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

Newly Described Poison Dart Frog Hatched for the First Time in Captivity

The first captive-bred Andinobates geminisae at the Gamboa Amphibian Research and Conservation Center at the Smithsonian Tropical Research Institute.

The first captive-bred Andinobates geminisae at the Gamboa Amphibian Research and Conservation Center. Photo by Jorge Guerrel, Smithsonian Tropical Research Institute.

Smithsonian Conservation Biology Institute (SCBI) and Smithsonian Tropical Research Institute (STRI) scientists working as part of the Panama Amphibian Rescue and Conservation Project hatched the first Andinobates geminisae froglet born in captivity. The tiny dart frog species only grows to 14 millimeters and was first collected and described last year from a small area in central Panama. Scientists collected two adults to evaluate the potential for maintaining the species in captivity as an insurance population.

“There is a real art to learning about the natural history of an animal and finding the right set of environmental cues to stimulate successful captive breeding,” said Brian Gratwicke, amphibian conservation biologist at SCBI and director of the Panama Amphibian Rescue and Conservation Project. “Not all amphibians are easy to breed in captivity, so when we do breed a species for the first time in captivity it is a real milestone for our project and a cause for celebration.”

Scientists simulated breeding conditions for the adult frogs in a small tank. The frogs laid an egg on a bromeliad leaf, which scientists transferred to a moist petri dish. After 14 days, the tadpole hatched. Scientists believe adult A. geminisae frogs may provide their eggs and tadpoles with parental care, which is not uncommon for dart frogs, but they have not been able to determine if that is the case. In the wild, one of the parents likely transports the tadpole on his or her back to a little pool of water, usually inside a tree or on a bromeliad leaf.

Andinobates geminisae egg

After the tadpole hatched, scientists moved it from the petri dish to a small cup of water, mimicking the small pools available in nature. On a diet of fish food, the tadpole successfully metamorphosed into a froglet after 75 days and is now the size of a mature adult.

Andinobates geminisae tadpole

Panama Amphibian Rescue and Conservation Project scientists are unsure if A. geminisae is susceptible to the amphibian-killing chytrid fungus. However, since it is only found in a small area of Panama and is dependent on primary rain forests, which are under pressure from agricultural conversion, they have identified it as a conservation-priority species.

The Panama Amphibian Rescue and Conservation Project breeds endangered species of frogs in Gamboa, Panama and El Valle, Panama. The Panama Amphibian Rescue and Conservation Project is a partnership between the Houston Zoo, Cheyenne Mountain Zoo, Zoo New England, SCBI and STRI. This study was supported by Minera Panama.

The Panama Amphibian Rescue and Conservation (PARC) project receives IX BBVA Foundation’s Biodiversity Conservation Award for Latin America

25 November, 2014, Madrid. The Panama Amphibian Rescue and Conservation (PARC) project was awarded the IX BBVA Foundation’s Biodiversity Conservation Award for Latin America, which recognizes projects, policies or actions that have had significant impact on the protection of habitats, species and ecosystems in Latin America.  Our project was chosen among approximately 30 nominations for its achievements in saving critically endangered amphibians in Panama and globally through research, rescue and outreach.

Roberto Ibanez BBVA Award

The award, made on Nov 25th at the Palacio del Marqués de Salamanca in Madrid, Spain includes a prize of 250,000 Euros (approx. $323,000).  The award was made to STRI’s Roberto Ibáñez, director of the Panama Amphibian Rescue and Conservation Project,  Heidi Ross, director and co-founder of the El Valle Amphibian Conservation Center, Sharon Ryan, STRI’s director of public programs and the Smithsonian Conservation Biology Institute’s Brian Gratwicke who is the international coordinator of the amphibian rescue project.

Panama Amphibian Rescue and Conservation Project LogoPARC was created in 2009 as a partnership of eight institutions, in response to the massive loss of Panama’s amphibian biodiversity due mainly to the chytrid fungus, and has several key goals: prevent species extinctions by establishing ex-situ assurance colonies of endangered amphibians threatened with extinction from a deadly fungus decimating amphibians worldwide; develop tools to mitigate the disease and lead to reintroductions in the wild; and, engage constituents to support conservation of amphibians and habitats.

The founding organizations include Africam Safari, Houston Zoo, Cheyenne Mountain Zoo, New England Zoo, Smithsonian Conservation Biology Institute, Defenders of Wildlife and  the Smithsonian Tropical Research Institute.

To date, we have invested $1.5 million to build two ex-situ facilities in Panama, bred more than 10 endangered amphibian species, including the Panamanian Golden Frog, a conservation flagship species now extinct in the wild. Our scientists actively monitor disease and frog populations in the wild, use the latest molecular tools to find beneficial skin bacteria to help frogs fight chytridiomycosis infections, research genetic mechanisms of chytrid resistance in Panamanian Golden Frogs, and develop assisted reproduction technologies to breed frogs in captivity and cryopreserve their gametes for future use.

Kids on a float in the Golden Frog Day Parade

The Panama Amphibian Rescue and Conservation Project’s education campaign generates extensive coverage in local and international press, raising awareness and support for this crisis, including: award-winning documentary aired on Smithsonian Channel in the USA, and coverage in major media outlets. Our annual Panamanian Golden Frog Festival drew 6,280 people in 2013 and had a media reach of nearly 500,000. Two exhibits, the Fabulous Frogs of Panama in Panama City and another at the El Valle Amphibian Conservation Center are visited by 200,000 Panamanians annually. More than 35,000 schoolchildren and 300 teachers participate annually in amphibian-related education programs.  Nearly 300 volunteers have supported the project since 2009.

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

Frog Friday: Toad Mountain Harlequin Frog

atelopus certus male calling

The toad mountain harlequin frog is all decked out in orange and black for Halloween! Atelopus certus is a biological treasure found only in Panama. This terrestrial species has a golden iridescent hue with spots like a giraffe. In its natural setting, the toad mountain harlequin frog can be found streamside or on mossy rocks in moist lowland and mountain forests.  We have a healthy population in captivity to safeguard against predicted chytridomycosis-related declines.

Atelopus certus  is endemic to Panama and identified only on a single mountain range in the south-western Darien region. Before 2011, this species was quite common within its small known range, and they were still abundant in 2013, but they are predicted to decline severely once chytridiomycosis arrives in that region. Thankfully, due to the swift action of the PARC project, Atelopus certus is being successfully reproduced in captivity. The Gamboa ARC has bred a very stable population 

Did you know? The Smithsonian Channel documentary “Mission Critical: Amphibian Rescue” recounts the valiant rescue effort to save this species from imminent extinction.

Post by Dara Wilson

Frog Friday: Granular Glass Frog

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cochranella granulosa

Cochranella granulosa is a beautiful species with grey-gold eyes and a dark green, granular dorsal surface gives the frog a “crystal-like” appearance. This arboreal species can be found in humid lowlands and mountainous forest. They are usually spotted in bushes and trees along forest streams. Though the granular glass frog ranges from Honduras to Panama, it is more regularly encountered in Costa Rica and Panama, while sightings are rarely recorded in Honduras and Nicaragua. These nocturnal frogs can also be found sleeping on the leaves of bromeliads during the day in the exhibit Fabulous Frogs of Panama at Punta Culebra Nature Center. Though the frog is listed as Least Concern on the IUCN Redlist due to its wide distribution, it still faces threats of severe habitat loss resulting from deforestation and water pollution.

Did you know? The word granulosa is derived from the Latin word granulum, which means small-grained or granular. This reference was drawn due to the granular bumps on the frog’s transparent skin. If you look at the frog from its underbelly, you can see right through its skin and get a great glimpse of its tiny organs!

Frog Friday: Red-Eyed Tree Frog

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Beauty is in the eye of the beholder! The bright red eyes, and blue striped sides of Agalychnis callidryas are a defense mechanism the frog uses to surprise potential predators, and avoid predation. During the day the red-eyed tree frog folds its legs at its sides, closes its eyes, and sleeps, effectively camouflaging itself on green leaves. You can observe this behavior in their natural habitat  of low to mid-elevation rainforests from the Yucatan to Colombia.  They are considered a species of least concern according to the IUCN, and are abundant throughout their range. We also have these beauties on display in our exhibit Fabulous Frogs of Panama at Punta Culebra Nature Center.

Did you know? Red-eyed tree frogs lay their eggs on plants overhanging the wáter, and when they hatch the tadpoles fall into the wáter.  Eggs in the trees can be eaten by wasps, snakes, or katydids, or killed by pathogenic fungus. Embryos can hatch early to escape from attacks by egg predators and pathogens, or in response to abiotic threats, but they typically hatch later if undisturbed. Thus tadpoles enter the water at different ages, sizes and stages of development. Tadpoles that were induced to hatch early are more likely to be killed by aquatic predators and less likely to survive to metamorphosis. After a month or more in the water, the tadpoles metamorphose into froglets. Metamorphs on land remain relatively inactive near the pond while they absorb their tails, then climb up into the trees and disappear. We know very little about their lives as juveniles.

 

Snake induced hatching of red-eyed tree frogs.

Video and information by Karen Warkentin Lab

Post by Hannah Arney

Frog Friday: Limosa Harlequin Frog

Atelopus limosus - Limosa Harlequin Frog

Cute, crafty, and brightly patterned with yellow and black chevrons! The Limosa harlequin frog is a terrestrial species endemic to Central Panama that live in moist, tropical Atlantic forests. In the wild, you could once spot this species among the dark rocks or mossy patches on the banks of streams. It is here, in the fast-flowing rainforest streams that enthusiastic males call and use their characteristic hand-waving behavior to signal females or warn competing males to steer clear. If successful, a male attracts a female and embraces her in a behavior known as amplexus. They laytheir eggs in fast-flowing streams for good reasons. The shallow, swiftly flowing water protects the eggs and tadpoles from potential predators and once they emerge as tadpoles the tadpoles cling tightly to rocks using the suction disks on their bellies while they graze on diatoms and other algae that grow on stones in these clear, oxygen-rich environments.

This species is listed as Endangered on the IUCN Red List. Atelopus frog species’ have been hit hardest by the growing chytridomycosis epidemic. While deforestation of habitats for agricultural use and infrastructural development, stream sedimentation, and water pollution are also serious threats to Atelopus limosus, much of their range is in the Chagres National Park and scientists attribute the recent disappearances of this species to the emergence of amphibian chytrid fungus. Thankfully, the Panama Amphibian Rescue and Conservation Project has brought the endangered species into captivity and successfully hosts a very fruitful breeding population – no small feat at all!

Did you know? The limosa harlequin frog has two color forms; the brown form with a yellow nose and yellow fingertips mostly found at lowland sites, while the green/yellow form with black chevrons is normally found in the highlands. If you’d like to get a look at one of these amazing creatures, you can see them on exhibit at the Punta Culebra Nature Center’s Fabulous Frogs of Panama!

by Dara Wilson, Panama Amphibian Rescue and Conservation Project Volunteer

Scientists discover new poison dart frog species in Donoso, Panama

Andinobates geminisae

A bright orange poison dart frog with a unique call was discovered in Donoso, Panama, and described by researchers from the Smithsonian Tropical Research Institute and the Universidad Autónoma de Chiriquí in Panama, and the Universidad de los Andes in Colombia. In the species description published this week in Zootaxa, it was named Andinobates geminisae for Geminis Vargas, “the beloved wife of [coauthor] Marcos Ponce, for her unconditional support of his studies of Panamanian herpetology.”

Every new species name is based on a representative specimen. The specimen for this species was collected Feb. 21, 2011, in the headwaters of the Rio Caño, in the district of Donoso, Colón Province, Panama, by Samuel Valdés, who was then the MWH Global Inc. environment office director, and his field assistant, Carlos de la Cruz. Additional specimens were collected between the Rio Coclé del Norte and the Rio Belen by biologists Marcos Ponce and Abel Batista, then a student at the Universidad Autónoma de Chiriquí. The specimens were deposited in the Museo de Vertebrados at the University of Panama, the Museo Herpetólogico de Chiriquí at the Universidad Autónoma de Chiriquí and in the Círculo Herpetólogico de Panamá.

“Abel Batista and Marcos Ponce were the first to note the presence of this species,” said Cesar Jaramillo, Smithsonian herpetologist. “They’ve known it was there for several years. However, they were not sure if it was only a variety of another poison dart frog species, Oophaga pumilio, which exhibits tremendous color variation. Based on morphological characteristics of the adult and the tadpole, I thought it might be a new species of Andinobates.”

Andrew Crawford, professor at Universidad de Los Andes and former STRI postdoctoral fellow, sequenced the DNA, confirming that this was a new species of Andinobates. Genetic information about this species is available in the Barcode of Life Data System and in GenBank. A recording of the call is available at AmphibiaWeb.org.

Because this new frog species appears to be found in only a very small area, habitat loss and collecting for the pet trade are major threats to its existence. The authors recommend the formulation of special conservation plans to guarantee its survival. A. geminisae is included in the captive breeding program of the Panama Amphibian Rescue and Conservation project, a consortium of six zoos and research institutions dedicated to saving amphibians from the chytrid fungal disease, which is decimating amphibians worldwide, and habitat loss.

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All collecting and export were done with permission from Panama’s Environmental Authority, ANAM. Financial support for this study came from MWH Global Inc. and Minera Panama. Funding for DNA sequencing was provided by a CBOL grant to barcode the vertebrates of Panama.

For more information about amphibian biodiversity in Panama, visit amphibianrescue.org or the Smithsonian’s new exhibit, Las Fabulosas Ranas de Panama, at Culebra Point Nature Center on Amador Causeway in Panama City.

The Smithsonian Tropical Research Institute, headquartered in Panama City, Panama, is a unit 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: http://www.stri.si.edu. Promo video: https://www.youtube.com/watch?v=M9JDSIwBegk

Batista, A., Jaramillo, C.A., Ponce, M., Crawford, A. 2014 A new species of Andinobates (Amphibia:Dendrobatidae) from west central Panama. Zootaxa 3:333-352 http://dx.doi.org/10.11646/zootaxa.3866.3.2

Frog Friday! Strawberry Poison Dart Frog

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Look, but don’t touch! Strawberry poison dart frogs (Oophaga pumilio) are known for their strikingly beautiful skin colors. Their bright color serves as a warning to predators that they are toxic. This type of “warning coloration” is called aposematism.  In the wild Oophaga pumilio gets its toxicity from its diet of ants and termites. The frogs we maintain in captivity in our exhibit Fabulous Frogs of Panama are fed small crickets and fruit flies. This change in diet eradicates any trace of poison in these frogs when they are raised in captivity

Strawberry poison dart frogs are generally a small species, about 0.75 to 1.5 inches (20 to 40 mm) in length. They are also mostly diurnal, and can be heard calling in the flooded forest.There are many poison frogs in the Dendrobatidae family with slightly different distributional ranges that can be found in Central America and northern South America. The species Oophaga pumilio are found in Mesoamerican countries and Panama.

Did you know? An Oophaga pumilio look alike was recently discovered in Panama by STRI scientist Cesar Jaramillo with  Abel Batista and Marcos Ponce (UNACHI) and Andrew Crawford (Universidad de los Andes). This new species Andinobates geminisae is still being described. Click here for more information.

Post by Dara Wilson