APPLIED MICROBIOLOGY, Jan. 1972, p. 174-176 Vol. 23, No.

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Copyright © 1972 American Society for Microbiology Printed in U.S.A.

Inoculation Technique for Fungus Cultures

RAMON M. FUSARO
Department of Dermatology, The University of Nebraska Medical Center, Omaha, Nebraska 68105
Received for publication 23 August 1971

A plastic straw and wood applicator stick serve as a simple, inexpensive, and
disposable inoculation unit for fungal studies. The method gives a uniform and
intact inoculum. The technique is especially useful because a large number of
agar plates can be inoculated rapidly.

To inoculate a large number of Sabouraud
agar plates with a fungus, the standard tech-
niques can be time-consuming and do not al-
ways give uniform (diameter) inoculations.
The following is a description of a simple, in-
expensive, and disposable method.
The materials consist of plastic drinking
straws (4 mm in diameter by 26 cm in length;
Carnival Straws Supra Plastic, National Soda
Straw Co., Chicago, Ill.), which were cut into
thirds, and 6-inch (ca. 15.2 cm) wood appli-
cator sticks. A single applicator stick is in-
serted into each straw and gas-sterilized
(ethylene oxide).
Prior to the inoculation, a hole is made in
the center of the agar with another plastic
straw without an applicator stick (Fig. 1). The
sterile agar core cut by the plastic straw will
remain within the tip of the straw when the
straw is withdrawn by sealing the other open
end of the straw with the finger tip (Fig. 2).
The same straw can then be used to remove 5
to 10 more sterile agar cores from other plates
as the entrapped agar cores act as suction
plugs.
When using the inoculating unit (plastic
straw with wood applicator stick), the middle
portion of the straw is grasped and squeezed to
hold the stick within the straw. The applicator
stick is pulled up into the tube so that its end
is approximately 1 to 2 cm from the distal end
of the plastic straw which will be inserted into
the fungus colony growing on Sabouraud
agar. After punching to the bottom of the plate
through the agar (Fig. 3), the straw is rolled
between the fingers to sever the core. The
straw is then tilted to break the adhesive force
of the agar plug to the petri plate. The unit is
lifted out and the tip of the plastic straw con-
taining the fungus plug is placed into the agar
hole in the recipient plate (Fig. 4). The appli-
cator stick is pushed gently down against the
agar plug while the plastic straw is lifted
slowly out of the agar hole, thereby placing the
inoculum in the agar hole (Fig. 5).
It should be emphasized that the inoculum
is uniform in diameter only, not cell number.
The time necessary to inoculate many plates is
minimal.

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VOL. 23, 1972 NOTES 175

FIG. 3. Inoculation unit cuts out the fungal inoc-

FIG. 1. Plastic straw without an applicator stick ulum by rolling the straw between fingers.
inserted into recipient agar.

FIG. 2. Agar core remains in the tip of the straw

as the index finger prevents entry of air at top of FIG. 4. Tip of the straw containing the fungal
straw. plug is inserted into the agar hole.
176 NOTES APPL. MICROBIOL.

FIG. 5. After pressing out plug and withdrawing

straw in one motion, the fungal inoculum remains in
the agar hole.