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1. MICROBIOLOGY PTACTICAL
REPORT
ASEPTIC TECHNIQUES PRACTICAL
NAME: NICOL
SURNAME: MALULEKE
STUDENT NO : 217013244

2. ASEPTIC TECHNIQUES: TRANSFER OF BACTERIA
INTRODUCTION
Many methods and tools used in the modern pharmaceutical microbiology laboratory
were derived in the nineteenth century. (Cappuccino & Sharma,2005). Microbiological
culture Is a method of multiplying microbial organisms by letting them reproduce in the
predetermine cultures media under controlled laboratory conditions. (Kransher,2013),
microbiological cultures are foundations and basic diagnostic methods used extensively
as a research tool in the molecular biology.
Aseptic technique (Slayers & Whith,2002) is a fundamental and important laboratory skill
in the field of microbiology. Microbiologists use aseptic technique for a variety of
procedures such as transferring cultures, inoculating media, isolation of pure cultures,
and for performing microbiological tests. Proper aseptic technique prevents
contamination of cultures from foreign bacteria inherent in the environment. For example,
airborne microorganisms including fungi, microbes picked up from the researcher’s body,
the lab bench-top or other surfaces, microbes found in dust, as well as microbes found
on unsterilized glassware and equipment, etc.
Flaming the loop: Holding the loop in the flame of the Bunsen burner kills all
contaminating organisms and it is sterilizing the loop. The loop should become red for a
few seconds. After flaming, make sure to slightly cool the loop before picking up
organisms from the culture that is to be transferred. (Brown,2005)
When transferring a culture from a plate (Greenword. at al,2017), cool the loop by
touching on the very edge of agar. When transferring from a broth, the red-hot loop will
make a sizzling noise as soon as you insert it into the culture. The loop will automatically
cool once it contacts the broth culture, but wait a one or two second before removing the
loopful of inoculum from the tube. The hot loop may create aerosols when it touches the
media containing microorganisms. It will cause some of the broth and bacteria to boil
briefly, creating a bacteria-containing aerosol. This airborne bacterium has the chances
of entering into the respiratory tract or into the body parts.

3. Flaming the Mouth of the Test Tube: when you put the mouth of a tube through the
flame of a Bunsen burner creates a convection current which forces air out of the
tube. This prevents airborne contaminants from entering the tube. The heat of the
Bunsen burner also causes the air around your work area to rise, reducing the chance of
airborne microorganisms contaminating your cultures. (Black,2015)
An agar plate (Parker,2017) is a Petri dish that contains a solid growth of microorganisms,
used to culture small organisms such as microorganisms. Sometimes selective
compounds are added to influence growth, such as antibiotics.
The purpose of a streak plate is to produce colonies of a certain organism. These colonies
are formed by encouraging the growth of only one organism, which allows the separation
of that organism from a sample containing multiple organisms. (Pommorville,2007)
AIM :HOW TO TRANSFER BACTEREA TO ANOTHER
MADEA

4. MATERIALS
Inoculation
Bunsen burner
Lighter or matches
Relevant solid cultures media in sterile Petri dishes e.g. nutrient ager
Relevant broth/solid culture
Incubator set at 37 C
METHODS
Following thorough decontamination of bench surfaces and observation of all laboratory
safety rules, Bunsen burner was ignited to obtain a flame for sterilizing a wire loop. When
using a broth, culture was being mixed thoroughly. The loop was sterilized over the flame
until it become red hot. The loop could cool down and cap was removed carefully from
the test tube. The mouth of the tube with the culture was flamed. The sterile loop was dip
into broth culture and transferred loopful of the culture onto sterile solid culture media.
Mouth of the culture was flamed and the cap was replaced. Petri dish lid was replaced.
Again the wire loop was fumed until it become red. The loop was cooled on the side of
media and made a pool by spreading culture in a small area of culture media. Loop was
flamed again. Base of Petri dish was rotated on the bench and cooled the loop on the
side of media. Four parallel lines was made at an angle from the pool to the circumference
of the plate. The plate was covered, the loop was flamed and rotated the plate same

5. direction as before cool loop on the side media and made four parallel lines to the
circumference at an angle as before. Again, the loop was flamed and the plate was rotated
and the loop was cooled on the side of plate one streak was made from the second set
of lines careful enough not to touch the original pool and the loop was flamed. Inoculated
culture media was putted into a thermos controlled incubator with the plate facing down
for 24 -48 hours. Lastly the surfaces were decontaminated and all the accessories were
returned to their proper storage areas and hands were washed.
RESULTS
With the use of aseptic techniques to transfer microorganism to one plate to another
helped us to see that it can also grow to another plate. the experiment was done and the
bacteria was incubated for 24 hours and after that time the bacteria growth was found in
new plate. on the plate there were colonies and some colonies were in the lines were
strap. On the nutrient ager on the line was strap the growth was align to the line .and the
nutrient slant the tube was blurry to show that the was growth.
.

6. Bleary vision (growth)
Growth
Heavy growth Colonies
Heavy confluent
growth