Duke Research Blog

Following the people and events that make up the research community at Duke.

Category: Biology (Page 1 of 27)

Marine Parasites — Little Guys That Make a Big Difference

If you’re anything like me, the first images that come to mind when you hear the words “marine biology” are singing whales, dolphins racing each other, sharks flying out of the water, maybe a swordfish brawl or two — all the big, flashy stuff.

Of all the things “marine biology” invokes, parasites are probably at the very bottom of my list.

Not so for Joe Morton, a PhD student at the Nicholas School of the Environment and self-taught expert on the parasites that inhabit marine organisms. In fact, Morton posits that parasites play one of the most important roles in all of ecology, by modifying the behavior of ecologically influential host species. And he’s got the research to back it up.

Once back at the lab, Morton takes his place behind the microscope to study his research subjects: marine parasites. Courtesy: Joe Morton.

Morton’s academic quest into the world of marine parasites began about six years ago when he was a master’s student at UNC’s Institute of Marine Sciences — just down the road from Duke’s own Marine Lab, where he’s now stationed. Having just read Carl Zimmer’s pop-science book Parasite Rex, Morton wondered whether the marsh periwinkle snails (Littoraria irrorata) he was studying could be infected.

“In my spare time, I would go into the lab at night with a hammer and crack open a bunch of snails to see what I would find,” Morton said. “I didn’t find anything in the literature at the time about Littoraria harboring parasites, which I thought was really unusual because they’re really well-known, important marsh gastropod.”

Morton began to systematically collect Littoraria from local salt marshes, determine their infection status, then examine how the parasites affected the behavior of infected individuals and, in turn, how these behavioral changes affected the ecological health of the salt marsh. This way, Morton figured out that Littoraria infected with digenean trematodes (a class of parasite) climbed and grazed on marsh grass less often than uninfected Littoraria. He also noticed that infected Littoraria congregated at salt marsh “die-off borders,” the edges where marsh grasses stop growing sparsely and start growing in healthy amounts.

A microsopic view of digenean trematodes, the parasites that infect marsh periwinkle snails. Courtesy: Joe Morton.

Based on these observations, Morton designed an experiment to test whether the prevalence of infection among Littoraria correlated with marsh grass health.

“I found that, even under drought stress conditions, parasites could effectively slow the rate at which the marsh died off and help maintain marsh ecosystem structure,” Morton said. “More structure means more nursery habitat for fish. It means more nursery habitat for fiddler crabs. Increased filtration rate of water into the sediment because of crab burrows. The point is, parasites help to increase ecosystem resistance to drought stress.”

Joe Morton traipses through the salt marsh on a windy day. Courtesy: Joe Morton.

Morton was the first to demonstrate this relationship between parasites and marsh health in a behavioral experiment. It’s been a major focus of his research ever since.

“Parasites constitute more than half the life on the planet, but until very recently, parasites were somewhat ignored by ecologists,” Morton said.

Indeed, Morton’s former advisor once told him “never study anything smaller than your thumb.” According to Morton, this was a very widely-held view in ecology up until the last few decades.

“That was very much the idea at the time: these are small things; they probably mean a lot to individual organisms, but they’re may not be important to ecosystems. And now we know that’s just not the case,” Morton said. “Almost everywhere we look, parasites are there; they’re ubiquitous. And they have an important role to play.”

Though parasites are a hot topic in ecology nowadays, Morton, a self-declared “lifelong contrarian,” has a very distinct memory of a childhood moment foreshadowing his current research focus.

“I remember sitting in a barber shop and reading Popular Science magazine, which has an annual list of the ten worst jobs in science. I remember right at the top of the list was ‘parasitic worm biologist.’ And something in my head was just like ‘yeah, I’ll do that,’” Morton said.

Post by Maya Iskandarani

3D Virus Cam Catches Germs Red-Handed

A 3D plot of a virus wiggling around

The Duke team used their 3D virus cam to spy on this small lentivirus as it danced through a salt water solution.

Before germs like viruses can make you sick, they first have to make a landing on one of your cells — Mars Rover style — and then punch their way inside.

A team of physical chemists at Duke is building a microscope so powerful that it can spot these minuscule germs in the act of infection.

The team has created a new 3D “virus cam” that can spy on tiny viral germs as they wriggle around in real time. In a video caught by the microscope, you can watch as a lentivirus bounces and jitters through an area a little wider that a human hair.

Next, they hope to develop this technique into a multi-functional “magic camera” that will let them see not only the dancing viruses, but also the much larger cell membranes they are trying breech.

“Really what we are trying to investigate is the very first contacts of the virus with the cell surface — how it calls receptors, and how it sheds its envelope,” said group leader Kevin Welsher, assistant professor of chemistry at Duke. “We want to watch that process in real time, and to do that, we need to be able to lock on to the virus right from the first moment.”

A 3D plot spells out the name "Duke"

To test out the microscope, the team attached a fluorescent bead to a motion controller and tracked its movements as it spelled out a familiar name.

This isn’t the first microscope that can track real-time, 3D motions of individual particles. In fact, as a postdoctoral researcher at Princeton, Welsher built an earlier model and used it to track a bright fluorescent bead as it gets stuck in the membrane of a cell.

But the new virus cam, built by Duke postdoc Shangguo Hou, can track particles that are faster-moving and dimmer compared to earlier microscopes. “We were trying to overcome a speed limit, and we were trying to do so with the fewest number of photons collected possible,” Welsher said.

The ability to spot dimmer particles is particularly important when tracking viruses, Welsher said. These small bundles of proteins and DNA don’t naturally give off any light, so to see them under a microscope, researchers first have to stick something fluorescent on them. But many bright fluorescent particles, such as quantum dots, are pretty big compared to the size of most viruses. Attaching one is kind of like sticking a baseball onto a basketball – there is a good chance it might affect how the virus moves and interacts with cells.

The new microscope can detect the fainter light given off by much smaller fluorescent proteins – which, if the virus is a basketball, are approximately the size of a pea. Fluorescent proteins can also be inserted to the viral genome, which allows them to be incorporated into the virus as it is being assembled.

“That was the big move for us,” Welsher said, “We didn’t need to use a quantum dot, we didn’t need to use an artificial fluorescent bead. As long as the fluorescent protein was somewhere in the virus, we could spot it.” To create their viral video, Welsher’s team enlisted Duke’s Viral Vector Core to insert a yellow fluorescent protein into their lentivirus.

Now that the virus-tracking microscope is up-and-running, the team is busy building a laser scanning microscope that will also be able to map cell surfaces nearby. “So if we know where the particle is, we can also image around it and reconstruct where the particle is going,” Welsher said. “We hope to adapt this to capturing viral infection in real time.”

Robust real-time 3D single-particle tracking using a dynamically moving laser spot,” Shangguo Hou, Xiaoqi Lang and Kevin Welsher. Optics Letters, June 15, 2017. DOI: 10.1364/OL.42.002390

Kara J. Manke, PhDPost by Kara Manke

Lemur Research Gets a Gut Check

Baby Coquerel’s sifaka

Clinging to her mom, this baby Coquerel’s sifaka represents the only lemur species at the Duke Lemur Center known to fall prey to cryptosporidium, a microscopic parasite that causes diarrhea that can last for a week or more. The illness wipes out much of the animals’ gut microbiome, researchers report, but fecal transplants can help them recover. Photo by David Haring, Duke Lemur Center.

DURHAM, N.C. — “Stool sample collector” is not a glamorous way to introduce oneself at a party. But in the course of their research, gut microbiologists Erin McKenney and Lydia Greene have spent a lot of time waiting for animals to relieve themselves.

They estimate they have hundreds of vials of the stuff, from a dozen primate species including lemurs, baboons and gorillas, sitting in freezers on the Duke University campus.

The researchers aren’t interested in the poop per se, but in the trillions of bacteria inhabiting the gastrointestinal tract, where the bugs help break down food, produce vitamins and prevent infection.

A few years ago, McKenney and Greene started collecting stool samples at the Duke Lemur Center to see how the microbial makeup of lemurs’ guts varies from birth to weaning, and as their diets change over the seasons. And what happens when they get sick?

Illustration of Cryptosporidium, a widespread intestinal parasite that causes diarrhea in people, pets, livestock and wildlife worldwide. Courtesy of the U.S. Centers for Disease Control.

Illustration of Cryptosporidium, a widespread intestinal parasite that causes diarrhea in people, pets, livestock and wildlife worldwide. Courtesy of the U.S. Centers for Disease Control.

Between 2013 and 2016, ten of the lemurs they were studying contracted cryptosporidium, or “crypto” for short, a waterborne parasite that causes diarrhea in people, pets, livestock and wildlife worldwide.

All of the infected animals were Coquerel’s sifakas — the only lemur species out of roughly 20 at the Duke Lemur Center known to fall prey to the parasite — and most of them were under five years old when they fell ill.

Animals that tested positive were moved into separate holding areas away from other animals and visitors. Keepers wore protective suits, gloves, face masks and booties while working in the animals’ enclosures to prevent infection.

All of the animals eventually recovered. Along the way, six of the affected animals were treated with antibiotics, and three were also fed a slurry of saline and feces from a healthy relative.

McKenney and Greene collected stool samples before, during and after infection for up to two months. They used a technique called 16S ribosomal RNA sequencing to identify the types of bacteria in the samples based on their genes, and compared the results with those of 35 unaffected individuals.

In a healthy gut microbiome, “good” bacteria in the gut compete with “bad” microbes for space and nutrients, and secrete substances that inhibit their growth.

The guts of sick and recovering sifakas are host to a very different assortment of microbes than those of unaffected animals, the researchers found.

Not surprisingly, both crypto infection, and antibiotic treatment, wiped out much of the animals’ gut flora — particularly the bacterial groups Bifidobacterium, Akkermansia, Succinivibrio and Lachnospiraceae.

Even after the infections cleared, most animals took another several weeks to stabilize and return to normal levels of gut biodiversity, with younger animals taking longer to recover.

The only animals that made a full comeback within the study period were those that received a fecal transplant, suggesting that the treatment can help restore gut bacterial diversity and speed recovery.

The patterns of gut recolonization following crypto infection mirrored those seen from birth to weaning, said McKenney, now a postdoctoral researcher at North Carolina State University.

The researchers hope their findings will help control and prevent crypto outbreaks in captive primates. Because lemurs are more closely related to humans than lab mice are, the research could also help scientists understand how the gut microbiome protects humans from similar infections and facilitates recovery.

“Thanks to bioinformatics and advances in sequencing, the microbiome gives us a window into the health of these animals that we’ve never had before,” said Greene, a graduate student in ecology at Duke.

They published their findings June 15, 2017, in the journal Microbial Ecology in Health and Disease.

Duke evolutionary anthropology professors Christine Drea and Anne Yoder were senior authors on this study. This research was supported by the National Science Foundation (1455848) and the Duke Lemur Center Directors Fund.

CITATION:  “Down for the Count: Cryptosporidium Infection Depletes Gut Microbiota in Coquerel’s Sifakas,” Erin McKenney, Lydia Greene, Christine Drea and Anne Yoder. Microbial Ecology in Health and Disease, June 15, 2017. http://dx.doi.org/10.1080/16512235.2017.1335165

Post by Robin Smith, science writer, Office of News & Communications

Scientists Engineer Disease-Resistant Rice Without Sacrificing Yield

Researchers have developed a way to make rice more resistant to bacterial blight and other diseases without reducing yield. Photo by Max Pixel.

Researchers have successfully developed a novel method that allows for increased disease resistance in rice without decreasing yield. A team at Duke University, working in collaboration with scientists at Huazhong Agricultural University in China, describe the findings in a paper published May 17, 2017 in the journal Nature.

Rice is one of the most important staple crops, responsible for providing over one-fifth of the calories consumed by humans worldwide. Diseases caused by bacterial or fungal pathogens present a significant problem, and can result in the loss of 80 percent or more of a rice crop.

Decades of research into the plant immune response have identified components that can be used to engineer disease-resistant plants. However, their practical application to crops is limited due to the decreased yield associated with a constantly active defense response.

“Immunity is a double-edged sword, ” said study co-author Xinnian Dong, professor of biology at Duke and lead investigator of the study. “There is often a tradeoff between growth and defense because defense proteins are not only toxic to pathogens but also harmful to self when overexpressed,” Dong said. “This is a major challenge in engineering disease resistance for agricultural use because the ultimate goal is to protect the yield.”

Previous studies have focused on altering the coding sequence or upstream DNA sequence elements of a gene. These upstream DNA elements are known as promoters, and they act as switches that turn on or off a gene’s expression. This is the first step of a gene’s synthesis into its protein product, known as transcription.

By attaching a promoter that gives an “on” signal to a defense gene, a plant can be engineered to be highly resistant to pathogens, though at a cost to growth and yield. These costs can be partially alleviated by attaching the defense gene to a “pathogen specific” promoter that turns on in the presence of pathogen attack.

To further alleviate the negative effects of active defense, the Dong group sought to add an additional layer of control. They turned newly discovered sequence elements, called upstream open reading frames (uORFs), to help address this problem. These sequence elements act on the intermediate of a gene, or messenger (RNA, a molecule similar to DNA) to govern its “translation” into the final protein product. A recent study by the Dong lab in an accompanying paper in Nature has identified many of these elements that respond in a pathogen-inducible manner.

The Dong group hypothesized that adding this pathogen-inducible translational regulation would result in a tighter control of defense protein expression and minimize the lost yield associated with enhanced disease resistance.

To test this hypothesis, the researchers started with Arabidopsis, a flowering plant commonly used in laboratory research. They created a DNA sequence that contains both the transcriptional and translational elements (uORFs) and fused them upstream of the potent “immune activator” gene called snc1. This hybrid sequence was called a “transcriptional/translational cassette” and was inserted into Arabidopsis plants.

When plants have snc1 constitutively active, they are highly resistant to pathogens, but have severely stunted growth. Strikingly, plants with the transcriptional/translational cassette not only have increased resistance, but they also lacked growth defects and resembled healthy wild-type plants. These results show the benefits of adding translational control in engineering plants that have increased resistance without significant costs.

The Dong group then sought to apply these findings to engineer disease-resistant rice, as it is one of the world’s most important crops. They created transgenic rice lines containing the transcriptional/translational cassette driving expression of another potent “immune activator” gene called AtNPR1. This gene was chosen as it has been found to confer broad spectrum pathogen resistance in a wide variety of crop species, including rice, citrus, apple and wheat.

The dry yellowish leaves on these rice plants are a classic symptom of bacterial blight, a devastating disease that affects rice fields worldwide. Photo by Meng Yuan.

The transgenic rice lines containing the transcriptional/translational cassette were infected with bacterial/fungal pathogens that cause three major rice diseases — rice  blight, leaf streak, and fungal blast. These showed high resistance to all three pathogens, indicating broad spectrum resistance could be achieved. Importantly, when grown in the field, their yield — both in terms of grain quantity and quality per plant — was almost unaffected. These results indicate a great potential for agricultural applications.

This strategy is the first known use of adding translational control for the engineering of disease-resistant crops with minimal yield costs. It has many advantages, as it is broadly applicable to a variety of crop species against many pathogens. Since this strategy involves activating the plants’ endogenous defenses, it may also reduce the use of pesticides on crops and hence protect the environment.

Additionally, these findings may be broadly applicable to other systems as well. These upstream elements (uORFs) are widely present in organisms from yeast to humans, with nearly half of all human transcripts containing them. “The great potential in using these elements in controlling protein translation during specific biological processes has yet to be realized,” Dong said.

Corresponding author Xinnian Dong can be reached at xdong@duke.edu or (919) 613-8176.

CITATION:  “uORF-Mediated Translation Allows Engineered Plant Disease Resistance Without Fitness Costs,” Guoyong Xu, Meng Yuan,   Chaoren Ai, Lijing Liu, Edward Zhuang, Sargis Karapetyan, Shiping Wang and Xinnian Dong. Nature, May 17, 2017. DOI: 10.1038/nature22372

 

Guest post by Jonathan Motley

Where Some Ski, Others Do Science

For most people, Lost Trail is a ski spot located at 7,000 feet in the Rocky Mountains on the border of Idaho and Montana. Skiers and snowboarders descend down steep slopes, past forests and alpine meadows that get more than 25 feet of snow each year. But for a team of researchers led by Duke biology professor Thomas Mitchell-Olds, buried beneath the snow is a hidden population of native plants on the cusp of dividing into two new species.

Molly Rivera-Olds shovels snow at Lost Trail Pass.

Studying a spindly North American wildflower called Boechera stricta, Mitchell-Olds and colleagues suspected that a process called chromosomal inversion — in which part of a chromosome breaks off and reattaches itself upside down — plays a central role in speciation. To test the idea, they planted Boechera stricta seedlings in a mountaintop meadow near the Lost Trail resort.

To reach the meadow, the researchers carried thousands of seedlings up the mountain in specially constructed backpacks. They also lugged up nine empty garbage cans and filled them with snow to water the plants throughout the summer.

Once the seedlings matured, the researchers measured flowering time, seed production, and survival. They found that plants with the chromosomal inversion had a leg up on the steep slopes of the Rocky Mountains. Eventually, the researchers say, this can lead to plants with the inverted DNA splitting off and forming a new species.

The findings were published April 3, 2017 in the journal Nature Ecology & Evolution.

# # #

CITATION:  “Young Inversion with Multiple Linked QTLs Under Selection in a Hybrid Zone,” Cheng-Ruei Lee, Baosheng Wang et al. Nature Ecology & Evolution, April 3, 2017. DOI:10.1038/s41559-017-0119.

Guest post by Molly Rivera-Olds

 

 

 

 

 

From Sunfish-Seeker to Planet-Saver: Dr. Tierney Thys

Marine biologist Tierney Thys believes that science make us superheroes. In her words, the tools of science are the superpowers that “allow us to explore worlds that are invisible to the naked eye.”

As a National Geographic Explorer, Research Associate at the California Academy of Sciences, and, in my humble opinion, one of the most effective, passionate science communicators out there, she may as well be a superhero already.

Dr. Tierney Thys snorkels with some aquatic research subjects. Photo credit: Tierney Thys.

Thys, an alumna of Duke’s Biology Department, presented at the Marine Science and Conservation Leaders’ (MSCL) inaugural Marine Science Symposium on Saturday, March 25. She was one of four featured speakers — all women in STEM— whose research interests range from marine biology to physical oceanography.

Though she discussed her own research and life story in depth, the main point Thys drove home was the importance of (and science behind) powerful science communication.

Like most marine biologists, Thys’ love for the ocean began when she was a child. She received her Ph.D. from Duke in 1998, an endeavor which, she said, “looked interminable while [she] was in the midst of grad school but, in retrospect, was just a blink of an eye.”

Among the many fun tidbits she has gleaned studying ocean science are the following:

  • As adults, humans retain a lot of characteristics from our fish-like time in the womb; e.g. “we can thank fish for washboard abs.”
  • Humans, for all our obsession with large brains, have nothing on the African elephantfish, which has a “higher brain weight to body weight ratio than any other vertebrate.”
  • Fish had the gender continuum “totally nailed” before it became trendy among humans, with fish of many species having the ability to change sex at will.

Thys, right, and her dissertation advisor, Dr. Stephen Wainwright, left. Photo credit: Tierney Thys.

Her most impactful lesson out of Duke, however, came from her dissertation advisor Stephen Wainwright, James B. Duke Professor emeritus of zoology. Wainwright is the founder of Duke’s Bio-Design Studio, an art studio within a scientific research laboratory employing a full-time sculptor “to create three-dimensional working models of biological systems for research,” as reported by Duke Magazine. Exposure to this unique melding of disciplines in the final stages of Thys’ education set her on what she said was “an eclectic career path” that would also seek to fuse the artistic and the scientific.

Thys’ research specialty out of graduate school is the Mola mola, more commonly known as the Ocean sunfish—the heaviest bony fish on the planet. According to Thys, sunfish can grow to “60 million times their starting weight,” the equivalent of a human child growing to the weight of six Titanic ships. The heaviest Mola ever caught weighed over 5000 lbs., though, surprisingly, jellies (what most folks would call jellyfish) comprise most of the adult sunfish’s diet.

Thys hailed pop-off satellite tags as the “superpower” of science that allows her to track sunfish through the world ocean, generating data that can improve environmental protection of the species.

A fun graphic Thys used in her presentation to explain the technology of pop-off tags for tracking Mola mola, pictured right. Photo credit: Mike Johnson.

“Studying the sunfish has eclipsed studying any other fish for me. [They’re ] a massive part of the bycatch in driftnet fisheries all over the world—[but] we need to keep our jelly-eaters intact. With data, we can figure out the [sunfish] hotspots, and work to protect those areas,” Thys said.

Thys has tackled this problem herself by adapting the discipline-blending approach of her advisor, Wainwright. She has primarily used filmmaking to bridge the gap between the arts and sciences, playing key roles in high-profile documentary projects meant to improve public understanding of marine science, technology, and conservation. These include the Strange Days on Planet Earth series with National Geographic, The Shape of Life series with PBS, and several short documentary films. She has also collaborated with dance companies to create conservation-oriented dance productions, K-6 schools for educational art projects, and prisons to improve inmates’ scientific literacy with nature imagery—all to widen the scope of her science-education efforts. Thys supports her creative ideas with science itself:

“One very large filter exists between our conscious mind and subconscious mind, she said. “Our conscious mind can only process a tiny amount of the information gathered by our subconscious mind.”

“A good story can cut through these filters and light up our brains in new ways,” Thys said “By using different forms of art to tell stories infused with scientific information, we can message in profound ways. We can reach people who might not otherwise be interested or receptive to science. The arts are not a luxury, but rather a powerful vehicle for helping message, teach and share our vital scientific findings,” Thys said.

A mural Thys made with students out of bottle caps at a California elementary school, one of Thys’ many efforts to spread public awareness of scientific issues. Photo credit: Tierney Thys.

As though she hadn’t already empowered everyone in the audience to save the world, Thys concluded with a compelling piece of advice: “Be a part of something much bigger than yourself.”

Post by Maya Iskandarani

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