Health & Habitat analysis of endangered Ohio rattlesnakes

Katie Backus, second-year veterinary student at Ohio State (left), acquires a blood sample from a massauga rattlesnake in northeastern Ohio in the summer of 2015.

Katie Backus, second-year veterinary student at Ohio State (left), acquires a blood sample from a massauga rattlesnake in northeastern Ohio in the summer of 2015.

This summer, second-year student at The Ohio State University College of Veterinary Medicine Katie Backus completed a comprehensive health analysis of massasauga rattlesnakes in partnership with the Ohio Department of Natural Resources. The data will be studied along with land use and habitat condition.

Eastern massasauga rattlesnakes dwell in northeastern regions of the U.S., including Ohio. The species has been a candidate for the Federal Endangered Species Act since 1999, according to the U.S. Fish and Wildlife Service.

The decline of massasauga rattlesnake populations is mostly due to human and agricultural activity as well as habitat loss or destruction. The snakes typically inhabit wetland areas.

The research team tracked and captured massasaugas to acquire blood samples, swab tests for fungal disease and height and weight measurements. They’re also reviewing data on water quality, vegetation density and land use in surrounding areas. Water that flows downstream from farms may contain chemicals associated with agricultural runoff, which has been observed to cause adverse effects in other species.

“Agricultural runoff could be weakening massasaugas’ immune systems,” Backus said, “Possibly predisposing them to fungal diseases and other health issues.”

Backus and her team, led by Greg Lipps, amphibian and reptile conservation coordinator and conservation biologist, were able to capture approximately 55 massasaugas in northeast Ohio. Around 20 were recaptures. The results from fungal disease tests are still processing.

To capture the massasaugas, Backus and her team would walk around in protective gear with snake tongs and bags. Once captured, they would guide the snakes head-first into a plastic tube to prevent biting (as they are venomous), to perform diagnostics. A stress measure – a hormone called leptin – was taken into account during the health assessment.

A massassauga rattlesnake in a wetland area in northeastern Ohio. The species is now endangered.

A massassauga rattlesnake in a wetland area in northeastern Ohio. The species is now endangered.

The results of the study will help the ODNR with conservation efforts, as not much is currently known about the relation between massasauga health and habitat. The ODNR will likely try different techniques on different plots of lands to see what works for the snakes, Backus said.

“I learned so much throughout this process,” she said. “Not only handling snakes, but about government wildlife departments, the history of Ohio’s environment and ecology in general.”

Veterinary medicine plays many roles, and conservation medicine is an important one. This study is an example of how veterinary research can intertwine factors of human, animal and environmental health.

Acute Lung Injury in influenza patients

Patients with severe cases of influenza sometimes develop Acute Lung Injury (ALI), a highly damaging condition that can be fatal. Treatment options are limited.

MicroRNA (miR) are non-coding RNA molecules that take part in the regulation of gene expression, and they’ve been observed to act abnormally in inflammatory diseases, some forms of cancer and more.

Third-year veterinary student at Ohio State, Leon Schermerhorn. August 6, 2015; Columbus, Ohio.

Third-year veterinary student at Ohio State, Leon Schermerhorn. August 6, 2015; Columbus, Ohio.

Last summer, third-year veterinary student at The Ohio State University Leon Schermerhorn studied the molecular structure of influenza-induced ALI lung cells and, with his team, was able to conclude that a single miR, miR-155, may play a direct role in the progression of the disease. The team, led by associate professor in Ohio State’s Department of Veterinary Biosciences Dr. Ian Davis, discovered this by determining all miR expression levels in ALI lung cells. The results showed that miR-155 was highly over-expressed by alveolar type II (ATII) cells , which are the primary site of influenza virus replication. As influenza increases in severity, miR-155 expression becomes greater. This is harmful because an upregulation of miR-155 seems to provoke a raise in several types of white blood cells and signaling proteins, causing lung inflammation.

Since miR-155 expression by ATII cells could be responsible for the progression of influenza-triggered ALI, Schermerhorn and Davis hypothesize that miR-155 may be a target for therapeutic intervention. Influenza-induced ALI becomes less severe in mice that ATII cell miR-155 expression has been blockaded, called miR-155-knockout mice.

This summer, Schermerhorn is testing a gene therapy method’s ability to delay onset or reduce severity of influenza-induced ALI in mice. The method involves inserting pieces of specially engineered DNA – in this case lipoplexes carrying antagomiRs – into mice that will target ATII cells and inhibit miR-155 expression. His results will give further data on the role of miR-155 in influenza development in general, as well as the efficacy of inhibiting ATII cell miR-155 expression with antagomiRs.

“Research in public health and infectious diseases has always interested me; I like to solve problems,” Schermerhorn said. “Since influenza is a high-consequence pathogen, this study was a good fit from the start.”

Understanding how influenza affects cystic fibrosis patients

An estimated 30,000 people in the U.S. are living with Cystic Fibrosis (CF), a fatal genetic disease, according to the Cystic Fibrosis Foundation patient registry.

CF can affect many parts of the body, but it primarily impairs lung function. The lungs in a person with CF are colonized and infected by bacteria from a young age due to poor mucus clearance, which results in chronic inflammation. This makes them susceptible to various infections, and seasonal influenza viruses in particular present a huge risk. When CF patients contract influenza, it can cause severe, sometimes life-threatening symptom exacerbations, making their lungs vulnerable to additional bacteria. Currently, winter CF exacerbations are difficult to treat.

Part of the problem may be an abnormal increase in activity of proteins called pattern-recognition receptors (PRRs) following infection. PRR signaling usually helps the immune system detect viral infection to begin fighting it off, but an excessive response to viruses can cause severe inflammation.

Young5Second-year student Sarah Young is working with Dr. Ian Davis, associate professor in the Department of Veterinary Biosciences, to examine the molecular processes behind influenza infection in CF patients, specifically abnormal PRR signaling in infected lung cells. Young and Davis hypothesize that irregular PRR signaling causes the excessive inflammatory response in CF patients with influenza. To determine if their hypothesis is correct, Young is infecting both regular and CF human airway cells with influenza A virus to examine how PRR signaling differs between groups. The cell cultures that she’s working with use cells from donor lungs, gathered by Dr. Mark Peeples of Nationwide Children’s Hospital.

Once complete the study will provide new information on the natural immune response of CF patients to influenza infection, said Young, who was born with the disease.

“It’s rare to get the chance to research the disease you actually have,” she said. “It has given me more interest and motivation.”

CF is only known to occur in humans, but the study utilizes Young’s knowledge in veterinary medicine nonetheless, Davis said.

“Veterinarians have a great role to play in human medical research,” Davis said. “We have a different perspective, and there are not enough of us involved in it.”

Measuring benefits of environmental enrichment on dairy calves

To prevent the spread of infection and disease, dairy cattle farmers separate calves from each other for approximately six weeks until they are weaned from milk. Though beneficial to health, this practice may hinder calves’ social and cognitive development as well as lead to abnormal habits.

Young calves sometimes display excessive sucking and licking behaviors, often directed toward inappropriate sources such as the enclosure or another calf. As they progress through life, the behaviors can become quite exaggerated and harmful, said Dr. Katy Proudfoot, assistant professor in Veterinary Preventive Medicine and extension specialist in animal welfare and behavior at The Ohio State University. Letting calves feed naturally rather than from a bucket can help reduce this habit, but many producers are hesitant to adopt the practice.

“Veterinarians were the ones who first came up with the practice of separating dairy calves, as a vital health precaution,” Proudfoot said. “But now we’re recognizing that creating a more complex environment could help calves later in life.”

Ohio State veterinary student Emily Cosentino works with dairy calves at Waterman Dairy Center in Columbus, Ohio, as part of the school's Summer Research Program.

Ohio State veterinary student Emily Cosentino works with dairy calves at Waterman Dairy Center in Columbus, Ohio, as part of the school’s Summer Research Program.

Hannah Manning and Emily Cosentino, second-year veterinary students at The Ohio State University, are working under Proudfoot’s direction and studying the impact of four environmental enrichments on individually housed dairy calves. They hope to see improved social skills and overall welfare in calves who are provided the enrichments, as well as decreased sucking behaviors. They include a slightly larger pen size, a brush to rub up against, an artificial teat and rings to suck on and a tube filled with molasses to stimulate licking behavior.

The team is looking at ten calves in each group – one with environmental enrichments and one without – at Ohio State’s Waterman Dairy Center. They are continually capturing video footage of the calves during their research so they can analyze it from start to end.

“I’ve never had experience with dairy cows,” Manning said. “Working with them this summer has taught me patience and shown me that there’s still a lot more we can figure out.”

Second-year Ohio State veterinary students Hannah Manning (left) and Emily Cosentino (right) at Waterman Dairy Center in Columbus, Ohio, in August 2015.

Second-year Ohio State veterinary students Hannah Manning (left) and Emily Cosentino (right) at Waterman Dairy Center in Columbus, Ohio, in August 2015.

The study is mostly preliminary, and will produce data about the relationship between calf housing and animal welfare. If the enrichments prove to be beneficial, the team thinks it would be realistic for dairy cow farmers to start providing similar elements to calves.

Raw pet food is growing in popularity, but is it safe?

Raw pet food has a growing consumer base, says Paulynne Bellen, third-year veterinary student at The Ohio State University College of Veterinary Medicine, but the food may not come without health risks.

One concern is that raw pet food can sometimes carry a bacterium called Listeria monocytogenes, among other foodborne bacteria, since it does not go through the cooking and steaming processes that dry pet food does.

If ingested, L. monocytogenes can cause more extreme symptoms than E. coli, such as infection, fever, gastrointestinal problems and sometimes death. The bacterium can survive in low temperatures and in the presence or absence of oxygen, which is how it likely gets into some raw pet food. L. monocytogenes poses a much higher risk for animals (including humans) that are immunocompromised.

Bellen and Dr. Thomas Wittum, research mentor, professor and interim chair of the Department of Veterinary Preventive Medicine at Ohio State, are testing 120 raw pet food samples for the presence of L. monocytogenes, E. coli and Salmonella. They are comparing the levels of foodborne pathogens found in each sample to the four standard processing methods used in raw pet food production: freeze drying, high-pressure pasteurization, raw freezing and dehydration.

Paulynne Bellen, third-year veterinary student at The Ohio State College of Veterinary Medicine, analyzes a sample of raw pet food for foodborne pathogens as part of a study funded by the college's Veterinary Scholar Summer Research Program

Paulynne Bellen, third-year veterinary student at The Ohio State College of Veterinary Medicine, analyzes a sample of raw pet food for foodborne pathogens as part of a study funded by the college’s Veterinary Scholar Summer Research Program

This does not mean that all raw food is unsafe for pets to eat or without its benefits, and some veterinarians regularly recommend it, Bellen said.

“As a scientist, you can’t be swayed either way without the facts,” she said. “We just want to determine if one processing method is better at keeping bacteria out than the others, which would allow us to guide pet owners toward the safest kinds of raw food.”

The FDA has issued recalls on several major raw pet food brands and companies in recent years, including three in 2015, brand names Primal, OC Raw Dog and Vital Essentials .

Bellen said that the study has improved her skills in the lab and in responding objectively to a controversial issue. The team expects to see several food samples test positive for foodborne bacteria, Wittum said.

Testing therapeutics on feline lung cancer

Dylan Burroughs, second-year student at The Ohio State University College of Veterinary Medicine, is working on some of the world’s first research on Feline bronchioloalveolar carcinoma (BAC), a form of lung cancer.

The best method for researchers to study BAC without running tests on a cat is to use a live model of the cancer, called a cell line. Worldwide, there is only one feline BAC cell line available, so he and his mentor Dr. Gwendolen Lorch, assistant professor at Ohio State in Veterinary Clinical Sciences, had to wait a while to acquire it for their research.

The team used the cell line to test different concentrations of drugs and how they work in controlling the cancer. They also measured gene and protein expression levels to know what cell regions should be targeted during treatment.

It is typical in human lung cancer for the epidermal growth factor receptor (EGFR), a protein that takes part in cell communication, to be overexpressed. The overexpression of EGFR and related genes leads to an overabundance of cells, which could play a large role in causing the cancer, Burroughs said. In an attempt to stem the excess cell growth, Lorch and Burroughs are testing the effect of EGFR inhibitors on the feline BAC cell line in addition to standard cancer-treatment drugs.

Dylan Burroughs, second-year veterinary student at Ohio State, works in the lab with a feline cell line. July 21, 2015; Columbus, Ohio.

Dylan Burroughs, second-year veterinary student at Ohio State, works in the lab with a feline cell line. July 21, 2015; Columbus, Ohio.

In cats, this type of cancer tends to metastasize – or spread – to the paws. This complicates the effectiveness of treatment options because the tumors are growing in complex patterns that are hard to recognize and target.

The results of the study could potentially help human lung cancer research, but the team is mainly working on establishing normal values for feline BAC that will guide future scientists. They are also focused on understanding the method in which the tumors are able to spread from the lungs to the paws.

Burroughs’ study is being funded by the National Institute of Health, and he presented his findings at the 2015 Merial-NIH Veterinary Scholars Symposium at UC Davis.

Work continues to eliminate rabies in Ethiopia

Up to five students in The Ohio State University College of Veterinary Medicine’s Summer Research Program are chosen each summer to be supported for international research to work on projects in various parts of the world.

Second-year veterinary student Sarah Waibel is spending her summer in Gondar, Ethiopia, to continue the “Rabies Elimination Outreach Project,” which has been funded by Ohio State’s Outreach and Engagement since 2013. Ethiopia has the world’s second-highest human rabies incident rate, which makes it a One Health global preference location to help address and eliminate the disease.

In the U.S. it is commonplace to have our pets vaccinated for rabies – an effort that has decreased the country’s human rabies cases to about 2 or 3 per year, according to the Centers for Disease Control and Prevention. In Ethiopia, there are an estimated 1,456 human fatalities per year according to Rabies and Infections of Global Health in the Tropics (RIGHT), a partnership between Ohio State, the CDC, University of Gondor and Ethiopian Public Health Institute. Dogs are the largest transmitters of the disease, accounting for nearly 95 percent of all human rabies cases in the world, according to the World Health Organization.

Although many wild animals can transmit rabies to humans, we tend to interact with dogs more, especially in Ethiopia where dogs are known to roam freely around communities. Therefore it’s believed that vaccinating the dog population will significantly reduce human rabies cases.

Two young locals relax with their dog, which often roams the community, in Gondar, Ethiopia. Photo taken in July 2014 by Dr. Wondwossen Gebreyes, professor of Veterinary Preventive Medicine and Director of Global Health Programs for The Ohio State University College of Veterinary Medicine

Two young locals relax with their dog, which often roams the community, in Gondar, Ethiopia. Photo taken in July 2014 by Dr. Wondwossen Gebreyes, professor of Veterinary Preventive Medicine and Director of Global Health Programs for The Ohio State University College of Veterinary Medicine

Last summer, the dog population in Gondar was estimated by the RIGHT team, which included another Ohio State student in the Summer Research Program Alexandra Medley. The 2014 team recorded the number of dogs observed on a representative sample of the streets by surveying people in 13 urban, semi-urban and rural areas of the city. Now that researchers have this information, Waibel will be working on surveillance and monitoring of rabies cases in Ethiopia to better understand what the effective vaccination rate of dogs is to create widespread immunity, she said.

Waibel will additionally be working with the CDC on surveillance monitoring of reported bites and rabies cases in Ethiopia. She’s also studying Mycobacterium bovis, a bacterium that causes tuberculosis in cattle. Humans can contract the disease by drinking unpasteurized milk, which is a common practice in Ethiopia. Her principal advisors are Dr. Wondwossen Gebreyes, professor of Veterinary Preventive Medicine and Director of Global Health Programs for The Ohio State University College of Veterinary Medicine and Dr. Jeanette O’Quin, clinical assistant professor in Veterinary Preventive Medicine.

“I’ve always had a fascination in finding out how and why things work,” Waibel said.

She will be discussing what she and her team found at the Third International Congress on Pathogens at the Human-Animal Interface (ICOPHAI) in Thailand this August.

The effort is shared by many partners, including the Ohio State Colleges of Veterinary Medicine, Food, Agricultural and Environmental Sciences, and Public Health; the University of Gondar; Ethiopian Public Health Institute; U.S. Centers for Disease Control and Prevention; Federal Ministry of Health and Agriculture, Ethiopia; Ethiopian Wildlife Conservation Authority and the Pan African Union Vaccine Institute.

Veterinary students study genes involved in cancer formation

Students at The Ohio State University College of Veterinary Medicine Marisa Maglaty (left) and Alexander Diedrich (right), pose in their lab at the Biomedical Research Tower in July 2015.

Students at The Ohio State University College of Veterinary Medicine Marisa Maglaty (left) and Alexander Diedrich (right), pose in their lab at the Biomedical Research Tower in July 2015.

Two Ohio State veterinary students, Alexander Diedrich and Marisa Maglaty, are inspecting genes that are scientifically correlated with the development of various cancers.

Irregular cell cycles, such as the loss of control over cell growth or cell death, underlie many human and animal cancers.

Maglaty is studying the gene E2F8, a member of a larger family of genes called E2F, which is vital to coordinating progression through the cell cycle. E2F8 differs in structure from other E2F genes, Maglaty said, and she and her team are marking its locations throughout the cell cycle using a mouse model. This technique allows for observation of which genes and proteins E2F8 directly interacts with. Understanding the specifics of these processes is yet another step in the fight to cure cancer.

“The more knowledge we gain through basic discoveries, the more we help humans and animals,” Maglaty said.

Diedrich’s research focuses on PTEN, a gene that is widely expressed throughout the body, and its role in endometrial (uterus lining) cancer. If functioning correctly, PTEN works as a tumor suppressor, possibly preventing cells from growing and dividing too rapidly. Mutations of the gene are believed to be a step in the formation of many cancers.

Cancer of the endometrium occurs more frequently in developed countries, and has the highest incidence rate of gynecologic cancers in U.S. women, according to the Centers for Disease Control and Prevention. Diedrich will gain a better understanding of the tumor-suppressing effects of PTEN in female mice by measuring the degree to which certain genes are being expressed during stages of the estrous cycle (a mouse’s menstrual cycle).

Diedrich and his team hypothesize that anything that changes the normal function of the estrous/menstrual cycle could be a risk factor for developing endometrial cancer, since PTEN is important in regulating these cycles. For example women who take tamoxifen, a drug that treats breast cancer, often develop endometrial cancer later in life, Diedrich said. They think this may be because tamoxifen alters hormone signaling.

“Oncology research tends to flow across species,” Diedrich said. “So although this study focuses on human medicine, it feeds back into veterinary medicine.”

Both of the students are working under Dr. Gustavo Leone, professor in the College of Medicine’s Department of Molecular Genetics .The research teams aim to evaluate how these genes perform in combination with mice’s normal physiology, as opposed to in vitro, Leone said.

Scientists have a good idea of how E2F8 and PTEN operate throughout cell and estrous cycles, and this study will give them concrete evidence to confirm existing hypotheses.

“We’re pretty sure of where the genes should be, and when,” Leone said, “But we haven’t actually seen them there.”