First-Time Breeding of Frog Suggests Hope for Critically Endangered Species

First captive-bred Craugastor evanesco photo by R. Ibanez.

When researchers discovered Craugastor evanesco in the rainforests of Panama, they called it the vanishing robber frog to signify just how quickly the deadly infectious amphibian disease chytridiomycosis had devastated its population. By the time the researchers had published about the new species in 2010, the vanishing robber frog had already disappeared from the park where they had discovered it.

Now, however, the vanishing robber frog may have a fighting chance at a future thanks to the Panama Amphibian Rescue and Conservation Project, which in December became the first program to breed the species in human care. After multiple attempts at breeding the species since 2015, a single pair has now produced one offspring—a success that has encouraged a cautious optimism that the rescue project can replicate the effort.

“A single individual doesn’t make a successful captive breeding program, but demonstrates that it can be done,” says Brian Gratwicke, an amphibian conservation biologist for the Smithsonian Conservation Biology Institute and rescue project international coordinator. “Every journey begins with the first step and this is a critical first step, not just for this species, but potentially for other endangered amphibians with similar reproductive needs.”

The rescue project, a world-class amphibian center run by SCBI and the Smithsonian Tropical Research Institute, currently has a founding population of 20 males and 20 females of the vanishing robber frog. Conservationists collected the frogs from a lowland site in central Panama where the rescue project is working with the support of Minera Panama S.A. to conserve amphibians in the area. But bringing a new and critically endangered species into human care requires learning its own unique husbandry and reproductive needs before it blinks out of existence—sometimes resulting in insurmountable challenges.

“Piecing together a species’ natural history with artificial systems, we can recreate to the best of our abilities an environment where the animals feel comfortable enough to breed,” said Heidi Ross, STRI’s director of El Valle Amphibian Conservation Center, whose expertise and persistence led to the successful first-time breeding of the species. “If we can get them to this point, to become sexually active in our artificial habitat, then we can simply tweak the system based on what worked, what did not work, and what materials are at our disposal. What we arduously do day in and day out is make sure we are providing the basic needs to the animals so that they help us help them from going extinct in the wild.”

The Craugastor group of frogs has a unique reproductive system called direct development—they bury eggs in wet sand and fully formed miniature adults hatch from the eggs. Understanding the frogs’ reproductive cues, special dietary needs and how to emulate the natural environment is essential to successful breeding, Ross says.

“Given the current difficult situation for amphibians in our region, this project represents scientific and biological hope, not only for this species of frog, but also for the recovery of Craugastor evanesco within its distribution range,” said Blanca Araúz, biologist and biodiversity superintendent of Minera Panamá. “As one of the species of interest for our project Cobre Panamá, its reproduction in captivity is important. Because the deadly infectious disease acts fast, experienced scientists can control the infection in these frogs and breed them under better conditions.”

Although scientists are still occasionally finding individual vanishing robber frogs in the field, they have not found a viable, self-sustaining population. Chytridiomycosis has been linked to dramatic population declines of amphibian species worldwide. This particular group of frogs in the Craugastor rugulosus series are particularly susceptible to chytridiomycosis with three closely related species in Panama having disappeared, putting extra pressure on ensuring the survival of Craugastor evanesco.

“It’s all a learning curve,” Gratwicke says. “I’m hopeful that we’ll be able to replicate this breeding event to develop a sustainable breeding program. If we can do that, we’ll be able to get this species back out in the wild as soon as we figure out how to safely do so. If we can do that, it’ll be time to celebrate.”

The Panama Amphibian Rescue and Conservation Project is a partnership between the Houston Zoo, Cheyenne Mountain Zoo, Zoo New England, SCBI and STRI.

Earth optimism: Frogs

What’s Working in Conservation

The global conservation movement has reached a turning point. We have documented the fast pace of habitat loss, the growing number of endangered and extinct species, and the increasing speed of global climate change. Yet while the seriousness of these threats cannot be denied, there are a growing number of examples of improvements in the health of species and ecosystems, along with benefits to human well-being, thanks to our conservation actions. Earth Optimism is a global initiative that celebrates a change in focus from problem to solution, from a sense of loss to one of hope, in the dialogue about conservation and sustainability.

The Smithsonian will host an Earth Optimism Summit in Washington DC April 21-23, 2017
Please register if you are coming in person or stream the presentations online here: https://earthoptimism.si.edu/calendar/summit Sunday is a free public day.

Dr Brian Gratwicke will present the Panama Amphibian Rescue and Conservation Project on Saturday April 22 5:15pm on the panel Science on the Edge

Love potion for frogs

Scientists at the Smithsonian Institution and partners have published a paper that optimizes sperm collection protocols from the critically endangered Panamanian Golden Frog Atelopus zeteki. It also improves our understanding of reproduction in endangered harlequin frogs. The research, to be published published 15 March 2017, in Theriogenology, was conducted by Dr. Gina DellaTogna, a Panamanian biologist who studied this charismatic animal at the National Zoological Park in Washington DC. The study characterizes the dose-response patterns for several artificial hormone treatments and describes the sperm morphology for the first time in this species.

Atelopus zeteki spermatazoan

“This study is important, because it contributes towards the basic understanding of reproduction of a highly endangered group of frogs in Latin America,” said DellaTogna, who performed the experiments for her PhD at the University of Maryland. “This study has already helped us to solve critical reproduction problems in captive Atelopus collections in Panama and allowed us to repeatedly collect high-quality sperm samples for genome resource banking at any time of the year, without harming the frogs.”

“Basic reproductive research is something that has yielded huge conservation dividends for the successful care and management of other endangered species like Pandas and Black Footed Ferrets,” said Pierre Comizzoli, a co-author of the paper and reproduction specialist at the National Zoo. “Gina’s research opens the door to develop methods like sperm freezing and storage to preserve the long term genetic integrity and diversity in small populations.”
The research is particularly relevant to current amphibian conservation efforts in Panama where the Panama Amphibian Rescue and Conservation Project has captive-breeding colonies of five species of Atelopus that are threatened with extinction from the deadly fungal disease chytridiomycosis.

Roberto Ibáñez, and Gina DellaTogna working on hormonal stimulation of frogs at the Panama Amphibian Rescue and Conservation Project

“Successful reproduction is key to any captive assurance program,” said Roberto Ibáñez, the director of the Panama Amphibian Rescue and Conservation project at the Smithsonian Tropical Research Institute in Panama. “Gina has already begun applying what she has learned to successfully help us to produce offspring from four other endangered harlequin frog species. I hope that she will eventually extend it to species with different modes of reproduction that are also difficult to breed”.

The research was made possible with assistance from the Maryland Zoo in Baltimore who manage the Golden Frog Species Survival Plan. Funding was provided from the Panamanian Government’s Secretaría Nacional de Ciencia y Tecnología (SENACYT), The WoodTiger Fund, the Smithsonian Endowment for Science and the University of Ottawa Research Chairs Program.

Della Togna G, Trudeau VL, Gratwicke B, Evans M, Augustine L, Chia H, Bronikowski EJ, Murphy JB, Comizzoli P. 2017 Effects of hormonal stimulation on the concentration and quality of excreted spermatozoa in the critically endangered Panamanian golden frog (Atelopus zeteki). Theriogenology. http://dx.doi.org/10.1016/j.theriogenology.2016.12.033 

More Good Reasons Not to Lick a Toad

A New Review of Chemicals Produced by the Toad Family, Bufonidae

Cane toad (Rhinella marinus)

As human diseases become alarmingly antibiotic resistant, identification of new pharmaceuticals is critical. The cane toad and other members of the Bufonidae family produce substances widely used in traditional folk medicine, but endangered family members, like Panama’s golden frog, Atelopus zeteki, may disappear before revealing their secrets. Smithsonian scientists and colleagues catalog the known chemicals produced by this amphibian family in the Journal of Ethnopharmacology highlighting this largely-unexplored potential for discovery.

“We’re slowly learning to breed members of this amphibian family decimated by the chytrid fungal disease,” said Roberto Ibañez, Panamanian staff scientist at the Smithsonian Tropical Research Institute (STRI) and in-country director of the Panama Amphibian Conservation and Rescue (PARC) project. “That’s buying us time to understand what kind of chemicals they produce, but it’s likely that animals in their natural habitats produce an even wider range of compounds.”

15 of 47 frog and toad species used in traditional medicine belong to the family Bufonidae. For millennia, secretions from their skin and from glands near their ears called parotid glands, as well as from their bones and muscle tissues have been used as remedies for infections, bites, cancer, heart disorders, hemorrhages, allergies, inflammation, pain and even AIDS.

Toxins of two common Asian toad species, Bufo gargarizans and Duttaphrynus melanostictus, produce the anticancer remedies known as Chan Su and Senso in China and Japan, respectively. Another preparation used to treat cancer and hepatitis, Huachansu or Cinobufacini, is regulated by the Chinese State Food and Drug Administration. In Brazil, the inner organs of the toad, Rhinella schneideri, are applied to horses to treat the parasite Habronema muscae. In Spain, extract from the toad Bufo bufo is used to treat hoof rot in livestock. In China, North and South Korea, ranchers use the meat of Bufo gargarizans to treat rinderpest.

Only a small proportion of the more than 580 species in the Bufonidae family have been screened by scientists. “In Panama, not only do we have access to an amazing diversity of amphibian species,” said Marcelino Gutiérrez, investigator at the Center for Biodiversity and Drug Discovery at Panama’s state research institute, Instituto de Investigaciones Cientificas y Servicios de Alta Tecnologia (INDICASAT),  “we’re developing new mass spectrometry and nuclear magnetic resonance spectroscopy techniques to make it easier and cheaper to elucidate the chemical structures of the alkaloids, steroids, peptides and proteins produced by these animals. We work closely with herpetologists so as not to further threaten populations of these species in the wild.” Their efforts to catalog chemicals produced by the Bufonidae included researchers from the University of Panama, Vanderbilt University, in Tennessee, U.S.A. and Acharya Nagarjuna University in Guntur, India.

Most of the chemicals produced by frogs and toads protect them against predators. Atelopus varius contains tetrodototoxin. Chiriquitoxin is found in Atelopus limosus, one of the first species that researches succeeded in breeding in captivity as well as in Atelopus glyphus and Atelopus chiriquiensis. An atelopidtoxin (zetekitoxin) from the Panamanian golden frog, Atelopus zeteki, appears to consist of two toxins. Toxins from a single frog skin can kill 130-1000 mice.

The golden frog, A. zeteki, Panama’s national frog, is the only species of the genus Atelopus that secretes zetekitoxins. Threatened by the chytrid fungal disease that infects the skin and causes heart attacks, with collection for the exotic pet trade and by habitat destruction, if golden frogs were to disappear, they would take this potentially valuable chemical with them.

More than 30 percent of amphibians in the world are in decline. Racing to stay ahead of the wave of disease spreading across Central America, Panama is leading the way in conservation efforts. The Smithsonian’s Panama Amphibian Rescue and Conservation project (PARC) identified several Atelopus species in danger of extinction, and are learning how to create the conditions needed to breed them in captivity. Not only do animal caretakers at their facilities in Gamboa and El Valle, Panama experiment to discover what the frogs eat, they also recreate the proper environment the entire frog life-cycle: egg laying, egg hatching and tadpole survival, to successfully breed Atelopus. Each species has unique requirements, making it an expensive challenge to create this Noah’s ark for amphibians.

The chemical building blocks amphibians use to create toxic compounds come from sources including their diet, skin glands or symbiotic microorganisms. Toads in the genus Melanophryniscus sequester lipophilic alkaloids from their complex diet consisting of mites and ants. Researchers found that toxins found in a wild-caught species of Atelopus could not be isolated from frogs raised in captivity: another reason to conserve frog habitat and to begin to explore the possibility of releasing frogs bred in captivity back into the wild.

Learn more about amphibians by visiting the PARC blog and the Panama’s Fabulous Frogs exhibit at the Smithsonian’s Culebra Point Nature Center in Panama.

The Smithsonian Tropical Research Institute, headquartered in Panama City, Panama, is a part of the Smithsonian Institution. The Institute furthers the understanding of tropical nature and its importance to human welfare, trains students to conduct research in the tropics and promotes conservation by increasing public awareness of the beauty and importance of tropical ecosystems. Website. Promo video.

# # #

Rodriguez, Candelario, Rollins-Smith, Louise, Ibanez, Roberto, Durant-Archibold, Armando, Gutiérrez, Marcelino. 2016. Toxins and pharmacologically active compounds from species of the family Bufonidae (Amphibia, Anura). Journal of Ethnopharmacology, doi:10.1016/j.jep.2016.12.021

2016 Golden Frog Festival Calendar of Events

Golden Frog Day Calendar 2016

EL VALLE DE ANTON

SATURDAY, AUGUST 13
FAMILY DAY #RANATICOS
Paseo El Valle, El Valle de Anton
11:00 AM – 3:00 PM
Activities: Games for children, exhibition of frogs, food and drinks, and the Golden Frog mascot.

SUNDAY, AUGUST 14
La Dorada Race
6:00 to 11:00 a.m.
Walk or run 5 to 15km to save a national treasure.
CLOSING PARADE
main streat of El Valle
2PM

GAMBOA
SUNDAY, AUGUST 21
OPEN HOUSE
Amphibian Rescue and Conservation Project
1:00 – 4:00 PM
Activities: Open house to visit the amphibian Ark and meet the scientists.

BOOK FAIR
18 -19 AUGUST
Atlapa Convention Center
9:00 a.m. to 9:00 PM
Activities: exhibition of science and live frogs, games, dynamic, book sales and more.

PUNTA CULEBRA
SATURDAY, AUGUST 20
FAMILY DAY #RANATICOS
Punta Culebra Nature Center,
Calzada de Amador, Panama
11:00 AM – 4:00 PM
Activities: Meet experts frogs, Restaurant Rana, face painting, many games, food and drinks.

Please remember that up until golden frog day, all donations to the amphibian rescue project will be generously matched by Golden Frog CLICK HERE TO DONATE

Update on Golden Frog Conservation Plan Implementation

I am pleased to distribute the Reporte del Taller de Conservación de las Ranas Doradas de Panamá in its official version in Spanish.

golden frog report

The Panamanian golden frog conservation group met in El Valle de Anton in November 2013 to identify and address critical issues necessary to preserve the golden frogs of Panama, foster collaboration and exchange of information, and develop an action plan to expand conservation efforts with a future view.

With the mission of “saving a national treasure, returning the golden frog back to nature”, the group has made significant progress in the management of captive populations, disease, habitat, population viability analysis (PVA) and communication and collaboration.

These are some of the most important achievements:
1. The management of captive populations of golden frogs in Panama and the United States has combined efforts to improve husbandry and increase breeding of their most valuable individuals. EVACC has had a substantial reproduction success in recent years. And the use of technologies of cryopreservation and hormonal treatment is being developed by our partners to improve breeding success.

2. Approximately ten research groups and disease management work in a wide range of projects that generate valuable information about the characteristics and epidemiology of Bd; virulence and transmission; natural defenses and immune response; and frog’s heritability of resistance and/or tolerance to disease, among many others.

3. Rescue missions and diseases monitoring in the wild continue at historic distribution sites of golden frogs. The Project Atelopus found four populations of Atelopus varius in nature and continue the search for other survivor populations.

4. The first PVA is being developed by members of the group under the same name. Leading scientists in natural history, behavior, habitat requirements, captive breeding, and disease modeling participated in a workshop for prioritizing research projects that will provide feedback to these programs.

5. For three consecutive years the group has had regular meetings where progress is reviewed, challenges are identified and solutions are discussed within each working group. They have produced documents published and distributed in scientific journals, major conservation websites such as the red list of endangered species of the IUCN, zoos and aquariums, decision-making organizations and outreach materials for schools and environmental education programs.

6. A strong education component has generated attention and awareness of the general public through activities such as The Golden Frog Festival and its many venues.

Amphibian conservation efforts in Panama are mainly led by Panamanians supported by international funding agencies and donors. One of the project aims is to get more Panamanian investment for conservation while continuing in-country capacity building and support for the reinforcement of conservation laws.

Angie Estrada, Coordinator of the Golden Frog Conservation Plan

Meet 9 of Panama’s ‘Lost Frogs’

The deadly amphibian chytrid fungus has caused much devastation to Panama’s native frogs, salamanders and caecilians. We have learned a lot about this disease in the last 10 years and we have been able to take stock of its effects. A recent survey of Panamanian frog experts revealed that of Panama’s 214 described amphibian species, about 100 species can still be reliably found even in places where the chytrid fungus is found, and experts consider these species less susceptible to the fungus. Approximately 80 species are very rare, and we simply do not have any idea about their susceptibility to chytridiomycosis, or their current population numbers. 36 species were considered highly susceptible to the chytrid fungus and were once reliably encountered but have experienced, or are predicted to experience, severe chytridiomycosis-related declines.
Unfortunately a number of these species have already completely disappeared in the wild and have not been seen in many years. We call these Panama’s ‘lost frogs’.

1) Atelopus chiriquiensis – Chiriquí harlequin frog
These attractive diurnal frogs were appealing research subjects and occurred in high numbers in highlands on the border of Costa Rica and Panama. There are many scientific papers about this species, and they were primarily studied for their highly toxic tetradotoxins in their skin as well as their unique signaling and aggressive mating behavior. A study by Dr. Karen Lips in the las Tablas reserve of Costa Rica reports that they occurred in high numbers – up to 20 individuals seen in 100m of stream on a single visit, but the frogs experienced a severe chytridiomycosis-related decline over a 5-year period and were last seen in 1996.

A pair of Atelopus chiriquiensis in amplexus. Photo (c) Marcos Guerra, Smithsonian Tropical Research Institute.

A pair of Atelopus chiriquiensis in amplexus. Photo (c) Marcos Guerra, Smithsonian Tropical Research Institute.

2) Atelopus zeteki – Panamanian golden frog
This is Panama’s national amphibian, a charismatic emblem of the environment and conservation. August 14th is a dedicated national day to honor the golden frog as a symbol for Panama’s incredible biodiversity heritage. Recognizing the chytridiomycosis threat, a conservation project called Project Golden Frog established a healthy breeding colony of golden frogs at the Maryland Zoo in Baltimore, another colony is maintained in Panama at the El Valle Amphibian Conservation Center. As predicted, Panamanian golden frogs experienced severe chytridiomycosis-related declines starting in 2006, and the last confirmed observation of Panamanian golden frog in the wild was in 2009. Project Atelopus continues to survey known golden frog sites for survivors, and a detailed conservation plan has been developed by stakeholders and facilitated by the IUCN Species Survival Commission for golden frogs in Panama. The plan aims to eventually reintroduce them to the wild.

One of 2,000 captive Panamanian Golden Frogs managed in captivity by the Golden Frog Species Survival Plan and the Maryland Zoo in Baltimore Photo: Brian Gratwicke Smithsonian Conservation Biology Institute

One of 2,000 captive Panamanian golden frogs managed in captivity by the Golden Frog Species Survival Plan and the Maryland Zoo in Baltimore Photo: (cc) Brian Gratwicke, Smithsonian Conservation Biology Institute

3) Craugastor obesus
This species was found in the spray zone on rocks, boulders in the Atlantic-facing slopes of Western Panama and Costa Rica. The species was last recorded from Costa Rica in 1984 rainforest. This species belongs to the Craugastor rugulosus group and all these closely related species of amphibians have all been have been decimated by the amphibian chytrid fungus.
4) Craugastor punctariolus
This semi aquatic species was found in mountainous streams of Central Panama. Rapid chytridiomycosis-related declines and disappearances were observed in the field in 2004-2008. This species belongs to the Craugastor rugulosus group and all these closely related species of amphibians have been decimated by the amphibian chytrid fungus. Genetic analysis revealed that it is likely a species complex. It has been maintained in captivity, and occasionally deposited eggs that were either infertile or did not develop fully and a viable captive population was not established.

Craugastor punctariolus, Bob's Robber Frog at the El valle Amphibian Conservation Center, photo (c) Kevin Johnson Amph

Craugastor punctariolus, Bob’s Robber Frog at the El Valle Amphibian Conservation Center (EVACC) photo (c) Kevin Johnson Amphibian Ark

5) Craugastor rhyacobatrachus
This species is found in premontane and lower montane southern slopes of the Talamanca Mountains of Costa Rica and western Panama. Despite extensive searches for this species in both Costa Rica and Panama, there are no recent records of this species. This species belongs to the Craugastor rugulosus group and all these closely related species of amphibians have all been have been decimated by the amphibian chytrid fungus.
6) Incilius majordomus
Males of this species were lemon yellow, and females were brown, the only other known toad of this genus that exhibited similar sexual dimorphism was Incilius peringelis—the famous Monte Verde Golden Toad of Costa Rica that is now extinct. Incilius majordomus is known only from the Pacific slope of Cerro Bollo, on the border between the provinces of Bocas del Toro and Chiriquí. This species was described in 2013 using a series of specimens collected in 1980. It has not been seen in the wild since 1980 despite extensive herpetological surveys in the area.

Incilius majordomus type specimen © Smithsonian Institution, National Museum of Natural History, Department of Vertebrate Zoology, Division of Amphibians & Reptiles

Incilius majordomus type specimen © Smithsonian Institution, National Museum of Natural History, Department of Vertebrate Zoology, Division of Amphibians & Reptiles

7) Isthmohyla calypsa
A  treefrog frog covered with spiny tubercles found in a small mountainous area on the border of Costa Rica and Panama where is used to be locally common. At las Tablas in Costa Rica, the species experienced severe chytridiomycosis-related declines between 1993 and 1998. Despite extensive recent survey efforts in Costa Rica and Panama, the species has not been seen recently and is possibly extinct. Many other stream breeding species in this genus have also experienced dramatic declines and are now extremely rare frogs.

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Isthmohyla calypsa in the wild, Photo (c) Marcos Guerra, Smithsonian Tropical Research Institute.

8) Ecnomiohyla rabborum – Rabbs’ fringe-limbed treefrog
Rabbs’ treefrog is thought to be endemic to the vicinity of El Valle de Anton, where it was always a rare frog difficult to find as they live high in trees and breed in tree holes. Experienced herpetologists could hear their calls reliably at some places, but the last individual was heard in El Valle de Anton in 2008. A few individuals of this species were collected for captive breeding efforts at the El Valle Amphibian Conservation Center and at the Atlanta Botanical Gardens, but captive breeding efforts were unsuccessful. As of 2015 only a single individual persists in captivity at the Atlanta Botanical Gardens.

Ecnomiohyla rabborum - Rabb's Fringe-Limbed Frog at the Atlanta Botanical Gardens. Photo (c) Brad Wilson

Ecnomiohyla rabborum, Rabb’s fringe-limbed  tree frog at the Atlanta Botanical Gardens. Photo (c) Brad Wilson

9) Oophaga speciosa – Splendid poison frog
This large, unmistakable bright red dart frog lives only in the mountains of Western Panama. It was once collected for the pet trade, and was exported as recently as 1992. This species has not been seen in the wild in many years, despite intensive searches. It is not known whether it still lives in captivity, but has probably disappeared from the wild.

Oophaga speciosa, the Splendid poison dart frog. Photo (c) Marcos Guerra, Smithsonian Tropical Research Institute

Oophaga speciosa, the Splendid poison dart frog. Photo (c) Marcos Guerra, Smithsonian Tropical Research Institute

If you have any recent records of these missing species please let us know, and consider uploading your record to the global amphibian bioblitz on inaturalist.

by Brian Gratwicke

*WE ARE SEEKING VOLUNTEERS TO HELP TRANSLATE OCCASIONAL AMPHIBIANRESCUE.ORG WEB PAGES INTO SPANISH, IF YOU ARE WILLING TO HELP US OUT OCCASIONALLY, PLEASE EMAIL Gratwickeb[AT]si.edu FOR MORE INFORMATION.

Surveying amphibian skin bacteria in Panamá

Amphibians are dying all over the world due to chytridiomycosis. This disease, caused by Batrachochytrium dendrobatidis (Bd), is responsible for dramatic amphibian declines and extinctions in the Neotropics, including Panamanian tropical forests.

We are a research team that is part of an NSF project investigating microbial diversity on frog skin in Panama. This team includes three principal investigators (Lisa Belden, Reid Harris and Kevin Minbiole), three postdoctoral fellows (Eria Rebollar, Myra Hughey and Tom Umile) and several graduate and undergraduate students from James Madison University, Virginia Tech, Villanova University and the Smithsonian Tropical Research Institute. We are interested in understanding how microbial communities from amphibian skin might contribute to the survival of amphibian species that have persisted in the wild despite the presence of Bd. Since 2012, we have collected samples from multiple sites in lowland tropical forests to describe the factors that shape skin microbial communities in tropical amphibians, including the presence of the pathogen Bd.

The author in Panama, 2013

The author in Panama, 2013

We  recently published a study on the skin microbiota of five species of tropical amphibians from one of the few sites in Panamá where amphibians have not been infected with Bd, called Serranía del Sapo, in the Darién Province. In the summer of 2012, Myra Hughey, Roberto Ibáñez and Daniel Medina collected skin swab samples from this lowland site of highly susceptible and less-susceptible species, including two highly threatened species: Atelopus certus and Strabomantis bufoniformis. When we analyzed the bacterial species present on the skin of these five amphibian species we found that amphibians had a unique microbiota on the skin that was very distinct from the bacterial communities in the environment. These symbiotic bacteria were not only different from the environment, but were also different among the amphibian species. Interestingly we found that the three less Bd-susceptible species that we studied (Craugastor fitzingeri, Espadarana prosoblepon and Colosthetus panamansis) had a common set of bacteria that was not present on the two highly susceptible species (A. certus and S. bufoniformis).

Atelopus certus, thought to be a species susceptible to Bd. Photo (c) Joel Sartore

Atelopus certus, thought to be a species susceptible to Bd. Photo (c) Joel Sartore

We think that the bacteria present in the less-susceptible species might be playing a defensive role against pathogens like Bd. If these bacteria indeed have antifungal properties, what are the factors determining the presence of these antifungal bacteria on the skin of less susceptible species? To pursue this idea, we compared the microbial communities of C. fitzingeri in the Darién region with skin communities from regions where the frogs were infected with Bd, in Colón and Panamá provinces (Mamoní, Soberanía and Gamboa). We found that the skin bacterial communities in the infected regions had an increased proportion of bacterial species like Pseudomonas and members of the Actinomycetes. Interestingly, these bacteria are known for their antifungal activities in other amphibians, and therefore it is possible that they might be playing an important role in Bd resistance. Since other factors could be influencing these communities, we are currently analyzing experimental data to determine if Bd infection is driving these changes in the skin microbiota.

How can we use the information we have gathered to protect amphibians that do not have high proportions of Pseudomonas and Actinomycetes on their skins? Can we culture these bacteria and study their antifungal properties? We are currently analyzing cultured microbes from less susceptible species to determine if they have anti-Bd properties in vitro. If these microbes are indeed effective against Bd, can we use them as skin probiotics for the highly susceptible species? Probiotics are a very promising avenue towards conservation of susceptible amphibians since the skin microbiota has already been manipulated in some species to protect them from Bd. However a lot of additional studies still need to be done to implement this approach successfully.

Publications

Belden LK, Hughey MC, Rebollar EA, Umile TP, Loftus SC, Burzynski EA, Minbiole KPC, House LL, Jensen RV, Becker MH, Walke JB, Medina D, Ibáñez R and Harris RN (2015) Panamanian frog species host unique skin bacterial communities. Front. Microbiol. 6:1171. doi: 10.3389/fmicb.2015.01171

Rebollar EA, Hughey MZ, Medina D, Harris RN,  Ibáñez R and Belden LK (2015) Skin bacterial diversity of Panamanian frogs is associated with host susceptibility and presence of Batrachochytrium dendrobatidis. ISME Journal. doi:10.1038/ismej.2015.234

by Eria Rebollar

New Amphibian Study Helps Smithsonian Scientists Prioritize Frogs at Risk of Extinction

graphical abstractScientists at the Smithsonian Institution and partners have published a paper that will help them save Panamanian frog species from extinction due to a deadly fungal disease called Chytridiomycosis (chytrid). The study, which was published Jan. 4 in Animal Conservation, draws on the expertise of amphibian biologists and scientists the Panama Amphibian Rescue and Conservation Project to mathematically determine which frog species have the best probability of escaping extinction with the rescue project’s help.
“We don’t want to arbitrarily decide which species lives and which species don’t, nor do we want to waste our time on species that don’t need our help,” said Brian Gratwicke, co-author on the paper and international coordinator of the rescue project out of the Smithsonian Conservation Biology Institute. “This study took into account the differences in opinions among amphibian experts in Panama and found consensus in a systematic away. This has allowed us to focus on the species where we have the best chance of making a difference.”

The study also found that eight Panamanian species are likely now extinct in the wild due to disease-related declines. About 80 of Panama’s frog species were too rare for conservationists to prioritize their need for help or the likelihood of successful rescue. The new prioritization scheme, however, will allow the scientists to adapt to new information as it becomes available.

“Over the years, several frog populations—and even species—have vanished or nearly vanished from Panama,” said Roberto Ibáñez, the in-country director of the rescue project at the Smithsonian Tropical Research Institute, “Unfortunately, it is impossible to save them all through conservation programs. With this study, we can focus our limited resources on those species that we are more likely to find in the wild and breed in captivity, while we simultaneously look for a way to manage chytrid.”

Since 2009, the rescue project has been building and maintaining insurance populations of frog species susceptible to chytrid, bringing small groups into captivity to breed as the species crashes in the wild. For each of Panama’s 214 known frog species, the paper’s authors asked amphibian experts to determine the probability that: 1) the rescue project could locate an adequate founding population (20 males and 20 females), 2) the rescue project could successfully breed the species and 3) without the rescue project’s help, the species would go extinct.

While most of the rescue project’s original priority species ranked high based on the new prioritization scheme, the conservationists have already started making some changes. They have determined that the likelihood of successfully breeding La Loma tree frogs (Hyloscirtus colymba) is low and they are instead shifting resources to the recently discovered Craugastor evanesco and the Rusty robber frog (Strabomantis bufoniformis), both of which came up as high priorities.

The Panama Amphibian Rescue and Conservation Project is a project partnership between the Cheyenne Mountain Zoo, the Houston Zoo, Zoo New England and Smithsonian Institution.