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

tting.pdf