Evans Jarrel C.

Dion 19-2DMT
Microbiology: Inoculating Materials
Definition of Terms:
*Inoculate- to introduce or insert a bacteria
into a medium by various means.
*Inoculum-the active material used in an
inoculation.
*Aseptic Techniques- practices to reduce
the likelihood of bacterial contamination.
Materials:
*Inoculation Loop
-also called a smear loop, inoculation wand,
or microstreaker.
-used to retrieve an inoculum from a culture
of microorganisms.
-Diameter: 5mm
-Platinum/Nichrome
*Serological Pipette
*Disposable Transfer Pipette
*Pasteur Pipette
*Disposable Inoculating Loop
*Spreading Rod
Basic Techniques:
*Flaming the Loop Wire
-Inceneration of an inoculating loop wire is
done by passing it through the tip of the
flames inner core of the Bunsen burner.
Begin at the wires base and continue to the
end making sure that all parts are heated to
a uniform orange color. Allow the wire to cool
before touching it or placing it on the
culture.
*Removing the Cap
-Keep the loop hand still. Separate the cap
and the tube with the hand not holding the
loop wire. Always hold the cap and never
place it anywhere else.
*Flaming the tube
-Hold the open tube at an angle to minimize
the chance that airborne microbes will drop
into it. Quickly pass the tubes mouth
through the flame a couple of times.
Specific Transfer Methods:
Obtaining the sample:
*From a Broth
-suspend the bacteria on the broth using a
vortex mixer or by agitating using hands.
-perform three basic techniques

-obtain sample. Keep the loop hand still. Do

not let the wire touch the tube lip to avoid
bacterial aerosols.
*From a Slant
-perform three basic techniques.
-With the agar surface facing upward, hold
the open tube at an angle to prevent
airborne contamination.
-obtain sample. Keep the loop hand still. Do
not let the loop wire touch the tube lip to
avoid bacterial aerosols.
*From an Agar Plate
-flame the loop.
-lift the lid of the agar plate, but continue to
use it as a cover to prevent contamination
from above.
-obtain a small amount of bacterial growth
by gently touching a colony with a wire tip.
Transferring to a Sterile Culture Medium:
*Broth tubes
-broth cultures are often used to grow
cultures for use when fresh cultures or large
number of cells are desired.
1. Remove the cap
2. Sterilize the tube
3. Hold the open tube on an angle to
minimize airborne contamination
4. With the agar surface facing upward,
carefully move the over the wire. Gently
touch the loop to the agar surface near the
base.
5. Beginning at the bottom of the exposed
agar surface, drag the loop in a zigzag
pattern(Fishtail) as the tube is withdrawn.
*Petri Dish-Fishtail Inoculation
Inoculation of Sterile Mediums:
*Put the medium in an incubator.
*Fix temperature:
-37 degrees Celsius (human body
temperature)
-25 degrees Celsius (bacteria from the
environment)
-lower incubation temperatures results to
possible pathogens (discourage growth)
Results:
-Cultures are usually examined after 24
hours of incubation
-Liquid media such as broth become cloudy
if bacteria are present. This could be the
result of only one bacterial cell originally

entering the medium, then dividing

repeatedly to produce millions.
-Bacteria on agar plates become visible as
distinct circular colonies. Each colony should
represent an individual bacterial cell, which
has divided repeatedly, but being kept in
one place, the resulting cells have
accumulated to for a visible patch.

Evans Jarrel C. Dion 19-2DMT

Quiz on Microbiology: Inoculating
Materials
1. The visible result of the
inoculating loop wire after
equally heating all its parts. A.
orange
B. Prussian blue
C. metallic black
2. The size or diameter of the
inoculation loop wire.
A. 3mm
B. 5mm
C. 8mm
3. These are often used to grow
cultures when fresh cultures are
desired.
A. Fishtail Inoculation
B. Agar Slants
C. Broth Cultures
4. Bacterias on agar plates
become visible as .
A. Cylindrical Colonies
B. Circular Colonies
C. Dome-shaped Colonies

5. This is generally used for

growing stock cultures that can
be refrigerated after incubation
for several weeks.
A. Agar Slants
B. Inoculums
C. Formalin
6. The visible result when different
kinds of bacterias are present in
a liquid media.
A. Cloudy
B. Turbid
C. Clear
7. The duration or time needed
before a media is examined.
A. 1 week
B. 24 hours
C. 6 months
8. This is to introduce or insert a
bacteria in the media.
A. Incineration
B. Insertion
C. Inoculation
9. The ff are Basic Techniques in
Inoculating Materials except.
A. Incineration of the
Microstreaker
B. Sterilization of the
Culture Media
C. Removal of the Cap
10.Temperature of bacteria needed
from the environment.
A. 37 degrees Celsius
B. 25 degrees Celsius
C. 14 degrees Celsius