Virology and Mycology
(MDU4307)
Virology and Mycology (MDU4307)
Yeast
R. S. R. Rajakulasooriya
Department of Medical Laboratory Sciences
R. S. R. Rajakulasooriya
Faculty of Health Sciences, The Open University of Sri Lanka
[Link]. in MLS (USJP)
Dept. of MLS, Faculty of Health Sciences
Open University of Sri Lanka
Date:13th Novmber 2023 1
Yeast
2
Agents
•The common yeasts found to cause infections in
human infections are,
• Candida species
• Candida albicans , Candida tropicalis, Candida
glabrata, Candida parapsilosis, Candida krusei
• Trichosporon species
• Trichosporon beigelii, Trichosporon capitatum
• Saccharomyces species
• Cryptococcus species
• C. neoformans and C. gattii
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Candida species
• Normal microbiota of oral cavity, gastrointestinal
tract and vagina.
• Usually cause mucosal, cutaneous or nail
infections.
• The commoner species are :
• Candida albicans , Candida tropicalis,
Candida
• glabrata, Candida parapsilosis, Candida krusei 4
Candida spp.
• Globose to elongate
yeast-like cells
(blastoconidia)
• Reproduce by narrow-based
multilateral budding
• Pseudohyphae and
occasionally true hyphae
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may also be present
Laboratory Candida spp.
Identification - Candida spp.
Specimens Skin, Nail same as for
: dermatophyte infections
Vaginal and oral swab dipped in sterile
swabs normal saline and
sampled
Blood, CSF, Bronchial sterile containers are
washings, BM used
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Direct
examination
•For skin, Nail and mucus
membrane– 10-30% KOH mount
• Single or budding yeast cells
• Pseudohyphae and true hyphae
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Direct
examination
•Deep seated or systemic infections
• Processing does not require KOH.
• Demonstration of candidal yeast cells and /or
hyphal forms in these specimens are of
importance for diagnosis.
• But presence of fungal elements in specimens
like
• sputum or urine may represent normal
microbiota
• Tissue biopsies can be stained by
histopathological techniques
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Culture - conditions
• Species identification is possible only by culture
• Sabouraud’s dextrose agar (SDS) with
chloremphenicol or cycloheximide
• some Candida species are sensitive to
cycloheximide
• Incubation 🡪 aerobically at 26°C or 37°C up to 2-5
days
• Colonies are smooth, dry, white to cream coloured
about 2-3 mm in diameter.
• The yeast colony is again subjected to a direct
• mount using lactophenol cotton blue
• to study the morphology of the yeast and
• to determine the purity of the isolate 9
Candida species
identification
•GERM tube test
• Presumptive identification of Candida
albicans
• Screening test
• Production of the germ tube within 2
hours in serum is indicative of Candida
albicans
• Germ tubes - initiation of hyphal growth,
arising directly from the yeast cell
• Parallel walls at their point of origin and
are not constricted
• About 30% of the cells should show
germ tube production to be positive 10
Rice Agar
(Corn Meal
Agar) test
C. albicans C. krusei C. tropicalis
• Terminal chlamydospores • Elongated yeasts, • Abundant
abundant pseudohyphae, pine
pseudohyphae forest arrangement,
(match stick like blastoconidia formed
appearance) at or in between
septa
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C. parapsilosis C. glabrata
• Giant hyphae, blastospores • Yeasts only
at nodes
Carbon Assimilation Test
Determines the pattern of sugar assimilation on yeast
nitrogen base (YNB)
11 different sugars
Presence of growth around a sugar is considered as
positive for that particular sugar
CHROMagar test
• Specific enzymes of the different species reacts
with the substrate
• Presumptive identification method 12
• A capsulated yeast
• Causes localized and disseminated
disease which can be an
acute or chronic infection.
• There are 2 types
• C. neoformans var. neoformans
Cryptococcus • C. neoformans var. gatti.
neoformans • Each of these can be divided into two
serological groups A and D and B and
C.
• The 2 varieties differ in their
biochemical properties.
• They do not differ significantly in their
morphological characteristics.
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Laboratory Candida spp.
Identification - Candida spp.
Specimens :
○ Cerebrospinal fluid (CSF) - the most common
○ Blood, sputum, urine, prostatic fluid, other fluids into sterile containers
○ All samples have to be sent to the laboratory as quickly as possible
○ Specially the CSF sample should be sent immediately and in the
laboratory it has to be processed immediately as well
○ CSF centrifuged deposit - for direct microscopy with India ink and for
culture.
○ The supernatant - for the cryptococcal antigen test
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Direct microscopy
•India ink stain
• direct mount of CSF sample in India ink
• The India ink stain mount will show
• encapsulated cryptococcus as a
negative stain.
• The background will be stained a black
colour with the India ink stain and the
• Capsule will be unstained showing the
capsule as a white halo around the yeast
cell
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Culture
• Cultures are on Sabouraud
dextrose agar with
chloramphenicol.
• The colonies are round,
white smooth and mucoid in
appearance.
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Cryptococcus • An important test to the diagnosis of
antigen test Cryptococcosis.
• Commercially available kits are used.
•Principle of the test
• A latex agglutination test that detects the antigen
using latex particles sensitized with anti
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cryptococcal antibodies.
• When the latex particles come in contact with the
polysaccharide antigen of C. neoformans in the
specimen, the two form a complex causing
visible agglutination.
• This test is very sensitive and specific.
• In all patients the antigen test is positive in over
90% of cases.
• False negative results are slightly more common
in non-immunosuppressed patients.
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