MYCOLOGY & VIROLOGY

Credits: MS. ABIGAIL MARIE PEREZ, RMT, MLS (ASCPi), MSMT (CAR)

OUTLINE OF THE LESSON (MYCOLOGY) MYCOLOGY OF TERMS

A. MYCOLOGY TERMS MOLD vs. YEAST

a. Mold vs. Yeast
b. Hyphae vs.Pseudohyphae MOLD YEAST
c. Septum vs.Spore
● Also called mycelia ● UNI-cellular fungi/
d. Mycelium vs. Saprophytes vs. ● MULTI-cellular fungi single celled fungi
Sporangium ● Appears either
B. THE FUNGAL ORGANISM spherical to ellipsoidal
a. Cell membrane (3 - 15um)
b. Cell wall
c. Capsule ● Multicellular colonies ● Reproduces by
d. Fungi vs. Bacteria composed of clumps budding & at a slower
C. FUNGAL STRUCTURE of intertwined rate than bacteria
a. Hyphae branching hyphae ● When buds do not
● Grows by longitudinal separate, they form
b. Condia
extension and long chains of yeast
c. Conidiophore produce spores cells
d. Sexual production (PSEUDOHYPHAE)
e. Asexual production
f. Monomorphic fungi ● Produces cottony or ● Produces moist,
g. Dimorphic fungi powdery colonies creamy, opaque and
h. Diphasic fungi ● Grows best at 25℃ pasty colonies
D. BASIC CLINICAL MYCOLOGY resembling bacteria
a. Specimen collection ● Grows best at 37℃
b. Specimen transport
c. Laboratory safety
d. Methods in Specimen Processing
e. Fungal Culture
f. Selection of Culture Media
g. Culture Considerations
h. Pathogen vs.Contaminant
i. Body sites and Possible fungal
pathogens
E. LABORATORY IDENTIFICATION
a. Direct examination methods
b. Tissue examination: Stains
c. Mold identification
d. Yeast identification
F. MEDICALLY IMPORTANT FUNGI

*Not same picture, but

similar

Lactophenol Cotton Blue

stain of Yeast

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MYCOLOGY & VIROLOGY
Credits: MS. ABIGAIL MARIE PEREZ, RMT, MLS (ASCPi), MSMT (CAR)

Gram Stain of Yeast

Take note, it looks similar to
bacteria (specifically
MYCELIUM vs. SAPROPHYTES vs. SPORANGIUM
that of the
Staphylococcus spp.,
hence it has to be MYCELIUM SAPROPHYTES SPORANGIUM
differentiated using
tests) ● Mass of ● Also known ● Sac-like
mat of as saprobes structure
hyphae ● Organisms where
● A mold capable of sporangiosp
HYPHAE vs. PSEUDOHYPHAE colony living on ores are
decaying formed
HYPHAE PSEUDOHYPHAE organic (Zygomycet
material to es)
● Threadlike, branching, ● Chains of elongated serve as
cylindrical tubules budding yeast cells or energy
composed of fungal blastoconidia source
cells attaching from
one end to end
● Grows by extending in
length from the tips of
tubules
● Separated by porous
cross-walls or septa THE FUNGAL ORGANISM

● Non motile
● Eukaryotic organisms
● Have definite cell walls
● Devoid of chlorophyll
● Reproduce by means of spores and conidia
● “Heterotrophic” (different nourishment)

CELL MEMBRANE
SEPTUM vs. SPORE
● Bilayered phospholipids
SEPTUM SPORE ● Contains sterols
○ Sterols are also found in the cell
● Hybrid cross-wall, ● The reproducing membranes of humans and genus
typically perforated body of molds
● Is what separates ● A specialized
Mycoplasma
hyphal cells propagule with ○ Ergosterol vs. Cholesterol
enhanced survival ■ Ergosterol - essential sterol in
value (ex. Resistance fungi
to adverse condition)
■ Cholesterol - essential sterol in
humans
● Functions:
○ Protects cytoplasm
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MYCOLOGY & VIROLOGY
Credits: MS. ABIGAIL MARIE PEREZ, RMT, MLS (ASCPi), MSMT (CAR)

○ Regulates intake of nutrients

THE FUNGAL STRUCTURE
○ Facilitates capsule and cell wall
synthesis
● The majority of antifungal agents work through HYPHAE
disrupting ergosterol by:
are long, branching filaments that come together to form
○ Amphotericin B & Nystatin - by the mycelium
binding to it and leaving holes in the
fungal cell wall
SEPTATE HYPHAE ASEPTATE HYPHAE
○ Azoles & Echinocandins - by
interfering with ergosterol synthesis ● Have cellular ● or Sparsely septate
separation or hyphae
cross-walls ● Contain few to no
CELL WALL cellular separations
● COENOCYTIC also
● ANTIGENIC refers to hyphae
lacking cross-walls
● Multilayered
○ Polysaccharides (90%)
○ Chitin
○ Proteins & glycoproteins (10%)
● Provides shape and rigidity to the cell

CAPSULE

● Polysaccharide
○ Much larger than bacterial capsule
● Antiphagocytic virulence factor PSEUDOHYPHAE TRUE HYPHAE
○ Cryptococcus neoformans
○ Encapsulated yeast cell ● Chains of elongated ● Appears as a short
○ Viewed under India ink stain budding yeast cells or lateral extension from
blastoconidia the yeast cell
● Does not have a
constriction (septum)
FUNGI BACTERIA where it meets the
yeast cell
NUCLEUS Defined nuclear No membrane -
membrane bounded nucleus
(EUKARYOTE) (PROKARYOTE)

MITOCHO Present Absent

NDRIA

CELL Composed of No glucose and

WALL glucose and mannose polymers
mannose polymers with muramic acid
(CHITIN); NO and teichoic acids
muramic and VEGETATIVE HYPHAE AERIAL HYPHAE
teichoic acids
● Functions for food ● Functions for
DIMORPHI Can exist in yeast None absorption supporting
SM and mold forms ● Extends below the reproductive structures
agar surface called conidia
CHROMOS More than 1 and Only 1 but not ● Extends above the
CHON associated CHON associated agar surface
OME

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MYCOLOGY & VIROLOGY
Credits: MS. ABIGAIL MARIE PEREZ, RMT, MLS (ASCPi), MSMT (CAR)

pushed through a small pore

in the parent cells

CONIDIA

sporelike, reproductive structures produced in an

asexual mode and are only formed by imperfect fungi

MICROCONIDIA MACROCONIDIA

● Single-celled, small ● Multicellular, large

conidia conidia

SEXUAL PRODUCTION

● Requires the formation of specialized fungal

structures called spores
CONIDIOPHORE ● Fungi that undergo sexual reproduction are
called as perfect fungi
structure that supports conidia
ASCOSPORES ● Sexual spore formed in a
ANNELLOCONIDIA ● Vase-shaped conidia sac-like structure after meiosis
● The remaining parent outer
cell wall takes on a
saw-toothed appearance as
the conidia are released

BASIDIOSPORES ● Sexual spores dormed in a

clubn-shaped reproductive
structure after meiosis
PHIALOCONIDIA ● Tube-shaped conidia that
can be branched

POROCONIDIA ● Conidia formed by being

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MYCOLOGY & VIROLOGY
Credits: MS. ABIGAIL MARIE PEREZ, RMT, MLS (ASCPi), MSMT (CAR)

OOSPORE ● Sexual spores resulting from BLASTOCONIDIA ● Conidia that form as a result
the fusion of cells from two of budding
different hyphae ● Budding of cell (mother and
daughter)

ZYGOSPORE ● Sexual spores resulting from

the fusion of cells from two
identical hyphae

ASEXUAL PRODUCTION

● Only involves division of the nucleus and

cytoplasm
● Fungi that undergo this are called imperfect
fungi
● Imperfect fungi are the ONLY fungal group to
produce conidia
CHLAMYDO- ● Result from terminal cells in
CONIDIA the hyphae that enlarge and
ARTHROCONIDIA ● Are conidia resulting from the have thick-walls
fragmentation of hyphae into ● Directly from hyphae
individual cells directly from (swelling)
hyphae my modification of ● These conidia can survive
cell wall (barrels) adverse environmental
conditions
● In molds, they are
chlamydoconidia; in yeast,
theta re found as
chlamydospores

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MYCOLOGY & VIROLOGY
Credits: MS. ABIGAIL MARIE PEREZ, RMT, MLS (ASCPi), MSMT (CAR)

○ Dermatologic samples requires dry

containers
○ NO transport media
● Processed within a few hours
● Specimens can be refrigerated at 4℃
○ ONLY IF DELAYED processing
○ BLOOD and CSF: 30℃ to 37℃
○ BLOOD and CSF: 15℃ to 30℃

MONOMORPHIC FUNGI
LABORATORY SAFETY
● Mold at both 25℃ and 37℃ (TRUE MOLD)
● Both Universal and Standard Precautions MUST
● Yeast at both 25℃ and 37℃ (TRUE YEAST)
be followed class 2 or 3 biosafety cabinets are
used
DIMORPHIC FUNGI ● Disinfectants are phenol-based in nature
● Biohazard containers should be available for use
● Appears both as a mold at room temperature (
25℃) and yeast at body temperature (37℃) and
in tissues METHODS IN SPECIMEN PROCESSING
● What form is infectious to man?
● DIRECT INOCULATION
○ Adding several drops of sample to the
DIPHASIC FUNGI media
○ For solid media, samples can be
● Appears as a mold at 25℃ and 37℃ and as streaked
yeast in tissues ○ Specimen types:
■ Bronchial brush/wash,
aspirates, CSF
BASIC CLINICAL MYCOLOGY
■ Swabs, body fluid, hairs, skin
“If a physician is dependent upon Microbiology scrapings
laboratory data for helping to save his patient, the ONE
WHO COLLECTS THE SPECIMEN may determine the ● CONCENTRATION TECHNIQUES
course of the patient’s recovery” - Albert Balows, PH.D. ○ Large volumes can be concentrated by
centrifugation
○ Specimen types:
SPECIMEN COLLECTION ■ CSF, body fluids, urine sample

● The following important factors should be ● MINCED/HOMOGENIZED

considered when isolating and identifying a ○ Some solid samples must be
fungal pathogens “destroyed” to expose a buried
○ Correct type of specimen pathogen to the media
○ Quality of specimen ○ Specimen types:
○ Rapid transport ■ Nails, tissue, biopsies
○ Use of appropriate culture media
● Processed within 2 hours
FUNGAL CULTURE

SPECIMEN TRANSPORT ● Petri dishes or tubes

● Proper oxygen requirements should be provided
● Sterile, leak-proof container ● Humidity

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MYCOLOGY & VIROLOGY
Credits: MS. ABIGAIL MARIE PEREZ, RMT, MLS (ASCPi), MSMT (CAR)

● Subculture is sometimes necessary

○ Temperature range
○ Dimorphism
○ Sexual and asexual development
structures
○ Get rid of bacterial contamination
● Teasing needles
○ Are used more than inoculating loops
● Electric incinerator is used
○ Because flame causes aerosols
● Culture is very important to identify the fungal CELLOPHANE TAPE METHOD
species present ● Used to transfer aerial hyphae from the colony
to a slide for examination

SELECTION OF CULTURE MEDIA

● For primary cultures

○ Test tubes are used
○ Less likely to be contaminated, less
drying involved
● For Subcultures
○ Petri Dishes are used SLIDE CULTURE METHOD
○ Larger surface area for growth ● Uses a block of agar overload with a coverslip
● Use of inhibitory substances may be required ● Fungal colonies are grown on the slide of the
○ Chloramphenicol, Gentamicin, agar block
Cycloheximide ● Cover slip is removed and used for microscopic
○ May encounter some fungal inhibition examination
● This method prevents damage to the fungal
structures
CULTURE CONSIDERATIONS

● Fungal Cultures are incubated at 30℃

● Growth requires from several days to several
weeks
● Cultures should be maintained in a
high-humidity environment
● Several techniques are used to obtain culture
material for slide prep:
○ Tease mount method
○ Cellophane tape method
○ Slide culture method

TEASE MOUNT METHOD

● A dissecting needle is used to pull apart fungal PATHOGEN vs CONTAMINANT
colonies, then placed on a slide
● The clinical picture
● This method may damage conidia
○ Are patients’ symptoms consistent with
● May take several attempts to obtain samples
fungal infection?
with intact conidia
○ Does this fungus normally cause these
symptoms?
● Laboratory findings
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MYCOLOGY & VIROLOGY
Credits: MS. ABIGAIL MARIE PEREZ, RMT, MLS (ASCPi), MSMT (CAR)

○ Fungal elements in tissue or other ● Modifies SDA (Emman’s)

specimen ○ Neutral pH, less glucose
○ Fungus grown in culture ○ Better supports fungal growth, less
○ More than one culture positive inhibitory for bacteria

BODY SITES AND POSSIBLE FUNGAL MYCOSEL AND MYCOBIOTIC

PATHOGENS
● SDA + chloramphenicol + cycloheximide
SITE FUNGAL PATHOGENS ● Selective recovery of dimorphs and
dermatophytes
BLOOD Candida spp.
Blastomyces dermatitidis
Histoplasma capsulatum BRAIN HEART INFUSION AGAR (BHIA)
Cryptococcus neoformans
● Enriched to enhance recovery of Cryptococcus
CSF Candida spp.
Coccidioides immitis
neoformans and dimorphic transitions in
Histoplasma capsulatum Sporothrix spp. And Paracoccidiodes spp
Cryptococcus neoformans ● In plates or tubes
● Broth + penicillin for Zygomycetes spp.
HAIR Microsporum spp.
Trichophyton spp.
● BHI + Gentamicin + Chloramphenicol for
Cryptococcus neoformans from contaminated
NAILS Aspergillus spp. samples
Epidermophyton spp.
Trichophyton spp.
BRAIN HEART INFUSION AGAR WITH BLOOD
SKIN Candida spp. (BHIB)
Microsporum spp.
Epidermophyton spp.
Trichophyton spp. ● Composed of brain heart infusion agar and
Blastomyces dermatitidis sheep’s blood
● Used for fungal culture of samples from sterile
LUNGS Candida albicans body sites
Aspergillus spp.
Rhizopus spp. ● Can be made selective for dimorphic fungi by
Penicillium spp. adding:
Blastomyces dermatitidis ○ Cycloheximide - inhibits saprophytic
Coccidioides immitis
fungi
Histoplasma capsulatum
○ Chloramphenicol - inhibits many G (+)
THROAT Candida albicans and G (-) bacteria
Geotrichum candidum ○ Gentamicin - inhibits G (-) bacteria
URINE Candida albicans
Candida glabrata SABOURAUD DEXTROSE + BRAIN HEART
INFUSION AGAR (SABHI)
GENITAL TRACT Candida albicans

● Strengths of both media

● Enriched medium for Cryptococcus spp. And
SABOURAUD'S DEXTROSE AGAR (SDA)
thermally dimorphic fungi
● General purpose, nutritionally poor medium ● Can be made selective for dimorphic fungi by
mildly selective for fungi adding:
● SDA with acidic pH of 5.6 = inhibits most ○ Cycloheximide
bacteria ○ Chloramphenicol
○ Gentamicin

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MYCOLOGY & VIROLOGY
Credits: MS. ABIGAIL MARIE PEREZ, RMT, MLS (ASCPi), MSMT (CAR)

CHROMAGAR CANDIDA MEDIA FOR SUBCULTURE

● Selective and differential for presumptive ● Potato Dextrose Agar (PDA)

identification of genus Candida from primary ● Potato Flake Agar (PFA)
plates ● Cornmeal Agar for yeast morphology
○ C. albicans - light to medium green ○ Recommends form promoting
○ C. tropicalis - light blue to metallic blue sporulation
○ C. krusei - light rose with a whitish ● Incubation:
border ○ Obligate filamentation: 25℃ or 37℃
○ Dimorphic: 25℃ and 37℃
○ Yeast: 25℃ or 37℃
SELECTIVE MEDIA
● Aerobic
○ 3 to 4 week incubation period
INHIBITORY MOLD DERMATOPHYTE TEST
AGAR (MIA) MEDIUM (DTM) LABORATORY IDENTIFICATION
● Contains ● Used to isolate ● Direct examination of clinical specimens
chloramphenicol and dermatophytes ○ Laboratory methods and tissue stain
gentamicin ● Contains ● Macroscopic/ microscopic evaluation
● Inhibits bacteria cycloheximide, ○ Colony features and hyphae/ conidia
● Specially formulated gentamicin and
morphology
to recover phenol red (pH
cycloheximide - indicator) ● Advanced methods
sensitive ● Dermatophytes ○ Exoantigen. DNA probes, DNA
Cryptococcus from heavily sequencing
contaminated
specimens (pink
to red color DIRECT EXAMINATION METHODS
change)
● SALINE WET MOUNT
○ Used to view fungal elements (hyphae,
DIFFERENTIAL MEDIA conidia, budding yeasts)
○ Limited use and is most commonly
● POTATO DEXTROSE AGAR (PDA) applicable for vaginal secretions to
○ Used to enhance candida development diagnose vaginitis
○ Enhances pigment development of
Trichophyton rubrum ● LACTOPHENOL COTTON BLUE WET MOUNT
○ Used to stain and preserve fungal
● BIRD SEED (NIGER SEED) AND CAFFEIC elements in culture isolates
ACID AGARS
○ For growth of C. neoformans ● POTASSIUM HYDROXIDE (KOH)
○ Forms block to brown colonies dues to ○ Used to dissolved non fungal materials
activity of phenol oxidase in skin scrapings, nail, hair, fluids
exudates and biopsy specimens
● CORNMEAL AGAR WITH TWEEN80 ○ Fungal elements (hyphae, yeast)
○ Used to differentiate Candida spp. ○ Needs reduced light or phase-contrast

● AGAR WITH RICE, CASEIN, & OTHER ● GRAM STAIN

NUTRIENTS ○ Used to view yeasts
○ Used to differentiate Trichophyton spp.

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MYCOLOGY & VIROLOGY
● HAIR BAITING
○ Hair is used as a bait for superficial and
cutaneous fungi
○ Study infected hair for:
■ Ectothrix - growth on the surface
of hair (Microsporum canis)
■ Endothrix - growth on the
surface and inside
(Trichophyton spp.)
● Presence of V-shaped
cracks in the hair
● WOOD’S LIGHT
○ Examines infected hair under Wood’s
lamp (UV light)
○ Hair infected with Microsporum
fluorescence while those infected with
Trichphyton does not
○ M. canis - yellow green
○ M. audouinii - apple green
● INDIA INK
○ To view encapsulated capsules
surrounding C. neoformans in CSF
○ However, due to low sensitivity, direct
antigen detection assays have generally
replaced the india ink wet mount
● CALCOFLUOR WHITE STAIN
○ Fluorochrome that stains chitin
○ Stain is NOT absorbed by human tissue
Credits: MS. ABIGAIL MARIE PEREZ, RMT, MLS (ASCPi), MSMT (CAR)
○ Slide is viewed using UV light
○ Fungi appears white-blue-green
○ Depending on wavelength of light
○ KOH can be added to clear cell debris
TISSUE EXAMINATION: STAINS
hair ● GIEMSA, WRIGHT-GIEMSA
○ Blood and bone marrow, Histoplasmosis
capsulatum (intracellular)
● HEMATOXYLIN AND EOSIN (HE)
○ Pink to pinkish blue appearance
● MEYER’S MUCICARMINE
○ Cryptococcus neoformans (rose red)
● GOMORI TECHNIQUE METHENAMINE
SILVER (GMS)
○ Black appearance
● PAPANICOLAOU STAIN
○ Pink to blue appearance
● PERIODIC ACID-SCHIFF (PAS)
○ Red to purple appearance
MOLD IDENTIFICATION
Advanced techniques
1. EXOANTIGEN TEST
a. Rapid information of immunoidentity
b. Extract soluble antigen from unknown
isolate then Concentrate
c. React with antiserum specific to known
fungi
d. Positive control necessary for definitive
ID
e. Test is read at 24 hours
f. For Blastomyces, Coccidioides,
Histoplasma
2. DNA PROBE
a. Rapid kits that use nucleic acid
hybridization to identify fungi in culture
b. Highly specific because it is based on
DNA sequences
c. Needs to be performed on cultures NOT
from samples
d. Specialized labs use DNA sequencing
to establish fungal ID
e. For Blastomyces, Coccidioides,
Histoplasma
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MYCOLOGY & VIROLOGY
Credits: MS. ABIGAIL MARIE PEREZ, RMT, MLS (ASCPi), MSMT (CAR)

3. UREA HYDROLYSIS
a. Detected on simple urea agar
YEAST IDENTIFICATION
b. Rapid, easy
Physiologic techniques c. Differentiate Cryptococcus from
1. Germ Tube Test Rhodotorula
a. Filamentous outgrowth from d. POSITIVE: pink color, C. neoformans
blastoconidia e. NEGATIVE: little to no color change, C.
b. Most basic and easiest to perform albicans
c. Requires the use of serum or plasma
d. Over Incubation and over inoculation 4. TEMPERATURE STUDIES
are bigger problems a. Cryptococcus spp. - weak growth at
e. Other agents can form germ tubes 35℃// NO growth at 42℃
f. NOT VALID if read after 2 hours b. Candida spp. - several can grow well
g. Candida albicans - “TRUE” germ tubes exceeding 45℃
h. No constriction at base, where tube ● Microscopic appearance
attaches to the mother cell ○ Saline wet mounts and Gram stains will
i. If constricted base is present - Candida show budding yeast
tropicalis ○ Yeast are discovered in routine
j. Other species that have germ tubes: urinalysis
i. Candida stellatoidea - sucrose ○ India ink preparations are used to
assimilation to differentiate from show the capsule surrounding
C. albicans Cryptococcus neoformans
ii. Candida dubliniensis - no ● Culture appearance
growth at 45℃ ○ Yeasts are grown on SABHI at 22℃ -
k. Positive and negative controls are 30℃
necessary ○ Yeasts will form cream-colored, mucoid
to smooth colonies within several days.
■ BAP - yeast colonies can
resemble Staphylococcus spp.
○ Cornmeal agar with Tween 80 -
differentiates Candida spp. By
enhancing the formation of hyphae,
psuedohyphae and conidia
○ C. aclbicans - has chlamydospores with
clusters of blastoconidia along the
hyphae
○ C. tropicalis - produces long-branched
pseudohyphae, blastoconidia are short
and does not produce chlamydospores
○ CHROMagars allow the yeast ID. this
2. FERMENTATION, ASSIMILATION media contains variety of substrates
a. Fermentation - carbohydrates use in resulting in production of colonies with
absence of O2 different colors
b. Assimilation - can be used as sole
Carbon source
c. API 20C - multi-strip test
d. Vitek - automated, cards with
biochemical tests

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MYCOLOGY & VIROLOGY
Credits: MS. ABIGAIL MARIE PEREZ, RMT, MLS (ASCPi), MSMT (CAR)

MEDICALLY IMPORTANT FUNGI cosis Paecilomyces,

trichosporon
and other
CATEGORY MYCOSIS GENUS hyaline mold
● Phaeohyphomy ● Cladophialophor
SUPERFICIAL ● Pityriasis ● Malassezia spp. cosis a bantiana,
versicolor Alternaria,
● Tinea nigra ● Exophiala Cladosporium,
palmaris werneckii Bipolar,
● White piedra ● Trichosporon Exserohilum,
spp. Dematiaceous
● Black piedra ● Piedraia hortae fungi
● Mucormycosis ● Rhizopus,
CUTANEOUS ● Dermatophytosi ● Microsporum, (Zygomycosis) Absidia,
s Trichophyton, Cunninghamella
Epidermophyton and other
floccosum Zygomycetes
● Candidiasis of ● Candida ● Penicilliosis ● Penicillium
skin, mucosa or albicans and marneffei
nails other Candida
spp.
CLINICALLY SIGNIFICANT YEAST
SUBCUTA- ● Sporotrichosis ● Sporothrix
NEOUS schenckii
● Chromoblastom ● Phialophora YEAST INFECTIONS CULTURE BIOCHEMI
ycosis verrucosa, CAL TEST
Fonsecaea
pedrosoi ● Oral thrush ● Grows on ● Positive
● Mycetoma ● Pseudallescheri ● Vaginal most agar, Germ Tube
a boydii, thrush SDA, EMB ● ** C.
Maduerella ● Diaper rash (spider-like dubliniensi
mycetomatis ● Onychomyco projections) s is also
● Phaeohyphomy ● Exophiala, sis (nails) ● Gram (+) GT (+) **
cosis Bipolaris, ● Paranychomy ● Blastoconidi ● Urease (-)
Exserohilum, cosis a w/o ● Inositol (-)
Candida (cuticles) constriction
Dermatiaceous albicans ● Endocarditis s
fungi ● Meningitis ● Cornmeal
● UTI Agar with
SYSTEMIC ● Coccidioidomyc ● Coccidioides ● Pulmonary Tween 80 -
osis immitis,Coccidio infections produce
ides posadasii ● fungenia chlamydosp
● Histoplasmosis ● Histoplasma ores
capsulatum
● Blastomycosis ● Blastomyces ● Most frequently isolated yeast
● Normal flora in GI tract, mucocutaneous areas
dermatitidis ● Serious infections in immunocompromised
● Paracoccidioido ● Paracoccidioide patients
mycosis s brasiliensis
● Lung ● Yeast cells ● Urease (+)
OPPORTU- ● Systemic ● Candida infection that are ● Phenol
NISTIC Candidiasis albicans and can surrounded oxidase (+)
other Candida disseminate by a thick ● Inositol (+)
spp. to the brain capsule ● Nitrate
● Cryptococcosis ● Cryptococcus ● Niger seed reduction
agar - (-)
neoformans and Cryptoco maroon to
Cryptococcus ccus brown-black
gattii neoforma colonies
● Aspergillosis ● Aspergillus ns ● Only forms
fumigatus and blastoconidi
other a
Aspergillus spp.
● Hyalohyphomy ● Fusarium, ● Found in bird, bat droppings, decaying vegetation

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MYCOLOGY & VIROLOGY
Credits: MS. ABIGAIL MARIE PEREZ, RMT, MLS (ASCPi), MSMT (CAR)

● Capsule seen in India ink

● Cryptococcal antigen test preferred
● Disseminated cryptococcosis common in AIDS
patients

● Uncommon ● Cornmeal
cause of agar -
wound hockey
Geotrich infections stick-shape
um ● Oral thrush d
candidu arthroconidi
m a
● No
blastoconidi
a

CLINICALLY SIGNIFICANT SUPERFICIAL FUNGI

YEAST INFECTIO APPEARA DIAGNOSI

NS NCE S

● Tinea ● Bottle-sha ● KOH

versicolor ped smear -
● Infection is budding from
common yeast lesions that
on upper ● Characteri have yeast
back stic collar like cells
chest, between and
shoulders, mother hyphae
arms and ● (spaghetti
Malassezia ● Systemic daughter and ● Tinea ● Light ● KOH
furfur infection in cells meatballs) nigra - brown, smear
children ● Short, ● Wood’s characteri branching
slightly lamp - zed by septate
curved lesions will Hortaea presence hyphae
septate fluoresce werneckii of brown and
hyphal yellow to black budding
chain seen non scaly yeast
in skin macules
scrapings on palms
and dorsa
● There may be an association between the of feet
disease and excessive sweating
● Other names: Exophiala werneckii,
Phaeoannellomyces werneckii, Cladosporium
werneckii

Page 13 | Mycology
MYCOLOGY & VIROLOGY
Credits: MS. ABIGAIL MARIE PEREZ, RMT, MLS (ASCPi), MSMT (CAR)

● White ● Microscopi ● Hair

piedra - c- treated in
fungal intertwined 10% KOH
infection of hyaline or 25%
facial, septate NaOH
pubic, hyphae solution
axillary breaking with 5%
Trichosporon hairs and into oval glycerol
spp. the scalp or ● Urease (+)
● Soft, rectangula ● Carbohydr
white, r ate
yellowish, arthroconi assimilatio
beige or dia and n (+)
greenish blastoconi
nodules dia
● Black ● Compact ● Submergin on the hair ● cream-col
piedra - masses of g hair in shaft ored ,
fungal dark, 25% smooth
infection of septate KOH/NaO colonies
the scalp, hyphae, H solution on solid
hair, less round to with 5% media in 1
frequently oval asci glycerol week
on beard, containing and
Piedraia mustache, aseptate heating ● Can be isolated from soil, animals and
hortae pubic, banana-sh humans
axillary aped ● T. ovoides - causative agent of scalp
hair ascospore infections
● Disease s ● T. inkin - causes most cases of pubic White
characteri piedra
zed by
presence
of hard,
dark
nodules
on hair
shaft

Page 14 | Mycology
MYCOLOGY & VIROLOGY
Credits: MS. ABIGAIL MARIE PEREZ, RMT, MLS (ASCPi), MSMT (CAR)

CLINICALLY SIGNIFICANT CUTANEOUS FUNGI

YEAST INFECTION APPEARA OTHER

S NCE CHARACT
ERISTICS

● Skin, hair ● MACROC ● T.

and nails ONIDIA - mentagrop
rare, hytes -
pencil-sha grapelike
ped, clusters of
multiseptat microconid
ed, ia & (+)
thin-walled hair
, smooth, perforation ● Skin, ● MACROC ● E.
borne test sometimes ONIDIA - floccosum
singly on ● T. rubrum - hair, rarely club-shap - invades
appearanc few or nails ed, nails, on
e numerous septate, KOH
● MICROCO macroconi thin-walled preparatio
NIDIA - dia, Epidermophyt ,smooth n, chains
predomina numerous on spp. ● MICROCO of
nt, club-shap NIDIA - arthroconi
spherical, ed absent dia can be
Trichophyton tear-shape microconid ● On SABHI seen.
spp d or ia borne at RT: Forms
clavate singularly; yellow tot khaki-yello
● On SABHI forms an reverse w colonies
at RT: buff deep red with tan
granular reverse on reverse
colonies, PDA (-)
rose to tan hair ● E. floccosum is the only species
colored, perforation
with a test
yellow, ● T.
brown or verrucosu
red m-
reverse. produces
White chlamydoc
fluffy onidia on
colonies SDA or
with PDA. on
colorless thiamine-e
to yellow nriched
reverse media,
elongated
rat-tail
microconid
ia are
produced.
(-) hair
perforation
test

● Hair infections endothrix or ectothrix

● Hair doesn’t fluoresce. May have nodular
bodies.

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MYCOLOGY & VIROLOGY
Credits: MS. ABIGAIL MARIE PEREZ, RMT, MLS (ASCPi), MSMT (CAR)

Microsporum ● Skin, hair, ● MACROC ● M.

spp. rarely nails ONIDIA - audouinii -
large forms
spindle or pectinate
cylinder (comb-like
shaped, ) septate
thick-walle hyphae
d, with
multi-sept terminal
ate, rough, chlamydoc
spiny. onidia with
Borne pointed
singly on ends.
short Grows TYPES OF TINEA INFECTIONS AND THEIR
conidiospo poorly on
res rice grains
CAUSATIVE AGENTS
● MICROCO ● M. canis -
NIDIA - forms ● TINEA PEDIS
few or numerous,
absent, thick-walle
○ or an athlete's foot. An infection of the
small, d, spaces between the toes.
club-shap spindle-sh ○ Caused by Trichosporon spp. And
ed aped
● On SABHI macroconi Epidermophyton spp.
at RT: dia with ○ Characterized by itching and scaling
colonies tapered
will be ends.
light tan ● M.
with a gypsum -
salmon-col forms
ored numerous
reverse thin-walled
elliptical
macroconi
dia

● Microsporum are very slow growers.

● M. audouinii - an anthropophilic species
(found in humans)
● M. canis - a zoophilic species (found in ● TINEA COPORIS
animals) ○ or ringworm. An infection of smooth skin
● M. gypsum - a geophilic species (found in
soil) all over the body.
○ Caused by Trichophyton spp. And
Microsporum spp.
○ Characterized by circular patches of
scaly skin

● TINEA UNGUIUM
○ or onychomycosis. An infection of the
nails
○ Caused by Trichophyton spp. And
Epidermophyton spp.

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MYCOLOGY & VIROLOGY
Credits: MS. ABIGAIL MARIE PEREZ, RMT, MLS (ASCPi), MSMT (CAR)

○ Characterized by discoloration,
thickening and progressive destruction
of the nails

CLINICALLY SIGNIFICANT SUBCUTANEOUS

FUNGI

● MYCETOMA
○ Granulomatous infection causing
● TINEA CAPITIS cutaneous abscesses
○ An infection of the scalp ○ Exudates contain red, yellow, or black
○ Caused by Trichosporon spp. And granules
Microsporum spp. ○ Causative agents: Pseudallescheria
○ Characterized by circular bald patches boydii, Exophiala, Acremonium,
on the scalp Madurella

● TINEA BARBAE
○ or barber’s itch. An infection of the
beard hair
● CHROMOBLASTOMYCOSIS
○ Caused by Trichosporon spp. And
○ Localized infection characterized by
Microsporum spp.
chronic, hard, tumor-like lesions
○ Characterized by skin lesion
○ Most infections involve the feet or lower
legs
○ Presence of sclerotic bodies
(copper-colored fungal cells) in lesions
○ Colonies are folded or heaped and are
gray to black
○ Causative agents: Phialophora,
Cladosporium, Exophiala, Wangiella

● TINEA CRURIS
○ or jock’s itch. An infection of the groin.
○ Caused by Trichosporon spp. And
Epidermophyton spp.
○ Characterized by itching and scaling of
the groin area

Page 17 | Mycology
MYCOLOGY & VIROLOGY
Credits: MS. ABIGAIL MARIE PEREZ, RMT, MLS (ASCPi), MSMT (CAR)

● PHAEOHYPHOMYCOSIS
○ Subcutaneous infection that can
become systemic
○ Resulting systemic infections can cause
CLINICALLY SIGNIFICANT SYSTEMIC FUNGI
endocarditis and brain abscesses
○ Causative agents: Bipolaris, Curvularia, ● Blastomyces dermatitidis
Phialophora, Alternaria ○ INFECTIONS:
■ North american blastomycosis
● SPOROTRICHOSIS (Gilchrist’s disease)
○ or Rose Gardener’s disease ○ MOLD PHASE
○ Lymph and pulmonary infection can also ■ White to gray-brown colonies
occur ■ Hyaline septate hyphae with
○ Infections come from rose thorns and small oval conidia borne singly
contact with sphagnum mass at tips of conidiospores.
○ Causative agents: Sporothrix schenckii - ■ “Lollipops”
is a dimorphic fungus ○ YEAST PHASE
○ Blood media at 35℃ - yeast cells inside ■ Round, thick-walled.
PMNs and are cigar-shaped ■ Single bud connected by wide
○ SDA or PDA at RT - mold form neck
○ Forms white to yellow colonies ○ OTHER
○ Clusters of pear-shaped conidia at tips ■ Endemic to Ohio & Mississippi
of conidiospores RIver alleys
○ “Flowers” ■ Usually in farmers who contract
○ Gram positive cigar shaped cells from soil
○ Usually not seen in direct smears unless ■ Begins in lungs then becomes
by immunofluorescence systemic
○ Found in Mississippi & Missouri River ■ Can be cultured from tissues or
valleys body fluids
○ Rose bushes, barberry bushes,
sphagnum moss. Mulch
○ Usually infects farmers or florists,
introduced by trauma
○ Cutaneous lesions spread along
lymphatics

Page 18 | Mycology
MYCOLOGY & VIROLOGY
Credits: MS. ABIGAIL MARIE PEREZ, RMT, MLS (ASCPi), MSMT (CAR)

● Paracoccidioides brasiliensis ■ More common in dark-skinned

○ INFECTIONS: men
■ South american blastomycosis ■ Spores are inhaled or
○ MOLD PHASE contaminate injured skin
■ Resembles blastomyces ■ Affects respiratory tract first then
○ YEAST PHASE spreads to other organs
■ Round, double-walled. ■ CAUTION: arthrospores are
■ Multiple buds pinched at highly infectious
attachment ■ As with all mold phase fungi,
■ “Mariner’s wheel” always use a BSC to prevent
○ OTHER spore inhalation
■ Mainly in Brazil, Argentina, ■ Serological tests are available
South Mexico
■ Acquired by spore inhalation or
ingestion
■ Begins in lungs
■ Can become systemic

● Histoplasma capsulatum
○ INFECTIONS:
■ Histoplasmosis
■ Spelunker’s disease
■ Darling’s disease
○ MOLD PHASE
■ Silky, white to gray-tan colonies
■ Thick-walled macroconidia,
surrounded by finger-like
● Coccidioides immitis projections
○ INFECTIONS: ■ “Tuberculate chlamydospores”
■ San Joaquin Valley fever ○ YEAST PHASE
○ MOLD PHASE ■ Round to oval
■ Fluffy or powdery white to ■ Intracellular in phagocytes of
gray-tan colonies blood or bone marrow
■ Barrel-shaped arthrospores ○ OTHER
○ YEAST PHASE ■ Endemic to Ohio & Mississippi
■ Spherule containing numerous RIver alleys
endospores ■ Soil saprophytes
○ OTHER ■ Found in manure of chickens,
■ Found in desert of Southwest pigeons, starlings and bats
U.S (guano)

Page 19 | Mycology
MYCOLOGY & VIROLOGY
Credits: MS. ABIGAIL MARIE PEREZ, RMT, MLS (ASCPi), MSMT (CAR)

■ Acute pulmonary disease that ○ MICROSCOPIC MORPHOLOGY

can be self-limited or spread ■ Hyphae: large, broad,
■ Predilection for RE system nonseptate
■ Tissue phase may be confused ■ Produces horizontal stolons that
with Leishmania spp attach by rhizoids
■ Serological tests and nucleic ■ Sporangiospores in clusters
acid probes available opposite rhizoid
■ Terminate in dark, round
sporangia containing
sporangiospores

CLINICALLY SIGNIFICANT OPPORTUNISTIC

FUNGI

● Rhizopus
○ CLASSIFICATION
■ Zygomycetes
○ SIGNIFICANCE
■ Common laboratory
contaminant
■ Propensity to attack vascular
system of immunocompromised ● Aspergillus
■ Rapidly spreading ○ CLASSIFICATION
■ Often fata ■ Hyaline
○ COLONIAL MORPHOLOGY ○ SIGNIFICANCE
■ Mature within 4 days ■ Common contaminant
■ “Lid litter” ■ Can cause invasive infection,
■ Front is dense cotton candy-like colonization, toxicosis, allergy
growth, white at first, turning ■ A. fumigatus is the most
gray or yellowish brown common spp.
■ Reverse is white ○ COLONIAL MORPHOLOGY

Page 20 | Mycology
MYCOLOGY & VIROLOGY
Credits: MS. ABIGAIL MARIE PEREZ, RMT, MLS (ASCPi), MSMT (CAR)

■Mature within 3 days ■ Large, canoe-shaped,

■Front is fluffy, granular, powdery multiseptated macroconidia
texture. White at first, then color ■ Small oval or cylindrical conidia
depends on species: in singles or cluster on simple
● A. fumigatus - white to conidiophores
blue-green
● A. niger - black
● A. flavus - yellow to
green
● A. terreus - tan to
cinnamon
● Reverse is white,
goldish or brown
○ MICROSCOPIC MORPHOLOGY
■ Septate hyphae, branching at
45॰ angle
■ Unbranched conidiophore arises
from foot cell
■ Expands into large, spherical
vesicle covered with phialides - END -
that produces chains of round
conidia

● Fusarium
○ CLASSIFICATION
■ Hyaline mold
○ SIGNIFICANCE
■ Can be contaminant
■ Can cause eye, skin, nail,
systemic infections
○ COLONIAL MORPHOLOGY
■ Mature within 4 days
■ Front is white & cottony
developing pink or violet center
■ Reverse is light
○ MICROSCOPIC MORPHOLOGY
■ Septate hyphae
■ Unbranched conidiospores

Page 21 | Mycology