micro prax 1- mycology
TRANSCRIPT
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Micro Prax 1- Mycology
Thanks to Renn, Lilia, Kate and Iami for the pictures
Some pictures are from past prax reviewer (batch 2013)Thank you!
Some I got from the internet thank you google!!!
Go Batch 2014!!!!
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Exercise 10- KOH Mount
Potassium hydroxide (KOH) mount
a rapid method for the demonstration of fungal forms in clinical material
facilitates the clearing of specimens for enhanced microscopic observationwithout altering the fungal elements
partially digests proteinaceous components (e.g. host epithelial cells) that are
collected along with the fungi leaves the polysaccharide-containing cell walls intact
***rapid because the specimen placed in a drop of KOH will dissolve at a faster ratethan the fungi
***proteinaceous components are partially digested by the alkali, leaving the cell
walls intact***the chitinous cell walls of fungi protects fungal elements from disintegration
10% KOH- for skin scrapings
20% KOH- for nail scrapings
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Upon potassium hydroxide (KOH)
examination, hyphae are visible and
grow into strands within clumps of
keratinocytes.
Thick-walled spores frequently occur
in grapelike clumps. Individual spores and short stubby
hyphae float in the clear areas
between clumps of keratinocytes.
Many of the short hyphae are
dystrophic.
hyphae
spores
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KOH Mount Skin Scrapings
Hyphal filaments Spores
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Ptyriasis Versicolor
Basidiomycota
Infectious forms: short
hyphae and yeast like
cells Can be visualized by
woods lamp
Culture not done
Scaly patches of
variable color
Mostly endogenous
Lipophilic
Also called as tinea
versicolor, tinea alba
and tinea flava Spread from person to
person
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Exercise 11- Microscopic Morphology
of Fungal CultureAccurate identification of filamentous fungi is based on the microscopic examinationof sporulating parts of a colony, since each species has a characteristic morphologyand arrangement of its spores and fruiting bodies.
Tease Mount Preparation
- traditional procedure used by most laboratories
Principle: To demonstrate conidia or other reproductive structures or morphologicalforms which might give information toward the identification of theorganism.
Slide culture or van Tieghem cell
- best method for preserving and observing the actual structure of a fungus
Principle: In an undisturbed state, important microscopic structures and morphologicdetails are demonstrated
Lactophenol Cotton Blue (LPCB)
- for very quick evaluation of fungal structures
Principles: It has 3 components: lactic acid which preserves fungal structures, cottonblue (an acid dye) which stains the chitin present in the cell walls, and phenol
which kills any live organisms suspended in the stain
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Candida albicans
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Tease Preparation
Candida albicans
Ovoid yeast cells
Note: budding yeast cells (arrow)
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Below:
Candida albicans in its hyphal form
Budding yeast (arrows)
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Phylum: Ascomycota
Representative Genus:Candida albicans
Brief Description:Opaque colored colonies with pasty consistencyAppear as budding yeast cells, psudohyphae, or acombination of both
Reproduction: no sexualAsexual spores chlamydiospores
Hyphae:pseudohyphae occurring in elongated chains
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Epidermophyton floccosum
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Tease Preparation
Epidermophyton floccosum
Smooth, thin-walled macroconidia
which are often produced in clusters
growing directly from the hyphae
Macroconidia
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Numerous chlamydoconidia are
formed in older cultures.
No microconidia are formed.
Chlamydoconidia
Hyphae
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Phylum: Deuteromycota Representative Genus: Epidermophyton floccosum Brief Description:
Colonies are olive green or khaki-colored with powdery surface that become folded with floccose patches as cultureages. Reverse is orange to brownish at times with yellow border
Reproduction: no sexual reproduction
Hyphae: septate Macroconidia:
best seen in young culture- smooth, club-shaped with rounded ends, contain 2-6 cells, found singly or in characteristic
clusters; transform to chlamydiospores Microconidia: absent
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Trichophyton mentagrophyte
Mi idi
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Trichophyton mentagrophyte
Brief Description: Colonies vary greatly. Surface may be buff
and powdery or white and downy;powdery form exhibit concentric andradiate margin
Reproduction: no sexual reproduction, only in anamorph
stage
Hyphae
Microconidia
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Phylum: Deuteromycota
Representative Genus/Species:
Trichophyton mentagrophytes
Hyphae:
coiled spiral hyphae maybe present
Macroconidia: Usually rare, not always present,cigar shaped thin-walled, narrowly attached tothe hyphae and contain 1-6 cells
Microconidia: numeous, borne in clusters alongthe hyphae
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Microsporum gypseum
Mi
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Microsporum gypseum
Brief Description: Colonies re flat and spreading with
powdery to granular surface, irregularly
bifringed border; buff at first, thenbecomes tan to cinnamon brown
Reproduction: No sexual reproduction
Macroconidia
Hyphae
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Phylum: Deuteromycota
Representative Genus: Microsporum gypseum
Hyphae: With septate
Macroconidia: numerous, symmetric with roundedends and having no more than 6 cells
Microconidia: club-shaped and usually seenamong hyphae
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Aspergillus sp.
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Slide Culture Preparation
Aspergillus sp.Vesicle
Conidiophore
Hyphae
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Swollen Vesicle
Conidiophore
Septated
Hyphae
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Phylum: Ascomycota
Representative Genus: Aspergillus sp.
Brief Description:
Smokey green color colonies,
Reproduction:
No sexual reproduction
Hyphae:
Wide, septated,dichotomously branchingwith conidial heads
Classification Accg. To Habitat:
Ubiquitous (soil, plants,organic debris)
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Aspergillosis
Infectious form:
Conidia
Tissue form: Hyphae
Most common species:
A. Fumigatus
Natural reservoir:
Soil and air
Occurs as:
Allergic form
Colonizing form
Disseminated
Mycotoxicosis
In culture:
Conidiophore with
swollen vesicle, rows of
phialides bearing radial
chains
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Penicillium sp.
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Penicillium sp.
Slide Culture Preparation
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Conidiophore
Hyphae
Metullae
Conidia
Metullae
Conidia
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Metullae
Conidiophore
Conidia
Hyphae
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Phylum: Deuteromycota
Representative Genus:Penicillium marneffei
Brief Description:For P. marneffei- the only species that is dimorphicFlat, powdery to velvety, and tan to reddish yellow colonies
Reproduction:no sexual reproduction
Hyphae: septate
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Penicillium
Produce red pigments
Reproduction:transverse fission/schizogony
Unknown habitat
Infectious form:
Conidia
KOH: small ovoid yeastcells with transverseseptum
Culture RT:conidiophores (flaskshape) branch intometullae with phialides
Culture at 37C- Roundyeast cells with crosswalls
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