Michel Wattiaux, GHI Advisory Committee member, follows dairy cows’ carbon footprints from barn to field

This story was originally posted on news.wisc.edu.

April 4, 2018 / By Bob Mitchell

Michel Wattiaux examines the contents of a cow’s stomach. UW-MADISON

Sometimes dairy scientist Michel Wattiaux approaches his research like a cop at a traffic stop. He uses a breath analyzer to check for problematic products of fermentation.

Last spring, the University of Wisconsin–Madison researcher began using a specialized device to measure the methane being exhaled or belched by a group of Holsteins and Jerseys. It was the first step in an ongoing study by dairy scientists, engineers and agronomists to see how a cow’s breed and forage consumption affect the greenhouse gases generated by her gut and her manure.

Greenhouse gases, which collect in the atmosphere and trap the sun’s radiation, are a big issue for the dairy industry. Methane is a concern because it’s particularly potent — it traps about 30 times as much radiation as carbon dioxide does — and a cow generates a lot of it in her rumen, the huge stomach chamber where microbes are fermenting as much as 200 lbs. of plant material. Also worrisome is nitrous oxide, another potent greenhouse gas that is emitted from manure during storage and after it’s spread in the field.

The U.S. dairy industry has set a goal of reducing its greenhouse gas emissions by 25 percent by the year 2020, and UW–Madison researchers are helping identify strategies to accomplish that.

Sampling the cows’ breath was the first in a sequence of experiments designed to measure greenhouse gas emissions at three critical points: from the cow’s breath, from her manure during storage, and from the field where her manure is spread. The researchers are looking at how three variables — breed of cow, type of silage fed, and relative levels of forage in the diet — affect greenhouse gas emissions at each point.

While versions of each of these experiments have been done on a standalone basis on the UW–Madison campus, Wattiaux says this is the first time the three have been integrated so that emissions originating from a cow and her manure can be tracked from barn to manure storage to field.

“This is the first time where we do the nutrition part, the manure storage part and the field application part sequentially, and then put it all together to give the Wisconsin dairy industry a solid number for how much methane and nitrous oxide comes out of their farms depending on the breed, the kind of diet and the amount of forage in the diet,” he says.

For the first experiment, which began in June and ran for four months, researchers fed 24 Holsteins and Jerseys a ration that included either alfalfa silage or corn silage, the two primary forages fed on Wisconsin dairy farms, along with some grain. Some cows were fed high levels of forage relative to grain, while others got less silage and more grain. Researchers periodically sampled each cow’s exhaled breath using the GreenFeed system, an analytical tool designed to determine daily methane emission.

“It drops a bit of sweet feed to entice her to stick her nose up to it,” Wattiaux explains. “The equipment sucks the air in, measures airflow, measures the concentration of methane and then estimates the amount of methane.”

In the second experiment, the manure from the cows was collected and held in barrels for two months to simulate manure storage on a dairy farm. Graduate student Elias Uddin collaborated with biological systems engineering professor Rebecca Larson to measure emissions of both methane and nitrous oxide from each barrel for 60 days.

The third experiment began at the end of October to simulate the post-harvest manure spreading typical of many Wisconsin farms. Researchers applied the stored manure to 24 field plots at the Arlington Agricultural Research Station. Under the supervision of agronomists Greg Sandford and Randy Jackson, a team of students began monitoring emissions from the plots last fall and will resume this spring.

Wattiaux believes that the findings from this research will be useful to scientists who create whole-farm decision models that producers use to predict the outcome of various management practices. He likens it to software such as Wisconsin’s SnapPlus, which farmers use to minimize soil and nutrient loss from their fields.

“In SnapPlus, you provide the field characteristics such as location and slope and crop management practices, and the model gives your ‘T’, your tolerable soil loss, so you can make sure you stay below that,” he says. “I think we’re going the same direction with this research. A model might calculate a tolerable level of greenhouse gas emissions and provide information on how to stay below that total by adopting new techniques in the field, new techniques in storage and new techniques in feeding.”

Ebola vaccine inches toward human clinical trials

A whole-virus vaccine to confront Ebola, the rare but often fatal hemorrhagic disease that periodically erupts in sub-Saharan Africa, may soon be one step closer to the clinic.

Yoshihiro Kawaoka

With the help of experts at Waisman Biomanufacturing, within the University of Wisconsin-Madison’s Waisman Center, UW-Madison School of Veterinary Medicine Professor Yoshihiro Kawaoka will lead a $3 million effort to produce as many as 1,000 doses of an experimental vaccine that has already been proven to work safely in monkeys.

“The goal is to produce a safe and effective vaccine against Ebola virus for people,” says Kawaoka, a world expert on Ebola and influenza. The vaccine is planned for use in a phase 1 clinical trial in Japan and is the only whole-virus Ebola vaccine candidate under development.

It will be produced at Waisman Biomanufacturing, a specialized facility whose mission is to help translate scientific discovery into early-stage clinical trials. The staff of the facility provides expert help with manufacturing processes, quality control and overall product development in addition to regulatory support.

Ebola virus swarms the surface of a host cell in this electron micrograph. Like most viruses, Ebola requires the help of a host cell to survive and replicate. Photo by Takeshi Noda, University of Tokyo.

“Waisman Biomanufacturing produces many different types of biopharmaceutical products, keeping our range of expertise broad in order to serve any University of Wisconsin investigator who has a biological that they wish to bring into the clinic,” says Carl Ross, the facility’s managing director. “We have made many prophylactic and therapeutic vaccines for use in human clinical trials.”

The technology behind the new Ebola vaccine was devised nearly a decade ago by Peter Halfmann, a research scientist in Kawaoka’s lab who is also an expert on the Ebola virus. It is known as “Delta VP30,” and is a form of Ebola virus that is noninfectious and safe to work with under routine laboratory conditions such as those at Waisman Biomanufacturing. The virus is missing a critical gene – one of only eight genes that make up the virus genome – that makes a protein the virus needs to reproduce in host cells.

Vaccines work by exposing the immune system to viruses or parts of viruses. The Delta VP30-based vaccine may offer better protection against Ebola virus than others in the pipeline, Kawaoka says, because it is a whole-virus vaccine. Other Ebola vaccine candidates use vector viruses to ferry a single Ebola protein, a surface antigen, to prime the immune system.

“Here, we have a whole-virus vaccine that presents all the viral proteins to the immune system, which may result in increased and broadened immune responses compared to vaccines that present only a single viral antigen to the immune system,” Kawaoka explains.

The need for an Ebola vaccine is acute. Periodic outbreaks of the disease in sub-Saharan Africa, including an epidemic between 2013 and 2016, caused major loss of life and serious economic disruption in the three countries where it occurred: Sierra Leone, Guinea and Liberia.

The new vaccine project will be the subject of an informational meeting to be held Feb. 27 at 4:30 p.m. at the Friends of the Waisman Center Auditorium on the first floor of the West Annex. The Waisman Center is located at 1500 Highland Ave. Free parking is available after 4:30 p.m. in Lot 82, behind the Waisman Center and accessible from Highland Avenue.

By Terry Devitt, University Communications/ February 21, 2018

This story was first published at news.wisc.edu.

Madison firm co-founded by GHI Advisory Committee member Yoshihiro Kawaoka advances human trials of revolutionary influenza vaccine

 

Yoshihiro Kawaoka, professor of pathobiological sciences in the School of Veterinary Medicine and a co-founder of FluGen, gives a slide presentation to a group of media representatives touring the Influenza Research Institute. PHOTO: JEFF MILLER

This story appeared first at news.wisc.edu.

By David Tenenbaum

Amid predictions that this year’s flu vaccine will offer limited protection, medical researchers are renewing their focus on a universal flu vaccine.

A universal flu vaccine would offer more broad protection than today’s vaccines, which must be targeted at viral strains deemed dangerous many months before flu season begins.

The problem is that the flu virus can change fast enough to evade those vaccines.

No versatile influenza vaccine is on the market, but one of the most promising is being developed by FluGen, a spinoff from the University of Wisconsin–Madison. The Madison startup has announced a plan to evaluate its innovative influenza vaccine in a trial of 100 people later this spring.

Yoshihiro Kawaoka PHOTO: JEFF MILLER

For almost a decade, the company has been exploring a genetically altered virus that can reproduce only once in the human body. “Our vaccine contains a live virus that infects you but cannot make you sick,” says CEO Paul Radspinner.

Viruses reproduce by hijacking cellular machinery. The essential technology for Flugen’s project deletes a gene the virus needs to reproduce more than once. The discovery was made in 2002 by company co-founders Yoshihiro Kawaoka, an internationally known influenza researcher at the School of Veterinary Medicine, and his colleague Gabrielle Neumann.

The principle that an infection can be powerful medicine, however, is much older, Radspinner says. “Our vaccine is based on the concept that if you get the flu this year, the odds that you will get it next year go down dramatically.”

Beyond creating a real but extremely limited infection, Flugen’s invention also uses a promising delivery route. Instead of being injected into the muscle, it will be squirted into the nose, which is far more faithful to a natural infection. “If you think how you get the flu. It’s usually through the nasal passages or the throat,” Radspinner says.

Intranasal delivery triggers immunity in the mucus been shown to “activates multiple immune systems in the body, while the traditional shot in the arm affects primarily the antibody-based immune system.”

Influenza is a global disease caused by a group of viruses that change rapidly and spread easily through coughs and sneezes. Worldwide, the disease kills between 250,000 and 500,000 per year. Due to the long lead time needed to produce vaccine in eggs, public-health authorities must choose vaccine for the northern hemisphere in February.

During the nine or more months before flu season begins, the virus undergoes “genetic drift” as it evolves and reasserts its genetics. Such drift is a major reason why, even in healthy adults, influenza vaccines are only 10 to 60 percent protective.

FluGen’s vaccine is made in mammalian cells, not eggs, so faster production should allow less time for genetic drift. Unlike the current vaccine, the virus is alive, and therefore more likely to trigger immunity. Already, FluGen has shown that the vaccine triggers a much broader immune response, affecting not just the mucus tissues but also B-cells, T-cells and antibodies.

“Our goal is to trick the body into thinking that it’s infected, so you get protected, without getting the symptoms that are the historic price of protection.”

Paul Radspinner

FluGen’s technology is protected by multiple patents held by both the Wisconsin Alumni Research Foundation and FluGen.

A test in healthy adults, completed in 2016, showed no warning flags related to safety, “and it told us we are hitting many sections of the immune system,” Radspinner says.

Those results helped to justify a $14.4 million grant awarded by the Department of Defense to test whether a vaccine based on a 2009 strain of flu can protect against the strains that circulated in 2014-15. “We’re aiming to prove effectiveness after six years of genetic drift,” says Radspinner. “That’s a big mismatch.”

Half of the 100-odd volunteers in the upcoming study will get a saline squirt in the nose; the others will get FluGen’s vaccine. Later, everyone will get a nasal dose of live flu virus. “They’ll be in ‘flu camp’ for 11 days in an isolation facility in Belgium,” Radspinner says. “We will measure how well our vaccine protected against the flu challenge, based on infection and symptoms, and we’ll look at a broad range of safety indicators as well.”

The goal is to have initial results by the end of the year.

Despite the dual benefits from faster vaccine production and broader immunity, regulatory approval is expected to entail safety-efficacy trials involving several thousand people. “With any vaccine, safety is the paramount issue, so the FDA [Food and Drug Administration] wants to see as many subjects as possible,” Radspinner says.

Depending on the scientific results, and regulatory approval, 2025 is a reasonable time for gaining FDA approval and reaching the market, he says.

As FluGen’s dozen employees prepare for the larger trials, the company has begun to use more outside resources, including specialized manufacturers that are not found locally. “We are a semi-virtual company,” says Radspinner.  “We have half of our employees doing basic research and development in the lab, and we coordinate external clinical research and manufacturing through outside organizations.”

Influenza can be extraordinarily dangerous, and flu researchers are haunted by the 1917-18 pandemic, which killed 50 to 100 million people worldwide. Public-health authorities continue to promote the benefits of flu vaccine, but the protection is better in some years than others, Radspinner says. “It may be too early to tell, but the early surveillance, an area where Wisconsin has long been a leader, clearly indicates that this is going to be strong year for flu.”

Even though getting the flu is your best protection against another bout, “Who wants to get sick for a couple of weeks?” he asks. “Our goal is to trick the body into thinking that it’s infected, so you get protected, without getting the symptoms that are the historic price of protection.”

In the heart of devastating outbreak, research team unlocks secrets of Ebola


a vial is labeled and prepared to hold blood from an Ebola patient in Sierra Leone. Researchers from the UW-Madison, the University of Tokyo and the University of Sierra Leone will compare blood from those who died of the virus to those who survived and those who never got sick to try and develop treatment. (Photo courtesy of Kawaoka Lab.)

This story appeared first at news.wisc.edu.

In a comprehensive and complex molecular study of blood samples from Ebola patients in Sierra Leone, published today (Nov. 16, 2017) in Cell Host & Microbe, a scientific team led by the University of Wisconsin–Madison has identified signatures of Ebola virus disease that may aid in future treatment efforts.

Conducting a sweeping analysis of everything from enzymes to lipids to immune-system-associated molecules, the team — which includes researchers from Pacific Northwest National Laboratory (PNNL), Icahn School of Medicine at Mount Sinai, the University of Tokyo and the University of Sierra Leone — found 11 biomarkers that distinguish fatal infections from nonfatal ones and two that, when screened for early symptom onset, accurately predict which patients are likely to die.

With these results, says senior author Yoshihiro Kawaoka, a virology professor at the UW–Madison School of Veterinary Medicine, clinicians can prioritize the scarce treatment resources available and provide care to the sickest patients. Kawaoka is also a member of the GHI Advisory Committee and received a 2017 GHI Seed Grant to catalog viruses circulating among West Africans with an eye to improving diagnoses, identifying new viruses and, potentially, preventing the next epidemic.

Studying Ebola in animal models is difficult; in humans, next to impossible. Yet, in Sierra Leone in 2014, a natural and devastating experiment played out. In September of that year, an Ebola outbreak like no other was beginning to surge in the West African nation. By December, as many as 400 Ebola cases would be reported there each week.

That fall, Kawaoka sought access to patient samples. He has spent a career trying to understand infectious diseases like Ebola — how do they make people sick, how do bodies respond to infection, how can public health officials stay at least a step ahead?

“Here, there is a major outbreak of Ebola. It is very rare for us to encounter that situation,” says Kawaoka, who is also a professor of virology at the University of Tokyo.

Yet blood samples were proving difficult to obtain and people continued to die.

Then, just weeks before Christmas, Kawaoka learned about a colleague in his very own department at UW–Madison, a research fellow from Sierra Leone named Alhaji N’jai, who was producing radio stories for people back home to help them protect themselves from Ebola. The pair forged a fortuitous partnership.

“He knows many people high up in the Sierra Leone government,” says Kawaoka. “He is very smart and very good at explaining things in lay terms.”

By Christmas, Kawaoka, N’jai and Peter Halfmann, a senior member of Kawaoka’s team, were in Sierra Leone.

“On the first trip, Alhaji took me to Parliament and we talked to a special advisor to the president, then the vice chancellor of the University of Sierra Leone,” says Kawaoka. “We got the support of the university, which helped us identify military hospitals and provided space. We went to the Ministry of Health and Sanitation and the chief medical officer and we explained what we hoped to do.”

Yoshihiro Kawaoka, professor of pathobiological sciences at the UW-Madison School of Veterinary Medicine, meets with Ekundayo Thompson, vice chancellor of the University of Sierra Leone, while in the African nation to establish a partnership to study and fight Ebola while improving the research capacity and infrastructure of the University of Sierra Leone. (Photo courtesy of the Kawaoka Lab.)

By February of 2015, Kawaoka and other select senior researchers on his team, including Amie Eisfeld, set up a lab in a military hospital responding to the outbreak in the capital city of Freetown (the researchers never entered patient wards). With the approval of patients and the government of Sierra Leone, health workers collected blood samples from patients after they were diagnosed with Ebola and at multiple points thereafter.

They obtained 29 blood samples from 11 patients who ultimately survived and nine blood samples from nine patients who died from the virus. The samples were transported to the lab where Kawaoka’s experienced and expertly trained team inactivated the virus according to approved protocols. Blood samples were subsequently shipped to UW–Madison and partner institutions for analysis.

For comparison, the research team also obtained blood samples from 10 healthy volunteers with no exposure to Ebola virus.

SIDEBAR: Video reaches ‘Spiderman’ audience with Ebola messaging

“Our team studied thousands of molecular clues in each of these samples, sifting through extensive data on the activity of genes, proteins and other molecules to identify those of most interest,” says Katrina Waters, a biologist at PNNL and a corresponding author of the study. “This may be the most thorough analysis yet of blood samples of patients infected with the Ebola virus.”

The team found that survivors had higher levels of some immune-related molecules, and lower levels of others compared to those who died. Plasma cytokines, which are involved in immunity and stress response, were higher in the blood of people who perished. Fatal cases had unique metabolic responses compared to survivors, higher levels of virus, changes to plasma lipids involved in processes like blood coagulation, and more pronounced activation of some types of immune cells.

UW-Madison’s Yoshihiro Kawaoka, Peter Halfmann and Alhaji Njai stand outside of a military hospital with Foday Sahr, a Sierra Leone military official and chair of microbiology at the University of Sierra Leone. Ebola patients are treated at many of the country’s military hospitals like the Joint Medical Unit.
(Photo courtesy of the Kawaoka Lab.)

Pancreatic enzymes also leaked into the blood of patients who died, suggesting that damage from these enzymes contributes to the tissue damage characteristic of fatal Ebola virus disease.

And, critically, the study showed that levels of two biomarkers, known as L-threonine (an amino acid) and vitamin D binding protein, may accurately predict which patients live and which die. Both were present at lower levels at the time of admission in the patients who ultimately perished.

“We want to understand why those two compounds are discriminating factors,” says Kawaoka. “We might be able to develop drugs.”

When Ebola virus leads to death, experts believe it is because of overwhelming viral replication. Symptoms of infection include severe hemorrhaging, vomiting and diarrhea, fever and more.

Kawaoka and his collaborators hope to better understand why there are differences in how patients’ bodies respond to infection, and why some people die while others live. The current study is part of a larger, multicenter effort funded by the National Institutes of Health.

“This may be the most thorough analysis yet of blood samples of patients infected with the Ebola virus.”

Katrina Waters

“The whole purpose is to study the responses of human and animal bodies to infection from influenza, Ebola, SARS and MERS, and to understand how they occur,” Kawaoka explains. “Among the various pathways, is there anything in common?”

In the current Ebola study, the team found that many of the molecular signals present in the blood of sick, infected patients overlap with sepsis, a condition in which the body — in response to infection by bacteria or other pathogens — mounts a damaging inflammatory reaction.

And the results contribute a wealth of information for other scientists aimed at studying Ebola, the study authors say.

Kawaoka says he is grateful to UW–Madison, University Health Services and Public Health Madison and Dane County for assistance, particularly with respect to his research team’s travel between Madison and Sierra Leone. Each provided protocols, monitoring, approval and other needed support during the course of the study.

“I hope another outbreak like this never occurs,” says Kawaoka. “But hopefully this rare opportunity to study Ebola virus in humans leads to fewer lives lost in the future.”

By Kelly April Tyrell, UW communications/ November 16, 2017

THE STUDY WAS FUNDED BY A JAPANESE HEALTH AND LABOR SCIENCES RESEARCH GRANT; BY GRANTS FOR SCIENTIFIC RESEARCH ON INNOVATIVE AREAS FROM THE MINISTRY OF EDUCATION, CULTURE, SPORTS, SCIENCE AND TECHNOLOGY OF JAPAN; BY EMERGING/RE-EMERGING INFECTIOUS DISEASES PROJECT OF JAPAN; AND BY AN ADMINISTRATIVE SUPPLEMENT TO GRANT U19AI106772, PROVIDED BY THE U.S. NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES, PART OF THE NATIONAL INSTITUTES OF HEALTH. SUPPORT WAS ALSO PROVIDED BY THE DEPARTMENT OF SCIENTIFIC COMPUTING AT THE ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI AND BY A GRANT FROM THE NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES (P41 BM013493). SOME ANALYSES WERE PERFORMED AT THE ENVIRONMENTAL MOLECULAR SCIENCES LABORATORY, A NATIONAL SCIENTIFIC USER FACILITY SPONSORED BY THE U.S. DEPARTMENT OF ENERGY OFFICE OF BIOLOGICAL AND ENVIRONMENTAL RESEARCH.