Emily Karwacki, who recently earned her biology degree from the 麻豆原创, didn鈥檛 set out to track the deadly pathogen Perkinsea, but after landing a research spot in Assistant Professor Anna Savage鈥檚 lab, she was set with the task of trying to test for the disease. Frogs, which are indicators of environmental changes, have been dying off in mass quantities. They are also an important part of the food chain. Without frogs, many other species would die, Savage said.

Scientists have narrowed down what鈥檚 most affecting frogs to three pathogens, including Perkinsea.

鈥淣ot a lot of people have studied Perkinsea because it has only recently been identified,鈥� Karwacki said. 鈥淚t鈥檚 different from other diseases because of the way it attacks the host.鈥�

The pathogen enters the frog through the skin or may be ingested through its mouth. Scientists know it goes straight to the liver, embedding itself, before moving onto the rest of the tissue. It spreads and then the frog dies.

Karwacki, along with Savage and doctoral student Matt Atkinson, suspected that Perkinsea was killing frogs in Central Florida, but the researchers needed a way to test for it first. Karwacki was tasked with creating the molecular test. The method is called qPCR, but because Perkinsea was newly discovered, there wasn鈥檛 enough genetic data to make a specific test. Karwacki had to create what鈥檚 called a primer pair, and match it with a DNA sequence of Perkinsea, to get the qPCR test to work.

鈥淭he test amplifies the DNA so you know if your pathogen is there or not,鈥� Emily said. 鈥淚 had to align a bunch of DNA sequences from our samples with others from around the word to create my primer set. It was four or five months before we had both the primers and the probe to create a successful test.鈥�

Karwacki was the first to do this for Perkinsea and her work was recently published in the journal Diseases of Aquatic Organisms.

Using Karwacki鈥檚 qPCR assay, the team of researchers found that 25 percent of the frogs they sampled tested positive for the pathogen. They sampled three sites in Florida: Gold Head Branch State Park in Keystone Heights, the 麻豆原创 Arboretum in Orlando, and the Archbold Biological Station in Venus. The area they found with the most prevalent infection was Gold Head Branch, which is the farthest north. Archbold, the farthest south, had no infection at all.

鈥淭here are only three papers on this disease that identify it specifically,鈥� Karwacki said. 鈥淚t has greatly been affecting amphibians in the southeastern United States and should be studied more. It鈥檚 most likely at least a co-factor in these extinction events we are seeing.鈥�

Karwacki鈥檚 method will now allow researchers all over the world to test for the disease. After graduating this summer, she is working on a new study, swabbing frog tissue samples at the Florida Museum of Natural History in Gainesville. She鈥檚 swabbed more than 900 samples, and has found that Perkinsea dates back to 1922. This proves the disease has been in frog populations before, and scientists are trying to figure out why it鈥檚 only now killing off large numbers of frogs.

鈥淣ow with my qPCR, people can test areas where they are trying to release frogs to rebound populations,鈥� Karwacki said. 鈥淪cientists can test water and soil to see if Perkinsea is there so we don鈥檛 send frogs out to die.鈥�

Karwacki is entering the nonprofit business management master鈥檚 program at 麻豆原创 before pursuing graduate school. She will continue her work as a research associate in Savage鈥檚 lab.

麻豆原创 assistant professor and biologist Kate Mansfield works on various sea turtle research projects and is now partnering with 麻豆原创 molecular ecologist Anna Savage to see if they can figure out how fibropapillomatosis or FP for short develops in turtles and why.

There is some existing work on FP, but Savage brings a unique approach to the 麻豆原创 team. She specializes in figuring out the role genes play in species resistance to viruses and bacteria. Most of her work has been with frogs and she had never worked with turtles, until now.

鈥淭hat鈥檚 why I鈥檓 so excited,鈥� Savage said. 鈥淭here鈥檚 been a lot of data collected and there is some literature on FP. But it鈥檚 a complicated disease. The belief is there are a lot of factors that play into the tumor formation. I鈥檓 taking a slightly different approach by looking at the genes related to turtle immunity to see if we can find a pattern that might lead us to some answers.鈥�

Preliminary data show that the tumors in the sea turtles Savage has examined carry a specific herpesvirus. But some turtles with no tumors also have biomarkers for the virus in their skin and blood. And there are some turtles that have no presence of the virus at all. Important clues may be found in the genes of turtles with the virus, but no tumors, Savage said.

She and Mansfield recently earned a $17,631 grant from the Sea Turtle Conservancy to take a deeper genetic look at what may be driving the tumor disease among green and loggerhead sea turtles. They have a deep data set to review thanks to the 麻豆原创 Marine Turtle Research Group that has been collecting sea turtle data since 1982. 麻豆原创 researchers and students have been monitoring the health and nesting patterns of sea turtles at the Archie Carr National Wildlife Refuge (with US Fish and Wildlife Authorization), the Indian River Lagoon and the Tidal Basin near Cape Canaveral. The Archie Carr is a one of the most important nesting area for sea turtles in the Western Hemisphere.

Mansfield and her students capture sea turtles in the Indian River Lagoon twice a month to collect data and monitor their health. The biologists check the turtles鈥� weight body condition, collect samples of blood, skin, and when present, tumors. When finished, all turtles are flipper tagged and released back into the water. Large numbers of juvenile green sea turtles visit the lagoon 鈥� one of the most important feeding areas for the species.

鈥淲e encounter on average 100-200 turtles in the IRL a year and in some cases we have samples going back a decade or two,鈥� Mansfield said. 鈥淪o we have a unique opportunity to leverage our long-term dataset to answer some long-standing questions, and to fill some needed data gaps in sea turtle conservation.鈥�

Courtesy of 麻豆原创 Marine Turtle Research Group

The 麻豆原创 Marine Turtle Research Group studies all life stages of these sea creatures. Sea turtles remain at sea for most of their lives, where it is difficult to observe and track them over time. 聽Turtles don鈥檛 reach maturity for 20-30 years. It is important to understand the threats and impacts to the health of the juveniles, like those found in the IRL, in order to ensure that future generations of sea turtles will continue to nest in Florida.

In addition to work on our local nesting beaches and coastal waters, Mansfield and her graduate students have been working in the Gulf of Mexico, and south Atlantic and Indian oceans to unravel the mystery of the sea 鈥渓ost years鈥� 鈥� examining where the youngest stages of sea turtles go and how they interact with their oceanic habitats.

Mansfield and the turtle team鈥檚 work helps state and federal managers better protect the endangered and threatened species for future generations.