A Jolt to the System

As a linebacker for the UW–Madison Badgers, Chris Borland made a name for himself as a hard-hitting tackler. His senior year, he was selected as a first-team All-American as well as the top linebacker and defensive player in the Big Ten Conference.

A third-round draft pick, Borland seemed destined for a headline career in the National Football League. But during a full-contact practice at the San Francisco 49ers summer training camp in August 2014, Borland got his “bell rung” by a 290-pound fullback during a routine exercise. Though Borland felt dazed, he played through—as he’d done dozens of times before.

Like many football players, Borland had endured his share of hard hits, including two diagnosed concussions. This particular hit, however, got him thinking seriously about the future, and about the negative effects that repeated collisions could have on his long-term physical and cognitive health. Even so, he went on to play a dynamite rookie year.

Then, after the season was over, Borland quit.

The announcement shocked the sports world. Borland was 24 years old and healthy, yet chose to walk away from a $2.3 million, four-year contract.

“I just honestly want to do what’s best for my health,” Borland explained on ESPN’s Outside the Lines. “From what I’ve researched and what I’ve experienced, I don’t think it’s worth the risk.”

With their repeated hits, football players—along with boxers—are at increased risk of developing chronic traumatic encephalopathy (CTE), a degenerative brain disease marked by memory loss, depression, suicidal thoughts, aggression and dementia. Of 91 brains donated to science by former NFL players, 87 have tested positive for CTE. It’s seen as a likely contributing factor to nine suicides by current and retired football players over the past decade.

Borland didn’t want to share that fate.

“To me, Chris Borland is a hero. He walked away before he made the big bucks and he was very explicit about why he quit—that it was not worth it to him,” says CALS genetics professor Barry Ganetzky, whose findings about the central nervous system in fruit flies are shedding light on what hard hits do to humans.

Ganetzky isn’t a sports guy, but he started paying attention to football-related brain injuries after the 2012 suicide of New England Patriots linebacker Junior Seau, intrigued by the biological processes driving this tragic phenomenon.

“I started wondering, what’s the link between a blow to the head and neurodegeneration 10 or 20 years down the line? When I started digging into the scientific literature, it became clear that we know very little,” says Ganetzky, who held the Steenbock Chair for Biological Sciences for 20 years. “And my usual response is, well, if we don’t understand something about the brain, then we should be studying it in flies.”

Fruit flies, officially known as Drosophila melanogaster, are a widely studied model organism, with a vast arsenal of genetic and molecular tools available to support that work. Flies reproduce rapidly and are easy to work with, enabling swift research progress. They are well suited for brain research because they have nerve cells, neural circuitry and a hard skull-like cuticle remarkably similar to our own, allowing scientists to conduct probing experiments that would be difficult in rodent models—and impossible in human subjects.

Fly models already exist to study Alzheimer’s, Parkinson’s and a number of other neurological diseases. Why not concussion? But there wasn’t a model available.

Then Ganetzky remembered work he’d done decades earlier.

“It occurred to me that I knew how to make flies have a concussion, and I had done it 40 years ago as a post-doc,” says Ganetzky. “I thought, ‘That’s it!’”

It was a simple thing: As a post-doctoral researcher at the California Institute of Technology, Ganetzky decided to see if any of his flies happened to be bang-sensitive mutants, flies that display seizures and paralysis after given a high-powered swirl on a vortex machine. But he didn’t have a vortex nearby, so he decided to just bang the vials against his hand.

“After a couple of sharp whacks, some of the flies were hanging out at the bottom of the vial, stunned. Others were on their backs, obviously knocked out. And after a few minutes, they all got up and started walking around again,” recalls Ganetzky.

He immediately knew the flies weren’t bang-sensitive—it’s an extremely rare mutation—but Seau’s death helped Ganetzky realize they had displayed symptoms “very similar in many respects to the empirical definition of a concussion.”

After developing and validating the new fly model, Ganetzky and UW genetics professor David Wassarman have been able to charge forward with brain injury research. The model has already been used to reveal key genes involved in the body’s response to brain injury. It’s also poised to help unlock medical applications, including a genetic test for high-risk individuals and an assortment of promising drugs and treatments.

In addition to helping athletes in contact sports, these advances will benefit the millions of Americans each year who experience traumatic brain injury due to falls, car accidents and violent assaults.

“At the most fundamental level, we just want to understand how traumatic brain injury works,” explains Ganetzky. “However, this is a major medical problem for which there are not many good—or any good—treatments or therapies or preventives, and so that is part of our motivation. If we can learn the genes and the molecules and the pathways, can we come up with interventions?”

Ganetzky was raised in a working-class neighborhood in Chicago by a candy salesman father and a homemaker mother. Growing up, he had an abundance of natural curiosity and asked a lot of tough questions—and often questioned the answers he received. While this trait caused him some problems as a youth, it came to serve him well in science.

At the University of Illinois in Chicago, he figured he’d become a chemist for the good career prospects. He ended up switching to the biological sciences, however, after a 10-week honors biology research experience in a Drosophila lab that expanded into a two-year project. From that point forward he stuck with flies, earning his doctoral degree at the University of Washington and then doing his post-doc work at Caltech.

In 1979, Ganetzky joined the University of Wisconsin–Madison, where he chose to focus his research program on exploring temperature-sensitive paralytic mutants, flies that behave normally at room temperature, but then start to tremble and twitch—or pass out—when things heat up. For each mutant he identified, he sought to uncover the faulty gene involved, and thus better understand how brain cells work.

Over the decades, this approach enabled Ganetzky’s team to discover a number of critical genes and molecular pathways involved in brain cell signaling, including those required for the release of neurotransmitters. That body of work established Ganetzky as one of the foremost leaders in neurogenetics. Some of his findings shed light on human genetic diseases and led to a test that’s now routinely used to assess the safety of new pharmaceutical drugs. For his contributions, Ganetzky was elected in 2006 to the National Academy of Sciences, the nation’s preeminent scientific society.

After Ganetzky’s “eureka moment” about fly concussions in spring 2012, he immediately reached out to colleague David Wassarman, a genetics professor in the UW–Madison School of Medicine and Public Health. Wassarman, who studies human neuronal disorders using fruit flies, had already been attending Ganetzky’s lab meetings for a few years after some of their research findings linking the innate immune response and neurodegeneration dovetailed.

“I did a demonstration of fruit fly concussion for David, and I remember his response very well,” says Ganetzky. “His jaw kind of dropped, and he said, ‘If you’re not going to study that, then I want to.’”

It was exactly the response that Ganetzky had been hoping for. With retirement looming on the horizon, Ganetzky needed a trusted and enthusiastic collaborator to help pursue the work—someone who would be willing to take on more and more as time went on. Wassarman was game.

“I wanted to put both feet in,” says Wassarman. “I said, ‘If we’re going to do it, let’s do it.’”

As a first order of business, Wassarman developed a tool capable of delivering a consistent “dose” of brain injury to flies. The result, known as the High-Impact Trauma (HIT) device, utilizes a metal spring to slam a vial of flies against a firm foam surface. In this setup, it’s important to note, the brain injury the flies experience is caused by the rapid acceleration and deacceleration of their bodies; it’s not necessarily about a direct hit to the head.

“Quite often, as with football players, it can happen because they are running fast and then meet an immovable object. The concussion is caused by a kind of whiplash, where the brain is ricocheting off the inside the skull, and that’s what’s causing the damage,” says Ganetzky. “That’s what we’re doing here with the flies.”

Ganetzky and Wassarman found that flies injured using the HIT device exhibit many of the classic symptoms of traumatic brain injury (TBI) seen in humans. As they reported in the Proceedings of the National Academies of Science in 2013, flies show temporary incapacitation and loss of coordination immediately after injury. Those that survive severe injury go on to develop long-term symptoms: activation of the innate immune response, neurodegeneration and early death.

These TBI flies have the potential to reveal much-needed insights—and medical interventions—for the millions of Americans who experience traumatic brain injury each year. According to the U.S. Centers for Disease Control and Prevention, TBIs cause around 2.5 million emergency room visits, 283,600 hospitalizations and 52,800 deaths each year. Top causes are falls, motor vehicle accidents, and blows or jolts to the head or body, including sports-related concussions. Bomb blasts can cause brain trauma in soldiers in combat zones. Across the country, as many as 6.5 million people are believed to be struggling with the consequences of TBI, and the total economic cost of this health issue is estimated to be $76 billion per year.

In a demonstration of the power of the TBI model, Rebeccah Katzenberger, a senior research specialist in Wassarman’s lab, subjected 179 genetically unique strains of flies to four strikes of the HIT device—meant to simulate a series of severe brain injuries—and then monitored them for death at 24 hours post-injury, a data point that serves as an easy-to-measure proxy for the various negative events unfolding inside the body.

The results revealed a huge diversity of responses, underscoring the fact that genotypes matter when it comes to TBI response. Some strains were particularly susceptible to death, losing as many as 57 percent of the flies in those first 24 hours, while others were much more resilient, losing just 7 percent. The team then identified the genes that possibly made a difference, publishing their findings in eLife in March 2015.

“Now we have these 100 genes, and scientists can start looking at them in more detail,” says Wassarman. “A lot of them are genes that had never really been implicated in traumatic brain injury before. I think this is going to be one of our big contributions.”

These findings, the researchers note, may help explain why people respond so differently to similar brain injury events, and may help lead to a genetic test to identify high-risk individuals.
“Once we understand those genetic links, we’ll be able to test people and tell them, ‘Look, you probably shouldn’t play football. You should play non-contact sports,’” explains Ganetzky.

After identifying the TBI genes, Ganetzky and Wassarman immediately noticed a handful of genes involved in tissue barrier regulation. Tissue barriers—such as the intestinal barrier and the blood-brain barrier—function as biological blockades keeping “bad” things out while allowing “good” things to pass through.

To explore the connection between brain injury and tissue barriers, the duo had Katzenberger conduct a simple, colorful experiment that involves adding bright blue dye to the flies’ food. Under normal conditions, when flies eat the blue-colored food, it stays in the gut, something that is readily observable through the fly exoskeleton. However, after exposure to brain injury—via the HIT device or by having their heads pinched with a forceps—they found that the dye leaks out of the gut and turns the entire body blue, a phenomenon called “smurfing” (after the blue Smurf cartoon characters).

Leaky tissue barriers have previously been observed in rodent models of brain injury as well as in human medical cases. “Somehow this injury to the brain is triggering a series of events that leads to the breakdown of the intestinal barrier,” notes Ganetzky. “So there’s some sort of cross-talk going on between the brain and the intestine, but we don’t fully understand it yet.”

Upon further exploration, Ganetzky and Wassarman were able to confirm that—along with the blue dye—glucose and bacteria were also crossing the intestinal barrier into the fly’s circulatory system, or hemolymph, after brain injury. Homing in on glucose, they found that it plays a causative role in fly death after TBI. “By simply withholding sugar, we were able to keep some of these flies alive, and by a substantial margin,” says Wassarman.

If the findings hold up in rodent models and in human trials, he notes, athletes may one day find themselves advised to avoid certain foods after experiencing concussion.

The bacteria that cross the intestinal barrier appear to be playing more of a long game. Ganetzky and Wassarman believe they are the culprits triggering the innate immune response observed in TBI flies. The innate immune response, also known as the inflammatory response, is the body’s natural reaction to microbial invasion and other stressors. If properly controlled—turned on and off at the right time—it protects the body. If left on, however, it can cause collateral damage throughout the body, including damaging brain cells.

“Here’s what we think is happening: Traumatic brain injury is causing increased intestinal permeability. That causes the bacteria to leak out, which turns on the innate immune response, and that is possibly leading to neurodegeneration down the line,” explains Wassarman.

Ganetzky and Wassarman are intrigued by a concept that is emerging from their work and related studies: that TBI accelerates aging. Some of the key physical outcomes of brain injury—problems with tissue barriers and increased inflammation—are also hallmarks of the natural aging process. More support for this idea came in summer 2015, with the release of a report describing signs of early aging in the brains of war veterans exposed to bomb blasts in Iraq and Afghanistan.

“Somehow a blow to the head is activating all of these pathways related to aging and speeding them all up. Biologically, I think that this is maybe one of the most fascinating things about the whole project,” says Ganetzky, noting that TBI flies are a great model for further exploration.

Even at this early stage, without fully understanding the basic scientific mechanisms involved, the model is already revealing some promising medical applications. As soon as Ganetzky and Wassarman realized that the inflammatory response might lead to neurodegeneration, a treatment suggested itself: Could a simple anti-inflammatory help? They tried giving TBI-injured flies some aspirin mixed in their food. It helped.

“Our studies show that there appears to be a window of time after brain injury when the flies are particularly susceptible to dying. And if we can prevent certain events from happening during this time, then we can prevent death,” says Wassarman. “That’s what we think aspirin is doing—by lowering the innate immune response.”

The next step is to look for drug candidates that work even better than aspirin. Ganetzky and Wassarman are in the process of screening a set of 2,400 compounds, and they’ve already found a handful of very promising ones that can now be tested in rodent models and, ultimately, in human clinical trials.

“It would be wonderful if someday it were possible to offer a simple intervention beyond surgery to help individuals who have suffered a severe traumatic brain injury,” says Wassarman.

There’s a lot left to learn, and Ganetzky and Wassarman are eager to pursue all that the model can tell them. With Ganetzky’s retirement set for early 2016, the work of securing the project’s first federal grant and conducting experiments will largely fall to Wassarman.

But Ganetzky won’t be out of the picture. He continues to keep up on brain injury medical cases and scientific discoveries, and is encouraged by the national conversation about sports and brain injuries that’s starting to gain traction—and by the NFL’s commitment to scientific research in this area.

Some of these advances can be attributed, in part, to Chris Borland, whose post-NFL journey has led him deeper into the world of sports-related brain injury. Borland has submitted to numerous brain scans to support research, and has also become a sought-after speaker, touring the country to raise awareness about the risks of concussion.

It’s that kind of dedication to public service on the part of Borland and many other athletes, along with the excitement of discovery, that’s keeping Ganetzky in the game. Despite his retirement, Ganetzky plans to keep a scaled-back version of his lab running for at least a few more years.

For the Birds

Slipping into a patch of woods in western Dane County, Jim Berkelman ignores the swarming mosquitoes and strains to sort through the early- morning chatter of warblers, robins and vireos and the nearby drum of a pileated woodpecker. “I’m hearing something I wouldn’t expect to hear,” says Berkelman, a lecturer in the Department of Forest and Wildlife Ecology at CALS and a volunteer contributor to the Wisconsin Breeding Bird Atlas II, a comprehensive, volunteer-powered survey of birds that nest in Wisconsin.

Experienced birders use their ears as much as their eyes to identify species, and Berkelman thinks he hears a northern parula, a small warbler that doesn’t typically nest this far south. Finding a bird, Berkelman explains, is only the start. The point of the Atlas, he notes, is to identify and map where birds in Wisconsin are courting, nesting, breeding and raising their broods.

To be sure of that, “atlasers,” as volunteer observers like Berkelman are called, must find tangible evidence that a species has actually taken up residence. A nest, of course, is the most obvious clue. But most birds are assiduously covert in their nesting and only conspicuous players like robins, herons, orioles, house wrens and bluebirds construct their nests in ways that make them easy to find and identify.

Other definitive hallmarks of breeding birds include observations of birds carrying nesting material or food for nestlings; distraction displays where birds seek to draw animals, other birds or humans away from a nest; and, of course, fledglings. Some bird species are fastidious as well and carry fecal matter away from occupied nests. Such an observation is also a telltale sign of breeding and can be used by an atlaser to confirm breeding activity and provide a new data point that science can ultimately draw on.

Following a rising wooded path to the top of a hill, Berkelman’s rounds on this warm June day encompass two different types of ecosystems: forest, and open fields and prairie. His block is designated as a “priority block,” a specified block within a six-block “quad” on a grid of more than 7,000 three-mile-by-three-mile blocks that covers Wisconsin. Within that grid are 1,175 priority blocks, each of which requires at least a year’s documentation of breeding birds within a five-year period to ensure that the state is uniformly surveyed for the new Atlas. In addition, there are 153 “specialty blocks” that have unique habitat, are of high conservation value or are of particular interest to ornithologists.

Today, Berkelman is recording his data the old-fashioned way: with pen and notebook. Later, he can plug his observations into Atlas eBird, an online checklist program that is a direct conduit to the database that is the bedrock of the Wisconsin Breeding Bird Atlas.

Data, of course, are the raw material of science. Astronomers gather it by measuring and parsing starlight. Molecular biologists get data by plumbing the sequence of the chemical base pairs that make up a gene or genome. Meteorologists numerically dissect the many variables of weather—temperature, precipitation, wind, clouds.

To be sure, most data collection is a laborious and numbing process—the antithesis of the eureka moment. Harvesting data can be very expensive, too, as the tools of modern science have become bigger, more complex and more powerful in their ability to see farther or smaller, drill deeper, or accelerate particles to higher energies. Indeed, much of what we hear about modern scientific discovery rests on the pillars of sophisticated technology. Think of the Hubble Space Telescope, the Large Hadron Collider, the IceCube Neutrino Observatory and the Human Genome Project as just a few examples.

But while technology is taking science to new heights, it’s also giving a boost to the age-old methods of data gathering like the ones Berkelman uses in his efforts to document the presence of breeding birds. The Internet and personal computing technology are being used like never before to crowd-source traditional observational data collected by a growing cadre of citizen scientists. Groups of people or individuals armed with laptops and app-laden smartphones are collectively logging everything from trash in the ocean and flying ants to cosmic rays and precipitation, giving working scientists access to oceans of new data and the revelations that come from subsequent analysis and interpretation.

In the realm of ecology, citizen science has gained a new standing as researchers have tapped into the potential of an interested public. Citizen science projects, mapping things like the presence and behaviors of bumblebees, manta rays, butterflies and bats, have fueled dozens of published studies.

It’s proven to be a powerful resource for Ben Zuckerberg, a professor of forest and wildlife ecology at CALS. North American birds and their distribution on a changing landscape are a primary focus of his research, a significant portion of which depends on data gathered by volunteer observers.

For instance, Zuckerberg and post-doctoral fellow Karine Princé drew on citizen science data to tell us that the cast of characters we see at our bird feeders in the winter is shifting, most likely due to climate change. Their study of wintering songbirds shows that some species, once rare during the Wisconsin winter, are shifting their ranges north, remaking the resident communities of birds that visit our backyard feeders.

The conclusions of the study rested on two decades of data gathered by thousands of citizen scientists through the Cornell University Laboratory of Ornithology’s Project Feederwatch.

“Birds have always been important environmental indicators,” Zuckerberg explains. Rapidly declining songbird populations in the 1950s and 1960s, he notes, were used to help ascertain the consequences of widespread use of the chemical insecticide DDT, which was subsequently banned, first in Wisconsin and then nationally.

The DDT story was famously informed by the unintended involvement of ordinary citizens who gathered baseline data in the form of bird eggs. In the 19th century, collecting bird eggs was a widespread hobby, an artifact of the Victorian obsession with the natural world. Many collections ended up in museums where, decades later, CALS ornithologist Joseph Hickey and his students used them to document the thinning of eggshells subsequent to the widespread introduction of DDT into the environment in the 1940s and ’50s.

Today the contributions of citizen scientists tend to be more directed, and the advent of personal computers and smartphones, in particular, are making participation easier, more immediate and more effective. And a prime example of that trend is the Wisconsin Breeding Bird Atlas, a collaborative project by the Wisconsin Department of Natural Resources (DNR), the Wisconsin Society for Ornithology, the Wisconsin Bird Conservation Initiative and the Western Great Lakes Bird and Bat Observatory.

This year, the group launched a second iteration of the Atlas. Zuckerberg and other scientists are working with Atlas coordinators and waiting in anticipation of a flood of new data from the project, which recruits volunteers statewide to survey thousands of designated blocks over a five-year period for evidence of breeding birds.

The first Wisconsin Breeding Bird Atlas featured data collected by nearly 1,600 volunteers between 1995 and 2000. As its name implies, the Atlas is a survey that documents the distribution and abundance of birds breeding in Wisconsin. It provides critical baseline information about bird species that live in our state and is an important benchmark in terms of assessing potential changes in bird populations over time due to things like habitat loss and climate change. It also helps document avian diversity, the state of endangered and rare bird species, and habitat needs in Wisconsin.

Such data, explains Zuckerberg, help scientists make sense of a world that involves players ranging from microbes to plants and animals, including birds. There are so many moving parts that capturing a wide snapshot of what exists where at a given point in time can give scientists insightful information about the dynamics, nuances and health of an ecosystem.

“Ecology is necessarily a messy endeavor,” Zuckerberg observes. “But at certain scales, it all becomes very clear.”

Drawing on things like Breeding Bird Atlas data, Zuckerberg and other scientists can get at the scales that matter: geography and time. As the Wisconsin Breeding Bird Atlas II effort gets under way, ecologists are laying the groundwork for analyzing the data by formulating hypotheses and ideas about what the data might show and how it will compare to data in the first iteration of the Atlas, which, according to the Wisconsin Society of Ornithology, “represented the largest coordinated field effort in the history of Wisconsin ornithology.”

Data collection for the Wisconsin Breeding Bird Atlas II began in 2015 and runs through 2019. In September the DNR released findings for the first Atlas season. Volunteers submitted nearly 24,000 checklists documenting the location and breeding activity of 229 species of birds. These early data show that wild turkeys are on the move, now populating nearly every corner of our state. And eight species of birds new to the Wisconsin breeding landscape since the last survey—including the iconic whooping crane—have cropped up in the new Atlas data.

“The stories that come out of the data are so robust,” Zuckerberg says. “We go in with our ideas of what we’re going to uncover, and some of the patterns just jump out at us.”

The major advantage of the Wisconsin Breeding Bird Atlas, according to noted ornithologist Stan Temple, a CALS emeritus professor in forest and wildlife ecology, is that it documents the relationship between birds and the places they require to successfully reproduce. “Habitat affinity is where the Atlas works best,” Temple explains.

Temple cites other long-standing citizen science efforts to document birds. The North American Breeding Bird Survey was officially launched in 1966. Conducted during the breeding season, volunteers traverse by car more than 3,700 randomly selected 24.5-mile road transects in the United States and Canada. Stopping every half-mile, volunteers document every bird seen or heard in a three-minute span before moving to the next observing station. The North American Breeding Bird Survey, Temple argues, is the gold standard for measuring population trends among birds.

A more recent citizen science effort—one that capitalizes on personal computing technology and helps inform the Wisconsin Breeding Bird Atlas—is the aforementioned eBird. Taking old-fashioned pen and paper checklists into the digital age, eBird is an online checklist linked to a central database. Used by amateur and professional birders, eBird logs millions of bird observations worldwide in any given month through a simple and intuitive web interface. The Wisconsin Breeding Bird Atlas II is the first state Atlas effort to employ it.

“We’re in the information age now,” explains Nick Anich, the Wisconsin DNR Breeding Bird Atlas coordinator. “We have eBird. We’re excited to use this new system. The developers have put an awful lot of effort into the checklist input, and they just launched the maps function. And the data update at least every 24 hours, so we can see things in real time.”

But can the information gathered by armies of citizen scientists be trusted? Can it help researchers predict the future of Wisconsin’s environment? How is it validated? Can scientists get over any qualms they might have about data collected beyond the strict parameters of controlled experiments and expert observation?

Zuckerberg, who has published on the use and value of crowd-sourced data, believes that many scientists are coming around to the idea that the data indeed represent an accurate picture of the natural world. “There has always been some skepticism about it in ecology. But studies show it is valuable data that are relatively accurate for picking up ecological patterns and processes,” Zuckerberg says.

“There are entire subfields of ecology dependent on these data. Theories in macroecology and how species respond to widespread environmental changes, such as pollution or climate change, for example,” Zuckerberg observes, referencing the study of relationships between living organisms and their environments at large spatial scales. “We wouldn’t be able to do anything like that without citizen science.”

That kind of insight is essential, Zuckerberg stresses, as broad-scale environmental change due to pollution, deforestation, reforestation and climate change will have significant and possibly lasting effects on birds in many different types of ecosystems.

According to Temple, the power of citizen science lies in the sheer numbers of observers. As a new CALS faculty member in 1976, Temple launched the Wisconsin Checklist Project. “The Wisconsin Checklist Project did in the predigital age what eBird does now,” Temple explains. “It is a rigorous way of engaging lots of bird-watchers in a very systematic way.”

For the most part, Temple says, the data are trustworthy. “Bird-watchers are used to keeping records, so you’re not asking them to do anything that already isn’t part of the culture. Mistakes in observing and recording happen, but it is safe to say those few errors become insignificant noise in comparison to the strength of the signal: the overwhelming number of accurate observations.”

For atlasers like Florence Edwards-Miller, a 31-year-old communications specialist from Madison, the chance to go into the field and gather data blends neatly with her deep-felt appreciation of the natural world.

Trekking through the prime birding habitat of Madison’s Nine Springs E-way on a rainy midsummer morning, Edwards-Miller is on a mission. An experienced birder, she knows she can confirm any number of breeding birds that use the settling ponds of Madison’s Metropolitan Sewerage District to raise their broods. And she is eager to contribute those little bits of data to the Wisconsin Breeding Bird Atlas effort.

“You can’t make good decisions unless you know what’s out there,” says Edwards-Miller. “I believe in science. I believe in the importance of the data.”

In a little more than an hour, she confirms the presence of breeding mallards, Canada geese and red-winged blackbirds—all pedestrian wetland species—by noting offspring and, in the case of the blackbirds, a cantankerous distraction display.

It takes a little longer to find the killdeer fledglings, but at the end of our circuit around the pond, there they are: little puffballs on stilts trailing behind their foraging parents. It’s a beautiful sight. And another valuable data point for the Wisconsin Breeding Bird Atlas.

Age-Old Traditions, New Media

There is no better place to begin this story than on an August morning in the remote reaches of the Bad River Ojibwe Reservation, afloat on Lake Superior’s shining Chequamegon Bay beneath an expansive, cloud-filled sky.

Several flat-bottomed boats are lined up gunwale-to-gunwale, bobbing in the gentle waves. They’re filled with students—a mix of UW–Madison undergraduates and tribal youth—on a field project run through UW–Madison’s Global Health Institute. They are listening to Dana Jackson and Edith Leoso, Bad River tribal members and elders, talk about wild rice and the windswept, watery landscape around them, the sloughs and the tamarack stands, the distant islands and the shimmering headlands.

It is all ancestral home to the Ojibwe, and Jackson and Leoso bring it to life with their words. They tell the Ojibwe creation story of how their tribal forebears came to the land so many years ago from the east, seeking, as they had been told in visions, a place where “the food grows on top of the water.” They speak of the chiefs who signed treaties to protect this homeland and of the warriors who fought to protect it and of the threats that come with modern times.

The students, armed with video cameras and recorders, soak it all up. The land seems to take on new depth and meaning, peopled now with the ghosts and the place names and shrouded in the mystery and the magic of the old stories.

It’s an ideal classroom for the CALS professor who is the guiding hand behind this floating, open-air lecture session.

Patty Loew, a professor of life sciences communication, has brought these students here to share with them the lives and the culture of a people she knows well.

Loew is a tribal member of the Bad River Ojibwe. She can trace her family back to ancestors who were among the tribal leaders signing the tribe’s historic treaties in the 1800s. When she looks out upon the waters of Lake Superior and the winding sloughs of the reservation, she sees her own family’s history. These places are as special to her as to any other member of the Bad River community.

Two years ago, in a column in the Wisconsin State Journal about the importance of this place to the Ojibwe, Loew wrote, “You won’t see any stained glass or church spires in the Bad River or Kakagon Sloughs, but those wetlands are as holy to us as any temple or cathedral.”

A noted television journalist and the author of several acclaimed books on Wisconsin’s Native Americans as well as an accomplished scholar, Loew could easily be resting on her many successes.

Instead, she is deeply involved in a number of teaching and media projects that are not only bringing the stories of Wisconsin’s Native Americans to life, but also are providing new ways for those stories to be shared by tribal members themselves. Since 2007, she has led efforts to teach tribal teenagers digital storytelling and technology skills. Working with colleagues as well as tribal leaders, she has helped young people create documentaries sharing Native American issues and culture. In a 2012 project, for example, eight St. Croix Ojibwe students created a tribal history told through the life stories of five St. Croix elders.

In this work Loew has also partnered with the UW–Madison Global Health Institute. She’s currently in the midst of a project—the one that has us floating on Chequamegon Bay—in which global health students from a wide range of majors work alongside tribal youth to bring the power of digital media to bear on reservation health issues such as nutrition and childhood obesity. The Bad River reservation has some of the highest diabetes and cardiovascular disease rates in the United States, according to a 2008 Wisconsin Nutrition and Growth Study.

Loew’s projects can already boast some impressive successes. In 2013, three 14-year-old Bad River participants in her tribal youth media workshops produced a documentary, Protect Our Future, that detailed the potential environmental threats posed by a proposed iron mine in the Penokee Range above the Bad River reservation.

The video was an award-winning hit. It played to large audiences at film festivals throughout the Great Lakes region and was screened at the Arizona State University Human Rights Festival. The teens were on hand to introduce their film, which they also shared at the nearby Salt River Tribal High School.

The project followed a unique blueprint developed by Loew that melds traditional knowledge from tribal elders and leaders with the use of digital media skills now being deployed by tribal youth.
It is, in effect, an artful and sensitive blending of the old and new. Loew, not one to think small, says she sees the work in the context of a larger and more powerful dream. Oblivious to the breeze and splashing water from Lake Superior, she speaks from her seat in one of the boats as it motors through the reservation’s famed Kakagon Sloughs. In between her answers to questions, she patiently works with students as they learn how to use video cameras. She helps one of them frame a shot and assists another who is figuring out how to program a video card.

“My ultimate goal,” Loew says as she works, “is to help Bad River become the media center for Indian Country. We want to combine really strong media skills with a really strong sense of culture.”

Loew’s work has drawn praise from many quarters, from tribal leaders to academic colleagues.

Joe Rose is an elder with the Bad River Ojibwe and has watched young tribal members embrace Loew’s teachings. He describes the pride that the video Protect Our Future brought to the reservation.
“We were fighting against the mine then,” Rose recalls. “That was a very serious threat to us. We were very concerned about our wild rice. That was exceptional work that Patty did with the young people. She taught them how to use the media, how to do the photography and the interviewing. They even did the music. And it was all done by students, only 14 or 15 years old.”

Don Stanley, a CALS faculty associate in the Department of Life Sciences Communication who specializes in social media, has worked alongside Loew on the reservation, served as her co-investigator, and, Loew says, sparked the original idea for much of their tribal youth media work.

There are few better examples of the Wisconsin Idea in action, Stanley says, when it comes to sharing the department’s communication expertise and scholarship with a broader audience.

And, in this case, that sharing is with a community that few can reach as effectively as Loew. Loew has the ability to connect in a special way, Stanley notes, because of her deep tribal roots and connections. People know her and see her knowledge and respect for tribal life and culture. That understanding and empathy is not always common among academics.

“A lot of time in academia, we don’t understand that,” Stanley says. “Researchers come in, extract what they want and leave. But people you are working with relate on a scale that is much more real and visceral when they’re dealing with somebody who gets it.”

And Loew gets it.

“She’s got incredible street cred,” Stanley says of Loew’s work on the reservation. “It’s a blast traveling with her up there. Everybody is a family member. Everybody is ‘Hey, Patty!’ and big hugs. I also think that because she doesn’t take herself so seriously, she’s really approachable.”

Indeed, Loew is quick to laugh, and a talker. She will enthuse equally about her work or a Green Bay Packer game (she is a devoted fan). She evokes laughter from her students when, passing by a reservation boat flying a Packer pennant, she says, casually, “Oh, look. The tribal flag!”

Loew is quick to point out an important caveat when it comes to her work with the Bad River community as it relates to the Wisconsin Idea. This is not about just transferring knowledge from the campus to the reservation, she says. In fact, she prefers the phrase “knowledge exchange.”

The tribes, Loew says, are a rich and unrecognized source of information about the natural world. The elders and others on the reservation have much to share, and that traditional knowledge can inform and extend science and natural resource management in the non-Indian world, notes Loew.

In the Ojibwe, Loew sees a people who have valuable lessons for us in how to combine culture with a respect for the natural workings of the planet.

“Over the past 25 years, I’ve seen a real need for scientific information that has cultural relevance,” Loew says. “Native communities may be poor in an economic sense but they are rich in natural resources. And the culture is attached to those resources in a way that can’t be separated.

“So it’s a two-way street,” Loew continues. “We don’t necessarily have the scientific capacity. But what we do have is storytellers and people who know and embrace the culture.”

Loew did not come to these understandings suddenly. They are the result of a slow and gradual awakening on her part to her own Native American heritage and a lifetime spent learning the communication skills that would one day allow her to bring the power of story to bear on sharing the history and culture and struggles of not only the Ojibwe but all of Wisconsin’s tribes.

Loew’s path has led her to a very professorial office in Hiram Smith Hall on the UW–Madison campus, home to the Department of Life Sciences Communication (LSC) and just a stone’s skip from Lake Mendota.
But Loew, as her colleagues will point out, seems to have trouble staying in that comfortable office. Everyone who works with her in Hiram Smith Hall has had the pleasant experience of meeting a wide-eyed Loew in the hallway and being greeted by the phrase “Hey! I have an idea I wanted to try out on you.”

It is more than a charming aspect of her character. It is how she works, bringing to life the cherished Wisconsin ideal of “sifting and winnowing.”

Loew is an idea factory. In recent months, her friends and co-workers have listened and watched as Loew has worried about the many employees who will be out of work when Oscar Mayer’s Madison factory closes. Perhaps, she muses as she talks with her colleagues, there is a way one of her video classes can help provide video resumes.

More often than not, those ideas become reality.

“She’s phenomenal at taking ideas and making them come to fruition,” says Stanley.

Professor and LSC department chair Dominique Brossard says Loew heightens the department’s effectiveness at giving students a more global perspective on the intersections of culture and science in the natural world. Her courses in ethnic studies and Native American issues and the media are very popular, she notes.

And with her extensive background in television and video production, Loew is a key player in achieving another of the department’s goals—providing foundational communication skills to students.
“She’s uniquely positioned to do this kind of thing,” Brossard says.

Loew has traveled a long road to reach this stage in her career. She grew up on Milwaukee’s north side, little aware of her Native American background and the important role it would play as her life unfolded.

“I didn’t know I was Indian until I was 13,” Loew recalls. “I was just a kid growing up in a housing project in Milwaukee.”

Looking back, Loew believes her mother, who was born on a reservation, and her grandfather, who lived with the family, were trying to shield her from the discrimination frequently faced by Native Americans. Her grandfather, Edward DeNomie, was raised in the Tomah Indian Boarding School. Life in such schools was harsh, and children were often punished severely for speaking their native language or clinging to other aspects of their culture.

Even so, Loew heard and relished the stories of her ancestors. And by the late 1960s, she had become well aware not only of her rich cultural heritage but also the ugliness of racial prejudice. She recalls a growing sense of outrage, especially in the 1970s as Native American rights became a prominent news story.

Loew pursued a career in broadcast journalism. She earned a degree from UW–La Crosse and started her broadcasting career working in the city as a TV and radio reporter.

Eventually Loew moved to Madison, where she worked her way up to the anchor’s desk at the ABC affiliate, WKOW–TV. Her awareness of Native American culture and her desire to tell the stories of Wisconsin’s tribes grew. In the 1980s, she earned awards and gained respect throughout the state for her coverage of the fierce legal battle and sometimes ugly boat-landing confrontations as the Ojibwe fought to reestablish off-reservation hunting and fishing rights that had been included in the treaties.

Loew would go on to make dozens of documentaries telling the stories and covering the struggles of Wisconsin’s Native American communities. After moving on to Wisconsin Public Television, she made reporting on the tribes a regular part of her job as host of the show Weekend.

In a 2006 interview in the magazine Diverse: Issues in Higher Education, Loew described the important connection between her rediscovered culture and her professional life.

“As a journalist, a researcher, you have questions,” Loew said. “You realize you are struggling for answers about yourself. So you want to be open, to make connections to people. You find yourself being very relational, and that’s very Native.”

That willingness to be up-front about her debt to her past, and to be outspoken about the indignities that Native Americans have had to endure, have sometimes landed her in interesting, if not difficult, positions.

After she gave a talk about some of the more unpleasant truths of the first Thanksgiving, she earned the ire of none other than radio talk show host Rush Limbaugh. He accused Loew of being part of a “multicultural curriculum which is designed to get as many little kids as possible to question the decency and goodness of their own country.”

Few of Loew’s documentaries received more attention than Way of the Warrior, an exploration of the role of Native American soldiers in the U.S. military that aired on PBS in 2007. During her research, she stumbled across a film about her grandfather’s World War I outfit. Her quiet Ojibwe grandfather, it turned out, had fought in seven of WWI’s major battles as part of the 32nd Red Arrow Division.

Later, in another serendipitous discovery, she would find his diary. She describes how touched she was and how she is still so taken by the idea of Edward DeNomie raising his hand to take the oath and enlist in the U.S. Army—even though he had been denied citizenship in the country for which he was willing to give his life. Native Americans were not granted citizenship in the United States until 1924.

The popular, eye-opening documentary told the stories of many such Native American soldiers. And, later, after earning her master’s and doctoral degrees in journalism and joining the Department of Life Sciences Communication, Loew would continue telling the stories of Wisconsin’s tribes and of her own people at Bad River. She’s written several popular books, including Indian Nations of Wisconsin: Histories of Endurance and Renewal—which has been adapted for children and is now widely used in public schools—and, most recently, Seventh Generation Earth Ethics, a collection of biographies about 12 Native Americans who were key figures in environmental and cultural sustainability.

Sitting in the stern of one of the boats winding through the reservation sloughs, Loew reflects on her storytelling past and connects it with the ancient tradition of the Ojibwe and other native cultures.

“We are oral storytellers,” Loew says. But she is lending a new twist to the revered tradition. By adapting digital media to the old stories, the power of their message is amplified and made more accessible, especially important when it comes to lessons regarding nutrition and health among tribal members.

For example, some of the young tribal videographers have scoured the reservation collecting information from elders about age-old gardening and cooking skills. They hope to use that information at some point, Loew explains, to create “teen cuisine” cooking shows focused on healthy eating.

It makes so much sense to combine the old and the new, Loew says. After all, she adds, by the year 2020, 80 percent of content on the World Wide Web is expected to be video.

“These are new tools to help us be who we are, to help us capture the essence of who we are,” says Loew. “It’s a way to preserve our stories and a really unique approach to documenting life on the reservation at this particular time in history.”

Students from the Global Health Institute class, traveling with Loew on weeklong field trips, have worked side by side with tribal youth to gather information for the health and nutrition project and to create videos.

Cali McAtee, a CALS biology major who went with Loew to Bad River in August, wrote in her journal about not only establishing close relationships with tribal young people, but also of gaining valuable insight into another culture. She recalls in her writings the feeling of traveling through a sea of rice at the edge of Lake Superior.

“I have seen a lot of wild rice in my life, but from far away. I probably assumed it was a field because you can’t really see the water in between,” wrote McAtee. “I liked hearing about the importance of rice to the Ojibwe because I don’t think I necessarily have anything as important or meaningful in my life as rice is to theirs.”

Loew has felt the power of story in her own life and in her own search for connections. Researching one of her books, Loew found herself reading the classic book Kitchi-Gami: Life Among the Lake Superior Ojibway, by Johann Georg Kohl. In the book she came across a story in which Kohl brings to life a meeting he had with a tribal elder.

That elder was none other than Loew’s great-great-grandfather, Loon’s Foot. Kohl wrote how, during his conversation with the old man, Loon’s Foot stepped back into his lodge and came out with a smoky, stained birchbark scroll. Unrolling it and speaking in French, Loon’s Foot showed Kohl the story of his family told on the scroll and the dots and lines that denoted the passing years and decades. The story reached back to the year 1142.

“Here I was just reading Kohl, and then holy smokes!” Loew recalls. “Not bad for an oral culture.”

Loew firmly believes it is possible to capture that same kind of magic today with new approaches to traditional storytelling.

Don Stanley has watched as Loew has found a way to navigate between two worlds—the quickly receding years of the elders and the fast-paced, media-rich present of the tribal young—to create a new way to tell and preserve story and tradition, and then apply their lessons to modern-day problems.

As an example, Stanley describes how, as part of the nutrition project, he has seen Loew work with Native middle school students, teaching them how to videotape an elder speaking about traditional foods and health. While Loew is helping the teens develop communication skills, she knows full well that she is also preserving the knowledge of that tribal elder for future generations.

No less an expert on Ojibwe tradition than tribal elder Joe Rose admires and respects Loew’s ability to bridge old and new worlds. He says that with the passing of the generation that experienced the assimilation policies of the boarding schools, it’s important that the young be able to hear the elders’ voices—to see their faces, lined and carrying the weight of the years, but still alive with the resilience and strength and wisdom of their ancient heritage.

“It is very important, since we do come from an oral culture,” Rose says of Loew’s task. “But you’ve heard the expression that a picture is worth a thousand words? Well, there’s truth in that, too.”
As for Loew, she says that the girl growing up in the Milwaukee projects has found her place.

“I’m doing what I was supposed to do,” Loew says. “I’m incredibly grateful that Don and I have found such a dedicated, caring community—our students, our volunteers, the Bad River kids and their families—with whom to pursue this work. They’re the ones who make it possible.”

Keeping Us Safe

It’s hard to believe now, but when the Food Research Institute (FRI) was established in 1946—two years prior to the founding of the World Health Organization—botulism and salmonellosis were poorly understood, and staphylococcal food poisoning was just beginning to be elucidated. Many otherwise well-known diseases were only alleged to be food-borne, and the causes of many known foodborne illnesses had yet to be established.

Now the oldest U.S. academic program focused on food safety, FRI moved from the University of Chicago to the University of Wisconsin–Madison in 1966 under the leadership of bacteriology professor Edwin “Mike” Foster.

And ever since, FRI has served as a portal to UW–Madison’s food safety expertise for food companies in Wisconsin, in the U.S. and around the world. Housed within CALS, the institute is an interdepartmental entity with faculty from bacteriology, animal sciences, food science, plant pathology, medical microbiology and immunology, and pathobiological sciences, drawing not only from CALS but also from the School of Medicine and Public Health and the School of Veterinary Medicine.

FRI offers a wealth of educational opportunities to both undergraduate and graduate students. Since 2011, FRI has coordinated its Undergraduate Research Program in Food Safety, which provides students with hands-on experience in basic science and applied investigations of food safety issues. FRI faculty and staff have trained hundreds of undergraduate and graduate students, post-docs, visiting scientists and research specialists throughout the years, and FRI alumni have gone on to hold positions in industry, government and academia across the country and abroad.

In keeping with the Wisconsin Idea, FRI’s reach extends well beyond campus boundaries through industry partnerships, especially with its 40 sponsor companies. The Applied Food Safety Lab and laboratories of FRI faculty collaborate with food processors to identify safe food formulations and processing techniques. The institute also provides outreach and training to both food companies and the greater scientific community through meetings, short courses, conferences and symposia.

“FRI is an outstanding example of how a public-private partnership can benefit the academic mission of UW–Madison and the needs of the Wisconsin food industry,” says FRI director Charles Czuprynski.

During the past 70 years, FRI has made many insights into the causes and transmission of foodborne diseases. Early on, FRI research established methods to identify and detect staphylococcal enterotoxins. Work conducted by FRI scientists pioneered understanding of the molecular mechanisms of botulinum toxin production and led to the harness of the toxin for biomedical uses. FRI faculty are leaders in mycotoxin research and have made important contributions to understanding the shedding of E. coli O157 by cattle, survival of Salmonella in stressful conditions and the role of Listeria in foodborne disease. FRI research also identified the health benefits of conjugated linoleic acid in foods of animal origin and conditions that might result in formation of undesirable components in processed foods.

Looking to the future, FRI research is investigating novel mechanisms to prevent food-borne pathogen growth in meat and dairy products, interaction of plant pathogens and pests with human food-borne pathogens, food-animal antibiotic alternatives, and the role of the microbiome in health and disease.

FRI will celebrate its 70th anniversary at its 2016 Spring Meeting May 18–19 at the Fluno Center on the UW–Madison campus. There’s also a reception on May 17 at Dejope Hall, near the grounds of the original FRI building. For more information about FRI and anniversary events, visit fri.wisc.edu.

Milk, Motherhood and the Dairy Cow

In the 1990s, dairy farmers were seeing a troubling trend in their herds. As cows produced more milk, their reproductive performance declined. This downward slope in reproduction, related to changes in the hormone metabolism of high-producing cows, spurred researchers into action. And CALS scientists found a solution—a reproductive synchronization system that could save Wisconsin dairy farmers more than $50 million each year.

“The development of these systems has been one of the greatest technological advances in dairy cattle reproduction since artificial insemination,” says Paul Fricke, a CALS professor of dairy science and a UW–Extension specialist. “It is highly, highly significant.”

For the past 20 years, Fricke has been working on the synchronization systems with fellow dairy science professor Milo Wiltbank. The systems, called Ovsynch, consist of treatments with naturally occurring hormones and are based on Wiltbank’s research into the basic biology of the cow reproductive cycle. The hormonal treatments synchronize the cycles so that farmers know when their cows are most likely to become pregnant.

Pregnancy rates in a herd are a product of two numbers: the service rate (the percentage of eligible cows that are inseminated) and the conception rate (the number of inseminated cows that become pregnant). Historically, farmers relied on visually recognizing when cows were in heat in order to time insemination—a tricky feat that often resulted in missed opportunities and low service rates.
“One of the biggest problems in dairy cattle reproduction is seeing the cows in heat,” says Fricke. “If you can proactively control the reproductive cycle, you can inseminate cows without waiting for them to show heat.”

Synchronization systems take the guesswork out of insemination, increasing service rates and pregnancy rates. Since the technology was first published in the mid-1990s, Fricke, Wiltbank and their colleagues have worked to optimize the systems. Researchers now see conception rates of more than 50 percent, and pregnancy rates of 30 percent or higher. Just 15 years ago, average conception and pregnancy rates were around 35 and 15 percent, respectively. A 30 percent pregnancy rate in herds producing high volumes of milk was unimaginable.

With impressive pregnancy rates and the safety of the system—the natural hormones used are short-lived and do not end up in food products—researchers and farmers alike are excited about further adoption of the technology. The payoff is substantial, considering the costs and benefits of breeding dairy cows, says Kent Weigel, professor and chair of the Department of Dairy Science.

“If we say that this technology will result in a 6 percent improvement in pregnancy rates, and we assume that it costs about $4 for each extra day that a cow is not pregnant, the technology could save Wisconsin dairy farmers about $58 million per year with just 50 percent of farmers using it,” explains Weigel. “This is a prime example of basic biology that turned out to have a practical application with huge economic benefits.”

PHOTO—Dairy scientist Paul Fricke has developed a way to inseminate cows before they show signs of being in heat.

Photo by Sevie Kenyon BS’80 MS’06

Bees and Beyond

Over the past 10 years or so, massive die-offs of the European honeybee—a phenomenon known as colony collapse disorder (CCD)—have sparked increasing concern about the fate of agricultural crops with the loss of these important pollinators. At the federal level, a White House Pollinator Health Task Force was formed and in May 2015 released a national strategy for pollinator protection.

In support of that effort, a number of states are following up with plans of their own. In Wisconsin, professor Claudio Gratton and postdoctoral research associate Christina Locke PhD’14 from the CALS Department of Entomology were invited to partner with the Wisconsin Department of Agriculture, Trade and Consumer Protection (DATCP) in leading a broad array of stakeholders to create a state pollinator protection plan.

The goal of the plan is to provide best management practice recommendations and educational materials for beekeepers, growers, pesticide users, homeowners and landowners who want to improve the health and habitat of managed and wild pollinators. A draft of the plan was open for public review as of this publication’s press time in early 2016, with the final report expected soon thereafter.

How bad is the bee situation in our state?

Locke: We have had very few reports in Wisconsin of colony collapse disorder, a phrase I don’t like to use because it refers to a collection of symptoms rather than a specific disease. One identifying characteristic of CCD is the disappearance of worker bees. Beekeepers go out to their hives and have a healthy queen and healthy brood cells, but the worker bees have somehow disappeared. That is not happening much in Wisconsin as far as we know.

What we do have are elevated annual losses and over-wintering losses in honeybee colonies. Wisconsin beekeepers averaged around a 60 percent colony loss for 2014–15, which is very high. Beekeepers will tell you that a sustainable loss is between 10 and 20 percent every year. These high losses are due to a combination of things. We’ve had a couple of really hard winters, and the honeybees aren’t necessarily adapted to our Wisconsin winters. So there are some efforts to breed queens that are cold-adapted.

The biggest thing that correlates with colony loss in the U.S. overall is the introduction of the Varroa mite in the 1980s. That correlates with steeper declines more than any other single factor we know of. The Varroa mite doesn’t just weaken honeybees, it also spreads pathogens that cause diseases. Those pathogens can spread from managed honeybees to wild bees, too, so it’s something we’re concerned about.

How are our wild pollinators faring?

Gratton: It’s really hard to track populations of our wild pollinators. We manage honeybees. We move them around, we keep track of numbers, we can open up the hive and see what’s going on. With the native bees, there are more than 500 species in Wisconsin. In any one system like apples or cranberries, we may have 100-plus different species that visit them. But many of them are solitary and sometimes rare. We haven’t really been tracking their populations very well. So to know if they are declining, we need a reference point and we don’t have one. As a consequence, we actually don’t know that much about how populations of the native bees are doing.

The few studies that do exist have looked at historical data and suggest that for the most part, most native bees probably haven’t changed that much over time. The few native species that we do have better data on are the bigger, more iconic pollinators like bumble bees. There is some good evidence that these species are declining in North America. And you can point to a couple of species that really have shown dramatic declines compared to midcentury distributions. There may be reasons for those declines—again, having to do with pathogen spread, competitors and declines in flowers in the landscape.

So, is this a crisis for wild pollinators? I think the jury is still out on that. I think there are lots of reasons to be concerned. But I’m not seeing the data out there saying that there is a massive die-off of native bees that we need to be immediately guarding against. This means we may have some time to start helping them out.

We think the way we have approached the plan is helpful because all of the things we talk about in terms of making life better for honeybees are also going to make life better for the native bees. As one example, reduction and judicious use of pesticides.

Also, when you talk to beekeepers and they say, “My bees back in the ’50s and ’60s used to give me 60 pounds of honey per hive every summer. Now I’m only getting 30”—there is not enough food in the landscape out there for honeybees. Food for honeybees—that is, flowers—is the same as food for the native bees. So all of our discussion about habitat management—getting more flowers out on the landscape, making sure those flowers are blooming throughout the entire summer—those are all things that are going to help native bees as well. I think the plan is going to be able to help a lot of other pollinators that can ride on the coattails of honeybees: bumblebees, butterflies and many of the solitary species that we never pay attention to.

What are some of the more surprising or important points in the plan thus far?

Gratton: You can do some relatively simple things and potentially have a big impact. It’s not like you need to transform the world in order to have an effect. Some really common-sense, small things can go a long way.

Locke: For example, in the agricultural recommendations there is a range of simple to more difficult practices. You can reconfigure your entire farm and make sure everything is really diverse and use blooming cover crops and all of that—and then at the other end of the spectrum, there are suggestions like leaving woody debris if a tree falls. Leave some wood so that bees can nest. That’s an example of a beneficial practice that only requires not doing something.

Based on your scientific expertise, what things would help the most?

Locke: For me, it’s habitat. We used to have a landscape in the Upper Midwest that was dominated by oak savanna and prairie. Now it’s not. That’s a lot of acres of habitat to compensate for.

Gratton: And second, as a home gardener or as a farmer, being judicious about killing bees through insecticides. I have to say that most of the farmers that we work with, cranberry and apple farmers, know this. They don’t want to kill off their bees. They are very sensitive to that, so they know the things to do to maintain their bee populations. Also, the beekeepers that they’ve rented bees from would get very mad if you sprayed insecticides during bloom. The farmers, especially of pollinator-dependent crops, know this. They are not necessarily the ones for whom we have to emphasize the importance of not spraying insecticides at especially sensitive times for bees.

What’s the overall hope in doing this work?

Gratton: I hope that people will read this and recognize that insects—in particular bees, but insects in general—play really important roles in our lives. And that, rather than follow our first instinct to squish them or want them to go away, we appreciate them and try to do things that encourage the beneficial ones in the environment. I hope even in a general sense that anyone can read the plan and say, “Wow, I didn’t realize that these little insects, these joint-legged things that fly around, do so much for us that we benefit from. And here are a couple of easy and practical things that I can do to make their lives a little better.” That’s my immediate goal for the plan.

You can view the protection plan at http://go.wisc.edu/pollinator

PHOTO—Entomologist Claudio Gratton and research associate Christina Locke in Gratton’s lab, examining part of a vast collection of pollinators. A new state plan they helped create is aimed at better protecting them.

Photo by James Runde/UW-Madison Wisconsin Energy Institute

A New Tool to Fight Cancer?

A New Tool to Fight Cancer?A study involving CALS researchers has linked two seemingly unrelated cancer treatments that are both being tested in clinical trials. One treatment is a vaccine that targets a structure on the outside of cancer cells. The other is a slightly altered human enzyme that breaks apart RNA and causes the cell to self-destruct.

The new understanding could help both approaches, says biochemistry professor Ronald Raines, who has long studied ribonucleases—enzymes that break apart RNA, a messenger with multiple roles inside the cell. In 1998, he discovered how to alter one ribonuclease to avoid its deactivation in the body. Soon thereafter, he found that the engineered ribonuclease was more toxic to cancer cells than to others.

Raines patented the advance through the Wisconsin Alumni Research Foundation (WARF) and, with fellow CALS biochemist Laura Kiessling, co-founded Quintessence Biosciences in Madison. They remain shareholders in the firm, which has licensed the patent from WARF and begun early-phase human trials with the ribonuclease at the UW Carbone Cancer Center and MD Anderson Cancer Center in Houston.

The current study began as an effort to figure out why the ribonuclease was selective for cancer cells. To identify which structure on the cell surface helped it enter the cell, Raines screened 264 structures using a specially designed chip. The winner was a carbohydrate called Globo H.

“We were surprised—delighted!—to see that, because we already knew that Globo H is an antigen that is abundant in many tumors,” says Raines. Antigens are molecules with structures that are recognizable to proteins called antibodies. “Globo H is under development as the basis for a vaccine that will teach the immune system to recognize and kill cancer cells,” he says.

Working with Samuel Danishefsky, who solved the difficult problem of synthesizing Globo H at the Memorial Sloan-Kettering Cancer Center in New York, Raines found that reducing the Globo H display on their surface made breast cancer cells less vulnerable to ribonucleases like those that Quintessence is testing. “This was exciting, as we now have a much clearer idea of how our drug candidate is working,” says Raines.

CALS biochemistry professor John Markley aided the research with studies of the structure of the molecules in question.

The picture that emerges from their work is of ribonucleases patrolling our bodies, looking for signs of cancer cells, Raines says: “We are working to demonstrate this surveillance more clearly in mice.”

As other scientists test whether using a vaccine will start an immune attack on Globo H, Raines says, “We are probing a different type of immunity. This innate immunity does not involve the immune system. It’s a way for our bodies to fight cancer without using white blood cells or antibodies—just an enzyme and a carbohydrate.”

PHOTO—Biochemistry professor Ron Raines is devising new ways to destroy cancer cells.

Photo by Sevie Kenyon BS’80 MS’06

The Future, Unzipped

John Ralph PhD’82 talks with the easy, garrulous rhythms of his native New Zealand, and often seems amiably close to the edge of laughter.

So he was inclined toward amusement last year when he discovered that some portion of the Internet had misunderstood his latest research. Ralph—a CALS biochemist with joint appointments in biochemistry and biological systems engineering—had just unveiled a way to tweak the lignin that helps give plants their backbone. A kind of a natural plastic or binder, lignin gets in the way of some industrial processes, and Ralph’s team had cracked a complicated puzzle of genetics and chemistry to address the problem. They call it zip-lignin, because the modified lignin comes apart—roughly—like a zipper.

One writer at an influential publication called it “self-destructing” lignin. Not a bad turn of phrase—but not exactly accurate, either. For a geeky science story the news spread far, and by the time it had spread across the Internet, a random blogger could be found complaining about the dangers of walking through forests full of detonating trees.

Turning the misunderstanding into a teachable moment, Ralph went image surfing, and his standard KeyNote talk now contains a picture of a man puzzling over the shattered remains of a tree. “Oh noooo!” the caption reads. “I’ll be peacefully walking in a national park and these dang GM trees are going to be exploding all around me!”

That’s obviously a crazy scenario. But if the technology works as Ralph predicts, the potential changes to biofuels and paper production could rewrite the economics of these industries, and in the process lead to an entirely new natural chemical sector.

“When we talk to people in the biofuels industry, what we are hearing is that creating value from lignin could be game-changing,” says Timothy Donohue, a CALS professor of bacteriology and director of the UW–Madison-based Great Lakes Bioenergy Research Center, where Ralph has a lab. “It could be catalytic.”

After cellulose, lignin is the most abundant organic compound on the planet. Lignin surrounds and shapes our entire lives. Most of us have no idea—yet we are the constant beneficiaries of its strength and binding power.

When plants are growing, it’s the stiffening of the cell wall that creates their visible architecture. Carbohydrate polymers—primarily cellulose and hemicelluloses—and a small amount of protein make up a sort of scaffolding for the construction of plant cell walls. And lignin is the glue, surrounding and encasing this fibrous matrix with a durable and water-resistant polymer—almost like plastic. Some liken lignin to the resin in fiberglass.

Without lignin, the pine cannot soar into the sky, and the woody herb soon succumbs to rot. Found primarily in land plants, a form of lignin has been identified in seaweed, suggesting deep evolutionary origins as much as a billion years ago.

“Lignin is a funny thing,” says Ralph, who was first introduced to lignin chemistry as a young student during a holiday internship at New Zealand’s Forest Research Institute. “People who get into it for a little bit end up staying there the rest of their lives.”

The fascination is born, in part, from its unique chemistry. Enzymes, proteins that catalyze reactions, orchestrate the assembly of complex cell wall carbohydrates from building blocks like xylose and glucose. The types of enzymes present in cells therefore determine the composition of the wall.

Lignin is more enigmatic, says Ralph. Although its parts (called monomers) are assembled using enzymes, the polymerization of these parts into lignin does not require enzymes but instead relies on just the chemistry of the monomers and their radical coupling reactions. “It’s combinatorial, and so you make a polymer in which no two molecules are the same, perhaps anywhere in the whole plant,” says Ralph.

This flexible construction is at the heart of lignin’s toughness, but it’s also a major obstacle for the production of paper and biofuels. Both industries need the high-value carbohydrates, especially the cellulose fraction. And both have to peel away the lignin to get to the treasure inside. A combination of heat, pressure, and caustic soda is standard procedure for liberating cellulose to make paper; bleach removes the remaining lignin. In the biofuels industry, a heat and acid or alkaline treatment is often used to crack the lignin so that it is easier to produce the required simple sugars from cellulose. Leftover lignin is typically burned.

The economic cost of these treatments alone is significant, and lignin pretreatment is at the heart of many of the more egregious environmental costs of paper. On the biofuels side, lowering treatment costs to liberate carbohydrates from lignin could change the very economics of biofuels. In these large-scale, industrial processes, saving a percentage point or two is often worthwhile, but the Holy Grail is a quantum jump.

“Because it’s made this way”—Ralph jams his hands together, crazy-wise, fingers twisted together into a dramatic representation of lignin polymerization—“there is no chemistry or biology that takes it apart in an exquisite way,” he says. “We actually stepped back and thought: How would we like to design lignin? If we could introduce easily cleavable bonds into the backbone, we could break it like a hot knife through butter. How much can you actually mess with this chemistry before the tree falls down?”

Ralph’s team had their eureka moment more than 15 years ago, and have been trying to bring it to life ever since.

With a background in forage production and ruminant nutrition, John Grabber, an agronomist at the USDA–Agricultural Research Services’ Dairy Forage Research Center in Madison, got pulled into lignin chemistry through the barn door. On his family’s dairy farm he grew up with lignin stuck to his boots, though he never knew it. But during graduate school he became interested in how plants are digested by cows. Cell walls are potentially a great source of digestible carbohydrates—most plants contain anywhere from 30 to 90 percent of their mass in their cell walls—but it is entangled with lignin. “You quickly find out that lignin is the main barrier to feed digestion,” he says.

Grabber began working on a model system to understand plant lignification—for corn in particular—in 1989. After meeting at a conference, Grabber joined Ralph and plant physiologist Ronald Hatfield at the Dairy Forage Research Center back in 1992. There were many projects ongoing, but Grabber remained interested in trying to fully understand the structural characteristics of the lignin: how it’s made and how to modify it. In his model system they could make any kind of lignin they wanted to study, and see how the changes affected utilization.

Ralph and Hatfield advocated for the work, helping to find funding and offering their expertise. “If I had worked for somebody else I probably wouldn’t be doing this work,” says Grabber. “John and Ron gave me freedom and support to do it.”

Around the same time, Fachuang Lu joined Ralph’s lab seeking a Ph.D. His journey into lignin chemistry was not, at first, his idea. A native of mainland China, he’d enjoyed a successful undergraduate career in Beijing studying chemical engineering, then found himself assigned by the college to a master’s program in lignin chemistry. Lignin is an ingredient in the slurry of chemicals used in oil drilling, and that was his specialty. In 1989 Lu left Beijing for a teaching position at Guangxi University, but three years later he decided to continue his education. Though he’d never met Ralph, he was fascinated by the chemistry and applied to study in his lab.

As Ralph, Grabber, Hatfield and Lu continued to tinker with lignin chemistry, momentum began to build in the lab. Though lignin created a snowflake universe of different molecules, there were rules of assembly. A complex chemical pathway enabled lignin construction, with a mechanism that remained constant across different families of plants, but with many potential building blocks.

Ralph and his colleagues were the first to detail what was happening to lignin as the controlling genes of the biosynthetic pathway were turned on and off, a task ably completed by a slew of outstanding collaborators worldwide with expertise in biotechnological methods—but who lacked the diagnostic structural tools to determine what the plant was doing in response.

Ralph’s team quickly learned that lignification was somewhat flexible. “We figured that we could engineer lignin well beyond the previously held bounds,” says Ralph. As various pathways and chemical possibilities danced in their heads, it struck them: What if, during lignification, they could persuade the plant to slip in a few monomers that had easily broken chemical bonds? If they did it right, lignin would retain its structural value to the plant, but be easier to deal with chemically.

“In the course of our conversation we realized that if plants could do this, it could really revolutionize how readily you could make paper,” recalls Grabber. Says Ralph: “It’s almost impossible to tell which one of us actually verbalized it first—it is one of those great outcomes of the group dynamic.”

Lu’s particular genius was synthesizing the various complex chemicals needed, particularly a novel monomer-conjugate called coniferyl ferulate. It was the key to the zip-lignin—the teeth of the zipper. “He’s got to be one of the best in terms of making molecules,” says Grabber.

They were thrilled by such a revelation, but, in retrospect, they soon realized it was sort of an obvious idea—one suggested by the underlying chemistry and biochemistry of a pathway that was becoming increasingly well understood. Yet it was a discovery of huge potential value. They dropped into stealth mode and began to work on it. They finished important research and stuck it in drawers—signature research, the kind that, when finally published, would capture journal covers. And yet they sat on it, quietly chipping away for nearly a decade.

It helped that there was a flurry of controversy in the field—what Chemical & Engineering News called “the lignin war.” “Part of the reason we could sit on it was that, at the time, making these kinds of molecules was so far-fetched,” says Grabber. “Probably if we had talked about it, people would have laughed at us.”

But as the idea for zip-lignin grew in principle, it became stronger. Lu, Hatfield and colleague Jane Marita MS’97 PhD’01 found that balsa trees and a fiber crop known as kenaf produced very small amounts of coniferyl ferulate. But even as the idea seemed more and more feasible, Hatfield and Marita couldn’t isolate the gene needed to manufacture coniferyl ferulate because of its very low expression in these plants.

And they got stuck. “At the beginning we were thinking that this is just a fantastic idea, but we really didn’t have that much confidence,” says Lu. “Maybe John [Ralph] had more confidence than me.” So they just kept at it. “Every step you think, yes, we are closer, closer, closer.”

In 2008 Ralph moved his work from the Dairy Forage Research Center into UW labs, with research projects under the recently formed Great Lakes Bioenergy Research Center (GLBRC). The center, launched with a $125 million grant from the U.S. Department of Energy that has since been renewed, was just one manifestation of the money and intellectual heft infusing biofuels research—and for zip-lignin it was a lucky move.

During the center’s first full meeting, Curtis Wilkerson, a plant biologist at GLBRC partner Michigan State University, was sitting in the audience when Ralph took his turn at the podium.

Wilkerson is a cell wall specialist. Though lignin is a third of the wall’s carbon and is essential to the way plants conduct water, he confesses he’d never given it much thought. In a room full of cell wall specialists, Ralph would “likely be the only person talking about lignin,” he says. “It just split that way a long time ago. People like myself had very little exposure to what John was thinking.”

It was this kind of academic silo that a place like GLBRC was supposed to breach. Ralph talked about putting ester bonds into lignins and his team’s long search for the elusive enzyme. Wilkerson saw a solution. Due to recent technical advances, the price of determining all of the expressed enzymes in a plant had become more refined and much less expensive. He offered to use these recent developments to try to find the missing enzyme to enable zip-lignin.

From the previous work, Wilkerson knew essentially the size and shape of the puzzle piece he was looking for. He began, quite literally with Google, trolling through the scientific literature looking for a plant that made a lot of coniferyl ferulate. The Chinese medicinal “dong quai” or Chinese angelica (Angelica sinensis) soon emerged as a candidate. Its roots contained about 2 percent coniferyl ferulate.

The team used knowledge about the likely type of enzyme they were searching for and successfully identified the gene and its enzyme that could produce coniferyl ferulate. The whole search took less than six months.

Would you believe an essential tool for the genetic engineering of poplars is a hole punch? That’s the word from Shawn Mansfield, a molecular biochemist at the University of British Columbia, who took the zip-gene from the Angelica and made it work in poplar, a popular tree in the biomass and forest products industry.

Working from Wilkerson’s gene, the first job was figuring out how to tag the new protein so that it fluoresced during imaging. While not necessary to the function of the genetically modified plant, it essentially allows the scientists to check their work: see where the protein is, how much is there, and if it is behaving as a protein should.
Mansfield’s lab also had to find a way to turn the gene on at the right time and place. It could make all the coniferyl ferulate one wanted, but if it wasn’t made at the right time and tissue, there would be no zip-lignin.

After perfecting these finer points, the gene is inserted into a special bacterium—and then the hole punch finally comes into play. Disks punched from poplar leaves are mixed with bacteria that have been inoculated with a special chemical that stimulates the bacteria to share their DNA around. Then the leaf disks are put in a special growth medium. As many as 12 shoots might emerge off of a single disk, but the lab would select and nurture only one shoot from each disk.

In the end they had about 15 successful transgenic candidates that they grew in the greenhouse and then shipped off to Wilkerson and Ralph for further study. Final selection was made based on the amount of fluorescent yellow the trees gave off, and from a newly devised analytical method developed by Lu and Ralph that was particularly diagnostic for the incorporation of the zip monomer into the lignin polymer.

The team knew that genetically modified organisms are not popular or easily talked about—never mind the exploding trees. The idea of reworking a fundamental building block of the plant world will breed resistance.

Ralph argues that this is already part of nature’s vocabulary: they’ve found their building blocks within the plant kingdom, including mutants that do similar things. And now that they know what they are looking for, Steven Karlen, a member of Ralph’s group, is continuing to find more evidence that Mother Nature is doing it herself. “We managed to persuade plants to do this,” Ralph says. “Chances are that nature has already attempted it and you could actually get there by breeding.”

It’s no surprise that Mansfield, who created the final transgenic tree, argues that there is a role for this kind of technology. “We as scientists should be wise in advocating for the proper use of it,” he cautions. “I would never force it on anybody. I would never try to sway people to think that it is the end-all or be-all for everything.”
But given the growing human population and rising CO2 levels, something like zip-lignin has a definite use in reducing the carbon footprint by reducing processing energy and chemical loads. “That means there are less environmental pollutants that need to be cleaned up afterwards,” Mansfield says.

“Our ecological footprint can be much reduced using these kinds of transgenic trees,” he argues. “The caveat is that we need to be very smart about where and how we plant them.”

Not many things in the natural world can take apart lignin, but any homeowner with a deck knows that fungi are up to the task. A recent analysis of mushroom genomes suggests that fungi evolved this ability about 300 million years ago. This is about the end of the Carboniferous era, when earth’s coal production began to slow down. Coincidence? Perhaps not. Now that wood could rot, it probably slowed the burial of organic carbon via tree trunks and other lignin-rich plants.

Could the discovery of zip-lignin signal another transition, and hasten our move away from fossil fuels laid down in the Carboniferous?

Tim Donohue likes to think so. He likens biofuels now to the early oil industry, when oil was simply being turned into liquid fuel while the by-products were burned or dumped. It took a few decades for inventors to capitalize on this now valuable stream of raw materials to build the modern chemical industry.

“Lignin is about 25 to 30 percent of carbon in the plant. So if we’re going to catalyze an industry that makes clean energy and chemicals from plant biomass, figuring out what to do with the lignin is going to be key,” Donohue says.

People in the industry used to joke that you could do a lot of things with lignin except make money from it. But that may be changing. “The economics and profitability of the industry will be very different if lignin can be turned into valuable compounds,” says Donohue.

One of the early efforts to make use of lignin was in Rothschild, Wisconsin, at a company now known as Borregaard LignoTech. When processed properly, lignin has many uses, from the manufacture of vanilla flavor to additives for concrete. There is even a small amount of it in the battery of your car that allows it to keep recharging.

Jerry Gargulak is research manager at Borregaard LignoTech, and learned about zip-lignin recently in his capacity as a scientific advisor to the GLBRC. Despite its many uses, Gargulak and his colleagues dream about a time when lignin can replace carbon black in tires and be used to build carbon fibers and structural plastics.

Zip-lignin and the ideas behind it could bring this day closer. “It gives us a technology that might yield a more interesting lignin-derived starting material,” Gargulak says. “It could potentially lead to a lot of innovation downstream in lignin technology.” But he emphasizes, “There are a lot of i’s to be dotted and t’s to be crossed.”

This story is just beginning. Zip-lignin has a patent and has excited industrial interest that could be worth significant dollars. Ralph and his colleagues continue working to further refine the process, increasing the percentage of zippable bonds in poplar and also inserting the gene into more plants, such as corn and Brachypodium, both grasses.
And in the basement of the shiny new Wisconsin Energy Institute building, where the GLBRC is based, two massive new nuclear magnetic resonance (NMR) spectrometers work 24/7, providing a level of detail into lignin that Ralph has never had before.

“We spend a lot of time looking at these Rorschach test–like figures,” Ralph says of the information generated from the NMR. “The detail in them is unbelievable. These things have been revolutionizing what we do.”

Bitten

There’s no ignoring it. Some of the students enrolled in this medical entomology class are far more attractive than others. They know it, their classmates know it, and so does Susan Paskewitz, professor and chair of the Department of Entomology.

Paskewitz describes herself as “relatively unattractive,” and she proceeds to prove it using the same test her students have just performed. She fills a small vial with warm water, rubs it between her palms to coat it with volatile compounds from her skin, then places the vial on top of a thin membrane stretched over the top of a plastic container akin to an economy-sized ice cream tub. She invites a visitor to do the same.

Waiting on the other side of that membrane are 20 blood-starved specimens of Aedes aegypti, commonly known as the yellow fever mosquito. Hungry as they are, the insects don’t show a lot of interest in Paskewitz’s vial. They hover near where it touches the membrane, but only two or three land. The visitor’s vial, on the other hand, is a busy spot. At least a dozen have landed and are testing the surface with their needle-like proboscises.

“Wow,” says Paskewitz. “You’re really attractive!”

In another context, those three words could make your day. But not here. Nobody wants this kind of animal magnetism. Nobody wants to be the person who’s cursing and slapping and reaching for the DEET while others are calmly eating their brats and potato salad.

If you’re that person, take heart. Paskewitz can tell you a little bit about why you might have more than your share of interspecies charisma and offer some suggestions on how to scale it back. But first, let’s talk about why this matters.

An average American adult outweighs an average-size mosquito by about 30 million to one. Ounce for ounce, that’s like the USS Nimitz vis-a-vis a good-size duck. But while it’s a safe bet that a 100,000-ton aircraft carrier won’t change course to avoid a six-pound mallard, it’s almost certain that, on a regular basis, you change your behavior to avoid being bitten by a 2.5-milligram mosquito.

Mosquitoes cause us to do things we’d rather not, like dosing ourselves with a repellent that’s sticky and smelly and comes with a sobering warning label (you can apply it to your kids’ skin, but keep the bottle out of their reach), or pulling on long pants, long sleeves, a hat and maybe a head net on a sweltering midsummer day.

Mosquitoes keep us inside when we’d much prefer to go out. In the summer of 2009, Paskewitz and environmental economist Katherine Dickinson, of the Colorado-based National Center for Atmospheric Research, asked a sample of Madison residents how they coped when mosquitoes got fierce.

The second-most-common answer (right after applying repellent) was to stay indoors. About two-thirds of the respondents said they had curtailed outdoor household activities—gardening, yard work, sitting on the deck—in the past month because of mosquitoes, especially in the evening hours, which, for working people, may be the only time available to get a little fresh air. About a third said they had avoided outings, and a similar share said they had avoided outdoor exercise.

Nobody wants to be outside more than John Bates, of Manitowish. An author of seven books about Wisconsin’s north woods and a naturalist by trade, Bates leads interpretive hikes year-round—except in June: “We just kind of throw the month out. The mosquitoes cause too much discomfort for people to listen to interpretation. All we can do is keep walking. People hire me because they want to learn more about the place than they knew before they came. If they can’t stop to listen, what’s the point?”

If we do venture out when mosquitoes are massing, we may not get the experience we were hoping for. Andrew Teichmiller, an outfitter of bikes and paddling gear in Minoqua, recalls mountain biking in 2014, arguably the area’s worst mosquito year ever. “You had to ride the complete trail without stopping, all the way back to the parking lot, and jump in the car, quick, because if you stopped there were 15 or 20 mosquitoes on you immediately.” As for camping: “It’s a different type of experience when you can’t sit by the fire at night and tell stories. You’re forced to run for your tent. It definitely affects the feel of the trip.”

But let’s be clear: A ruined camping trip is far from the worst possible consequence of a mosquito bite.

Mosquitoes transmit diseases that kill nearly a million people every year and sicken hundreds of millions. Tropical and subtropical areas bear the brunt of this, but no place is immune, including Wisconsin. Malaria plagued the immigrants who settled in Wisconsin in the 1800s, and various types of encephalitis are diagnosed on a regular basis.

But today the biggest concern is West Nile virus (WNV). Wisconsin has been relatively lucky since the first case arrived here in 2002, with a total of 230 cases reported through 2014. But all four adjacent states have had bigger outbreaks—notably Illinois, with 2,093 cases total and 884 in its worst year, most of them just across the border in the Chicago area. Wisconsin’s worst year brought 57 cases.

Most cases of WNV bring no symptoms, according to the Centers for Disease Control, but about one in five can involve a fever, headache, body aches, vomiting and a fatigue that can last for weeks or months. Fewer than 1 percent of WNV victims display severe neurologic symptoms, including disorientation, coma, tremors, seizures or paralysis, and of those, about 1 percent die.

Nevertheless, Wisconsin residents are bothered much more by the nuisance of biting mosquitoes than they are worried about West Nile virus. The Madison residents responding to Katherine Dickinson’s 2009 survey said they’d be willing to pay an average of $149 for a hypothetical program to control nuisance mosquitoes, but wouldn’t pay anything for one targeted at mosquitoes carrying WNV when risks were as low as they were at the time (about one case per year in Madison with a population of 250,000).

It’s not surprising to find that attitude in Wisconsin, where mosquito-borne disease is relatively rare, but Dickinson says that people tend to think the same way in places where mosquito bites are often fatal. She observes that in Tanzania, biting mosquitoes were a major factor motivating people to use bed nets. “It was a similar situation to ours,” she says. “Some mosquitoes are more noticeable and more of a nuisance, but those that transmit malaria are kind of sneaky; people don’t feel them biting as much. In areas where mosquitoes were more of a nuisance, people used the bed nets more.”

Biting-wise, there’s an important distinction between nuisance mosquitoes and the ones that transmit WNV. The former come at us aggressively, in such staggering numbers that they’re impossible to ignore. They remind us to protect ourselves. Culex pipiens, the WNV vectors, are more subtle and harder to notice.

Nuisance mosquitoes and the WNV carriers also show up at different times. The most annoying biters—Aedes vexans in particular—are floodwater species that breed after a stretch of wet weather. Culex breed in water that stagnates during a dry spell.

“When it’s been really dry, the water just sits in the stormwater catch basins that are the biggest sources of the WNV vectors,” says Paskewitz. “There’s not enough rain to flush them. Things get more fetid, stinkier. That’s the year when we see a ton of Culex.”

The take-home message: If you only grab the DEET when the biting is so bad that you can’t stand to be without it, you’re not protecting yourself against West Nile virus.
“You need to protect yourself against bites even if you’re not getting a lot of them,” says John Hausbeck, director of environmental health services for Dane County and the City of Madison. “We’ll see summers where it’s really dry and the floodwater mosquitoes are very limited, but we still have plenty of small pools that the Culex can breed in.”

That “biting pressure” is something that Hausbeck needs to stay on top of, and Paskewitz helps with that. She and former grad student Patrick Irwin PhD’10 were able to characterize the types of sites where Culex are most likely to breed and identified alternatives for treating them—for example, introducing fathead minnows to feed on Culex larvae. She and her students analyze the mosquitoes trapped in the area to see how many are Culex and whether they’re carrying WNV. Their data tell Hausbeck whether he needs to issue a public alert.

It’s important to remain vigilant. “When West Nile first came into the country, people doubted it would make it through the first winter,” Paskewitz says. “Well, it did persist, and in a very short period of time it whipped across the whole country. We’ve had a lot of cases in new places. First it was really bad in North and South Dakota. Then Colorado and Arizona. Then Texas, Illinois. It’s really hard to predict. And given the vagaries of climate, we just don’t know whether the next year it might be Wisconsin.”

Maybe WNV hasn’t changed Wisconsin residents’ ideas about why to guard against mosquito bites, but it certainly has spurred a lot of questions about how. There is a seemingly endless list of products and strategies, that, according to somebody, will eliminate mosquitoes or repel them—and since WNV arrived, Paskewitz has been getting questions about pretty much all of them.

“They call me to ask, ‘Would this work or wouldn’t it?’ There is a lot of misinformation out there and not many good sources of information, so I realized I needed to get a better idea of what the science was behind these things,” Paskewitz says.

As she comes up with answers, she posts summaries online. Her website, http://go.wisc.edu/mosquitoes, gets plenty of visits (55,000 last year) and triggers a lot of calls from media from across the nation.

A few of her findings:

• Repellents can be very effective, but comparing them is tricky. There are lots of products with varying active ingredients offered in different concentrations and combinations. Generally speaking, DEET, Picaridin, IR3535, and oil of lemon eucalyptus have good track records. There are also a number of other plant-based compounds—garlic, catnip oil, vanilla and oil of cloves, for example—for which there’s less research and conflicting results. The website sums all this up and gives links to more information.
Yard traps get a thumbs-down. “We tested those and didn’t get any positive outcome,” Paskewitz says. Yard traps lure mosquitoes by releasing C02, light or octenol, a compound contained in our breath and sweat. Sure, they can catch mosquitoes by the hundreds, Paskewitz says. But does this significantly reduce the numbers that bite you? Properly controlled studies say “no.”

• “Sonic” devices—wristbands, smartphone apps, etc.—do better at extracting your money than keeping mosquitoes off your deck. “You can test them yourself,” Paskewitz says. “Sit at the picnic table and count how many mosquitoes land on you, then turn on the device and count again. Or you can trust the research and save your money.”

• Bats are busted. The idea that a colony of bats can consume millions of mosquitoes per night came from a study in which someone put a bat in a room full of mosquitoes and estimated how many it ate. The question is, given the choice, is that what bats eat in the wild? Researchers who examined the stomach contents and fecal pellets of bats have found bigger insects, like butterflies, moths and beetles, but very few mosquitoes. “Bat houses are great for conserving bats,” Paskewitz says, “but not for mosquito control.”

• Avoiding bananas—When she first heard the idea that eating bananas makes you more attractive to mosquitoes, Paskewitz raised her eyebrows. “I thought, okay, we’ll debunk that,” she says. She was teaching medical entomology at the time with 24 students—enough for a robust sample—so she made it a class project. For several weeks, each student ate a banana and then performed an attractiveness assay at prescribed intervals. “We were really intrigued. It did look like we were getting an increase a couple hours after eating the bananas.”

Paskewitz repeated the trial the next two times the course was offered, with a few tweaks to the methodology: Half the students ate bananas, the other half grapes. “The third trial was the best of all—the strongest statistical evidence and the most repeatable,” Paskewitz says. “We did it three times and saw a strong difference between the groups. Grapes didn’t matter, bananas did. At that point I was convinced. I think it’s real,” she says. Does that mean you if you leave bananas out of your picnic fruit salad, you can skip the bug spray? Probably not, Paskewitz says.

Because “less attractive” is not the same as mosquito-proof, Paskewitz gets plenty of mosquito bites, probably more than her share, because she spends a lot of time around mosquitoes—in the woods doing field research, in her garden, and in her lab. When you’re a mosquito researcher, getting bitten comes with the job.

What Makes You Attractive?

It sounds like the topic of an article in Seventeen magazine—and, interestingly, some of the same general categories apply whether you’re talking about your appeal to a mosquito or to a certain someone of your own species.

Your breath. If you breathe, you’re mosquito bait. Every breath adds to a plume of carbon dioxide (CO2 levels in your breath are 100 times that of the atmosphere) emanating from where you stand. “That’s the big signal,” says entomology professor Susan Paskewitz. “Insects are very sensitive to chemical cues. They’ll zigzag to pick up the chemical as it gets stronger and stronger, circling to narrow in on you.”

Your aroma. Once they find you, mosquitoes use chemical cues to decide whether to land and dig in. They have a lot to sort through: You emit roughly 400 different compounds from your skin and 200 in your breath. Many mosquito species won’t land on humans, even if they’re starved for blood. Others will bite us in a pinch but prefer other hosts, Paskewitz says.

Your genes. Perhaps you were born to be bitten. A pilot study at the London School of Hygiene & Tropical Medicine found that identical twin sisters were significantly more alike in their attractiveness to mosquitoes than were non-identical twins. Since identical twins are closely matched genetically, this suggests that some of your Culicidae charisma is inherited. Some volatile compounds on our skin are produced by skin cells (others are produced by bacteria), which would be gene-regulated, the study’s authors note.

Your jeans. What color you wear matters. This is based on a series of studies in which researchers draped different colors of cloth on human volunteers or on robots heated to simulate human body temperatures, then counted mosquito landings. For the most part, darker colors were more attractive. White was least attractive, followed by yellow, blue, red and black.

Your smelly feet. “The malaria mosquito is really attracted to the smell of funky feet,” Paskewitz says. “It’s a classic story in medical entomology. The compound that makes feet smell funky and attractive to mosquitoes is the same one that causes Limburger cheese to smell the way it does.” That compound is produced by bacteria that can accumulate in the moist spots between your toes, and are kin to those used to culture Limburger.

Your drinking habits. A number of researchers speculate that drinking alcohol makes you more attractive to mosquitoes. A team in Japan put this to the test. They asked some volunteers to drink 350 ml of beer while a control subject did not. The percentage of mosquito landings after alcohol consumption increased substantially. Why this happens is unresolved, although some have speculated that people who have been drinking are easier targets because they move more slowly.

Getting Under Your Skin

Maybe you don’t get more mosquito bites than other people. Maybe your body just makes a bigger deal of it. The swelling, redness and itching are signs of your immune system kicking into gear, explains Apple Bodemer, an assistant professor of dermatology at the UW–Madison School of Medicine and Public Health. And some people’s immune systems kick harder than others.

A mosquito bite involves give and take. Before drawing out up to .001 milliliters of your blood, the mosquito injects a bit of its saliva, which contains anticoagulants to prevent clotting. You can spare the blood, but the saliva is a problem. That’s how disease gets transmitted. And the saliva contains foreign proteins, or antigens, that spur your immune system to create antibodies, Bodemer explains. “When antibodies bind to the antigens, it initiates an inflammatory response, which includes the release of histamine, which causes the blood vessels to dilate, which brings the swelling and redness and the inflammatory mediators that are responsible for the itching.”

This doesn’t happen the first time you’re bitten. It’s the second time, when your body has built up the antibodies, that your immune system engages. If you get bitten enough times by the same strain of mosquito, you may become desensitized and have either a very mild reaction or no reaction at all to the bites. “People often have more vigorous immune responses early in the season and then, as the summer goes on, they don’t have as much swelling and redness and itching,” Bodemer says. “But when you go a winter without any exposure, you often become resensitized.”

For the same reason, younger kids tend to have more aggressive reactions. Once they’ve had several years of mosquito exposure, their response tends to die down, Bodemer says.
As for scratching? Doctor’s orders: Don’t! “Scratching really promotes the full inflammatory reaction. It causes more irritation, causing the blood vessels to be more dilated and further dispersing the inflammatory mediators. It initiates a cycle of swelling, redness and itching. If you can avoid scratching, a lot of times the bumps will disappear.”

Antihistamines can ease the itching, she says, or you can try a home remedy: “I paint a little clear nail polish on the mosquito bite. That will stop the itching to some degree and allow the inflammation to clear up more quickly,” Bodemer says. “Some people cover the bite with Scotch tape for two to four hours. The tape stops you from scratching and when you peel it off, it removes some of the mosquito saliva.”

Wisconsin’s Pestilent Past

Wisconsin’s 19th-century settlers knew that mosquitoes were biting them, and they knew that something was making them sick—but they didn’t put the two together.

Their doctors blamed the ailment on “malarial vapors” emitted by decaying vegetation in the swamps, according to Peter T. Harstad, a UW–Madison educated historian who authored several articles on the health of Midwestern settlers. Harstad used reports by military and civilian doctors as well as immigrants’ diaries and letters to chronicle the devastation caused by what was sometimes called “intermittent fever” because the symptoms—chills, aches and a general fatigue—often recurred over a period of months or years.

“I became sick as soon as I came here and have been sick for eighteen months with malarial fever, which is very severe and painful and sometimes fatal,” reads one letter excerpted by Harstad, written in 1941 by a resident of Muskego. “My wife and I are now somewhat better, but far from being well. This year seventy or eighty Norwegians died here … Many became widows and fatherless this year.” About 13 percent of Muskego’s population died that year, Harstad estimates. The town was hard hit because of an abundance of marshes, a relatively warm climate, and the fact that Norwegian immigrants had no resistance to the disease.

Soldiers also suffered. Harstad cites army reports of malaria outbreaks as far north as Ft. Snelling, near present-day St. Paul. Hardest hit was Ft. Crawford, located amid miles of Mississippi River wetlands at Prairie du Chien. In the fall of 1930, there were about 150 cases reported among the 190 soldiers stationed there. To treat the disease, army surgeons were directed to “extract from twelve to twenty ounces of blood, an operation which it is sometimes required to repeat once or twice.” Wisconsin was mostly malaria-free by the end of the 19th century, as farmers drained wetlands and better housing shut out mosquitoes.

Stealth Entry

Many human diseases—including cancer—are caused by protein malfunctions. Those malfunctions, in turn, are caused by damaged DNA that gets translated into the damaged proteins. While many clinicians and scientists are trying to treat those diseases by fixing the DNA, Ron Raines is taking a different approach—he’s looking to replace the proteins directly.

“Our strategy is to do gene therapy without the genes,” explains Raines, a professor of biochemistry. “We want to skip the genes and go right to the proteins.”

The strategy is intriguing, but there’s a problem. Proteins have a hard time getting into cells where they would do their work. The lipid bilayer of a cell membrane serves as a barrier that keeps the inside of the cell in and the outside out. That membrane stops potential intruders—including uninvited proteins—from entering.

Raines and his team have found a way around this in what amounts to a kind of biochemical calling card. They can attach “decorations,” using what is called an ester bond, to the protein to change its characteristics. The ester bonds link the protein to a “moiety,” a molecule that gives the protein a desired attribute or function.

“Moieties could encourage cell entry, which is one of our major goals,” says Raines. “But moieties could also enhance the movement of the protein in an animal body. Or they could be agents that target the protein, for example, to cancer cells specifically.”

Modifying proteins to give them these attributes has been done using other approaches, but those changes are permanent and can cause problems. The modified protein might not function normally, or the immune system might see the protein as foreign and mount an attack.

Raines’ strategy avoids these problems by using reversible modifications. Because the moieties are added using ester bonds, they are removed once inside a target cell. Naturally occurring enzymes in the cell—called esterases—sever the ester bonds and break off the moieties. What’s left is the normal protein without any decorations. That protein can then do its job.

“We don’t have the problem of damaging the function of the protein or of an immune response because what we ultimately deliver will be the wild-type protein, the protein as it’s naturally found in cells,” explains Raines.

The strategy is promising, and the Wisconsin Alumni Research Foundation (WARF) already has patent applications for it on file. Raines’ lab is now working to make adding the decorations as straightforward and user-friendly as possible. That way, scientists and clinicians could add a moiety of their choosing and get the protein to perform its desired function.

Raines sees innumerable possibilities.

“We’re very excited about this because it has a lot of potential,” he says. “We can now decorate proteins reversibly with pretty much any molecule you can imagine. We are exploring the possibilities to try to bring something closer to the clinic.”

Second Life for Phosphorus

Phosphorus, a nutrient required for growing crops, finds its way from farm fields to our food and eventually to our wastewater treatment plants. At the plants, the nutrient causes major problems, building up in pipes or going on to pollute surface waters.

Brushite bounty: Phil Barak displays brushite produced during trials at the Nine Springs Wastewater Treatment Plant of the Madison Metropolitan Sewerage District. Each jar contains brushite harvested from 30 gallons of anaerobic digest. Photo courtesy of Phil Barak

Brushite bounty: Phil Barak displays brushite produced during trials at the Nine Springs Wastewater Treatment Plant of the Madison Metropolitan Sewerage District. Each jar contains brushite harvested from 30 gallons of anaerobic digest.
Photo by Rick Wayne

But soil science professor Phil Barak has an idea about how to retrieve the nutrient from wastewater in a valuable form—and it started from a basic lab experiment. “I was doing some work on crystallizing phosphorus, just out of pure academic interest,” explains Barak. “That led me to crystallize a mineral called struvite. Then I realized it was forming in wastewater treatment plants as a nuisance.”

If he could form crystals in the lab, he reasoned, why couldn’t it be done in the wastewater treatment plants in a controlled way? It could. And, even better, if he collected the phosphorus early on in the treatment process in the form of a mineral called brushite, he could harvest even more of it.

Beyond removing phosphorus from wastewater, brushite can serve as a nutrient source for growers. While Barak will do further testing to prove its utility, brushite is a phosphate mineral that’s actually been found in agricultural fields for years.

“When conventional phosphorus fertilizers are added to soil, brushite forms. I maintain that we’ve been fertilizing with brushite for decades, but nobody’s been paying attention to it,” says Barak.

Being able to remove phosphorus from wastewater and supply it back to growers is a win-win situation, Barak notes. “We’re collecting phosphorus where it’s localized, at really high concentrations, which is the most economical place to collect it,” says Barak. “This works out in just about every dimension you can consider, from the treatment plants to the cost of recycling phosphorus as opposed to mining it new.”

Graduate students in Barak’s lab suggested that he commercialize the technology and start a company. After the Wisconsin Alumni Research Foundation (WARF) passed on the patent, Barak and his students sought help from the UW Law and Entrepreneurship Clinic. They received two federal Small Business Innovative Research grants, and, with some additional funds from the state, including the Wisconsin Economic Development Corporation, their efforts have turned into a spinoff company: Nutrient Recovery & Upcycling, LLC (NRU).

The company’s next step was a big one. This summer, a phosphorus recovery pilot plant is being implemented in a wastewater treatment plant in Illinois. The pilot project will test the research ideas on a larger scale.

Additionally, the NRU team will participate in the Milwaukee Metropolitan Sewerage District’s granting system to determine if a pilot project would be a good fit in Milwaukee. They hope to start collecting and analyzing data from Illinois by September, using that pilot system to lay the groundwork for others in Milwaukee and beyond.

Class Act: Keven Stonewall

Some researchers first find success late in their careers. And then there’s Keven Stonewall.

Now a rising junior majoring in biology, Stonewall made news with research he did while still in high school. A headline in the New York Daily News declared, “Meet the Chicago Teen Who May Cure Colon Cancer.”

Stonewall’s research, which he conducted as an intern at Rush University while he was a senior at the Chicago High School for Agricultural Sciences, revealed that an experimental colon cancer vaccine effective in younger mice did not work in older mice. Stonewall won numerous awards for his work and was selected as a finalist for the Intel International Science and Engineer Fair in 2013.

Stonewall, the child of two public school teachers, had always loved science, but while in high school, a close friend’s painful experience losing an uncle to colon cancer made Stonewall determined to fight the disease. “It motivated me to say, ‘Enough is enough, I want to step up and do something about it,’” he says.

More recently Stonewall’s interest has moved toward curing cancer in children. He spent his sophomore year as a student researcher in the lab of Christian Capitini, a pediatric oncologist with the UW–Madison School of Medicine and Public Health. There he worked with mice to study the use of natural killer cells to treat neuroblastoma, a cancer frequently seen in children.

“He has a very advanced understanding of immunology and the immune system,” Capitini says of Stonewall. “He understood the concepts of the project from the beginning, so he could get his hands dirty a lot faster than the typical student.”

And this summer he’s interning with AbbVie, a research-based biopharmaceutical company, at its North Chicago headquarters.

Stonewall is in cancer research for the long haul, and he wants to pursue it as a physician. “My goal is to go to medical school, and I am thinking of going into pediatric oncology afterward,” he says.