Golden frogs with unique communities of skin bacteria survive exposure to frog-killing fungus  

Chytridiomycosis is an amphibian disease that has wiped out populations of many frog species around the world, including the charismatic Panamanian golden frog, which now exists only in captivity in the United States and Panama.

Research published this week in the Proceedings of the Royal Society found unique communities of skin bacteria on golden frogs that survived chytridiomycosis. The original experiment was designed to test the idea that antifungal probiotic bacteria may be used to prevent chytridiomycosis in captive golden frogs. Approximately 25 percent of the golden frogs eventually cleared infection, but their survival was not associated with the probiotic treatment, rather it was associated with bacteria that were present on their skin prior to the start of the experiment. In fact, the probiotic antifungal bacteria did not appear to establish on the golden frog skin at all.

Study authors Matt Becker and Shawna Cikanek work to inoculate frogs with beneficial bacteriaMatthew Becker, a fellow at the Smithsonian Conservation Biology Institute who conducted the experiment as part of his PhD research at Virginia Tech University, says it is unclear why the microbes did not linger on the skin, but he thinks that the way he treated the frogs – with a high dose of bacteria for a short duration – may be part of the reason.

“I think identifying alternative probiotic treatment methods that optimize dosages and exposure times will be key for moving forward with the use of probiotics to mitigate chytridiomycosis,” Becker said.

Brian Gratwicke, amphibian conservation biologist at the Smithsonian Conservation Biology Institute where the experiment was conducted, says that he was disappointed that they did not find a ‘silver bullet’ to cure chytridiomycosis in this species, but noted that the results do advance our understanding of this disease.

“Previous experiments found that golden frogs are highly susceptible to chytridiomycosis, so any survival is cause for hope,” said Reid Harris, director of disease mitigation at the Amphibian Survival Alliance. “The tricky piece is figuring out the survival mechanism, and this exciting research gives some new insights in that direction.”

This research also provides additional support for the importance of symbiotic microbes, or the ‘microbiome,’ for the health of their hosts, ranging from sponges and corals to humans.

“In all multi-cellular organisms, we have suites of microbes performing critical functions for their hosts, and the same appears to be true for golden frogs,” said Lisa Belden, who supervised the study at Virginia Tech University.

The team, led by Becker, now plans to determine if this study is repeatable by investigating whether the golden frog’s skin microbiota can predict the susceptibility to chytridiomycosis. They will also investigate whether the bacteria associated with the surviving frogs from this study can be used as a probiotic treatment to prevent infections of golden frogs without a ‘protective’ microbiota.

“The ultimate goal of this research is to identify a method to establish healthy populations of golden frogs in their native habitat, despite the presence of chytridiomycosis in the environment,” Becker said.

Citation: Matthew H. Becker, Jenifer B. Walke, Shawna Cikanek, Anna E. Savage, Nichole Mattheus, Celina N. Santiago, Kevin P. C. Minbiole, Reid N. Harris, Lisa K. Belden, Brian Gratwicke (2015) Composition of symbiotic bacteria predicts survival in Panamanian golden frogs infected with a lethal fungus. Proc. R. Soc. B: 2015 282 20142881; DOI: 10.1098/rspb.2014.2881. Published 18 March 2015

The Recently Discovered Salamander-Devouring Fungus and Reasons for Concern for the Future the Salamander Biodiversity in the United States

appalachian salamandersEnigmatic Fire salamander (Salamandra salamandra) declines in the Netherlands have been attributed to the recently described fungal pathogen Batrachochytrium salamandrivorans (Bs). Since 2010, the S. salamandra population at Bunderbos, Netherlands has decreased by 96%. An Martel et al’s recent Science paper showed that some US salamander species are highly susceptible to Bs, confirmed its occurrence in the pet trade, and noted that it has not yet been detected in the US. Large numbers of live salamanders are legally imported into the US each year for the pet trade. In the first 6 months of 2014, for example, 3,445 fire salamanders imported into the US, mostly from Slovenia.

The genus Batrachochytrium, which before the discovery of Bs solely included Batrachochytrium dendrobatidis (Bd), has gained an infamous reputation for global amphibian declines. Biologists believe that we are witnessing the sixth mass extinction in part because of the virulence and global spread of Bd among the world’s amphibians. The discovery of this new pathogen and our improved understanding of the ravaging effects of emerging wildlife disease raise concerns that US salamanders could share the same fate.

The US is a biodiversity hotspot for salamanders

Appalachia is a global salamander biodiversity hotspot (Source: http://www.biodiversitymapping.org/amphibians.htm)

Appalachia is a global salamander biodiversity hotspot (Source: http://www.biodiversitymapping.org/amphibians.htm)

The Appalachian Mountains are a renowned biodiversity hotspot for salamanders. The potential threat of this emerging pathogen in the US is therefore magnified, and it is imperative that we keep this disease out of the US. Salamander genetic diversity in the Appalachians is the highest in the world with 72 salamander species that are mostly endemic. The United States is home to nine out of ten salamander families and four of the ten extant salamander families are endemic to the United States including amphiumas, Pacific giant salamanders, torrent salamanders and sirens. Mole salamanders are also found in Canada and Mexico, but nearly all of their biodiversity is contained with U.S. borders. Giant salamanders are a primitive lineage of giant salamanders with three extant species, located in the U.S., Japan, and China. The hellbender is one of the giants and has found refuge in the Appalachian Mountains since amphibians originated, some 360 million years ago.

The ecological role of salamanders, the smaller majority, can often go unnoticed, but consider this biomass assessment of salamanders in Appalachia. One classic mark-recapture study in the eastern US noted “The biomass of salamanders is about twice that of birds during the bird’s peak breeding season and is about equal to the biomass of small mammals” (Burton and Likens 1975). With densities this high, a novel salamander-specific pathogen to which these animals have never been exposed have the potential be able to spread like wildfire, much like Bd spread through naïve Neotropical amphibian populations.

Immediate action is needed

We should immediately halt the importation of salamanders from any overseas sources, unless they can be certified free from Bs and Bd. In May 2008 the OIE, which is the organization created to mitigate zoonotic diseases (i.e., anthrax, mad cow disease, etc.), recognized Bd as a notifiable disease. Stricter trade regulations recommended by OIE would substantially reduce the spread of both Bs and Bd, however the OIE changes have not been adopted by the US Department of Agriculture and Interior and until doing so there are no legal means to reject infected shipments. A joint statement from the Amphibian Specialist Group and Amphibian Survival Alliance calls for immediate policy actions to stop the further spread of devastating wildlife diseases, and this time it is not too late to do something about it.

by Blake Klocke

Researchers develop new method to test an amphibian’s susceptibility to the deadly amphibian chytrid fungus.

This new method could help us to test out new probiotic therapies and predict a captive-bred frog's survival from exposure to chytrid fungus, without ever having to expose them to it.

This new method could help us to test out new probiotic therapies. It can predict a captive-bred frog’s survival from exposure to chytrid fungus, without ever having to expose them to it experimentally.

Researchers at the University of Boulder Colorado, University of Zurich and Copenhagen University have developed a new method to predict how susceptible an amphibian is to a frog-killing fungus wiping out amphibians all over the world. The test looks at the antifungal properties of skin mucus that contains skin bacteria and chemicals secreted by the frog itself. Together the interactions between the skin bacteria and chemical secreted from glands on the frog skin are the frog’s first line of defense against skin disease.

Their paper, just published in PLOS One, sampled 8,500 frogs across Europe. They found that antifungal properties of the mucus were related to the prevalence of amphibian chytrid infection in natural populations. They found that when they experimentally exposed frogs to the chytrid fungus in a lab that they could predict survival of frogs based on an independent mucus sample. The researchers also found that when they added beneficial skin bacteria to the frogs that the anti-fungal properties of the skin were improved.

This study may help us to develop tools that we could use to reintroduce frogs back into areas affected by the frog-killing fungus, including Panama. “We have all these amphibians in captivity now, like the golden frog in Panama, a really beautiful species that is now extinct in the wild,” said Douglas Woodhams, a postdoctoral researcher at CU-Boulder and lead author of the paper. “We want to be able to reintroduce them, but the pathogen that attacked them is still out there,” he said. “Now we can determine what probiotic treatment might work best to protect the frogs without infecting them with the pathogen and seeing how many die.”

http://www.plosone.org/article/info%3Adoi/10.1371/journal.pone.0096375

Mission Critical: Amphibian Rescue

This award-winning documentary featuring our race to find a cure for a deadly amphibian disease and to build an amphibian ark in Panama is now available for FREE.  Watch the trailer below and download the full feature if you would like to see more on the itunes store for a limited time only.

CLICK HERE to download the full episode of Smithsonian Networks Mission Critical: Amphibian Rescue film. FREE FOR A LIMITED TIME ONLY!

Rescue Project Successfully Breeds Endangered Frog Species

 

Limosa harlequin frog (Atelopus limosus) baby on a U.S. quarter.

Limosa harlequin frog (Atelopus limosus) baby on a U.S. quarter. (Photo by Brian Gratwicke, Smithsonian Conservation Biology Institute)

The limosa harlequin frog (Atelopus limosus), an endangered species native to Panama, now has a new lease on life. The Panama Amphibian Rescue and Conservation Project is successfully breeding the chevron-patterned form of the species in captivity for the first time. The rescue project is raising nine healthy frogs from one mating pair and hundreds of tadpoles from another pair.

“These frogs represent the last hope for their species,” said Brian Gratwicke, international coordinator for the project and a research biologist at the Smithsonian Conservation Biology Institute, one of six project partners. “This new generation is hugely inspiring to us as we work to conserve and care for this species and others.”

Nearly one-third of the world’s amphibian species are at risk of extinction. The rescue project aims to save priority species of frogs in Panama, one of the world’s last strongholds for amphibian biodiversity. While the global amphibian crisis is the result of habitat loss, climate change and pollution, a fungal disease, chytridiomycosis, is likely responsible for as many as 94 of 120 frog species disappearing since 1980.

Between its facilities at the Smithsonian Tropical Research Institute in Gamboa, Panama, and the El Valle Amphibian Conservation Center in El Valle, Panama, the rescue project currently cares for 55 adult limosa harlequin frogs of the chevron-patterned form and 10 of the plain-color form. The project has had limited success breeding the plain-color form of this species, and has successfully bred other challenging endangered species, including crowned treefrogs (Anotheca spinosa), horned marsupial frogs (Gastrotheca cornuta) and toad mountain harlequin frogs (A. certus).

Each species requires its own unique husbandry to thrive and breed. The project’s animal care team and scientists learn husbandry techniques as they work with a limited number of individuals. Jorge Guerrel, conservation biologist at the Smithsonian Tropical Research Institute, arranged rocks in the breeding tank to create the submerged caves that appear to be the preferred egg deposition sites for limosa harlequin frogs. Like other Atelopus species, tadpoles require highly oxygenated, gently flowing water between 22 and 24 degrees Celsius. The tadpoles’ natural food is algal film growing on submerged rocks, which Guerrel and his colleagues re-created by painting petri dishes with a solution of powdered spirulina algae, then allowing it to dry.

The mission of the Panama Amphibian Rescue and Conservation Project is to rescue amphibian species that are in extreme danger of extinction throughout Panama. The project’s efforts and expertise are focused on establishing assurance colonies and developing methodologies to reduce the impact of the amphibian chytrid fungus so that one day captive amphibians may be reintroduced to the wild. Current project partners include Cheyenne Mountain Zoo, Houston Zoo, Smithsonian’s National Zoological Park, Smithsonian Tropical Research Institute and Zoo New England.

Lindsay Renick Mayer, Smithsonian’s National Zoo

Frog Poetry and the Washington Post

Washington Post

On Dec. 30, the Washington Post ran a front-page story about the rescue project.

The year ended on a high note for the Panama Amphibian Rescue and Conservation Project. William Booth, a science writer for the Washington Post, joined rescue project researchers on a field expedition and his story about the rescue project came out on the front page of the Post on Dec. 30. The story inspired one reader, Tim Torkildson, to share a lovely poem about frogs and the disease that is wiping them out.

Booth also did this NPR interview about his recent trip to Panama.

If you saw the story and are interested in making a donation to the rescue project, please follow this link to the National Zoo’s website.

THE FROG
by Tim Torkildson

The frog is an amphibian
Who thrives most ev’rywhere,
From the dry Namibian
To just off ol’ Times Square.
The ones who have a bumpy skin,
With warts and pits and nodes,
Are the closest Phylum kin;
We simply call them toads.
The bullfrogs in the early spring
give ponds reverberation
With their raucous verbal fling,
Attempting procreation.
The have a courtship ritual
that’s called, I think, amplexus,
Which gives them fits conniptual
Between the two odd sexes.
A little boy will manage to
Corral a tadpole, yes,
And give it quite a slimy view
Right down his sister’s dress!
And did you know the urine from
a pregnant lady will
cause some frogs to lay a scum
of eggs, with no male thrill?
And so they’re useful critters,
As the French will tell you so;
Their legs taste good in fritters,
Are mistaken for turbot.
And what of cane toads, mind you,
Where, if you lick the skin,
The psychedelics blind you
To sorrow, grief and sin?
But frogs, those little gargoyles,
Which are funny in cartoons,
Are engaged in lethal broils
That leaves their lives in ruins.
A fungus known as “Bd” kills
The frogs down in Belize,
Then jumps the valleys and the hills
So others it may seize.
The Costa Rica Golden Toad
Is now extinct, alack.
More are headed down that road,
Since habitat is slack.
Toxins give some frogs three legs,
Which doesn’t help them jump.
Instead they are like clumsy kegs
Who in the water flump.
Scientists preserve some frogs
In habitats in labs.
Dressed in their starched, stiff white togs
They keep meticulous tabs.
To save the frogs, oh please donate
A dollar or a yuan,
So the polliwog birth rate
Will someday be a shoo-in!

Using genetic tools to understand and manage chytridiomycosis

Lowland leopard frog (Lithobates yavapaiensis)

The lowland leopard frog is the focus of SCBI postdoc Anna Savage's work looking at the relationship between genes and chytrid resistance.

Chytridiomycosis, the disease wiping out frog species around the world, was described in 1999 by a team of researchers at the Smithsonian and the University of Maine. Today, in addition to creating an insurance population for various Panamanian species, the Smithsonian Conservation Biology Institute aims to find a way to manage this disease. One of the most promising solutions to stopping the killer may rest in genetics. SCBI scientists are looking to identify genes within frogs that provide resistance to the pathogen or that make them more susceptible to it.

Researchers pursuing this approach are optimistic that genetics could provide different answers than those offered by probiotics, which SCBI is also pursuing. The genetic approach is one based on natural selection.

“If you look across species, including humans, diseases rarely wipe out an entire species without any evidence of resistance, regardless of how virulent they are,” said Anna Savage, an postdoctoral fellow in SCBI’s Center for Conservation and Evolutionary Genetics studying the genetics of chytrid. “Immune systems are so complex that there’s a strong possibility for the development of resistance. The probability that a species develops no resistance to a disease is rather small.”

Genetics research within the last decade has identified frogs with varying degrees of resistance to chytrid. Savage is focusing her research on the lowland leopard frog (Lithobates yavapaiensis), which exhibits an intermediate level of resistance to the pathogen. Some individuals demonstrate resistance while others of the same species die if infected with chytrid. The identification of genetic variations between the individuals holds the potential of being the answer between life and death. One of the new frontiers for genetic research is the examination of a frog species’ immune system genes.

“If we can identify the genes responsible for resistance, we can breed the animals in captivity to ‘spread’ this genetic resistance and give adaptation a headstart,” Savage said. “This area of research holds the potential for creating populations of self-sustaining frogs that confer resistance to the rest of the population through reproduction.”

Anna Savage with bullfrog

Savage's research could hold the key to putting a stop to a deadly pathogen killing frogs around the world.

In order to understand a frog’s response to chytrid, the focus must be broadened to understanding how immune system genes interact with one another and which ones are being expressed during resistance. Savage is using this approach to understand chytrid susceptibility in lowland leopard frogs that are being raised at the Smithsonian’s National Zoo. This approach could potentially help scientists identify genes responsible for chytrid resistance. While this type of research shows considerable potential, researchers are only just beginning to scratch the surface.

“Disease outcomes from chytridiomycosis can depend on several factors,” said Brian Gratwicke, SCBI wildlife biologist and Panama Amphibian Rescue and Conservation Project international coordinator.  “Some frogs appear to have innate resistance to the disease and produce antimicrobial peptides from glands in their skin. It is possible that some might acquire resistance through exposure, while other frogs have beneficial anti-fungal skin bacteria or a behavioral preference for temperatures that are bad for the fungus. As the fight against chytrid continues, we remain hopeful that the answers are out there and that Dr. Savage’s work will give us some insight into how this system really functions.”        

–Will Lazaro, Smithsonian’s National Zoo

(Frog photo by Jared Grummer; photo of Anna Savage by Dennis Caldwell)

Celebrating tadpole diversity

Tadpoles

The ability of certain Panamanian species to survive will depend on the ability of the rescue project to perfect specialized care for the individual species. (Photo by Lindsay Renick Mayer, Smithsonian

When we talk about frog diversity, we always mention how many species there are, or how they are found all over the world. We say how much frogs vary in size, or how colorful they can be. This is all true: there are more than 4,900 frog species, found on every continent, ranging from ½-inch long to more than a foot, that come in every color of the rainbow. It makes sense, then, that frogs are just as diverse during their other stages of the life cycle. Tadpoles, in fact, are among the most diverse vertebrates on Earth and are themselves morphologically and behaviorally unique from their adult counterparts.

Special Adaptations

Depending on the species, tadpoles can stay in the larval stage from eight days to two years, and vary in length from 1 to 4 inches. Overall, there is greater tadpole diversity in the tropics, but variations occur within habitats, as well. Even tadpoles with the same feeding habits can have diverse mouth shapes or behaviors. For example, tadpoles of the Asian horned frog (Megophrys montana) have an upturned mouth because they eat from the surface. Recently, however, scientists have observed a Honduran tadpole called Duellmanohyla soralia that also eats from the surface, but has a mouth in the more traditional spot. Instead, the Duellmanohyla soralia turns its body upside-down to reach the surface.

“We see tadpoles solving the same problems of survival in different ways,” says Dr. Roy McDiarmid, an amphibian zoologist and tadpole expert at the National Museum of Natural History. “This is where their diversity derives from.”

These variations show how tadpoles’ features are shaped largely by their surrounding environments. For example, tadpoles seem especially good at responding to strong predator presence. Over time tadpoles can grow their tails longer and deeper if there are numerous predators, allowing them to swim faster and look bigger. For European common frog (Rana temporaria) tadpoles, longer tails increase their chances of escape from predators up to 30 percent, according to the Institute of Zoology.

Like tail length, other adaptations can protect tadpoles from being eaten. While the majority of tadpole species have brown or faded coloration, several are multicolored. Contrary to its name, Cope’s gray tree frog (Hyla chrysoscelis) tadpoles grow red tails in response to the presence of dragonflies. Called aposematism, vibrant colors make these tadpoles appear larger or distasteful to their predators.

Researchers have only recently discovered other survival mechanisms that tadpoles have developed. In 2006, scientists discovered that if attacked, red-eyed tree frog (Agalychnis callidryas) embryos can hatch themselves within seconds and escape into the water below. These embryos can interpret vibrations on the water with astonishing precision.

“It turns out that when a snake bites into a gooey mass, all the embryos try to wiggle free,” Karen Warkentin, a biologist working at the Smithsonian Tropical Research Institute in Panama, told National Geographic.  “A wasp’s more focused attack prompts only neighboring eggs to hatch. And a rainstorm triggers nothing at all.”

Other recent research has looked at tadpole sensory input, such as smell and sound. In 2009, researchers at the University of California Davis determined that wood frog (Rana sylvatica) tadpoles can “smell” their primary predator, the salamander. The “odor” of a salamander in the water caused tadpoles to freeze. The strength of the scent determined how long the tadpoles remained still. In 2010, Dr. Guillermo Natale discovered the first evidence of aural larva communication. Natale heard tadpoles of the Bell’s horned frog (Ceratophrys ornate) “screaming” underwater when a threat was present.

Discovery

Tadpole

The amount of tadpole diversity rivals that of their adult life forms. (Photo by Brian Gratwicke, Smithsonian Conservation Biology Institute)

New and unpredictable tadpole discoveries continue to go on worldwide. For example, biologists at the Australian Museum in Sydney recently discovered that tadpoles of the vampire flying frog (Rhacophorus vampyrus) have small black fangs, instead of traditional mouthparts. In general, scientists estimate that more than 1,000 frog species have yet to be discovered, not to mention all the intricacies of their life stages.

Tadpoles are also essential for understanding the dramatic decline in frog populations. Chytrid, the epidemic that has affected 30 percent of the world’s amphibian population, is the lead cause of this decline. In tadpoles, chytrid only infects the keratin around their mouths. However, as they metamorphose into frogs, chytrid fatally spreads throughout their bodies. By studying tadpoles, we can better understand how frogs contract and carry chytrid.

“When frog species are disappearing like they are, you would want to know what’s going on at each stage of the life cycle, egg to larva to juvenile to adult–everything,” McDiarmid says.

The Challenges of Breeding

For the Panama Amphibian Rescue and Conservation Project, every tadpole means the chance for a species to survive. But because many of the rescue project’s priority species have never been kept or bred in captivity before, rearing tadpoles can mean a steep learning curve.

“Breeding frogs isn’t just about putting a male and female together and hoping for eggs,” said Lindsay Renick Mayer, spokesperson for the rescue project. “It’s about specialized husbandry for each individual species among a diverse array and unfortunately for some of these species we’re learning those skills even as the species dwindles down to just a few remaining individuals. Whether these species are one day returned to the wild depends on the rescue project’s success in perfecting the variety of care in a short period of time.”

Nadia Hlebowitsh, Smithsonian’s National Zoo

New experiment may offer hope for frogs facing chytrid

Probiotics bath

The golden frogs were given a bath in one of four probiotic solutions. (Photo by Brian Gratwicke, Smithsonian Conservation Biology Institute)

We usually think of bacteria as bad for us, but that isn’t always the case. For us humans, the most common examples of helpful bacteria, or probiotics, live in yogurt. Now, scientists believe amphibian probiotics may be the key to fighting chytridiomycosis, the fungal disease devastating frogs around the world.

A few years ago, Reid Harris, a biology professor at James Madison University, discovered that local salamanders that could survive chytrid played host to bacteria in their skin. Now, Brian Gratwicke, a research biologist at the Smithsonian Conservation Biology Institute, is collaborating with a team from Virginia Tech, James Madison, Villanova and Vanderbilt Universities in an experiment to see if similar bacteria can protect the Panamanian golden frog, which he calls “the poster-child for amphibian conservation.”

The first step is to find a probiotic that will stick to the golden frogs. In early December, the team began giving golden frogs baths using four different types of bacteria. Researchers gathered the potential probiotics from frogs in Panama in 2009. The finalists were chosen based on their ability to prevent chytrid growth in lab tests, with a preference for bacteria that are common in close relatives of Panamanian golden frogs.

Every two weeks, each frog is swabbed to check whether its probiotic has made itself at home. The tests take some time, so a month and a half in, the team is still waiting for results to see which probiotics are sticking. But they do have some good news already.

“The bacteria haven’t been causing any problems with the frogs and they all look healthy,” said Gratwicke, who emphasizes how important it is to use only beneficial bacteria. In addition to tracking weight gain and other visible characteristics, Shawna Cikanek, a student at Kansas State College of Veterinary Medicine is using frog poop to study stress hormones to get a better picture of the animals’ overall health and whether the bacteria are causing any stress.

The probiotics that stick to the frogs for a full three months will move on to the next round of tests, when bacteria-shielded frogs will be infected with chytrid to check for any adverse effects.

“Hopefully, the bacteria are going to do their thing and protect these little guys,” said Matt Becker, a PhD candidate from Virginia Tech who is conducting the experiment. Whatever probiotics make the cut will be tested again on golden frogs bred in Panama before scientists develop a final plan.

So far, chytrid has defied attempts to stop it. Scientists may be able to selectively breed frogs resistant to chytrid, but there has been very little work done so far in that direction. But there are high hopes for probiotics’ potential to protect frogs. “It’s a long shot, but it’s our best shot,” said Gratwicke.

Becker hopes that one day, probiotics will allow Panamanian golden frogs to return to their homes. “These guys are really neat and it’s so sad not to see them in the wild,” he said. “We have a moral obligation since indicators are pointing to humans as major spreaders of the disease through the frog trade.”

Meghan Bartels, Smithsonian’s National Zoo

Smithsonian scientists survey frogs in the Peruvian Andes

Acancocha water frog (Telmatobius jelskii)

Last December two researchers from the Smithsonian Conservation Biology Institute spent two weeks in Peru surveying the Acancocha water frog (Photo by Jessica Deichmann, SCBI)

Imagine measuring the tail of a squirmy, inch-long tadpole. Now imagine doing that where the air is thin enough to make you dizzy, a hail storm is about to start and you just spent 45 minutes up to your elbows in a freezing cold stream.

Last December, Jessica Deichmann, a research scientist with the Smithsonian Conservation Biology Institute, and Ed Smith, a biologist at the Smithsonian’s National Zoo’s Amazonia Exhibit, spent two weeks doing just that to complete a survey of the Acancocha water frog (Telmatobius jelskii). They were participating in a frog surveying trip for SCBI’s Center for Conservation Education and Sustainability’s Peruvian Biodiversity Monitoring and Assessment Program (BMAP). The program lends Smithsonian scientific expertise to gas and oil companies to assess the effects of development projects on local plants and animals. This information is used to improve restoration work, and reduce environmental impacts of future development. This was the third survey along the path of a natural gas pipeline constructed in 2009 that runs from the Amazon over the Andes to the Pacific Ocean—about 250 miles.

The Acancocha water frog is found only in the cold clear streams of the high-elevation Peruvian Andes. The frog is one of about 40 species researchers are surveying around the construction of this particular pipeline. Scientists chose this species because although historically it has been fairly abundant, it lives in a relatively small area with precise habitat requirements. When individuals of a species are clustered together, it’s easier to lose the entire species.

“You always worry about frogs with small geographic ranges—not just frogs, but any species,” Deichmann said.

When you think of the Peruvian Andes, you probably think of Macchu Picchu, a lush green mountainside where, as Smith jokes, “if you sit too long you have orchids growing on you.” Smith and Deichmann, however, were in a very different ecosystem called puna and sometimes nicknamed equatorial tundra. Puna is found high in the Andes, where trees no longer grow, rain is scarce, and nights are freezing. The team spent much of its time about 15,000 feet above sea level, where the air is so thin that breathing is hard until you get used to it. “For us lowland landlubbers, that alone was an exhausting business,” Smith said.

Peruvian Andes

Smith and Deichmann were in a very different ecosystem called puna and sometimes nicknamed equatorial tundra. (Photo by Jessica Deichmann, SCBI)

The team—Deichmann, Smith and two Peruvian biologists—took samples at twelve sites where the pipeline crosses mountain streams. At each site, they spent about 45 minutes upstream, elbow-deep, feeling for frogs and tadpoles. For each tadpole, first, the team swabbed its mouth for a sample for chytrid testing. The team also recorded the tadpole’s body length, tail length, mass, and developmental stage, and photographed each sample before the animal was returned to the stream.  They also took measurements of the stream itself. Then the team repeated the whole process downstream of the pipeline. Any one site could take five or six hours.

But Deichmann is clear that the process was worth it. “Just to find frogs and tadpoles was exciting,” she said. “Especially adults—after hours and hours in freezing cold water and your arms are purple, when you find your first frog it’s just so exciting.”

The team hasn’t processed the data yet, but their initial measurements suggest some good news: so far, they have seen no obvious effects as a result of the pipeline. However, although the team found hundreds of tadpoles, they found only 10 adult frogs in all the sample areas.

Unfortunately, that suggests chytrid is playing a role in shaping the population structure. The skin fungus follows skin keratin proteins, the amphibian equivalents of those found in human skin calluses, hair and nails. Tadpoles usually survive chytrid infection because they have keratin only around their mouths. As they develop their keratin “suit of armor,” frogs are left vulnerable to the disease that has decimated them in less than half a century.

“From metamorphosis, something called the ‘chytrid clock’ starts ticking,” Smith said. In adults, “chytrid interferes with water balance, usually in a lethal way.” Because of this, chytrid-struck populations often consist of many tadpoles and a few adults.

And according to the previous surveys, “at pretty much all the sites where frogs and tadpoles were present, chytrid was present,” Deichmann said.

Mountain stream

At each of 132 sites, the team spent about 45 minutes upstream, elbow-deep, feeling for frogs and tadpoles. (Photo by Jessica Deichmann, SCBI)

But until they have the results from lab tests on the mouth swabs the team took, they won’t have the full picture about how the disease is affecting these populations.  And chytrid isn’t the only danger these frogs face. “In a lot of our streams we were, not surprisingly but disappointingly, finding trout,” an invasive predator, Smith said.

Deichmann and Smith agree that the data they collected will be useful for conservation efforts. “Monitoring populations now gives us a baseline without which we can never know what’s changed,” Smith said. The team has a tentative follow-up survey scheduled for the dry season of 2012, during the northern hemisphere’s summer. Deichmann hopes that the data from this December’s trip will allow the program to modify the survey protocol to make sure future trips are gathering the most helpful data possible.

And of this expedition? “It was an amazing trip,” Deichmann said. “The habitats are stunning, the scenery is stunning. You’re at the top of the world.”

Meghan Bartels