ABSTRACT

In our daily life, bacteria are virtually ubiquitous. They are everywhere. They are in air we
breathe, the food we eat and the water we drink. The objective of the experiment is
determining the effects of various environmental exposures to the sterile bacteriological
media. The methodology employed is swabbing the sample and transferring in to the agar
medium. At the end of the experiment, it is observed that the bacteria are most abundant on
the sole of the shoes compared to the body sample. There are many colonies of bacteria with
difference sizes, shapes and margin in the sole of the shoes. In conclusion, bacteria are
present everywhere.

INTRODUCTION
Microorganism or microbe is an organism that is microscopic which mean too small to be
seen by the naked eyes however visible under the microscope. For instance bacteria, fungi
and protists are classified as non-living microorganisms but not viruses and prions. The term
ubiquitous means found everywhere, therefore the ubiquity of microorganisms is the concept
that microorganisms can be found everywhere. Besides, microorganisms can be found in
wide variety of environment like in air, on land, plants, animals and in fresh or salt water
environments. Bacteria able to live everywhere because they are all well adapted to all
different type of environment and can colonize that particular environment. For example,
according to Oracle Think Quest Education Foundation, it stated that Cyanobacteria Nostoc
lives on the land and forms in filaments of hyphae that hold the microbial mat of lichen
together.
In microbiology lab, lots of time and energy is spent in order to culture the desired microbes.
Therefore to obtain pure cultures from these mixed colonies in the environment, we used
streak plate technique to obtain pure culture from sole in foot. Microorganism from mouth is
obtained from coughing at the surface of agar plate and at the shoe sole. There are two forms
of bacteriological media will be used to culture the microorganisms which are agar medium
and broth medium. The only difference between those two is the presence or absence of the
complex polysaccharide called agar, a solidifying agent purified from red algae. Agar-agar is
not a nutrient because it contain 1.52% agar for the solidified media so giving a solid surface
for the bacteria to grow on and could not be utilized by the organisms. Meanwhile the
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presence of bacteria will produce turbidity in broth culture. A mixed culture from most of
specimen can be seen based on different colony sizes, shapes, colours and many more.

OBJECTIVE
1. To compare the growth of bacteria from different environments.
2. To identify the ways that bacteria grow on and in bacteriological media.
3. To become accustomed with different type of media.

THEORY
The term ubiquity of microorganisms can be defined as everywhere. That means,
microorganisms can be found nearly everywhere on the planet. Microorganisms include
bacteria, archaea, algae, protists, protozoa and viruses. According to Hans G. Schlegel and
Holger W. Jannasch (2006), the three major factors that contribute to the ubiquity of
microorganisms are their small size that can be dispersed easily by air and water, their
metabolic versatility and flexibility, and their ability to tolerate the unfavorable conditions.
For example, some archaean can live in hot spring rich in Sulfur.
Besides, microbes especially bacteria are widely distributed in the human body and on
surfaces of any materials and tools. Therefore, to understand the concept of ubiquity of
microorganisms, an experiment should be conducted by taking the specimen from the body
and from the environment. In this experiment, the body sample was in the form of a cough,
and the environmental sample was taken from the sole of a shoe. Bacteria can be spread
through coughing, because according to Marshall Cavendish (2008), they are often present in
saliva, in the mucus from the nose and lungs which can be sent through an airborne manner.
On the other hand, there were many bacteria which were picked by shoes when walking. This
is because, the bacteria that is already present on the soil, toilet floor tiles and on the road or
any surfaces that we step on are transmitted to the shoe soles as we are walking on them.
Culture media Trypticase Soy Agar (TSA) and Trypticase Soy Broth (TSB) are used to isolate
and observe the growth of bacteria. These media contain casein and soy products that support
the growth of almost all microorganisms, and is a general purpose agar of sorts.
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After incubation for 24 hours, the colonies of microorganisms are visible to naked eye.
Therefore, the general form of the colony and the shape of margin of the colony could be
determined by looking at the top of the colony while the elevation of the colony could be
determined by looking the view side of the colony at eye level as below:

Adopted from Microbiology (First Edition), 2003

Figure 1: Variation in microbial colony seen with naked eye

APPARATUS AND MATERIALS

2 bottles of Trypticase Soy Broth (TSB)
2 Petri dish of Trypticase Soy Agar (TSA)
Sterile Cotton Swab
Laminar Flow Cabinet
Incubator

PROCEDURE
Note: Aseptic techniques were applied throughout the process of this experiment. (Procedure
was performed in the laminar flow cupboard.)
A. Isolation of Bacteria from the Environment.
Firstly the bottom of TSA plates and also TSB bottles were labeled with our group name and
source of our specimen.

Secondly, the environmental specimen which was from the sole of the shoe was collected
using a sterile swab by rolling it around the sole.

Next, the swab was rubbed on an agar plate in a zig-zag pattern. The Petri dish was closed.

The swab was then placed into the TSB and swirled around. The bottle was recapped.

The agar plate and universal bottle containing the sample was incubated for 24 hours.

Lastly, the result obtained was recorded and analyzed.

B. Isolation of Bacteria from the Human Body.

Firstly, the bottom of TSA plates and also TSB bottles were labeled with our group name and
source of our specimen.

Next, one of our group member coughed on the surface of agar plate and also into the
universal bottle.

The agar plate and universal bottle was incubated for 24 hours, along with the environment sample.

The results were recorded.

RESULTS

Sample

Agar Plate

Body - Cough

Environment- Shoe Sole

Agar Broth

DISCUSSION

Figure 2: Types of colony morphology

After landing on the agar plate, a bacterial cell will divide and continue to divide until it
forms a mound or pile of bacteria known as a bacterial colony. It is known that different
species of microorganisms are represented by different colonies with different visual
characteristics. Colony morphology includes the size, color, overall shape, elevation and the
margin of the colony (Engelkrik, 2008).
In our experiment, the sample from the body was a cough. In the results, there was no growth
on the solid agar surface, however very small particles were seen to be suspended in the broth
medium. We pin this on the fact that our sample was not collected efficiently as we did not
cough hard enough for any bacteria to be released.
There was no sign of any growth on the agar plate whatsoever. This does not correlate with
the theory that bacteria is spread every time a person coughs. However, it is believed that the
method for collecting the sample was not carried out correctly. The collection of the sample
involved placing the Petri dish in front of the face, and coughing several times into the plate.
It might have also been the fact that none of the samples taken were from an ill person,
therefore less bacteria was able to be transferred.

The sample from the environment used in our experiment was a sample from the sole of a
shoe. The resultant agar dish showed various types of colonies of bacteria, with different
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shapes, sizes and margins. The results showed colonies with that were circular, irregular and
also rhizoid shaped. The margins for the colonies were entire for circular colonies, lobate for
irregular colonies and filamentous for rhizoid colonies. Some colonies flowed into each other
and were a combination of either irregular and smooth, or irregular and rhizoid. There were
also colonies which had a very cloudy and unclear margin, and this was chosen to be
classified under irregular colony shape, and undulate margin. The elevation of the colonies
were not visible, as all the colonies looked to be only slightly raised compared to the agar
surface, and no colonies had a distinct elevation type to be characterized by it. The different
types of colony suggest that there are many types of bacteria present in the agar plate, as
mentioned before, different bacteria give rise to different colonies, and thus have different
morphologies. The distribution of the colonies in the plate was random, and had no structure
within the entire plate, where colonies with similar characteristics being situated at opposite
ends of the plate being observed.
The reason both agar plate and agar broth were used was to observe the difference in the
bacterial growth between the two media. What was observed in the agar plates for the sample
from the sole of the shoe was that the colonies were relatively large and non uniform, as there
were many different bacteria present. However, in the case of the sample from the sole of the
shoe in the agar nutrient broth, the bacterial colonies were very small and plentiful, as well as
suspended in the broth. We suspect this is due to the fact that much of the bacteria from the
sample was already spread on the agar plate, and there was not many bacteria left of the
sterile swab, when we immediately dipped it into the nutrient broth. However, we did get a
positive result for microbial growth, indicating that bacteria do inhabit the soles of our shoes.
CONCLUSION
From the results of the experiment, it can be concluded that bacteria are ubiquitous. The
bacteria are most abundant on the sole of the shoes compared to the body sample. There are
many colonies of bacteria with the different shapes, sizes and margins on the sole of the
shoes, which is expected as shoes are used to walk in many places. Different places have
difference kind of bacteria. In the plates, colonies are more distinct and are much clearer and
more visible than the bacteria that grow in the broth medium. The objectives of the
experiment were met.

RECOMMENDATION
Firstly, the correct technique of coughing is needed to improve so that we can get the
bacteria. Secondly, the technique of swapping the sole of the shoes must be correct, swap all
the area of the sole. Thirdly, the procedure needs to be improved. For the body part, put in the
incubator while for the environment, put in outside the incubator.

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