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

Studying behavior and hormones to improve amphibian care

Shawna_CikanekBefore beginning my research at the Smithsonian, I knew surprisingly little about the status of the world’s amphibians. I was fortunate to be awarded an internship through Kansas State University College of Veterinary Medicine based at the Smithsonian Conservation Biology Institute in Front Royal, VA. Once I arrived in Front Royal, I learned quickly about the plight of amphibians around the globe and I remember listening in awe when I was first told about the Amphibian Arks in Gamboa and El Valle in Panama and how they hold the last remaining individuals of some of Panama’s most precious amphibian species. They had been brought into these assurance colonies because of the threat of a devastating fungal pathogen Batrachochytrium dendrobatidis (Bd) that is responsible for the disappearance of many species around the world. Initially, the harlequin frogs in the Amphibian Arks in Panama were housed individually, because when placed together keepers observed that males would fight and they were worried the fighting would unduly stress these precious animals. However, this quickly led to a space shortage as the amphibian ark filled up. The Maryland Zoo in Baltimore, which helps coordinate the Species Survival Plan for Panamanian Golden frogs in the USA, suggested that we group the animals together in same-sex groups. They noted that this was working out just fine for frogs reared in captivity, but we wanted to evaluate whether wild-collected harlequin frogs would adapt to this group-housing situation.

Ethogram showing different types of aggressive interactions

Ethogram showing different types of aggressive interactions

My collaborators in Panama conducted a behavioral study of frogs placed in groups and monitored the number of aggressive interactions between the frogs, making sure that there were no injuries. Back in Front Royal, I worked with my colleagues in the endocrine labs to measure the amount of cortisol, a steroid hormone that the frog produces and can be detected in its poop. We did this by adapting existing stress hormone monitoring methods that the SCBI uses for many other wildlife species, like pandas and elephants. We found that, initially, the frogs housed together would interact aggressively towards one another. This aggression was mirrored by an elevation in stress hormones in the feces during the first week of the study. However, after the first 2 weeks, the frequency of aggressive behaviors declined dramatically, and the concentration of cortisol dropped back down to normal. Based on these results, we determined that wild caught harlequin frogs could be safely housed together in same-sex groups over the longer term, and this study helped us to greatly reduce space constraints in our ex-situ collection of amphibians. We hope that our new method may be useful to others wanting to evaluate husbandry issues in captive amphibian collections. Read the full paper in PLoS One here By Shawna Cikanek, Kansas State University

The Move to Gamboa

November was the culmination of a year of incredibly hard work for us in Panama. We finally moved into our beautiful our beautiful new frog conservation facility in Gamboa. Maersk Line generously donated 7 shipping containers that that once ferried ice cream and frozen vegetables around the world, but they now house a most precious collection of endangered Panamanian frogs. The new Gamboa ARC (Amphibian Rescue Center) is an incredible leap forward enabling us to more effectively tackle the amphibian conservation crisis in Panama.

We are incredibly grateful to the Summit Municipal Park, who have been our generous hosts for the first 4 years of our project, and to our project partners Cheyenne Mountain Zoo, Houston Zoo, Zoo New England, the Smithsonian Conservation Biology Institute and the Smithsonian Tropical Research Institute. We have relied intensively on each other for help over the last 4 years and it has truly been a team effort! Generous grants from USAID and Minera Panama were the primary source of funds used for the construction of phase I and thanks to them, we now have a world class amphibian conservation facility. We have essential back-up systems such as an emergency generator power, and backup air-conditioning so that frogs can be kept in simulated tropical montane forest environments, even in the event of a power failure. We are now getting ready to break ground on phase II, a new NSF-funded amphibian research lab, quarantine and office building that will be the hub of our new research facility for the conservation of endangered Panamanian amphibians.  

Golden Frog Successfully Bred in Captivity in Panama

Juvenile Panamanian golden frog, reared at the El Valle Amphibian Conservation Center

Juvenile Panamanian golden frog, Atelopus zeteki, reared at the El Valle Amphibian Conservation Center

Now in its fourth year, Panama’s Golden Frog Day, August 14, is a salute to Panama’s cultural and ecological heritage with the golden frog, one of the most iconic symbols of Panama. The national legislation promotes species preservation and maintains an objective to promote conservation and protection of this amphibian species. This year the country can celebrate the successful breeding of the Panamanian golden frog at the El Valle Amphibian Conservation Center (EVACC), located inside the Níspero Zoo in El Valle de Antón. The egg clutch laid on November 24, 2012 successfully developed into tadpoles and were raised to form a group of 42 healthy young golden frogs.

“Bringing wild animals into captivity is only the beginning of the work that we do in our facility. Fusing applied technology, available resources, and human innovation to create Mother Nature, inside, is the challenge, “ said Heidi Ross, Director of EVACC. “Learning from our past experiences we focused a lot of energy on diet, and as the saying “you are what you eat” applies to humans, it also is essential for amphibians”.

EVACC director, Heidi Ross with a box of juvenile captive-reared golden frogs (Atelopus varius).

EVACC director, Heidi Ross with a box of juvenile captive-reared golden frogs (Atelopus varius).

“We are extremely proud of our conservation team in Panama,” said Peter Riger, director of conservation programs at the Houston Zoo, and one of the principal sponsors of this project. “EVACC has successfully bred both golden frog species in captivity and they have aggressive population management goals to grow the captive population to at least five hundred individuals for each species that I’m sure they will meet.”

The EVACC facility forms part of the Panama Amphibian Rescue and Conservation Project. The Project collects frogs in areas threatened by the devastating chytrid fungal disease that has decimated amphibians worldwide. The hope is to learn to raise these animals in captivity until enough is known about the disease to allow researchers to release amphibians into the wild once again. Project partners include the Smithsonian Tropical Research Institute, the Smithsonian’s National Zoological Park, the Cheyenne Mountain Zoo, the Houston Zoo, and Zoo New England. To learn more about the project please visit the project’s website.

Contact: Beth King kingb@si.edu 202-633-4700

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)

Things we are thankful for: Amphibian baby boom at EVACC and Summit Zoo

Reflecting on our achievements this last year, we would like to thank you, our generous supporters. You have provided moral support; social media likes and retweets generously; more than 40 of you have given us your time to help us save frogs; and even more have sacrificed your hard-earned money to help us achieve this important mission. Thank you! In particular I would like to recognize the extraordinary dedication and above-and-beyond service from our devoted conservation staff seen above proudly displaying the the captive-bred products of their hard work, including a second generation of captive-bred frogs at EVACC, and our first grown-up generation of captive-born frogs at the Summit Zoo. We salute you all.

Brian Gratwicke – International project coordinator for the Panama Amphibian Rescue and Conservation Project, Smithsonian Conservation Biology Institute

Food for Frogs at EVACC: What it takes to raise the inverts served for dinner

Big frog, little frog, golden frog, marsupial frog, endangered frog, and common frog; what do all of those guys have in common? They all need to eat.  There is no call-up home delivery in Panama, so here in El Valle at EVACC, we have had to roll up our sleeves and become invertebrate breeders (adding to an eclectic list of “other skills” needed in this particular conservation project).

Lisandro Vasquez

Lisandro Vasquez, rolling up his sleeves in the cricket room at EVACC.

The thing that most people do not think about when thinking about an amphibian conservation project is food for the animals. It would be incredibly difficult to have a successful breeding/conservation project without being able to feed the subjects at hand. At EVACC we think about insects, and other invertebrates, on a very regular basis. And we think about what they eat, in a captive setting, just as much.

ingredients

Some of the ingredients that go into preparing insect diets.

What do our frogs eat?  We have quite a few different species we look after, so we have quite a few invertebrates to offer to them.  For the most part we delineate food items to specific species based on food size, and the mode in which the frog eats.  The smallest food we have to offer is springtails, from the insect order Collembola.  They go through simple metamorphosis, and the different size nymphs can be sifted and fed to different size amphibians.

Springtail colony

Springtail colony.

Working our way up through the sizes we have the two different kinds of fruit flies; Drosophila melanogaster and Drosophila hydei.  The two fruit flies at EVACC do not fly and the smaller one, D. melanogaster, does not have wings. They have both been genetically modified to possess these traits. Fruit flies, with vitamin powders, are fed alternately to our diurnal species, and to some of the nocturnal species at different stages in their life. These two species of Drosophilago go through complete metamorphosis.

Drosophila melanogaster in fly cup

Drosophila melanogaster in fly cup.

Drosophila hydei colony

Drosophila hydei colony.

Domestic crickets (Acheta domesticus) make up a large portion of the diet for many species at EVACC.  This captive food colony requires quite a bit of time and space, but well worth it.  This insect goes through incomplete or simple metamorphosis as well, making it a food item for the smallest of frogs to large ones, as well.

cricket box

Cricket box crawling with one-month-old nymphs.

Cricket breeding room

Cricket breeding room, boxes upon boxes of crickets stacked up at EVACC.

Woodlice, sow bugs, or roly-poly bugs are also on the menu at EVACC. These invertebrates are not insects, but a crustacean from the family Oniscidea.  Leaf litter frogs love these calcium-packed treats.

Woodlice

Woodlice at EVACC.

The super worm (Zophobas morio) is a larva of a species of darkling beetle. Only the larvae are fed out to amphibians, as the adult are not preferred foods for frogs. This insect goes through complete metamorphosis.

We also have a colony of earthworms. Our colonies are not thriving at the moment, so most of the time we harvest from our own backyard. The other food item that we are currently not breeding, but do feed out, is the Neoconocephalus saturatus, a type of cone-headed katydid. We rely on a local family to help us out by collecting these katydids for us.

Last, but certainly not least, is the newest food item on the menu at EVACC; Blaberus discoidalis, a very large cockroach.

Cockroach

Cockroach on the menu.

Larvae pupa and adult stages

Larvae, pupa, and adult stages of Zophobas morio

Heidi Ross, director of El Valle Amphibian Conservation Center

(All photos courtesy of EVACC)