Professional Documents
Culture Documents
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
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
MYCOSES
I. SUPERFICIAL MYCOSES
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)
green
on woods light
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)