Candida albicans

Candida albicans (Yeast)

(Note: For Candida dubliniensis, scroll to bottom of this post.)

I thought I’d start off my return from ‘sick leave’ by checking out the lab’s new digital camera.
Just back, I have no “exotic” isolates to play with and explore so I’ve just retrieved a common,
everyday Candida albicans. I’ve often, been disappointed in photographs found in most
textbooks as they are small in size, black & white, poorly focused and often scrunched between a
few lines of description. This yeast deserves a bit more respect as it is the most common cause of
candidiasis, an infection which can range from acute, sub-acute or chronic. It may be present,
yet unrecognized or have devastating, life threatening consequences. C.albicans may be present
as commensal flora in the normal mouth, the skin and in the stool. Problems arise when, for what
ever reason, the microbial balance is upset and Candida is allowed to overgrow other organisms
which may assist in keep it in check. Antibiotic therapy may lessen normal bacterial flora
allowing Candida to flourish. A change in the pH of the healthy vagina may result in a yeast
infection. Immunosuppressive therapy may predispose a patient to infections by yeast.

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.
 ***

Candida albicans retaining crystal

violet stain from routine gram stain taken from SAB Agar
(click on photos to enlarge for better viewing) X1000

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.

Refractile Chlamydospores and smaller 'grape-

like' Blastoconidia (Blastospores) produced on Oxgal Agar and photographed through the
microscope cover slip with the agar plate placed on the microscope stage.
(click on photo to enlarge for better viewing)

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)

Below are a few photos of Candida albicans as seen in actual specimens;

Candida albicans in a vaginal

swab seen in the form of a pseudohyphae and a few individual oval cells. (Gram Stain X1000)
(click on photo to enlarge for better viewing)
 Candida albicans seen as a
pseudohyphae at lower left and individual yeast cells at upper right.
(Gram Stain of Sputum specimen X1000)
(click on photo to enlarge for better viewing)

Candida albicans pseudohyphae

and individual cells seen in an aspirate from a kidney.
Depending of various factors, individual yeast cells, pseudohyphae, or both may be present in
any given specimen.
(click on photo to enlarge for better viewing)

New: April 01, 2012

An interesting photo of a fungus as it appeared in a direct blood culture gram stain. Identification
proved it to be Candida albicans. Just had to share my beautifully bizarre photo - hard to imagine
this spiny ball of spikes bouncing around inside someones blood vessels!
 Candida albicans in direct smear
of Positive Blood Culture.
(Usually seen as the round to oval cellular form and not as these clumps of pseudohyphae)

* * *
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.

• Both yeasts are virtually identical in their physiological characteristics

• Carbohydrate assimilation is virtually identical ( some variation +/- vs -/+ of little use in
identification or differentiation)

• Glucose fermentation – both positive

• Chlamydospore production – both positive

• Germ tube production – both positive

• Urease – both negative

• Nitrate – both negative

• Cyclohexamide (0.04%) – both positive for growth.

• Growth at 30oC & 37oC -both positive

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.

Aspergillus niger on Sabouraud-

Dextrose Agar 72 hrs at 30C
 Black pigment from conidia maturing from center of colony
(click on photo to enlarge for better viewing)

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.

Large vesicle at end of broken

conidiphore bearing metulae & phialides with black pigmented conidia already dispersed. Adhesive tape
preparation was too disruptive in capturing structures intact. (LPCB X400)

Ditto (LPCB X400)

(Click on photos to enlarge for better viewing)
 Aspergillus niger (LPCB X400)

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)

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