End of Summer Final Report

According the World Health Organization, approximately one-third of the human population is infected with tuberculosis.  This statistic is rather large in comparison to the percent of the population that individuals believe are infected with tuberculosis.  Due to many ethical guidelines that the medical community has established, researchers have been limited in the ways to study this disease. The way in which I have chosen to study this disease is rather new to the scientific community. My research thus far has been to find a way to utilize the nematode Caenorhabditis elegansin order to study tuberculosis in vivoin a successful manner to set a bases for further studies.

With the great amount of help that I have received from my mentors this summer, I was able to be very resourceful while figuring out how to go about this project.  The major point at hand was to make plates in which can allow both the bacteria and the worms to survive.  Through the extensive background that Dr. Marcy Kelly has on tuberculosis and BCG, I was already given a protocol for plates that allow BCG to grow. These plates consisted of 7H11-based media. The worms were found to thrive on cholesterol-based media. With many trials we were about to utilize and combine both protocols in order to create a plate consisting of 7H11- and cholesterol-based media.  Another aspect of trying to manipulate C. elegansis understanding their food choice. The worms’ preferred food choice was stated to be E. colithroughout much of the literature I read, and through the expertise of Dr. Marcello. Through many trial runs, we were not able to determine the food choice of the worms. In order to determine this, we put equal amounts of E. coli and BCG.

To summarize, C. elegansseems to have the potential to be an exemplar model for studying tuberculosis infection.  Even though C. elegans has not demonstrated a preference for BCG as a food source, there is data demonstrating that the worms will ingest the bacteria.  We have also established a media that allows the growth of both the bacteria and the worms.

I have experienced many small accomplishments throughout this research. To begin, I have gained a large understanding of both BCG and C. elegans. Of course, I plan to learn more and more every day and get a greater knowledge on the topic.

Overall, I have gained great patience through this experience. I know understand that I have to work around bacteria’s time frame, and it was challenging.  Fortunately, I was able to gain great time management skills. I also had to deal with a lot of experiences with research not cooperating well with me. This tested by patience tremendously, but in the end, it made me a better researcher.  The experience in the lab with Dr. Kelly and Dr. Marcello was incredible.  Being able to work one on one with them was an honor that I will continue to cherish. I hope more of their knowledge gets passed on to be throughout time.

My researchers were very understanding and helpful resources.  Not only did they dedicate their time to me, but they were willing to help with questions all hours of the day.  Having a demanding schedule really challenged me this summer, and the compassion they showed me was incredible.  They also taught me so much in such a small amount of time. I will forever be grateful for this opportunity that the provost grant and my mentors have provided me.

Determining the Gene Expression of Latent BCG in infected Caenorhabditis elegant

Throughout this summer have spent a lot of time learning about both BCG and C.elegans. Going into this I had already known that both organisms were studied to grow on different food sources, which provided a bigger challenge than I have ever expected. Data I have collected demonstrated that C. elegans had preferred to eat E. coli over BCG.  After some future studies, I would like to distinguish if this is a marker to state BCG as toxic to the worms.

With the help of professors at Pace University, I was able to make the agar of the plates that we ran our experiments on.  We were able to combine the food sources of both organisms in order to make the agar. BCG is usually raised in a 7H9 rich media, while C. elegans is more cholesterol based.  We have found a media that may allow both to live, but I will need to perform further tests in order to be more certain about this statement.

Working out the logistics has been a major goal with starting up this project during this summer.  Research thus far has shown that I need to be very patient with deal with the bacteria because at times it tends to behave erratically.  Research has also allowed me to grasp a more in depth foundation for both organisms.  Therefore, the grant gave me the opportunity to adjust my approach for the future and prepare me for the next three years of undergraduate research.

Determining the Gene Expression of Latent BCG in infected Caenorhabditis elegans

The World Health Organization (WHO) recently reported that approximately one-third of the world’s population is thought to be infected with Mycobacterium tuberculosis.  At least half of those infected individuals are infected with drug/multi-drug resistant strains. A major setback in researching tuberculosis is the limited options for in vivo experimenting. This project focuses on evaluating tuberculosis infection in the nematode worm, Caenorhabditis elegans, to try to determine if the worm can be used as a new model to study the disease.

The title of this project is “Determining the Gene Expression of Latent BCG in infected Caenorhabditis elegans.”  The worm C. elegans has an immune system in which is similar to that of humans. Along with attempting to prove that this worm is an ideal model for studying TB, the purpose of this project is to determine the gene expression of latent BCG in vivo. More specifically, we hope to pin point certain genes which are upregulated. From there, we may be able to identify the upregulated genes as the genes that force BCG into dormancy.

Through this project I expect to better understand the gene expression in latent BCG in comparison to the gene expression in active BCG.  Through this understanding, I hope to be able to isolate specific genes and correlate them to the latency phase. This sort of research does not have many research articles to base experiments off of, however, my research mentors and I were able to create a food source that both the BCG and C. elegans are able to grow on. From there we will allow C. elegans to feed on BCG for a week. In order to ensure that the bacteria infected the worm, we will be utilizing fluorescent BCG. This will give us a view of where the bacteria are in the worm. We will then lyse open the worms and extract the BCG. We will finally run an RNA sequence test on the bacteria to determine the genes in which are upregulated.