AIRBORNE CONTAMINANTS CULTURE

ALTERNARIA

ASPERGILLUS

BOTYRIS

CHAETOMIUM

CEPHALOSPORIUM

CURVULARIA

FUSARIUM

MORPHOLOGY

Rapid-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

DESCRIPTION

Large, 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 CULTURE

GELASINOSPORA

GEOTRICHUM

GLIOCLADIUM

HORMODENDRUM

NEUROSPORA (MONILIA)

NIGROSPORA

PAECILOMYCES

MORPHOLOGY

Dark 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

DESCRIPTION

Squashed 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 CULTURE

PENICILLIUM

PULLULARIA (Aureobasidium)

RHODOTORULA

RHIZOPUS

SACCHAROMYCES

SCOPULARIOPSIS

SEPEDONIUM

MORPHOLOGY

White colony at first but developing blue to green color

Brown to black, flat, greasylooking 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

DESCRIPTION

Small, 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 footlike 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

AIRBORNE CONTAMINANTS CULTURE

SPOROBOLOMYCES

STEMPHYLUM

STREPTOMYCES

SYNCEPHALASTRUM

VERTICILLIUM
SUPERFICIAL Dermatophytoses – dermatophytes •Genus Microsporum •Genus Epidermophyton •Genus Trichophyton Pityriasis Versicolor – Malazzezia furfur

A light tan yeast colony MORPHOLOGY

Brown to black, wrinkled fuzzy colony

Dry, 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.

Tinia nigra – Cladosporium werneckii Piedra •white – Trichosporum(beigelli) cutaneum •black – Piedraia hortai

MICROSCOPIC VIEW

Otomycosis •Aspergillus •Mucor •Penicillium •Rhizopus Keratitis •Fusarium •Aspergillus •Curvularia •Penicillium •Cephalosporium •Candida species

DESCRIPTION

Yeast 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 FERRUGINEUM

MICROSPORUM GYPSEUM

MICROSPORUM NANUM

MICROSPORUM VANBREUSEGHEMII

CULTURE

MORPHOLOGY

Cultured 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

DESCRIPTION

No distinguishing spores will not grow on rice

Highly diagnostic large, thickwalled, 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 EQUINUM

TRICHOPHYTON MENTAGRPPHYTES

TRICHOPHYTON RUBRUM TRICHOPHYTON SCHOENLEINII TRICHOPHYTON TONSURANS

CULTURE

DERMATOPHYTES

SUBCUTANEOUS

CULTURE CULTURE

TRICHOSPORON BEIGELII (room temperature cultures) SPOROTHRIX SCHENCKII (Room Temp)

EPIDERMOPHYTON FLOCCOSUM SPOROTHRIX SCHENCKII (37 C cultures)

MALASSEZIA FURFUR FONSECAEA PEDROSOI

PIEDRAIA HORTAI (room temp cultures) FONSECAE COMPACTA PHIALOPHORA VERRUCOSA

SUBCUTANEOUS Sporotrichosis – Sporothrix schenckii Chromomycosis or CLADOSPORIM CARRIONI Chromoblastomycosis RANARUM BASIDIOBOLUS Phialophora verrucosa Fonsecae pedrosoi Cladosporium carrioni Mycetoma or Maduromycosis Actinomycotic Actinomadurae Nocardia The colony has a dark surface, flat Streptomyces Colonies are flat, yellowish gray with slightly raised center. It is to creamy gray, glabrous Eumycotic(true gray, covered with velvety dullfungi) becoming radially folded and Allescheria(or covered by a fine powder, white gray green or purplish brown, short Petriellidium) napped mycelium. Reverse is Madurella surface mycelium black. Phialophora Entomopthoromycosis Basidiobolus ranarum Conidiobolus coronatus Lobomycosis – Lacazia loboi(Loboa loboi) Rhinosporidiosis – Rhinosporidium seeberi Phaeohypomycosis Wangiella spp The hyphae is septate, dark with There is the presence of large Exophialia spp lateral and terminal conidiophores vegetative hyphae forming Cladosporium spp. of varying size. conidiophores numerous round, smooth, thick produce long branching chainsspp. Hormodendrum of walled zygospores with two brown, smooth walled, oval, Dreschlera spp. closely appressed beaklike
pointed conidia which have dark scars of attachment

PITYRIASIS VERSICOLOR

MORPHOLOGY MORPHOLOGY

MICROSCOPIC VIEW MICROSCOPIC VIEW

DESCRIPTION DESCRIPTION

One observes clear septate hyphae which may break up Clear, septate hyphae with into individual cells.

spores (3 to 6 um) in “daisylike” clusters

Large, club-shaped spores with 2 to 5 septa, often form in Round, oval, or “cigar-shaped” pairs.

yeast cells, 1 to 3 um x 4to 10 um

10% KOH with Parker ink mount showing characteristic spherical yeast cells and short The hyphae is elements typical pseudohyphal septate, branched and brown. The conidia is dark of the lungs

PIEDRA

Cultured on Sabouraud The colonies are smooth to medium on 2 antibiotics for 1-3 slightly wrinkled (not fuzzy) Dark, greasy-looking culture White to tan yeast colonies after and 1 to weeks incubation on weeks at3room temp. Yellow toon brain afterwhite2to tan in color 1 colored surface; green greento weeks incubationto Sabouraud medium heart infusion brown underside. agar.

Colonies in Dixon’s agar are cream is dark green, gray or Surface to yellowish, smooth or lightly wrinkled, silvery, velvetblack, covered byglistening or dull, with the It is usually flat, like mycelium. margin being either entire a lobate then develops or cone shaped protrusion in the center. Reverse is black.

Surface is dark green to black. tinge. They are very compact, The coloniesraised center and are have a are heaped, brittle with irregular indented borders. slightly fuzzy. There are brownish hyphae on the surface One observes dematiaceous hyphae containing numerous septa

The colonies are dark brown and may have a metallic green 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.

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

appendages

SUBCUTANEOUS

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

EXOPHIALA (WANGIELLA) DERMATITIDIS

CONIDIOBLOUS CORONATUS (25-30 C)

EXOPHIALA JEANSELMEI

PSEUDOALLESCHERICHIA BOYDII (SEXUAL)
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.

CULTURE

MORPHOLOGY

Slow growing, dryappearing colonies on Sabouraud medium. Morphology varies between etiologic agents. Use biochemical tests.

Colonies are slow growing, initially black and yeastlike, 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

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

MICROSCOPIC VIEW

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

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 hairlike appendages called villae.

elongated, tubular and with a tapered, narrow end. Conidia are smooth, thinwalled, 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 suspected IDIUM SEEBERI (RT AND 37 C)

DEEP SEATED Histoplasmosis – Histoplasma capsulatum Coccidioidomycosis – Coccidioides immitis South American Blastomycosis – Paracoccidioides brasiliensis North American Blastomycosis – Blastomycosis dermatitidis

DEEP SEATED

HISTOPLASMA CAPSULATUM ROOM TEMP

37 C

COCCIDIODES IMMITIS

PARACOCCIDIOIDES BRAZILIENSIS RT

37 C

CULTURE

MORPHOLOGY

Whtie, 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

DESCRIPTION

Characteristic 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 um Grows 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 CULTURE

BLASTOMYCES DERMATITIDIS RT 37 C

OPPORTUNISTIC Candidiasis – Candida albicans Cryptococcosis – Cryptococcus neoformans

CANDIDA ALBICANS RT AND 37 C

CRYPTOCOCCUS NEOFORMANS

ASPERGILLUS SP RT / 37 C

RHIZOPUS

MORPHOLOGY

Colony 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

Aspergillosis – Aspergillus fumigatus Zygomycosis (Mucormycosis) Phycomycosis Rhizopus Mucor Absidia Basidiobolus Conidiobolus

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

MICROSCOPIC VIEW

DESCRIPTION

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

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

Geotrichosis – Geotrichum candidum Penicillosis – Penicilium spp.

Chlamydospore agar, 1 to 2 days incubationl Note yeast cells, pseudohyphae and chlamydospores Usually cultured at RT. Some species grow at 37 C and have morphology similar to that of organisms grown at RT

India ink preparation showing numerous heavily encapsulated yeast cells 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 structures Optimal incubation temp is 30 C. morphologically similar at any temp

OPPORTUNISTIC

BASIDIOBOLUS RANARUM

CONIDIOBLOUS CORONATUS (25-30 C)

GEOTRICHUM

PENICILLIUM

CULTURE

MORPHOLOGY

Colonies 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, rapidgrowing fungus

White colony at first but developing blue to green color

MICROSCOPIC VIEW

PHOT SANDTEXTPROVIDE BY MICROBIOL GYDEPARTMENT –USTFMS‘04-’05formatedby:AudsMartinez(SEC- )©

DESCRIPTION

There 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 “brushlike” formations

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

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

Coccidioides immitis.

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