Research project on Eastern mountain lion uses ancient DNA sample from 'Original Nittany Lion'
Penn State Schreyer Honors College Scholar Maya Evanitsky found a way to blend her major with her interests and is using DNA samples from the past with the hopes of helping to preserve the future for American mountain lions. Spending part of her afternoon on Monday, April 13, with the “Original Nittany Lion” at Penn State’s All-Sports Museum, Evanitsky came away with a sample of the more than 150-year-old artifact for a genome research project that could have a major impact on conservation and outreach for the species labeled as one of the most endangered mammals in the U.S.
Evanitsky latched onto the opportunity to combine her major, biochemistry and molecular biology in Penn State’s Eberly College of Science, with a lifelong interest in anthropology. That interest turned into an academic minor after Evanitsky took George (PJ) Perry’s class and joined his ancient DNA lab a year later. Perry suggested the Eastern mountain lion research project for several reasons, including its status as the beloved “Nittany Lion” mascot of Penn State.
As a Schreyer Scholar, Evanitsky not only found her topic for the required honors thesis, she also was excited by the possibility of working with one of the most-recognized icons of her University.
“This is my own mark on Penn State history,” said Evanitsky, a junior from Cranberry Township, Pennsylvania. “This is definitely exciting. I will be able to say, ‘I sequenced the Nittany Lion.’ It really reinforced my connection to the school. This is the first (sample) I have physically collected, so it feels really good to have at least one sample and have something to work with.”
Both of Evanitsky’s parents are Penn State alumni, and while their daughter discusses the technology and science behind her project with ease, she takes a different approach when explaining it to her family or others who aren’t familiar with ancient DNA and the questions it may answer for future generations.
“Our overall goal is to sequence the ancient DNA, analyze it and compare it to genetic data from other mountain lion populations in Florida and the western U.S.,” said Evanitsky. “We want to try and find out where the Nittany Lion fits in the phylogeny or evolutionary history, figure out which group was more genetically diverse and learn what we lost in its (regional) extinction.”
“Another aspect of this project is to help raise awareness and bring attention to the processes of extinction,” said Perry, an assistant professor of anthropology and biology. “Through the technology we have in our ancient DNA lab, it is possible for us to study our mascot even though our mascot is extinct (regionally). What we can learn about the Nittany Lion in relationship to other mountain lions across the country, we can apply to other potential conservation uses, and stress the value of conserving what we do still have.”
Evanitsky will extract and sequence the mitochondrial DNA, or mtDNA, which is found in energy-producing structures that are inherited only from the mother. With ancient DNA, where the DNA is damaged and extremely limited in quantity, the mtDNA is the easiest to study because there are many mitochondria in each cell.
Evanitsky’s preparation for her thesis project began well before the collection date. In addition to undergoing two years of training in the meticulous methods required for ancient DNA collection and lab work, she also prepared the information to help acquire the funds to complete her research project. The “Nittany Lion Genome Project" is seeking $12,000 through Penn State University’s USeed website to cover the cost of the mtDNA sequencing, undergraduate student researcher wages and laboratory supplies.
The “Original Nittany Lion,” which stands guard near the entrance of Penn State’s All-Sports Museum, has not always held such a prominent place on campus. Shot in 1856 in northeastern Pennsylvania by farmer Samuel E. Brush after being treed by dogs, the Nittany Lion was turned into a stuffed plaything for Bush’s grandchildren before coming to campus to be put on display in Old Main.
Loaned to the city of Chicago for the 1893 World’s Fair, he was returned to campus and stored in the basement of Watts Hall with the remains of Coaly the mule, another Penn State legend. The next time this Nittany Lion appeared was in Pittsburgh in the 1950s as part of a display with other endangered species. That was followed by a year in Harrisburg before he was returned to the Carnegie Museum, and stored, and where his “on loan” status was forgotten.
More than four decades later, Penn State Professor of Wildlife Conservation Richard Yahner heard about the lion in storage at the Carnegie Museum and arranged to have it brought back to central Pennsylvania. Lack of proper care had left the specimen in such poor shape that it required careful restoration from a skilled conservator .
Evanitsky, aware of the work done on the Nittany Lion during its 1992 restoration and of patchwork repairs made to preserve the look of the fur, collected her sample from one of the original skin areas. She was also careful to maintain the structural and visual integrity of the Nittany Lion. After determining that some of the more easily accessible areas felt synthetic and were probably materials used in the taxidermy or restoration processes, she was able to remove a 2-inch by half-inch skin and fur sample from his inside hind leg.
“Whenever you do sampling of historical artifacts, you always want to be careful both in terms of procedure and thinking through the value of taking the sample versus preserving the integrity of the item,” said Ken Hickman, director of the Penn State All-Sports Museum. “In this case, the project had sufficient merit to warrant proceeding with that kind of sampling. The Nittany Lion is one of the oldest mascots in the country and it is certainly symbolic of both the University and the area in a great number of ways. It will be fun to be able to blend the University tradition with hard science and see what the results are going to be.”
The sample from the “Original Nittany Lion” will sit in a chemical overnight to remove the DNA from the cells. An enrichment process to prepare RNA complementary to the Nittany Lion’s DNA will allow Evanitsky to separate the critical DNA before washing the rest of the solution away along with bacterial DNA, residue and other contaminants.
Next, the DNA will go to the Penn State Huck Institutes of the Life Sciences Genomics Core Laboratory for sequencing with an Illumina HiSeq sequencing instrument, using methods similar to those that have recently been used to sequence the genomes of Neanderthals, mammoths, and other extinct species. With the advances in sequencing technology, Evanitsky will receive massive amounts of data, analyze it and compare it to other sequencing data from studies of the California and Florida mountain lions.
“Maya has taken the lead on this independent project and she is a shining example of a Penn State undergraduate student who is taking advantage of the opportunities to conduct research,” said Perry. “Her dedication definitely makes her exceptional. She is doing publication-quality scientific research that also has a major community outreach component while taking a full load of undergraduate classes and working a part-time job. She has invested hundreds of hours in the lab and computer coding training to be able to do the ancient DNA lab work and bioinformatics analysis component of the study.”
Schreyer Honors College promotes academic excellence with integrity, the building of a global perspective and creation of opportunities for leadership and civic engagement. Schreyer Honors Scholars, including Gateway Scholars admitted after their first year of enrollment, total more than 1,800 students at University Park and six Commonwealth campuses. They represent the top 2 percent of students at Penn State University who perform well academically and lead on campus.
Additional information for this article was obtained from an August 1996 article “Lion’s rugged good looks restored” by Gary Cramer from the archives of Penn State’s Public Information Office and from “All Things Nittany” on the AskPSU section of the University’s website.