a l T e s t

Sp ec i
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for f
Germ tube
and hair ba
technique iting
Objectives Familiarize with the special tests used
to help identify fungi species

Understand the principles of the

special tests
Germ Tube test
To identify Candida albicans from other fungal species
 Principle
Formation of germ tube is associated with increased synthesis of protein and
ribonucleic acid. Germ Tube solutions contains tryptic soy broth and fetal bovine
serum, essential nutrients for protein synthesis. It is lyophilized for stability. Germ tube
is one of the virulence factors of Candida albicans. This is a rapid test for the
presumptive identification of C. albicans.
Candida albicans
Candida albicans is part of our natural microflora — or the microorganisms that
commonly live in or on our bodies. It can be found in the GI tract, the mouth, and the
vagina.

Most of the time it causes no issues, but it’s possible for overgrowths and infections to
happen.
Candida albicans is the most prevalent cause of fungal infections in people. Its species
name, albicans, comes from the Latin word for “white.” The yeast appears white when
cultured on a plate. And in the case of certain infections, like thrush, it can create white
patches.
Candida albicans
is the most
common cause of
genital yeast
infections.
Normally, a type of bacteria called Lactobacillus keeps the amount of Candida in the
genital area under control. However, when Lactobacillus levels are disrupted in some
way, Candida can overgrow and cause an infection
You can also develop a Candida genital infection after participating in certain sexual
activities, particularly those that involve oral-genital contact.
Although otherwise ● People that have taken
antibiotics recently
healthy individuals can ● People with uncontrolled
diabetes
get genital Candida ● Immunosuppressed individuals
● Pregnant women
infections, the ● People that are taking oral
contraceptives or who are on
following groups are at hormone therapy

an increased risk:
 Oral thrush
Despite being a normal part of the microflora of your mouth, Candida albicans can
cause infections if it overgrows. The infection may not be limited to just your mouth. It
can spread to your tonsils and the back of your throat as well.

Severe infections may spread to the esophagus.

People that are at an increased risk for developing oral thrush include:

• those taking antibiotics or corticosteroid drugs

• someone with undiagnosed or uncontrolled diabetes
• immunosuppressed individuals
• those who wear dentures, particularly upper dentures
Oral thrush
 Mucocutaneous candidiasis
Candida species can also infect your skin and mucus membranes.

Candida albicans is most often the cause of a fungal skin infection, although other
Candida strains can also cause it.
Areas that are warm, moist, or sweaty provide good environments for yeast to thrive.
Examples of such areas include the armpits, groin, the skin between your fingers and
toes, the corners of your mouth, and the area under your breasts.
Other risk factors for developing a Candida skin infection include:

• wearing tight or synthetic undergarments

• having poor hygiene or changing undergarments infrequently, including
infrequent diaper changes for infants
• taking antibiotics or corticosteroid drugs
• having diabetes
• having a weakened immune system
An example of a mucocutaneous candidiasis
infection
Procedure of
Germ Tube Test
1. Put 0.5 ml of sheep or human serum into a small tube.
2. Note: Fetal bovine serum can also be used instead of human serum.
3. Using a Pasteur pipette, touch a colony of yeast and gently emulsify it in the
serum.
4. Note: Too large of an inoculum will inhibit germ tube formation.
5. Incubated the tube at 37°C for 2 to 4 hours.
6. Transfer a drop of the serum to a slide for examination.
7. Coverslip and examine microscopically under low and high power objectives.
 Results and
Interpretation of
Germ Tube Test
Positive Test: A short hyphal (filamentous) extension arising laterally from a yeast cell,
with no constriction at the point of origin. Germ tube is half the width and 3 to 4 times
the length of the yeast cell and there is no presence of nucleus.
Negative Test: No hyphal (filamentous) extension arising from a yeast cell or a short
hyphal extension constricted at the point of origin
Hair baiting
technique
It is used to distinguish between isolates of dermatophytes, particularly Trichophyton
mentagrophytes and its variants.
 Principle
The technique makes use of the isolation of keratinophilic fungi from soil.
Keratinophilic fungi are those fungi which inhabit soils rich in keratin. These fungi
grow keratin substrates and degrade them. Keratin is found in hair, nails, hooves, skin
and horns etc.
 Trichophyton Mentagrophytes
Trichophyton mentagrophytes is primarily a zoophilic dermatophyte that often attacks
humans and may also survive saprophytically in the soil.
This fungus is known to cause a skin infection known as Dermatophytosis or
Ringworm which appears on a person’s skin as an inflamed circular pattern. The
invasion of the skin, hair, and nails can cause diseases such as tinea pedis or athlete’s
foot.
 Tinea pedis
Athlete's foot (tinea pedis) is a fungal skin infection that usually begins between the
toes. It commonly occurs in people whose feet have become very sweaty while
confined within tight-fitting shoes.
Signs and symptoms of athlete's foot include an itchy, scaly rash. The condition is
contagious and can be spread via contaminated floors, towels or clothing.
 Ringworm
Ringworm is a common skin infection that is caused by a fungus. It’s called
“ringworm” because it can cause a circular rash (shaped like a ring) that is usually red
and itchy. Anyone can get ringworm. The fungi that cause this infection can live on
skin, surfaces, and on household items such as clothing, towels, and bedding.
Procedure
1. Collect soil in sterile polythene bag.
2. Bring it to the laboratory.
3. Half fill the Petri plates with soil.
4. Moisten it with sterile distil water.
5. Place sterile strands of hair on the soil
6. Leave it covered in paper or dark place for 4
7. Weeks
8. Observe for fuzzy white mycelium growing on pieces of hair, record colonial
morphology.
9. Microscopic examination of some pieces using a wet mount smear Lactophenol
Cotton Blue