-brown and later the center becomes matted black, reverse is light or dark

Bipolaris sp. culture

Alternaria sp.

DARK hyphae, CYLINDRICAL four or five celled POROCONIDIA along a BENT KNEE CONIDIOPHORE (#power rangers outfits)

Bipolaris sp. microscopic

SABHI room temp - 7 days, powdery heaped and folded dark gray-green with black reverse.

Cladosporium sp. culture

DARK septate hyphae, short chains of BLASTOCONIDIA with distinct SCAR at point of attachment. Bourne from SHIELD CELLS

Cladosporium sp. microscopic

SABHI room temp - cottony, white, light pink, orange, or green, with brown reverse

Curvularia sp. culture

Dark septate, BENT KNEE CONIDIOPHORE with POROCONIDIA that have an OVER ENLARGED CENTRAL CELL (#orange slices)

Curvularia sp. microscopic

SABHI room temp - light gray woolly, with age becomes dark gray/black with a black reverse

Exserohilum sp. culture

DARK septate, long CYLINDRICAL POROCONIDIA with 6 to 14 cells and distinct, protruding TRUNCATE HILA(points of attachment)

Exserohilum sp. microscopic

SABHI room temp - colonial rings of yellow, orange, and brown- pigments of the same color may diffuse in agar

Epicoccum sp. culture

Thick clusters of SPORODCHIA(short conidiophores) support DARK round conidia with unconstricted HORIZONTAL and VERTICAL SEPTA.

Epicoccum sp. microscopic

SABHI room temp - White woolly colony with black reverse, fills up plate

Nigrospora sp. culture

Dark hyphae. SHORT,FAT CONIDIOPHORES support SINGLE oval BLACK CONIDIA at tips

Nigrospora sp. microscopic

SABHI room temp - wrinkled, white/gray/rose, reverse is colorless/pale yellow/pinkish

Acremonium sp. culture

LIGHT septate hyphae, unbranched TAPERING CONIDIOPHORES support closely packed BALLS of sickle shaped CONIDIA

Acremonium sp. microscopic

Colony is rugose ad velvety, many different colors depending on dense conidia production: blue, green, yellow, black, and white

Aspergillus sp. culture

Light septate, unbranched conidiophores with FOOT CELL support a large VESICLE, which supports flask shaped PHIALIDES in a SINGLE or DOUBLE ROW, which produce CHAINS of PHIALOCONIDIA

Aspergillus sp. microscopic

UNISERIATE, phialides usually only on upper 2/3 of vesicle, parallel to axis of conidiophore

Aspergillus fumigatus microscopic

BISERATE, phialides cover entire vesicle, form "radiate" head

Aspergillus niger microscopic

Conidiophore is ROUGH, PITTED, SPINY
Uniserate and Biserate,phialides cover entire vesicle, point out in all directions

Aspergillus flavus microscopic

SABHI room temp - heaped, velvety, buff colored, with white/yellow/reddish brown reverse (looks like a brain)

Chrysosporium sp. culture

Light septate, SINGLE round to club shaped CONIDIA perch on top of SHORT CONIDIOPHORES, poorly differentiated from vegetative mycelium

Chrysosporium sp. microscopic

SABHI room temp - white at first, woolly/cottony, later becomes LAVENDER, or yellow/orange with a light reverse

Fusarium sp. culture

Light septate, MICROPHIALOCONIDIA are one celled and occur in BALLS on conidiophores. MACROPHIALOCONIDIA are two to five celled BANANA or CYLINDRICAL SHAPED "fusiform"

Fusarium sp. microscopic

SABHI room temp - fills the plate with a GREEN LAWN, reverse is white

Gliocladium sp. culture

Brushlike conidiophores with flask shaped PHIALIDES which produce masses of PHIALOCONIDIA are held together in a LARGE BALL by a matrix

Gliocladium sp. microscopic

SABHI room temp - powdery,velvety, or cottony colony, OLIVE TAN color, may be shades of violet or brown

Paecilomyces sp. culture

Light septate, penicillus type ELONGATED PHIALIDES bear CHAINS of hyaline or pigmented OVAL CONIDIA

Paecilomyces sp. microscopic

SABHI room temp - Powdery BLUE GREEN with a WHITE PERIPHERY and colorless reverse

Penicillium sp. culture

Light septate, PENICILLI bear flask shaped PHIALIDES which support CHAINS of ROUND PHIALOCONIDIA

Penicillium sp. microscopic

SABHI room tem - finely wrinkled surface and DARK RED pigment that diffuses in agar.
Blood agar 37C - Yeastlike colonies, grayish white, waxy and attached to surface.

Penicillium marneffei culture

Conidiophores bear up to 5 METULAE (branched conidophores) with WIDE PHIALIDES that taper to narrow apices with CHAINS of smooth, lemon shaped PHIALOCONIDIA

Penicillium marneffei microscopic

SABHI room temp - velvety, rugose and white, later becoming light tan or brown with a tan reverse

Scopulariopsis sp. culture

Light septate, ANNELLOPHORES bear bowling pin shaped ANNELLIDES with CHAINS of large LEMON SHAPED ANNELLOCONIDIA, with age, become spiny

Scopulariopsis sp. microscopic

SABHI room temp - waxy/white becomes velvety and lemon colored with a peripheral fringe, white reverse

Sepedonium sp. culture

SINGLE or clustered, THICK WALLED, SMOOTH TO ROUGH MACROCONIDIA form at ends of conidiophores, not well differentiated from vegetative mycelium.
May be confused with H. capsulatum

Sepedonium sp. microscopic

SABHI room temp - BLACK, YEASTY colony with short OLIVE-GRAY mycelium

Exophiala werneckii culture

Caused by Exophiala werneckii
Brown to black nonscaly patches form mainly on palms

Tinea nigra

Yeast portion: DARK one to two celled blastoconidia
Mold portion: blastoconidia form along hyphae in large clusters, or in small clusters, or from ANNELLIDES with CLUSTERS or chains of ANNELLOCONIDIA

Exophiala werneckii microscopic

Caused by Malassezia furfur
Asymtomatic skin infection, scaly patches of different colors: reddish brown, brown, white. Fluoresce under wood's lamp

Pityriasis versicolor

Will not grow on routine media. Needs lipid (olive oil) and 37C.

Malassezia furfur culture

In KOH stains, thick round to oval cells in clusters with short, angular hyphae SPAGHETTI AND MEATBALLS

Malassezia furfur microscopic

SABHI room temp - slowly - compact, greenish black, heaped, glabrous colony

Piedraia hortai culture

DARK, THICK WALLED HYPHAE with SWELLINGS, asci with ascospores may be present

Piedraia hortai microscopic

Caused by Piedraia hortai.
Firmly attached, black nodules around the outside of scalp hairs.

Black piedra

SABHI room temp - cream colored, wrinkled, glabrous colony

Trichosporon beigelii culture

Caused by Trichosporon beigelii.
Light brown, soft nodules around beard and mustache hairs.

White piedra

Hyaline hyphae with BLASTOCONIDIA nd ARTHROCONIDIA. Additional biochemical tests needed.

Trichosporon beigelii microscopic

Microsporum
Epidermophyton
Trichophyton

3 genus of dermatophytes

Microsporum
Epidermophyton
Trichophyton

Dermatophytes that infect skin

Epidermophyton
Trichophyton

Dermatophytes that infect nails

Microsporon
T. mentagrophytes
T. rubrum
T. verrucosum

Dermatophytes that infect Ectothrix hair

T. tonsurans
T. schoenleinii
T. violaceum

Dermatophytes that infect Endothrix hair

Microsporon

Dermatophytes that fluoresce under a Wood's lamp

M. canis
M. gypseum
E. floccosum
T. mentagrophytes

Dermatophyte intermediate growers

M. audouinii
T. rubrum
T. schoenleinii
T. tonsurans
T. verrucosum
T. violaceum

Dermatophyte slow growers

Penicillium sp.

Scopulariopsis sp.

Microsporum canis

Microsporum gypseum

Epidermophyton floccosum

Trichophyton mentagrophytes

Trichophyton rubrum

SABHI room temp - matted to velvety light tan, with a REVERSE of SALMON
Poor growth on RICE GRAINS, other Microsporum grow well

Microsporun audouinii culture

Raquet hyphae, nodular bodies, and TERMINAL, POINTED VESCILES are most commonly found

Microsporum audouinii microscopic

SABHI room temp - white, woolly with a buff to brown center and bright yellow periphery, REVERSE is BRIGHT YELLOW to YELLOW ORANGE
Contracted from cats/dogs commonly

Microsporum canis culture

Numerous ROUGH, THICK WALLED, SPINDLE SHAPED MACROCONIDIA with 6 to 15 cells

Microsporum canis microscopic

SABHI room temp - powdery BUFF to CINNAMON colony with tan reverse
Contracted through soil

Microsporum gypseum culture

Numerous ROUGH, THIN WALLED, ELLIPTICAL MACROCONIDIA with 4 to 6 cells

Microsporum gypseum microscopic

SABHI room temp - velvety, KHAKI YELLOW colony with tan reverse
Causes athlete's foot.

Epidermophyton floccosum culture

CLUB SHAPED, smooth, thin walled, MACROCONIDIA with 2 to 4 cells. NO MICROCONIDIA!

Epidermophyton floccosum microscopic

SABHI room temp - fluffy white with colorless/tan reverse OR granular buff/rose-tan with brown/red/yellow reverse
Most common cause of athlete's foot

Trichophyton mentagrophytes culture

ROUND MICROCONIDIA in GRAPE LIKE CLUSTERS

Trichophyton mentagrophytes microscopic

SABHI room temp - granular/fluffy white with pink periphery and DEEP RED REVERSE
Produces red pigment in agar

Trichophyton rubrum culture

Numerous smooth walled, pencil shaped macroconidia with 3 to 8 cells. Numerous CLUB SHAPED MICROCONIDIA borne singly along hyphae.

Trichophyton rubrum microscopic

SABHI w/yeast extract 37C - heaped, waxy, white/bright yellow with yellow reverse
Usually acquired from contact with cattle.

Trichophyton verrucosum culture

On SABHI, only chlamydoconidia are produced.
On thiamine enriched, RAT TAIL MACROCONIDIA with 3 to 5 cells

Trichophyton verrucosum microscopic

SABHI room temp - waxy, heaped, light yellow/buff with colorless/yellow orange reverse
Causes severe scalp disease called FAVUS

Trichophyton schoenleinii culture

FAVIC CHANDELIERS are most prevalent feature, No macroconidia

Trichophyton schoenleinii microscopic

Numerous MICROCONIDIA with a GREAT SIZE and SHAPE VARIATION

Trichophyton tonsurans microscopic

SABHI room temp - waxy/suede VIOLET,heaped colony with violet reverse

Trichophyton violaceum culture

CHLAMYDOCONIDIA IN CHAINS and hyphal swellings observed. No macroconidia.

Trichophyton violaceum microscopic

Clamydospores: terminal, THICK WALLED
Blastoconidia: CLUSTERS AT SEPTA of Pseudohyphae

Candida albicans microscopic morphology on Corn-meal Tween 80 agar

Clamydospores: terminal THIN WALLED
Blastoconidia: SINGLE or short chained anywhere along Pseudohyphae

Candida tropicalis microscopic morphology on Corn-meal Tween 80 agar

Blastconidia: Oval in chains from the septa of thin Pseudohyphae OR clusters of numerous blastos at septa of short pseudohyphae

Candida guilliermondi microscopic morphology on Corn-meal Tween 80 agar

Blastoconidia: Tree like branching from the septa of elongated Pseudohyphae
"CROSS MATCHSTICKS"

Candida krusei microscopic morphology on Corn-meal Tween 80 agar

Blastoconidia: Few single or small clustered blastos at or between THIN CURVED Pseudohyphae

Candidia parapsilosis microscopic morphology on Corn-meal Tween 80 agar

Blastoconidia: Chains of elongated blastos at septa of BRANCHED Pseudohyphae
"LOGS IN A STREAM"

Candida pseudotropicalis microscopic morphology on Corn-meal Tween 80 agar

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