FUNGAL SMEAR AND CULTURE

Preparation of KOH from the following specimen:

Blood Wright’s or Giemsa’s stain; India ink or direct antigen test for
Cryptococcus; 5 ml blood to 20 ml broth
Bone marrow aspirate 0.5 ml of marrow with sterile syringe in heparin
anticoagulant

CSF Collect in sterile tubes. If volume is more than 2 ml,

centrifugation and membrane filtration are recommended.
Use sediment for slide and plating if volume is less than 2 ml,
prepare smears and plates from uncentrifuged specimen.

Cutaneous hair Pluck by roots with sterile forceps. Select hairs that fluoresce
or are broken and scally

Skin scraping Clean skin with 70% alcohol. Scrape outer edge of ring in
cases of suspected ringworm; scrape area of active infection if
no ring. Place in sterile petri plate, & examine microscopically
using KOH.
Prepare a glass slide, a few drops of KOH will be
added to the skin sample, then a coverslip will be placed over
the top. Examine microscopically.
o 10% KOH – soft tissues (skin scrapings)
o 20% KOH – hard tissues (hairs, nails)

Nail Clean nails with 70% alcohol. Scrape area; dispose of outer
layer and collect inner, infected mail; may also collect using
nail clipper. Place nail in sterile petri plate. Cut nail into small
pieces and examine microscopically using KOH
Sputum, bronchial Should be collected during early morning on 3 consecutive
washings, morning days. Avoid 24-hour collections because of
transtracheal contamination and overgrowth. Expectorated sputum. Collect
aspirates in sterile saline aspirate.
Throat Collect with 2 sterile swabs. Scrape off and collect material
with tongue depressor
Urine Clean catch midstream of first morning void into a sterile
container. Centrifuge and use sediment for microscopic
examination. Process within 2 hours or refrigerate to avoid
bacterial overgrowth
Vaginal, cervical Collect with 2 swabs and place in transport media. Prepare
slide from one swab, and examine microscopically.

SITE BLOOD BORNE PATHOGENS

Blood/bone marrow Histoplasma capsulatum, Cryptococcus neoformans, Candida
albicans, Blastomyces dermatitidis,
Malassezia furfur
CSF Cryptococcus neoformans, Coccidioides immitis, Histoplasma
capsulatum, Candida spp
Hair Trichophyton, Microsporum
Skin Candida, Trichophyton, Microsporum, Epidermophyton
Nail Trichophyton, Epidermophyton, Candida, Aspergillus
Sputum, bronchial Candida, Aspergillus, Rhizopus, Mucor, Penicillium, Blastomyces
washings, dermatitidis, Coccidioides immitis,
transtracheal Paracoccidioides brasiliensis, Histoplasma capsulatum, Sporothrix
aspirates schenckii
Throat Candida albicans, Geotrichum candidum
Urine Candida, Blastomyces dermatitidis, Coccidioides immitis,
Histoplasma capsulatum, Cryptococcus
neoformans
Cervical, vaginal Candida

Common structures in Mycology

 Hyphae – tubelike structures; fundamental units of fungus; many hyphae join to
form the mycelium, which forms the colony of the fungus
o Septate Hyphae – contains cross-walls or septa
o Aseptate Hyphae – lacks cross-walls
 Yeast cells – clusters of round cells, budding forms
 Arthoconidia – presence of spores, asexual spores formed by fragmentation of
mycelia into rectangular, barrel-shaped or cask-shaped, thick walled spores
 Fusiform – spindle-shaped conidium that is wider in the middle & narrows
toward either end
 Pseudohyphae – chain of cells produced by budding that may resemble true
hyphae. Constricted at the septa & form branches that begin at the septation.
Mass of pseudohyphae is pseudomycelium
 Ascopores – small spores produced in a round, saclike ascus that contains 2-8
ascospores
 Germ tube – no constriction. 3-4 times the length of the original yeast cell
 Piriform – pear-shaped conidia
 Spherule – large, round, thick walled
 Tuberculate – with knoblike projections