Lab 5

LAB 5 - MICROBIOLOGY
( ASEPTIC TECHNIQUE )
INTRODUCTION
The microorganisms group are substantially form of bacteria, yeast and fungi as well
as protozoa and viruses. These are most commonly found microorganisms in the surrounding,
capable of settling and living in various environments such as soil, water, plant and animal
tissues also human body. Some simple methods include cultivation and microscopic
techniques could be applied to identify and isolate most of microbiological culture. When
working in a microbiology laboratory, aseptic technique is fundamental to the safety and
success of an experiment. Generally, aseptic techniques performed by scientist under sterile
conditions to ensure that microbial contaminants not doing harm to the colleagues, suplies or
experimental cultures. One of the aseptic technique is streak plate. The streak plate method is
a rapid qualitative isolation method commonly used for isolation of discrete colonies that
require the number of organisms in the inoculums be reduced. Inoculation is performed from
a sector in order to isolate a pure culture. Also in this lab, we are going to determined the
effect of handwashing on bacteria on thumb. People used their hands for variety of activities
everyday so it is extremely easy to come in contact with microbes. Handwashing is thought to
be effective for the prevention of pathogens transmission. However, there is no conclusion
that handwashing with soap is more effective at reducing bacteria contamination than using
water only.
OBJECTIVE
 To produce isolated colonies of an organisms ( bacteria ) on an agar plate using streak
plate technique
 To determine the effect of handwashing using water, handsoap and hand sanitizer on
bacteria.
METHODOLOGY
Part A
1. Firstly, the inoculating loop was steriled by putting the loop into the flame until it is
red hot.
2. Next, lid of the petri dish were raised just enough to pick up isolated colony (S. Aures)
and spread it over the firest quadrant on separated agar plate.
3. Then, the inoculating loop were flamed again until it is red hot.
4. The agar plate were turned aroud 45° and streak the loop into the nest quadrant.
5. Step 4 and 5 were repeated until the fourth quadrant.
6. Lastly, the agar plate were sealed with parafilm and invert it then incubate at 37°C for
24 hours.
Part B
1. Firstly, the agar plate were divided into 4 sections ( control, water, hand sanitizer and
soap )
2. Then, using aseptic technique I pressed my thumb on the control section.
3. Next, I washing my hand with tap water and press my thumb on water section.
4. I repeated step 3 using hand sanitizer and soap.
5. Lastly, the agar plate were sealed with parafilm and invert it and incubate at 37°C for
24 hours.
RESULT
Image 1 : streak plate technique

DISCUSSION
Part A
For this experiment, I choose S. aures as my microbiological culture. Aseptic
techniques are performed to prevent contamination of the growth medium. As we know,
bacteria often occur in mixed populations and very rare to find a single species of bacteria.
So, it is vital to divide the bacteria from the other species that originate in the environment so
that we can study the cultural, physiological and morphological characteristic of the
individual species. The streak plate method is commonly used as isolation method to grow
bacteria on a media surface ( agar plate ) so that individual bacterial colonies are isolated and
can be sampled. When the selected culture media is inoculate using a single isolated colony,
the resulting culture will grow from the selected single colony. This techniques is useful when
we need to separate an organism in a mixed culture. It is also useful when we need to study
the colony morphology of an organism because biochemical test are only valid when
performed on pure culture. In this lab, we must use sterile tool because the crucial aspects is
to maintain a sterile work area surrounding us. Before we start the experiment, it is important
to disinfect all of our work surface using 70% ethanol or any other disinfectant reagent.
Sterile technique is absolutely necessary in many laboratory practical. It is a method that
prevents foreign cells from getting into our plates and cultures. Generally, the key to
succesfull experiment is to work quickly and efficiently otherwise, we will get a contaminant
resulting culture.
Part B
For part B, we going to determined how handwashing can effect the reducing of
bacteria in our hands. In theoretical, handwashing with non-antibacterial is much more
effective in reducing or removing bacteria than only handwashing with water. It is prove
through the policy of many current hand hygiene campaigns promoting the use of soap. There
are many products in the market claim to be able to kill the germs. But, how far is this
statement is agreed to our experimental result ? So, we are going to compare the amount of
bacteria resulting in the 4 section of agar plate, ( control, handwashing with water, hand
sanitizer and soap ). The highest colony of bacteria was found in the control agar plate as
expected. Then, unexpectedly, the water section show the least amount of bacteria present.
Meanwhile, the hand soap show the largest amount of bacteria compared to hand sanitizer.
It is maybe because the soap we are using contained the foreign or contaminant bacteria cells.
Handwashing with water is more likely to be effective for reducing or removing the bacteria
from our hands. It is because the tap water is impossible to be affected or have foreign cell
into it. Actually, the way and the times we take to wash our hands are crucial aspects that we
need to pay attentions. If we want to minimize the bacteria, we must wash our hands
thouroughly and use the best products because the presence of bacteria can be detrimental to
human health and growth.

CONCLUSION
We can obtain the individual colonies from a bacterial culture using streak plate
method. This is ideal for experimentation because the colonies are genetically identical.
When doing a microbiological laboratory, there are many aspects that we need to pay
attentions. Firstly, it is very crucial to maintain a sterile work area surrounding us. This
requires common sense and a continuous practices.
REFERENCES
1. https://microbeonline.com/streak-plate-method-principle-purpose-procedure-results/
2. http://www.austincc.edu/microbugz/streak_isolation.php
3. http://vlab.amrita.edu/?sub=3&brch=73&sim=212&cnt=1
4. https://www.amherst.edu/system/files/media/0782/2010_Manual_3_Sterile_Micropipe
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LAB 5 - MICROBIOLOGY

working in a microbiology laboratory, aseptic technique is fundamental to the safety and

success of an experiment. Generally, aseptic techniques performed by scientist under sterile

everyday so it is extremely easy to come in contact with microbes. Handwashing is thought to

be effective for the prevention of pathogens transmission. However, there is no conclusion

 To produce isolated colonies of an organisms ( bacteria ) on an agar plate using streak

and spread it over the firest quadrant on separated agar plate.

5. Step 4 and 5 were repeated until the fourth quadrant.

2. Then, using aseptic technique I pressed my thumb on the control section.

4. I repeated step 3 using hand sanitizer and soap.

Image 1 : streak plate technique

For this experiment, I choose S. aures as my microbiological culture. Aseptic

techniques are performed to prevent contamination of the growth medium. As we know,

Sterile technique is absolutely necessary in many laboratory practical. It is a method that

bacteria in our hands. In theoretical, handwashing with non-antibacterial is much more

human health and growth.

requires common sense and a continuous practices.

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