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Macroscopic Morphology:
Candida albicans grows rapidly in culture, reaching maturity in as little as three days. Colonies
are cream coloured, raised, entire, smooth & butyrous. On enriched media such as Blood Agar,
or Chocolate Agar, the colonies may develop small striations or outgrowths often referred to as
“feet” which are indicative of the Candida albicans species.
Candida albicans on Sabouraud-
Dextrose Agar at 48 hours at 30C
(click on photo and illustration at right to enlarge for better viewing)
Microscopic Morphology:
A smear made from colonies taken from Sabouraud-Dextrose Agar, or blood agar will appear as
round to oval cells about 4 to 8 µm. Though the cell wall structure differs from that of gram
positive bacteria, yeast cells retain the crystal-violet stain of the routine gram stain and therefore
appear purple. The yeast cell divides by budding. On primary media (reduced nutritionally) the
budding can create elongated cells which when lined up along the dividing plane, mimic the
appearance of a hyphae however these inline individual cells are referred to as a pseudohyphae
(false hyphae). Some true hyphae may also be formed.
Along side of the pseudohyphae, Candida albicans develops blastoconidia around the area of
the ‘septa’ (division). These appear as smaller round ‘grape-like’ clusters.
***
Another structure is the thick walled, rather refractle chlamydospore which usually develops at
the end of the pseudohyphae (terminal).
Refractile Chlamydospore
production on Corn Meal Agar at 2 hrs at 30C
(click on photo to enlarge for better viewing)
Technique: Chlamydospores production is best induced using the Corn Meal Agar (CMA) or
Oxgall Agar. Inoculate the plate by picking up a sample of the Candida albicans colony with a
straight wire and scratching the surface of the agar with it. With the surface inoculated, cover the
scratched area with a microscope cover slip. This has a two-fold purpose. The glass cover slip
reduces the atmospheric tension under the surface and protects the microscope objective from
damage when viewing the plate after incubation. A filter paper moistened with sterile water and
incubation at room temperature in the dark, may further encourage the production of
chlamydospores After incubation (24-48 hours) remove the petrie dish plate cover (and
microscope stage if you wish) and view the growth by carefully lowering the objective over the
cover-slipped agar (X100-250). Viewing the growth around the edge of the glass cover slip
should yield the best results.
Yet another characteristic of Candida albicans is that it has the ability to produce 'Germ Tubes'
when placed in a nutritionally rich horse serum. Inoculate about 1 to 2 ml of horse serum and
incubate at 37C for about 2 to 3 hours (too long may result in the formation of pseudohyphae,
mimicking germ tubes) . Examine under the light microscope for a protrusion growing out from
the yeast cell. Germ tubes appear as outgrowths from the side of the yeast cell and although
characteristic of Candida albicans, be aware that the closely related Candida dublinensis can
also produce germ tubes. Other characteristics which won't be discussed here can easily be used
to separate these two species. Germ tubes protrude from the originating cell with no "pinching"
seen at the point where they extend. If there is evidence of pinching, this may not be a germ tube
but rather the beginning pseudohyphal growth.
Candida albicans germ tube
production in Horse Serum at 37C after 3 hours incubation.
Note; there is no constriction/pinching where the germ tube leaves the originating cell indicating
that this is indeed a germ tube and not the beginning of a pseudohyphae. (wet prep X400)
(click on photo to enlarge for better viewing)
* * *
New: April 18th, 2015
Candida albicans vs Candida dubliniensis
Okay, you think you have isolated Candida albicans. But is it really?....and why should you
care? Well, Candida albicans & Candida dubliniensis are virtually indistinguishable using
common physiological tests (see list which follows). However, there may be one significant
difference which should be considered. Candida dubliniensis may exhibit increased resistance to
the anti-fungal agent Fluconazole. This is of particular concern when this species is isolated
from sterile sites such as blood cultures where delayed or failure of treatment may be fatal.
• Carbohydrate assimilation is virtually identical ( some variation +/- vs -/+ of little use in
identification or differentiation)
While some commercial identification systems claim they can distinguish between the two
species, this is a costlier solution to a problem that can be solved using incubation temperature
alone.
C.dubliniensis
• Growth at 42oC to 45oC = -/w
• Fluconazole –may exhibit increased resistance.
C.albicans
• Growth at 42oC to 45oC = +/-
• Fluconazole – generally considered susceptible.
Sunday, 13 June 2010
Aspergillus niger
(Fillamentous fungi)
Subgenus: Circumdati, Section: Nigri
This fungus was a bit of a challenge to photograph as it matured so quickly that conidia were often well
dispersed when I was prepared to photograph the culture. It was difficult to find really good examples of
the vesicles bearing metulae, phialides bearing intact conidial spores. Both adhesive tape preparations
and slide cultures were studied.
Ecology; Ubiquitous, worldwide distribution, commonly found in mesophilic environments. One of the
most common of the fungi in soil, rotting fruit & plant matter as well as many indoor environments.
A.niger may be found as a common laboratory contaminant.
Macroscopic; Rapidly growing on Saboraud-Dextrose Agar starting with a white to yellowish felt-like mat
of mycelia, quickly turning black as conida develop the pigment aspergillin during maturation. Reverse
remains white to pale in colour.
Microscopic; Septate, hyaline (clear) hyphae. Conidiophores (Stipes) are long (400-3000 µm) with
spherical vesicles at the apex measuring 30-75 µm. Aspergillus niger is biserate - metulae just about
cover the entire surface from which the phialides extend. Conidia are globose, brown to black in colour,
measure 3.5-4.5 µm in diameter and have a rough surface.
Pathogenicity; not tremendously pathogenic however has been implicated in aspergillosis particularly in
immunocompromised patients. Has also been isolated from ear infections (otomycosis), often as a
secondary invader, establishing itself after/on top of a bacterial infection.
Industry; A.niger produces a number of useful enzymes which have been utilized by industry in the
production of a variety of products such as citric acid. The possibility of A.niger being capable of
producing mycotoxins remains controversial.
Treatment; Studies on the susceptibility of A.niger to antifungal agents remain inadequate. Isolates
should be tested individually if therapy is warranted. Voriconizole as been shown to be effective in
documented cases as well as Itraconazole and Amphoterecin B.
Intended as Aspergillus niger
computer wallpaper (1024 X 768) when posted.
(click on photo to enlarge for better viewing)