CSU student Caitlin Lozano discovers new herpes virus in wild cats

While other students were writing papers or studying for exams, Caitlin Lozano spent her sophomore year finding new herpes viruses in wild cats.

In a research lab, headed by Sue VandeWoude, in the Department of Microbiology, Immunology, and Pathology, Lozano and her mentor, research scientist Ryan Troyer, have been examining whether cats have a gamma herpes virus.

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The project started back in spring of 2012 when Dr. Julia Beatty, a veterinarian from Sydney, Australia, visited the lab looking for herpes viruses in domestic cat samples that she had collected. While Beatty was here no viruses were found, but once she left, the lab decided to continue the project.

“After she left (we started thinking) we should take these techniques and see if we can find anything in pumas and bobcats,” Troyer said. “And so that is what started the project, without her coming and doing the sabbatical, we probably would not have even thought to do any of this.”

In August 2012, Lozano picked up the project, screening bobcat and puma DNA samples to see if the gamma herpes virus was present. They decided to work on wild cats because they had a previous project in the lab funded by the National Science Foundation to study the ecology of diseases in domestic cats, bobcats, and puma, according to Troyer. When the ecologist captured the wild cats to put radio collars on them, they would collect a blood sample and send it to the lab.

Troyer and Lozano started performing Polymerase Chain Reaction tests to try to detect the gamma herpes viruses. According to Troyer, because they knew the genetic sequences of other viruses in different species, they were able to use techniques that they thought would detect these viruses.

During that fall, Troyer found a new gamma herpes virus in a puma sample. Wanting to characterize the virus further, Troyer gave Lozano the project to begin screening more samples.

“Ryan was able to sequence the virus in order to create a test that was more specific in detecting it, but in order for him to get a test that was really good, he had to find a longer sequence,” Lozano said. “While he was trying to get this longer sequence he gave me the job to practice the test and gave me a couple bobcat samples. I ended up getting positives for those samples and when we sequenced it, it was different than the sequence that he had found.”

Troyer and Lozano determined that Lozano had found a second new gamma herpes virus, but in bobcats. After that, she was able to find a third new virus in bobcats.

According to Troyer, the characteristics of the three discoveries contrasted. One sample infected just bobcats, while the other was only found in pumas and the third inhabited both species. It showed the scientists that the virus didn’t have the same distribution in the three cats.

They were able to give them basic names that consist of the species name, followed with GHV — for gamma herpes virus — and one or two, Lozano said.

For Troyer and Lozano, the discovery of new viruses was exhilarating.

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“It was really exciting when we got the sequence and queried it against the sequence data base and when that came back and showed that it was clearly a herpes virus that hadn’t been described,” Troyer said. “I have been working in virology for 15 years and I always thought it would be cool to discover a new virus so that was just a fun experience.”

In order to detect and determine that it is a new virus, they had to copy and expand the sequences to get fragments to submit for sequencing to find the genetic makeup from a program called GenBank. This program, according to Troyer, is a repository for all the genetic sequence formations that are you there, so the two were able to simply type in their DNA sequence and compare it to what is already discovered.

“(GenBank) finds things that are closest to (the virus sequence), so that’s how we knew pretty much instantly that we had a new herpes virus because all the hits that came up were other herpes viruses,” Troyer said. “But none of (the other known virus sequences) were 100 percent identical to the sequence we were submitting, suggesting what we were putting in was a new sequence and a new virus.”

Since this discovery, Lozano has been working on developing a new test, the Quantitate Real Time PCR, which will specifically detect the gamma herpes virus. With this test, they will be able to screen hundreds of cat samples and determine what portion of them has the viruses.

“The test will be able to tell us what cats are infected, also in addition it gives us a measurement of how much herpes virus DNA is present in the cells, so it gives us some measurement of the level of infection,” Troyer said.

The two have not published their findings yet, but Lozano will be using her research and discovery to present at the Celebrate Undergraduate Research and Creativity symposium held at CSU.

Once they can get the Quantitative Real Time PCR and screen their samples, Troyer hopes to publish their findings in a journal the near future to share their findings.

Lozano and Troyer are continuing to try to sequence a larger portion of the genome for the viruses in order to prove that they exist and to further characterize them. As of now, their research helps contribute to the understanding of potential pathogens that are in the wildlife.

Collegian Writer Taylor Pettaway can be reached at news@collegian.com.