During the Summer of 2014 I did research in the lab of Dr. Amal Amer. Dr. Amer works with Legionella pneumophila, a pathogen that causes Legionnaire’s disease in those who have compromised immune systems and the elderly. My project deals with Legionella biofilm. Biofilm is an extracellular polymeric matrix with which bacteria can thrive in environments that have limited available nutrients. Legionella biofilm is the leading cause of infection and biofilm builds up in moist areas such as air conditioners or spa filters. Once a biofilm has developed, it is extremely difficult to eliminate because it is resistant to antibiotics. Because biofilm is a leading cause of infection, we are trying to learn more about Legionella biofilms and find a significant factor that leads to its development. Once a factor has been identified it can lead to new treatments that can target the factor and eliminate the biofilm. In order to identify a significant factor I grew biofilm for JR32 Legionella pneumophila Wildtype (WT) Strain, or non-mutated strain, and JR32 Legionella pneumophila mutant strains.
For each experiment I grew biofilm for JR32 WT and one JR32 mutant strain for 6 days and performed a Crystal Violet Assay at the end of the growing period. The Crystal Violet was used to quantify the amount of biofilm grown by the bacteria. Because we know that JR32 WT does not have any mutations and has the ability to grow biofilm. I used this as the control variable for my experiment. What I hoped to find was at least one mutant strain that was not able to grow healthy biofilm. This would signify that this specific mutant lacks a factor that is significant to biofilm growth. For the summer, I worked with mutants that lacked specific effector proteins and contained mutations in the structure of specific lipopolysaccharides.
During the summer I was able to grow and quantify the biofilms for five different effector protein mutants and three different LPS mutant colonies. Unfortunately, I was not able to identify an effector protein mutation or a LPS mutation that was not able to grow a proper biofilm.
In the future I plan to continue growing various other mutant strains and hope to find a factor that is significant in the production of Legionella biofilm. Once I find a mutation that is unable to grow healthy biofilm, the next step will be to show specifically how the Wildtype biofilm and mutant biofilm are different from each other. For example, I will take pictures of each biofilm with a Confocal Microscope in order to show visually how the two are different in structure. I will also work with a Reometer in order to test the bond forces of each biofilm and compare the two strains.
I chose to do this experience because it was a way for me to experience a possible career path and an interest. I have always been interested in doing research and was finally able to have a first-hand experience in implementing the scientific process in a professional setting. Initially I had high hopes for my experiments and it was definitely tough when I came to the realization that my experiments would not produce perfect results or, at times any results, at all. Sometimes they may need to be repeated due to a simple mistake or sometimes you may not even know what needs to be fixed. The most important thing was to just keep thinking of new ideas, making observations, and trying to solve the problem at hand. I also learned that I will not always know everything that is going on, which is fine, but it is my job to learn. Doing experiments is definitely a trial and error process and when something does not work, you need to adjust and keep trying.
In addition I learned that science is a very communicative subject. Yes, you do your own experiments but it is talking through discussion with others that allows you to learn or think of even more new ideas or solutions.
Overall, I very much enjoyed my experience. Working in Dr.Amer’s lab is a privilege and I am excited to continue working here.
As stated before, one of my main goals for the STEP experience is to find something I am interested in and experience possible career paths. I am a biochemistry student and graduate school has always been my goal. Through this experience I was able to see what type of work would be entailed as a graduate student. Though my decision is not final yet, I have decided that I do enjoy research and I hope to continue to do it whether it be through graduate school or working in a lab.
In addition, another goal for my STEP experience was to learn more about what I would like to do research on in the future. I have always been more interested in the biology aspect of biochemistry but I wasn’t sure whether or not I preferred microbiology or macrobiology. During my experience, I was able to learn more about microbiology and I now see it as a possible career path.