AIRBORNE CONTAMINANTS ALTERNARIA ASPERGILLUS BOTYRIS CHAETOMIUM CEPHALOSPORIUM CURVULARIA FUSARIUM

CULTURE

MORPHOLOGYRapid-growing colonies, grayish to black to brown; underside jet black

Flat, compact colonies, white at first then becoming black, green, bluish or yellow

Soft looking tan to gray colony Slow growing, flat, white, yellow, tan or brown colony

White to tan to rose-colroed colony, eventually developing White aerial hyphae.

Dark brown to black colony on both sides ragged in appearance.

Fast-growing colony. At first, white and cottony but develping rose to red color on both sides

MICROSCOPIC VIEW

DESCRIPTIONLarge, hand grenade-shaped spores with both longitudinal and transverse cross walls. Borne singly or in chains. Septate, dematiaceous fungi.

Small one-celled spores irradiating out from swollen base (see arrows)

Colorless, one celled spores borne in clumps

Large, dark, central structure is perithecium that contains ascospores (sexually produced)

Single celled, clear, elliptical spores held together in a ball unless broken loose

Large, bent spores with 3 to 5 cells. Similar to Helminthosporium spp. Brown, septate hyphae.

Largest spores are sickle-shaped and may contain several cells. Small spores with one to two cells have more rounded ends.

AIRBORNE CONTAMINANTS GELASINOSPORA GEOTRICHUM GLIOCLADIUM HORMODENDRUM NEUROSPORA (MONILIA) NIGROSPORA PAECILOMYCES

CULTURE

MORPHOLOGYDark colony with numerous small black dots which are perithecia (sexual organs for Ascomycetes)

White to tan, flat or fluffy, rapid-growing fungus

Flat, rapid-growing colony. White at first, then developing dark green central portion

Green to gray to black colony on both sides. Often wrinkled and grows flat

White at first but grows rapidly filling the entire Petri dish in a few days and becoming a salmon to brown color. Mycelium may hang from the lid of the Petri dish.

Rapid growing, producing abundant fluffly, aerial hyphae, gray to black on both sides. Resembles Mucor or Rhizopus

Flat, rapid-growing, tan-colored colony resembling Aspergillus spp.

MICROSCOPIC VIEW

DESCRIPTIONSquashed perithecium showing many asci, each containing 8 sexually produced ascospores

Note hyphae breaking into arthrospores. May be confused with Coccidioides immitis.

Numerous small spores held together in a clump. Similar to Penicillium spp. Except for the clumping of spores.

Dark brown septate hyphae bear branching chains of elongate to ovate spores that often contain a small black dot at the end. Spore bearing structures look tree-like

Clear septate hyphae with large masses of ovate spores which are air-dispersed

Large, clearly visible jet black spores.

Similar to Penicillium spp. except small spores are produced on very long, slender structures

AIRBORNE CONTAMINANTS PENICILLIUM PULLULARIA (Aureobasidium) RHODOTORULA RHIZOPUS SACCHAROMYCES SCOPULARIOPSIS SEPEDONIUM

CULTURE

MORPHOLOGYWhite colony at first but developing blue to green color

Brown to black, flat, greasy-looking colonies

A pale yellow to vivid orange yeast

Gray to brown to black colony filling a Petri dish in 2 to 3 days. Similar to Mucor spp.

White to tan yeast colonies Powdery, light brown, wrinkled colony resembling Penicillium spp. Except for color.

Fluffy, white colony resembles Histoplasma capsulatum

MICROSCOPIC VIEW

DESCRIPTIONSmall, round spores borne in “brush-like” formations

Mixture of dark, thick-walled, large hyphal cells and clear spores which seem to be budding

Long, slender yeast cells, usually budding, but may appear to form pseudohyphae.

Similar to Mucor spp. except foot-like structures (rhizoids) at base of spore bearing hyphae (see arrows). Spores in sporangium clear, coenocytic hyphae

Typical yeast cells that reproduce by budding.

Resembles Penicillium spp. except spores are larger and form unbranched-chains

Large, rough-walled spores that resembles Histoplasma capsulatum

SUPERFICIAL

Dermatophytoses – dermatophytes•Genus Microsporum•Genus Epidermophyton•Genus Trichophyton

Pityriasis Versicolor – Malazzezia furfur

Tinia nigra – Cladosporium werneckii

Piedra•white – Trichosporum(beigelli) cutaneum•black – Piedraia hortai

Otomycosis•Aspergillus•Mucor•Penicillium•Rhizopus

Keratitis•Fusarium•Aspergillus•Curvularia•Penicillium•Cephalosporium•Candida species

AIRBORNE CONTAMINANTS SPOROBOLOMYCES STEMPHYLUM STREPTOMYCES SYNCEPHALASTRUM VERTICILLIUM

CULTURE

MORPHOLOGYA light tan yeast colony Brown to black, wrinkled fuzzy

colonyDry, flat, leathery colony grows close to the medium

Gray to brown to black fluffy colony that may fill a Petri dish in a few days. Similar to Mucor and Rhizopus spp

Powdery to pinkish brown colony that looks similar to Penicillium spp.

MICROSCOPIC VIEW

DESCRIPTIONYeast cells that may form long slender tubes. Reproduces by budding

Hyphae are brown and strongly septate: huge multicelled terminal spores that may either be smooth or rough

Very fine (1um) branching hyphae that readily break apart

Broad, clear, nonseptate hyphae, spores in many slender sac-like structures (sporangia) adhere to a swelling on the terminal end of hypha.

Single-celled elliptical spores borne in clusters that are arranged in whorls

DERMATOPHYTES MICROSPORUM AUDOUINII MICROSPORUM CANIS MICROSPORUM DISTORTUM MICROSPORUM FERRUGINEUMMICROSPORUM GYPSEUM MICROSPORUM NANUM

MICROSPORUM VANBREUSEGHEMII

CULTURE

MORPHOLOGYCultured on Sabouraud plus 2 antibiotics at room temp. for 2-3 weeks. Fluffy white colony with slight yellow underside

Cultured on Sabouraud medium plus 2 antibiotics at room temp. for 1-2 weeks. White on top with bright yellow underside.

Culture on Sabouraud medium plus 2 antibiotcs at room temperature for 3 weeks. Similar to Microsporum canis but with less pigmentation.

Cultured on Sabouraud medium plus 2 antibiotics for 3 weeks at room temp. White to intense orange yellow strains; often sectors

Cultured on Sabouraud medium plus 2 antibiotics for 5-10 days at room temp. Grows rapidly, producing a cinnamon to brown colored flat colony.

Cultured on Sabouraud medium plus 2 antibiotics at room temp. for 1-3 weeks. White to buff surface: bottom often yellow red brown.

Cultured on Sabouraud medium plus 2 antibiotics for 1-2 weeks at room temp. Cottony white surface, may develop pink to tan coloration; bottom often colorless to yellow.

MICROSCOPIC VIEW

DESCRIPTIONNo distinguishing spores will not grow on rice

Highly diagnostic large, thick-walled, rough spores containing more than 6 septa.

Large spores similar to Microsporum canis but distorted and bent in shape.

No distinguishing spores. Prominent septa, giving term “bamboo hyphae”.

Numerous, characteristic, Large spore; thin-walled, pointed ends with 2-5 septa.

Egg-shaped, thin- walled; large spores with 1-3 septa

Huge, long, thick rough- walled spores with more than 8 septa.

DERMATOPHYTES TRICHOPHYTON VIOLACEUM TRICHOPHYTON CONCENTRICUM TRICHOPHYTON EQUINUMTRICHOPHYTON MENTAGRPPHYTES

TRICHOPHYTON RUBRUMTRICHOPHYTON SCHOENLEINII TRICHOPHYTON TONSURANS

CULTURE

SUBCUTANEOUS

Sporotrichosis – Sporothrix schenckiiChromomycosis or Chromoblastomycosis

Phialophora verrucosaFonsecae pedrosoiCladosporium carrioni

Mycetoma or MaduromycosisActinomycotic

ActinomaduraeNocardiaStreptomyces

Eumycotic(true fungi)Allescheria(or Petriellidium)MadurellaPhialophora

EntomopthoromycosisBasidiobolus ranarumConidiobolus coronatus

Lobomycosis – Lacazia loboi(Loboa loboi)Rhinosporidiosis – Rhinosporidium seeberiPhaeohypomycosis

Wangiella sppExophialia sppCladosporium spp.Hormodendrum spp.Dreschlera spp.

SUBCUTANEOUS SPOROTHRIX SCHENCKII(Room Temp)

SPOROTHRIX SCHENCKII (37 C cultures) FONSECAEA PEDROSOI FONSECAE COMPACTA PHIALOPHORA VERRUCOSA CLADOSPORIM CARRIONI BASIDIOBOLUS RANARUM

CULTURE

MORPHOLOGYDark, greasy-looking culture after 1 to 2 weeks incubation on Sabouraud medium

White to tan yeast colonies after 1 to 3 weeks incubation on brain heart infusion agar.

Surface is dark green, gray or black, covered by silvery, velvet-like mycelium. It is usually flat, then develops a cone shaped protrusion in the center. Reverse is black.

Surface is dark green to black. The colonies are heaped, brittle with irregular indented borders. There are brownish hyphae on the surface

Surface is dark greenish, brown to black with a close matlike, olive to gray mycelium. Some strains are heaped, granular or flat with a matted appearance. Reverse is black.

The colony has a dark surface, flat with slightly raised center. It is covered with velvety dull gray, gray green or purplish brown, short napped mycelium. Reverse is black.

Colonies are flat, yellowish gray to creamy gray, glabrous becoming radially folded and covered by a fine powder, white surface mycelium

MICROSCOPIC VIEW

DESCRIPTIONClear, septate hyphae with spores (3 to 6 um) in “daisy-like” clusters

Round, oval, or “cigar-shaped” yeast cells, 1 to 3 um x 4to 10 um

The hyphae is septate, branched and brown. The conidia is dark measuring 1.5-3.0 x 2.5-6.0 um.

Hyphae is septate, brown, branching and bear Cladosporium of conidiophores. Outstanding features are flasklike shape conidia and compact arrangement of conidial chain.

The hyphae is brown, branched and septate with vase-shaped Phialophora type conidiophores. The conidiophores are single or multiple, lateral or terminal and bear easily disrupted masses of oval conidia

The hyphae is septate, dark with lateral and terminal conidiophores of varying size. conidiophores produce long branching chains of brown, smooth walled, oval, pointed conidia which have dark scars of attachment

There is the presence of large vegetative hyphae forming numerous round, smooth, thick walled zygospores with two closely appressed beaklike appendages

SUBCUTANEOUS N BRASILIENSIS (TOP)/N MADUARAE(BOTTOM)-RT

EXOPHIALA (WANGIELLA) DERMATITIDIS CONIDIOBLOUS CORONATUS (25-30 C) EXOPHIALA JEANSELMEI PSEUDOALLESCHERICHIA BOYDII (SEXUAL)

CULTURE

MORPHOLOGYSlow growing, dry-appearing colonies on Sabouraud medium. Morphology varies between etiologic agents. Use biochemical tests.

Colonies are slow growing, initially black and yeast-like, becoming suede-like, ovilaceous grey and mould like age.

Colonies are flat, cream-colored, glabrous becoming radially folded and covered by fine powdery white surface mycelium and conidiophores.

The colonies are brown or green-black, moist and glistening. With age they become covered with velvety grayish hyphae. The reverse is black

MICROSCOPIC VIEW

The colonies have a cottony surface that is white to gray-brown in color and gets darker with age of the culture. The reverse is also white turning brown with age.

GRAPHIUM, asexual state of P. BOYDII (sexual state). The coremia or Synnemata(conidial structures) of the Graphium state of P.boydii have terminal hyaline conidia, club-shaped or cylindrical, approximately 6 x 3 um. In the sexual state(P.boydii), large, 50-200 um in diameter, round, brown cleistothecia are found containing ascospores

SCEDOSPORIUM APIOSPERMUM (ASEXUAL STATE OF BOYDII) The Scedosporium type of conidia of P. boydii may rise directly from the septate hyphae or from the tip of conidiophores, appear truncated at the base, and sometimes resemble the conidia of Blastomyces dermatitidis. The hyphae are long and slender, branch at acute angles and thus may resemble aspergilli.

Variations in acid fastness In new culture, oval and The hyphae have few septa. The The conidiophores are

DERMATOPHYTESTRICHOSPORON BEIGELII(room temperature cultures)

EPIDERMOPHYTON FLOCCOSUM

CULTURE

MORPHOLOGYThe colonies are smooth to slightly wrinkled (not fuzzy) and white to tan in color

Cultured on Sabouraud medium on 2 antibiotics for 1-3 weeks at room temp. Yellow to green colored surface; green to brown underside.

MICROSCOPIC VIEW

DESCRIPTIONOne observes clear septate hyphae which may break up into individual cells.

Large, club-shaped spores with 2 to 5 septa, often form in pairs.

MALASSEZIA FURFUR

Colonies in Dixon’s agar are cream to yellowish, smooth or lightly wrinkled, glistening or dull, with the margin being either entire or lobate

10% KOH with Parker ink mount showing characteristic spherical yeast cells and short pseudohyphal elements typical of the lungs

PIEDRAIA HORTAI(room temp cultures)

The colonies are dark brown and may have a metallic green tinge. They are very compact, have a raised center and are slightly fuzzy.

One observes dematiaceous hyphae containing numerous septa

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DESCRIPTION help to determine between some etiologic agents. Delicate hyphae, 1 um. All are members of the Actinomycetes.Recommended room temperature incubation, although morphology is similar at both temp

round budding yeast-like cells are formed. Subsequently these cells produce septate hyphae with flask-shaped to cylindrical phialides found at the tip of the phialide and also along d hyphae.

conidiophores are unbranched forming solitary terminal conidia. The conidia are spherical, single-celled and have a prominent papilla. It may also produce hair-like appendages called villae.

elongated, tubular and with a tapered, narrow end. Conidia are smooth, thin-walled, and ellipsoid and can gather in clusters around the conidiophores and at points along the septate hyphae

RHINOSPOR This organism has never been cultured; its existence as a water saprophyte or fish pathogen is suspectedIDIUM SEEBERI (RT AND 37 C)

DEEP SEATED

Histoplasmosis – Histoplasma capsulatum

Coccidioidomycosis – Coccidioides immitis

South American Blastomycosis – Paracoccidioides brasiliensis

North American Blastomycosis – Blastomycosis dermatitidis

DEEP SEATEDHISTOPLASMA CAPSULATUMROOM TEMP 37 C COCCIDIODES IMMITIS

PARACOCCIDIOIDES BRAZILIENSISRT 37 C

CULTURE

MORPHOLOGYWhtie, innocent looking (but dangerous) colony after 1-3 weeks incubation on Saboraud medium. Note color of underside

White to tan yeast colony after 1 to 3 weeks incubation on brain heart infusion agar.

Innocent looking, but dangerous white fluffy colony on Sabouraud medium after 1 to 3 weeks incubation. Underside may develop darker color

White, slow growing, nondescript colony on Sabouraud medium after 2 to 4 weeks incubation

Rough, dry yeast colonies after 2 to 4 weeks incubation on brain heart infusion agar

MICROSCOPIC VIEW

DESCRIPTIONCharacteristic large tuberculated macroconidia (10-20 um) and occasional smaller, infectious microconidia

Numerous, small (2 to 4 um), oval, or round, budding yeast cells

Hyphae forming characteristic arthrospores 4 x 8 umGrows sparsely at37, same morphology at 37 and RT

Fine, septae, clear hyphae lacking spores

Large yeast cells (10 to 60 um) with numerous, small buds 2 to 5 um

DEEP SEATED BLASTOMYCES DERMATITIDISRT 37 C

CULTURE

MORPHOLOGYColony after 1 to 3 weeks incubation on Sabouraud medium. White colony, brown to tan underside

Rough, dry, heaped-up yeast colony after 1 to 3 weeks incubation on brain heart infusion agar

MICROSCOPIC VIEW

DESCRIPTIONDelicate, septate hyphae bearing oval to pyriform spores, 4 to 8 um

Yeast cells, 10 to 15 um. Note broad-based budding

OPPORTUNISTIC

Candidiasis – Candida albicans

Cryptococcosis – Cryptococcus neoformans Aspergillosis – Aspergillus fumigatus

Zygomycosis(Mucormycosis) Phycomycosis Rhizopus Mucor Absidia Basidiobolus Conidiobolus

Geotrichosis – Geotrichum candidum

Penicillosis – Penicilium spp.

CANDIDA ALBICANSRT AND 37 C CRYPTOCOCCUS NEOFORMANS

ASPERGILLUS SPRT / 37 C RHIZOPUS

White colonies incubated on Sabouraud medium for 5 to 7 days

Off white, mucous colonies on Sabouraud medium after 5 to 10 days

Upper left: green colony after 5 to 10 days on Sabouraud medium. Colorless underside

All etiologic agents rapidly fill Petri dish after 2 to 4 days incubation. Becomes tan or gray

Chlamydospore agar, 1 to 2 days incubationl Note yeast cells, pseudohyphae and chlamydospores

India ink preparation showing numerous heavily encapsulated yeast cells

Usually cultured at RT. Some species grow at 37 C and have morphology similar to that of organisms grown at RT

Usually cultured at 37 C to inhibit contaminants

Typical

Usually cultured at room temperature. Some species grow at 37 C and have morphology similar to that or organisms grown at room temperature

Characteristic features – sporangium, coenocytic hyphae and rootlike structuresOptimal incubation temp is 30 C. morphologically similar at any temp

OPPORTUNISTIC BASIDIOBOLUS RANARUM CONIDIOBLOUS CORONATUS (25-30 C) GEOTRICHUM PENICILLIUM

CULTURE

MORPHOLOGYColonies are flat, yellowish gray to creamy gray, glabrous becoming radially folded and covered by a fine powder, white surface mycelium

Colonies are flat, cream-colored, glabrous becoming radially folded and covered by fine powdery white surface mycelium and conidiophores.

White to tan, flat or fluffy, rapid-growing fungus

White colony at first but developing blue to green color

MICROSCOPIC VIEW

DESCRIPTIONThere is the presence of large vegetative hyphae forming numerous

The hyphae have few septa. The conidiophores are unbranched forming solitary terminal conidia. The conidia are spherical, single-celled and have a

Note hyphae breaking into arthrospores. May be confused with

Small, round spores borne in “brush-like” formations

PHOTOS AND TEXT PROVIDED BY MICROBIOLOGY DEPARTMENT

– UST FMS ‘04-’05 -formatted by: Auds Martinez (SEC-C) ©

round, smooth, thick walled zygospores with two closely appressed beaklike appendages

prominent papilla. It may also produce hair-like appendages called villae.

Coccidioides immitis.