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Research Proposal

Andrew Choe
Independent Research GT
Title: The War on Bacteria

Overview of Research:
The general purpose of the research conducted this past year is on the application of
bacteriophage to kill bacteria, more specifically against antibiotic-resistant bacteria. Overall
ideas include background on bacteriophage, how phage infect host bacteria cells, the isolation
process of phage, and other applications of bacteriophage. An experiment was completed to
isolate possible Klebsiella, Acinetobacter, and E. coli bacteriophage. Now, this research and
information can be shared with others to inform them of the processes of isolating phage, and
hopefully, the presentation can spark interest in students to go out and continue the research of

Background and Rationale:

Bacteriophage, or phage for short, are viruses that are literally everywhere, and they are
viruses that eat bacteria. This process can help fight the war on bacteria that have resistance to
antibiotics; these bacteria are also known as superbugs. There are many cases where patients
suffer from illness caused by superbugs where even very strong antibiotics do not seem to kill
the bacteria. The most dangerous type of superbug is the carbapenem-resistant
Enterobacteriaceae (CRE), a group of bacteria that are normally found in the stomach.
However, a build up of CRE can cause life-threatening blood infections, and are resistant to most
available antibiotics. Examples of Enterobacteriaceae are E. coli and Klebsiella.
Since bacteriophage are everywhere, they can be isolated in samples from anywhere. To
treat patients with illnesses caused by bacteria, doctors and scientists can use phage therapy in
order to treat the patient. Phage therapy is the process of taking bacteriophage specific to
bacteria that is causing illness to a patient and allowing the phage to kill the bacteria in the
patient. Since most antibiotics are not effective in killing superbugs, bacteriophage are an
alternative solution to the problem, and can open new opportunities for scientists and those
interested in bacteriophage to go out and isolate phage. Bacteriophage therapy was a common
practice previously, but the invention of antibiotics caused phage therapy to decline. However,
recently, there have been more cases where bacteria have grown immune to antibiotics.

Research Methodology:
Research Question:
Can one possibly isolate bacteriophage from simple environmental samples?
Research Thesis:
Taking random dirt, water, and soil samples from the environment can supply
bacteriophage specific to the host bacteria.
Before going to a lab to isolate the bacteriophage from the environmental samples, 6
samples were collected at a house, park, and pond. There were two water, two soil, and two dirt
samples. Since phage are everywhere, and higher in places where bacteria are present, samples
were collected at the park, in a sewer, in a pond, and in front of a house. With the six samples,
bacteriophage for Klebsiella, E. coli, and Acinetobacter can be isolated.
Research Design Model:
The type of research method used in this research was quantitative. Experimental
research was conducted in order to fully understand the process of isolating phage from
environmental samples. Independent variables included six environmental samples tested on
three different types of pathogenic bacteria. The dependent variable is the bacteriophage that is
specific to one of the three types of bacteria out of the six samples.
This model of research was selected for hands on understanding of the process of phage
isolation. If going to the lab was not an option, learning how phage can be taken from any
environmental samples would be difficult to fully understand.
Data Collection:
The experiment was conducted with the six dirt, water, and soil samples. The samples
were centrifuged for ten minutes to suspend the phage in solution, and the bacteriophage were
further filtered to separate most other contaminants. After filtering, the six samples were each
mixed with Klebsiella, Acinetobacter, and E. coli bacteria that were grown overnight. All
together, there were eighteen plates with possible bacteriophage for the specific bacteria. The
bacteriophage mixed with bacteria were left overnight to incubate, and were examined the next
After letting the phage and bacteria incubate overnight, they were examined for clearings
where the bacteriophage killed the bacteria. These clearings are known as phage plaques. Only
one out of eighteen plates had a clearing that is most likely a phage plaque.
At a research lab, the ability to isolate phage from the environment was possible. This
research lab was chosen, for the scientist working there is a mentor and expert in the field of
bacteriophage. After being able to isolate phage from random environmental samples, it was
clear how simple it is to isolate phage, and how finding solutions to fighting antibiotic resistant
bacteria is a possibility through the isolation of bacteriophage for phage therapy.

Product Objectives:
The topics about bacteriophage learned over the year will be put into a
presentation/lesson for a high school GT Biology class. During one of the biology classes, a
presentation on the background of bacteriophage, the process of isolation, and the uses of
bacteriophage will be given. This presentation will have detailed information of the process of
bacteriophage isolation and uses of phage in order to inform and spark interests in students who
would enjoy this area of interest in order to help the community.
GT Biology students were chosen because they are studying in a class that is learning
similar topics to those in the presentation. Almost none of the students will know what
bacteriophage is, and after sharing the process of how easy it is to isolate phage, hopefully, some
students will be encouraged to later take on this field of interest in order to make the community
better. Students will learn that the application of phage does not just stop in the medicine field,
but can also be applied in fields such as agriculture.
The lesson on bacteriophage will be given through a PowerPoint presentation with
detailed facts and visuals. Also, a hands-on model of a phage will be available in order for
students to fully grasp the anatomy of the phage.

Logistical Considerations:
The resources needed to make the product possible are a presentation of some sort and a
class to teach. This presentation will most likely be a PowerPoint presentation along with visual
aids to fully understand the topic at hand. Special considerations to think about are the audience,
time, and place to share the product.
Class time would be the most crucial step in the product, for if there isn’t a class
available, then the presentation cannot be shared. The audience and place work together, for a
biology class full of high school students is the preferred audience and environment to share the
presentation with.
Fortunately, special permission was given by a GT Biology teacher allowing the use of
one of her classes to share a presentation on the topic of bacteriophage.
A timeline will be added that outlines the data collection, product development, and
audience distribution.


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Student Signature GT Resource Teacher Signature Mentor/Advisor Signature