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MYCOLOGY

MYCOLOGY-study of fungi

CULTIVATION
-incubated @ 30C, 40-50% relative humidity for
30 days
-all media should contain antibacterial agents
(except CSF)

FUNGI
-eukaryotic
-achlorophylous
-mostly aerobic

1. nucleus
2.mitochondri
a
3. antibiotic
susceptibility

CULTURE MEDIA

FUNGI

BACTERIA

defined
nuclear
membrane
present

no membrane

sensitive to
griseofulvin &
amphotericin
B

sensitive to
penic,
tetracycline &
chloramphenic
ol
only 1 ,not
CHON
associated
pH 7.2-7.6

4.
chromosome

>1, CHON
associated

5. cultivation

pH <6

absent

1. Saboraud dextrose agar -1 isolation of fungal


cultures
-pH 5.6 inhibits bacteria
2. mycosel/mycobiotic agar- 1 recovery of
dermatophytes
3. dermatophyte test medium- indicator:phenol
red
4. Cornmeal agar- stimulates spore formation
(especially for
C. albicans)
5. Czapecks agar- for aspergillus species
6. Niger seed agar- for C. neoformans (brown
pigment)
7. Cottonseed agar- for B. dermatidis
8. Rice medium- differentiates M. canis (+) from
M. audounii (-)
9. Urea agar- fro Cryptococcus sp.

2 morphologic forms of fungi:


1. YEAST
-unicellular & spherical/ellipsoidal in
shape
-colonies: moist, creamy, opaque & pasty
2. MOLDS
-multicelluar & filamentous
-colonies: fluffy, cottony, wooly &
powdery
DIMORPHISM
-most fungi grow only as yeast or molds.
-some may exhibit thermal dimorphism
@ 37C- yeast (parasitic/invasive/tissue
form)
@ 25-30C- molds

structure of molds:
1. HYPHAE
-basic structural units of molds
-branching cylindrical tubules
2. MYCELIUM
-mass of intertwined hyphae
a. vegetative -penetrates media
-absorbs nutrients
b. aerial projects above the surface
-produce asexual spores

types of spores:
1. SEXUAL SPORES
-formed as a result of nuclear fusion
-found in perfect fungi
a. ascospores
b. basidiospores
c. zygospores
d. oospores
2. ASEXUAL SPORES
-formed w/out nuclear fusion
-found in fungi imperfecti
a. thallospores
b. conidia

DIRECT MICROSCOPIC
EXAMINATIONS
1. Temporary mounts
a. 10-20% KOH
-RT for 20-30 minures
-dissolves keratin (epidermal cells)
b. Lactophenol cotton blue
-contains lactic acid 9preserves fungal
structures), phenol (killing agent) & aniline
blue (stain)
c. India ink or Nigrosin
-detection of C. neoformans
-demonstrate capsules through negative
staining
d. Calcoflourwhite stain
-preffered mehod
-fluorescence microscopy (CHO in fungus
wall
stains purplish red)
2. Permanent mounts
a. PAS stain
b. Gram stain
-Hucker modification (CV & ammonium
oxalate)
-all fungi are gram positive
c. Acid fast stain (Kinyoun)
-for Nocardia & B. dermatidis
d. Acridine orange
-for Tinea versicolor
-fungal elements: green fluorescence
epithelial cells: orange
e. Giemsa & Wrights
-for h. capsulatum
f. Gomori Methenamine silver stain & PAP stain
-for histologic examination

SPECIAL TEST
1. Hair Baiting/Perforation test
-(+) conical perforation of hair shaft
-(+) for T. mentagrophytes & (-) for T.
rubrum

2. Exoantigen test
-gel immunodiffusion precipitin test
3. Rapid Urease test
-same as urease test
-(+) control= C. neoformans
(-) control= C. albicans
4. Rapid Nitrate Reductase test
- benzalkonium chloride dissociates
yeast cell wall
-(-) results checked w/ zinc dust
5. Levodopa-Ferric Citrate test
-rapid ID of c. neoformans 9phenol
oxidase)
6. Germ Tube test
-(+) only to C. albicans

palms

h. T. unguinum

DERMATOPHYTID (ID REACTION)


-allergic response to fungal Ags wherein an
infection in one
area elicits an allergic rxn
elsewhere on the body
-(-) microscopic & culture on reaction site &
finding of
dermatophytes elsewhere on the
body

III. SUBCUTANEOUS MYCOSES

MYCOSES
I. SUPERFICIAL MYCOSES

-affects the hair & superficial layers of the skin


(stratum corneum)
-asymptomatic
1. Tinea verscolor
-uneven pigmentation of skin
-caused by Malassezia furfur
-microscopic: spaghetti & meatballs
pattern
-grows on Saborauds overlayed w/ olive
oil
2. Tinea nigra
-faded silver nitrate like stain in palms &
soles
-caused by P. werneckii
3. Piedra
-fungal infection of hair
a. Black piedra
- cuased by Piedraia hortai
-brown black crust outside hair shaft
b. White piedra
- caused by Trichospora beigelii
-light brown nodules in beard

II.CUTANEOUS MYCOSES
-infections involving the superficial keratinized
tissues: skin, hair & nails
-caused by DERMATOPHYTES
-includes 3 genera
1. Epidermophyton-affects skin & nails
a. E. floccosum2. Microsporum- affects skin & hair
a. M. audouinii (antrophilic)
fluoresce
yellow

b. M. canis (zoophilic)

c. Tinea barbae barbers itch


d. Tinea corporis
e. Tinea cruris- jocks itch
f. Tinea pedis- athletes foot
g. Tinea manus- interdigital areas &

green

on woods light

c. M. gypseum (geophilic)- doesnt


fluoresce on
woods light
3.Trichophyton- affects skin, hair & nails
a. T. rubrum- (-) HBT & urease
-teardrop microconidia
b. T. mentagrophytes- (+) HBT & urease
-spiral hyphae
c. T. tonsurans- growth enhanced by
thiamine
-ballon forms microconidia
d. T. verrucosum- rat tail macroconidia
e. T. schoenleinii- favic chandelier
hyphae
(antler type)
-infection usually reffered to as TINEA
(ringworm)
a. Tinea capitis
b. Tinea favosa

-involve the skin & subcutaneous tissue that


may rarely become systemic
1. Sporotrichosis
-aka Rose gardeners disease
-caused by dimorphic Sporothrix schenkii
RT: mold (flowerette conidia)
37C: yeast (cigar-shaped)
2. Chromoblastomycosis
-caused by dematiaceous fungi
(imperfect fungi
that produce melanin-like
pigments)
-on 10% KOH: scerotic bodies (copper
pennies that
exhibit transverse septations)
-cuased by Fonsecaea, Phialophora,
Cladosporium
3. Phaehyphomycosis
-characterized by presence of darkly
pigmented,
septate hyphae in tissues
4. Mycetoma
-aka Madura foot or maduromycosis
-granulomatous tumor involving lower
extremities
-2 types:
a. Actinomycotic- caused by
Nocardia &
Actinomycetes
b. Eumycotic- cuased by true
fungi,
Exophiala & P. boydii
-Pseudoallescheria boydii- most common
cause of
mycetoma
5. Rhinosporidoiosis
-polypoid masses of the nasal mucosa
-caused by Rhinosporidium seeberi
6. Lobomycosis
-chronic infection of humans & dolphins
-caused by Loboa loboi

IV. SYSTEMIC or DEEP MYCOSES

-involve any of the internal organs


-caused by inhalation of dimorphic fungi
-exoantigen test is the most conclusive method
for dimorphic fungi
1. Blastomycosis
-aka North American
blastomycosis/Gilchrist dis.
-caused by Blastomyces dermatidis
-RT: lollipop conidia
-37C: oval w/ single broad base body
-ExoAg test: specific A band
2. Paracoccidioidomycosis
-aka South American blastomycosis
-cuased by Paracoccidioides brasiliensis
-RT: chlamydoconidia
-37C: yeast cell w/ multiple buds
: pilot/mariner/ship wheel!
-ExoAg test:

3. Histplasmosis
-aka Darlings disease
-caused by Histoplasma capsulatum
-RT: tuberculate/pyriform macroconidia
-37C: small budding yeast cell w/in
macrophages -ExoAg test: H &/or M bands
4. Coccidiomycosis
-aka San Joaquin Valley fever/ desert
rheumatism
-caused by Coccidiodis immitis
-major biohazard to lab personnel
-RT: arthroconidia
-37C: spherule thick walled structure
filled w/
endospores (not a yeast)

V. OPPORTUNISTIC MYCOSES
-usually do not cause disease
-occur only in immunocompromised patients
1. Candidiasis
-causes trush(oral), moniliasis
(vaginal),esophagitis,
etc. may
also cause systemic infection
-caused by C. albicans & other Candida
species
-C. albicans= (+) for germ tube (hyphal
extension
w/out constriction)
2. Cryptococcosis
-aka Torula meningitis
-caused by C. neoformans (an
encapsulated yeast
cell in bird & fat
droppings)
-urease & inositol (+)
-nitrate (-)
-brown pigment when cultured on
birdseed/niger
seed/staibs
:media=SDA w/o cycloheximide
3. Aspergillosis
-commonly caused by A. fumigates
followed by
A. niger & A. flavus
-fungus ball in lungs
3. Zygomycosis
-caused by Zygomycetes (Mucor,
Rhizopus &
Absidia)