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 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 shoes compared to the body body sample. There are many colonies colonies of bacteria with difference sies, shapes and margin in the sole of the shoes. In conclusion, bacteria are present everywhere.
INTRODUCTION
!icroorganism or microbe is an organism that is microscopic which mean too small to be seen by the naked eyes however visible under the microscope. "or 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. $esides, microorganisms can be found in wide variety of environment like in air, on land, plants, animals and in fresh or salt water environments. $acteria able to live everywhere because they are all well adapted to all different type of environment and can colonie that particular environment. "or example, according to %racle Think &uest 'ducation "oundation, it stated that (yanobacteria )ostoc 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. !icroorganism 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 gar, a solidifying agent purified from red algae. Agar#agar is not a nutrient because it contain *.+- agar for the solidified media so giving a solid surface for the bacteria to grow on and could not be utilied by the organisms. !eanwhile the 1
presence of bacteria will produce turbidity in broth culture. A mixed culture from most of specimen can be seen based on different colony sies, shapes, colours and many more.
OBJECTIVE
*. To compare the growth of bacteria from different environments. .
To identify the ways that bacteria grow on and in bacteriological media.
. To become accustomed with different type of media.
THEORY
The term /ubiquity0 of microorganisms can be defined as everywhere. That means, microorganisms can be found nearly everywhere on the planet. !icroorganisms include bacteria, archaea, algae, protists, protooa and viruses. According to 1ans 2. 3chlegel and 1olger 4. 5annasch 67789, the three major factors that contribute to the ubiquity of microorganisms are their small sie that can be dispersed easily by air and water, their metabolic versatility and flexibility, and their ability to tolerate the unfavorable conditions. "or example, some archaean can live in hot spring rich in 3ulfur. $esides, microbes especially bacteria are widely distributed in the human body and on surfaces of any materials and tools. Therefore, to understand the concept of /ubiquity0 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. $acteria can be spread through coughing, because according to !arshall (avendish 677:9, they are often present in saliva, in the mucus from the nose and lungs which can be sent through an airborne manner. %n 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. (ulture media Trypticase 3oy Agar 6T3A9 and Trypticase 3oy $roth 6T3$9 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. 2
After incubation for ; 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 !icrobiology 6"irst 'dition9, 77
"igure *< =ariation in microbial colony seen with naked eye
APPARATUS AND MATERIALS
bottles of Trypticase 3oy $roth 6T3$9
>etri dish of Trypticase 3oy Agar 6T3A9
3terile (otton 3wab
?aminar "low (abinet
Incubator
3
PROCEDURE
)ote< Aseptic techniques were applied throughout the process of this experiment. 6>rocedure was performed in the laminar flow cupboard.9 A. Isolation of $acteria from the 'nvironment.
"irstly the bottom of T3A plates and also T3$ bottles were labeled with our group name and source of our specimen.
3econdly, the environmental specimen which was from the sole of the shoe was collected using a sterile swab by rolling it around the sole.
)ext, the swab was rubbed on an agar plate in a ig#ag pattern. The >etri dish was closed.
The swab was then placed into the T3$ and swirled around. The bottle was recapped.
The agar plate and universal bottle containing the sample was incubated for ; hours.
?astly, the result obtained was recorded and analyed.
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$. Isolation of $acteria from the 1uman $ody. "irstly, the bottom of T3A plates and also T3$ bottles were labeled with our group name and source of our specimen.
)ext, 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 ; hours, along with the environment sample.
The results were recorded.
RESULTS
3ample
Agar >late
$ody # (ough
'nvironment# 3hoe 3ole
5
Agar $roth
DISCUSSION
"igure < 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. (olony morphology includes the sie, color, overall shape, elevation and the margin of the colony 6'ngelkrik, 77:9. 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. 4e 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. 1owever, it is believed that the method for collecting the sample was not carried out correctly. The collection of the sample involved placing the >etri 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 6
shapes, sies and margins. The results showed colonies with that were circular, irregular and also rhioid shaped. The margins for the colonies were entire for circular colonies, lobate for irregular colonies and filamentous for rhioid colonies. 3ome colonies flowed into each other and were a combination of either irregular and smooth, or irregular and rhioid. 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 characteried 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. 4hat 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. 1owever, 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. 4e 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. 1owever, we did get a positive result for microbial growth, indicating that bacteria do inhabit the soles of our shoes.
CONCLUSION
"rom 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, sies and margins on the sole of the shoes, which is expected as shoes are used to walk in many places. @ifferent 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.
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RECOMMENDATION
"irstly, the correct technique of coughing is needed to improve so that we can get the bacteria. 3econdly, 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. "or the body part, put in the incubator while for the environment, put in outside the incubator.
REFERENCES
*. 'dited by !artin @workin, 3tanley "akow, 'ugene osenberge, Barl#1ein 3chleifer and 'rko 3tackebrandt. 778. The >rokaryotes 6Third 'dition9. A 1andbook on the $iology of $acteria< 3ymbiotic Associations, $iotechnology, Applied !icrobiology 6=olume *9. >rokaryotes and Their 1abitats. >.*C. 3ingapore. 3pringer#3cience !edia. Inc. . !arshall (avendish (orporation. 77:. @iseases and @isorder. Infections, bacterial. >.;8;. Tarrytown, )ew Dork. !arshall (avendish (orporation. . ). >. 3axena and @. B. Awasthi. 77. !icrobiology 6"irst 'dition9. !ethods for (ulturing !icroorganisms. >.C;. !eerut. BI31)A >rakashan !edia 6>9 ?td. ;. ?ab !odule *< Ebiquity of !icroorganisms. etrieved )ovember +, 7*, from the 4orld 4ide 4eb< httpaul 2. 'ngelkrik, 5anet ?. @uben#'ngelkrik. 77:. ?aboratory @iagnosis of Infectious @iseases< 'ssentials of @iagnostic !icrobiology. ?ippincott 4illiams 4ilkins. p*H:
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