Photo right: One of the three potentially new species appears to be a robber frog, genus Craugastor, shown here. The unique skin folds on its arms and feet distinguish it from other closely related species. Robber frogs are especially susceptible to chytrid. (Photos by Brian Gratwicke, Smithsonian Conservation Biology Institute)

“It is disturbing to witness the disappearance of species that some of us only recently described and even more devastating to lose those we know are probably new species,” said Roberto Ibáñez, local director of the project and a scientist at the Smithsonian Tropical Research Institute, one of nine project partners, including the Smithsonian’s National Zoo. “Scientists are just starting to investigate the ecological impact of the loss of amphibians, and while we’re aiming to preserve some of these species, we already know it will be impossible to save them all.”

Nearly one-third of all amphibian species globally are at the risk of going extinct. The rescue project aims to save more than 20 species of frogs in Panama, which is 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, the deadly amphibian chytrid fungus is likely at least partly responsible for the disappearances of 94 of the 120 frog species that are thought to have gone extinct since 1980.

Photo left: Two of the three potentially new species is a rain frog from the genus Pristimantis. The species pictured here has a bright red stomach that is uncharacteristic for rain frogs, earning it the nickname “red tomato.”

Although it can take years to determine that a species is new to science, project researchers have identified some telltale signs indicating that the three species found in eastern Panama are, indeed, new. The first two are rain frogs from the genus Pristimantis. One of these species has a bright red stomach that is uncharacteristic for rain frogs, earning it the nickname “red tomato.” The second species is much larger than any known Pristimantis in the region. The third frog species appears to be a robber frog, genus Craugastor, but unique skin folds on its arms and feet distinguish it from other closely related species. Robber frogs are especially susceptible to chytrid.

A new study by Andrew Crawford, a STRI research associate, and colleagues reveals that many frog species at a site in western Panama have gone extinct before researchers knew they existed. The project’s three potentially new species are evidence of the same story playing out right now in the mountains of eastern Panama. Researchers have brought a handful of animals of each species back to the Summit Municipal Park in Panama City, Panama, where the project has turned used shipping containers into amphibian rescue pods.

“We are doing our best to salvage what we can, but we are in urgent need of funding to build capacity in Panama to house all of these chytrid refugees,” said Brian Gratwicke, a National Zoo research biologist and the international coordinator for the Panama Amphibian Rescue and Conservation Project. “The species is the basic unit of conservation, so these discoveries are rewarding, but that comes with the daunting responsibility of deciding how we look after them. We already have a huge job, and it just gets bigger with every discovery.”

Now project scientists will use collections of frogs from the same region at the Smithsonian’s Natural History Museum and elsewhere to determine if these species are genuinely new or if they have already been discovered (or “described”) elsewhere. The project has also collected tissue sample to use DNA testing to map out the animals’ closest genetic relatives.

“Finding a new species is like discovering a new Pablo Picasso,” said Gratwicke. “Each species is a priceless creation painted with the brushstrokes of natural selection on the canvas of DNA and has something of value to offer. We might not know how they’re valuable to us right now, but if they go extinct, we lose the opportunity to learn what secrets they hold.”

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 re-introduced to the wild. Project participants include Africam Safari, Autoridad Nacional del Ambiente, Cheyenne Mountain Zoo, Defenders of Wildlife, El Valle Amphibian Conservation Center, Houston Zoo, Smithsonian’s National Zoological Park, Smithsonian Tropical Research Institute, Summit Municipal Park and Zoo New England.

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Japanese giant salamanders live in cold, fast-flowing streams in Japan. Their numbers have been greatly reduced over the years because of agricultural development and habitat modification.

Photo left: Rick Quintero (left), the primary Japanese giant salamander keeper at the Smithsonian’s National Zoo, feeds the Zoo’s new juvenile salamanders for Japanese Ambassador Ichiro Fujisaki (right). Fujisaki was at the Zoo on July 22 to help celebrate the arrival of the salamanders, a gift from the City of Hiroshima Asa Zoological Park. (Mehgan Murphy photo)

“In conserving salamanders, we conserve the ecosystems in which they live,” said Ed Bronikowski, senior curator at the Zoo. “People share those same ecosystems, so what is good for the salamanders is good for many species, including us. We hope our visitors will learn from this generous gift to embrace our own diverse native salamander populations and protect healthy ecosystems for all.”

The National Zoo has experience caring for Japanese giant salamanders since as early as 1940, but with this gift, the Zoo hopes to become the first in the United States to successfully breed this species, which has not been bred outside of Japan in at least 100 years.

During the donation ceremony on July 22, kids from Great Falls Elementary School in Great Falls, Virginia were present to help name one male salamander. The students were asked to choice between two names selected by the ambassador – Hiro, derived from Hiroshima, the salamanders’ home in Japan and Asa, of the City of Hiroshima Asa Zoological Park. Hiro won the student’s vote! –Jessica Porter

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View Muddy Creek and the Rhode River in a larger map

To survey the animals swimming up and down Muddy Creek, researchers use a fish weir—an expanse of nets, gates and boardwalks—that temporarily blocks aquatic traffic. Once a week, the researchers close the weir, set out the nets and identify and count all the species that get trapped. They began collecting data in 1983.

This type of fine-scale surveying, done on a weekly basis, is rare. It’s even more unique to have such long-term data. Many ecological studies are funded for just a few years at a time. These short time frames make it difficult for scientists to observe changes and patterns in species populations and composition.

In honor of the 2010 U.S. Census, staff at the Smithsonian Environmental Research Center have created this slide show of a recent spring survey. The salinity on this April day was fairly low and nearly a dozen golden shiners (a freshwater minnow) were caught along with several estuarine-resident and a few diadromous (fish that migrate between fresh and saltwater) species. Among the highlights: a sizeable snapping turtle, many white perch in spawning condition, juvenile American eels and a parasite.

Human activity and environmental conditions can affect which species are swimming in Muddy Creek. The water is brackish and salinity levels change seasonally and from year to year. During winter and early spring, when freshwater flow is usually the highest, researchers will generally catch more freshwater species like bluespotted and banded sunfish–-two protected species in Maryland. During periods of high salinity, researchers can catch many species indicative of the higher saline lower Bay such as red drum, spotted sea trout and Spanish mackerel. –Tina Tennessen

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Photo right: The limosa harlequin frog (Atelopus limosus) is one of 54 species that Amphibian Ark has identified as a priority rescue species for the Panama Amphibian Rescue and Conservation Project. (Click to enlarge)

“Each container provides us with critical space to house animals that may represent the last chance for the survival of their species,” said Brian Gratwicke, a National Zoo research biologist and the international coordinator for the Panama Amphibian Rescue and Conservation Project. “The containers are now self-contained ‘amphibian rescue pods’ that have been specially modified to control the climate and keep diseases out.”

The rescue pods will be part of the project’s Amphibian Rescue Center at Summit Municipal Park, which will also include a lab with a quarantine facility. After frogs are collected in the field, they will be quarantined for 30 days before being moved to the rescue pods that will serve as their new home. In addition to the two containers that are now in Panama, Maersk Line has agreed to donate two containers per year for the next two years to the project, for a total of six. Shipping company APL has also donated one container this year. Each container offers 995 cubic feet of space to house these animals. The seven together will more than double the amount of captive space the project currently has in Panama to safeguard endangered amphibians.

Photo left: Shipping company Maersk Line has agreed to donate up to six used shipping containers similar to this one to the Panama Amphibian Rescue and Conservation Project. The containers will serve as rescue pods for endangered amphibians.

“Maersk Line’s support of the amphibian rescue project is aligned with our long-term focus on sustainability,” said Mike White, head of Maersk Line’s North American organization. “Although we are pleased to donate these containers, the more valuable contribution is our expertise and resources. Our team’s assistance with documentation and transportation allows Brian’s group to concentrate on the overall effort.”

Amphibian Ark, an organization that mobilizes support for ex-situ (“out-of-the-wild”) conservation, has identified 54 amphibian species as rescue species for the Panama Amphibian Rescue and Conservation Project. At least 198 amphibian species live in Panama, of which 70 are listed as “critically endangered,” “endangered” or “data deficient” by the International Union for Conservation of Nature. Amphibian Ark estimates that about 500 amphibian rescue pods are needed to save the world’s 500 critically endangered amphibian species. Buying, outfitting and installing a single container costs about $50,000.

Photo right: Each shipping container offers 995 cubic feet of space to safeguard endangered species. (Photos by Brian Gratwicke)

“This requires an amount of resources that is insurmountable for the amphibian rescue community,” said Kevin Zippel, Amphibian Ark’s program director. “With a relatively small investment, the shipping industry has made a huge impact on one of the greatest conservation challenges that humanity has ever faced. We are currently seeking additional contributions of this kind.”

The mission of the Amphibian Rescue and Conservation Project is to rescue amphibian species that are in extreme danger of extinction from amphibian chytrid disease sweeping through Panama. The project’s focus is on developing appropriate technologies to control the amphibian chytrid fungus, so that one day captive amphibians may be reintroduced to the wild. Project participants include Africam Safari, ANAM (Autoridad Nacional del Ambiente), Cheyenne Mountain Zoo, Defenders of Wildlife, Houston Zoo, Smithsonian’s National Zoological Park, Smithsonian Tropical Research Institute, Summit Municipal Park and Zoo New England.

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Photo: A strawberry dart frog at the National Zoo

“Most frogs put out a tremendous amount of eggs, let them go and see what happens. The tadpoles swim off and there’s no one looking after them,” said keeper Justin Graves. “But not these guys. These guys are sticking around. Because the females only lay a few eggs, they put a greater amount of care and energy into looking after them.”

The strawberry dart frog’s reproductive process starts in thick foliage close to the ground, where the female lays about six eggs—each no bigger than a pea—in a moist place after mating. For about the next 10 days, the male frog will protect and water the clutch by emptying his bladder on it. When the eggs hatch, the female carries each tadpole on her back, one at a time, up the side of a tree in search of a small pool of water, less than a thimble’s worth, where she can deposit them. This can be a tree cavity, but many frogs use pools in bromeliad plants, which often grow high up on trees.

For the strawberry dart frogs at the National Zoo, finding such a nursery usually means a journey of about two feet up to the clusters of bromeliads in the Amazonia terrarium where they live. But researchers have witnessed frogs in the wild carry a tadpole nearly 40 feet—a trek that takes about an hour. Each tadpole gets its own pool, increasing the chance that at least a few of them will survive to become frogs. 

Photo: Strawberry dart frogs are among the Amazon’s smallest amphibians, growing to be no larger than the diameter of a quarter (compared to a dime here).

This entire process distinguishes these frogs from other species, such as the female American bullfrog, which can lay as many as 20,000 eggs and then abandons them.

The female spends the next six to eight weeks feeding the tadpoles by going back and forth between them and laying unfertilized eggs in the pools. This is how these frogs got their Latin genus name, Oophaga, which refers to the fact that the tadpoles are egg eaters; they get the nutrients they need to become frogs by eating unfertilized eggs.

Strawberry dart frogs are one of more than 100 species of poison dart frog and live primarily in the rainforests of Central and South America. Though not endangered, habitat loss and a disease known as amphibian chytrid fungus continue to reduce their numbers. Their natural predators include birds, reptiles and other amphibians.

Photo: An orange strawberry dart frog at the National Zoo.

Adult strawberry dart frogs get no bigger than the diameter of a quarter and can be one of 30 different color variations. The Zoo’s frogs primarily eat crickets, but in the wild they eat ants, beetles and other invertebrates, which give them their toxins. The Zoo’s strawberry dart frogs, which arrived this fall, share a home in a beautiful four-foot hexagonal terrarium with glass and lemur frogs, which have also successfully bred recently for the first time in the Zoo’s history.

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Photo: A female red-eyed treefrog deposits a clutch of eggs on a leaf overhanging a pond. 

 Recently, researchers from Boston University and the Smithsonian Tropical Research Institute (STRI) in Panama have been taking a closer look at the vibrations that red-eyed treefrog embryos use as cues to trigger early hatching. The researchers—Michael Caldwell and J. Gregory McDaniel of Boston University and Karen Warkentin of both Boston University and STRI—embedded tiny recording devices into red-eyed treefrog egg clutches and recorded the low-frequency vibrations caused by snakes as they ate the eggs and also by tropical rain storms. They played back these vibrations to egg clutches in a laboratory and found the embryos hatched in response to the snake-generated vibrations and not to the rain vibrations.

This infrared video recording show a parrot snake (Leptophis ahaetulla) attacking a red-eyed treefrog egg clutch. Some of the treefrog embryos escape the snake by hatching early.

This experiment showed that the embryos were responding to the vibrations and didn’t need chemical or visual cues from snakes, Warkentin says, and that the embryos could differentiate between snake vibrations and rainstorm vibrations. 

What puzzled the researchers was that the low-frequency vibrations that triggered the embryo hatchings were in some ways very similar to vibrations caused by many benign forest stimuli—such as rainfall, wind or non-predatory animals. “Hatching early in response to benign stimuli would be a serious error” Warkentin explains, “since premature hatchlings are vulnerable to predators in the pond. We reasoned that if a defense—such as hatching early—is costly for prey, then mechanisms should be in place to avoid false alarms, just as mechanisms exist to recognize and defend against predators.”

 Photo: The device in this egg clutch is an accelerometer, used to record vibrations. The vibration recording from a snake attack on an egg clutch is shown below.

In recent laboratory work the scientists improved their understanding of how these false alarm mechanisms work in regard to rainfall. The low-frequency vibrations generated by tropical rainstorms are accompanied by two elements that snake-feeding vibrations do not have: high frequency vibrations and an initial buildup of intensity. When these two features were removed from rainfall recordings and the edited recordings were played back to embryos in the lab, many of the embryos hatched. The intensity buildup and high frequency vibrations played simultaneously with low frequency vibrations indicative of danger suppressed the hatching response.

 “Although we don’t know the precise mechanism—whether, for instance, the high-frequency vibrations mask perception of the low-frequency vibrations that indicate danger, or whether the embryos perceive high frequency vibrations as indicative of safety—we do know that the embryo behavior is affected by these two features of rain vibrations in a way that benefits them,” Warkentin says.

 A paper on this research: “Is it safe? Red-eyed treefrog embryos assessing predation risk use two features of rain vibrations to avoid false alarms,” was published in the journal Animal Behavior at the Web address:  www.elsevier.com/locate/anbehav

Photo: A male red-eyed tree frog (Photos and video by Karen Warkentin)

 The Web site for the Warkentin Lab at Boston University is: people.bu.edu/kwarken/

—John Barrat

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Photo: Frog legs for sale at a market in Indonesia. (Photo by A. Roselli)

Amphibians are rapidly declining worldwide. More than one-third of the nearly 6,000 amphibian species are threatened with extinction—disease is one of the main causes. Among the known amphibian pathogens, the parasitic fungus Batrachochytrium dendrobatidis, also known as amphibian chytrid (KI-trid), is a top concern. The fungus, which attacks keratin proteins in the skin of amphibians, including frogs, causes respiratory and neurological damage and eventually death.

“Amphibian chytrid is an unusual example of a disease that is a primary cause of extinction in amphibian species,” said Brian Gratwicke, biologist at the Smithsonian’s National Zoo and lead author of the team’s paper. “In fact, amphibian chytrid has been listed as a likely threat in 94 cases out of the 159 extinct and potentially extinct amphibian species. There are several hypotheses about how amphibian chytrid has spread around the world, but the trade in amphibians for food, bait, pets and laboratory animals has been identified as the most likely mode of spread.”

Photo: The American bullfrog is one of the many species used in the frog leg trade. (Photo by Mehgan Murphy)

Although consuming amphibian legs is familiar to many people as a culinary curiosity, the global extent of the international trade is unknown. The team of scientists’ research focused on 1996 through 2006, during which more than 100,000 metric tons of frog legs were imported from both wild and farmed sources, at a net value approaching half a billion dollars. One kilogram of frog legs averaged about $4 over this period—a kilogram of frog legs requires 10 to 40 individual animals translating to approximately 100 to 400 million animals per year.

 The scientists found no recorded cases of the extinction of a frog species caused by collection for food. However, given the growing importance of aquaculture to supply frog legs to global markets, the team stresses that the risk of disease spread through poorly regulated amphibian trade is probably an even greater risk to amphibian biodiversity than the direct population effects of overharvesting.

In countries such as Indonesia, which exports about 45 percent of all frog legs, the majority of animals are thought to be wild-caught and there is little to no effort to monitor this food source for disease pathogens. “Any trade in live frogs or fresh, un-skinned frog legs presents a substantial risk of the spread of amphibian chytrid,” said Gratwicke. “The implementation and enforcement of some key amphibian trade policies could be a cost-effective conservation tool to mitigate disease risks associated with the trade.”

Graphic: Global exporters and importers of frog legs 1996 – 2006. Click graphic to enlarge. (Data from the UN Commodity Trade Statistics Database)

The exact origin of amphibian chytrid is unknown, but one theory is that it originated in Southern Africa and was distributed worldwide in the 1950s through the trade of the African clawed frog for pregnancy-testing and other amphibian trade. Amphibian chytrid has been detected in many parts of the United States, but some species are apparently resistant to the fungus, and it is not always associated with amphibian declines. The most dramatic declines have been observed in mountainous parts of Central and South America and Australia where it is responsible for the disappearance and probable extinction of many species.

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Photo: An endangered Shenandoah salamander, left, and a redback salamander, right in Shenandoah National Park. Competition between the two species is believed to confine the Shenandoah’s territory to just a few kilometers.

Its entire habitat is limited to just a few kilometers, a refuge surrounded by vast numbers of the more abundant redback salamander (Plethodon cinereus). How the Shenandoah salamander has managed to survive, and whether they can continue to do so, are questions that have drawn Jennifer Sevin, a biodiversity conservation specialist at the Smithsonian’s National Zoological Park, to the Virginia mountains.

Both Shenandoahs and redbacks are members of a group known as woodland salamanders. Air-breathers who lack lungs, they take in oxygen and expel carbon dioxide directly through their skin. Almost all woodland salamanders live under damp leaves, logs and rocks. At night and on rainy days they come out to prowl for mites, springtails, and similar small fare.

Once more widely distributed, the Shenandoah salamander likely retreated to its higher elevation habitat at the end of the Pleistocene (~10,000 years ago) as Earth’s climate warmed.  “Climate change is believed to have allowed the redbacks to expand,” Sevin says. Found in forests from Canada to North Carolina, redbacks also command the wooded areas of moist soil that surround the Shenandoah salamander’s talus patches. Competition between the two species may help confine Shenandoah salamanders to the rocky slopes.

Photo: Jennifer Sevin searches for Shenandoah salamanders on the moist, leaf strewn, forest floor of the Shenandoah National Park in Virginia.

Redbacks are not the only problem for Shenandoahs. Present-day climate change was ranked as the number-one threat to Appalachian salamanders by scientists attending a salamander conservation workshop last May at the National Zoo’s Conservation and Research Center, in Front Royal, Va.

In Shenandoah National Park, construction of the celebrated Skyline Drive in the 1930s may have further isolated Shenandoah salamanders, possibly cutting off gene flow between newly separated populations. Sevin has spent much of spring, summer, and fall the last two years scrambling over the talus slopes of Stony Man, Hawksbill, and the Pinnacles. She is collecting data that will shed light on the competition between salamanders on the three mountains. A nest she found last August containing one mother, six eggs and two hatchlings (the first Shenandoah salamander nest seen by any scientist) was a high point in an intensive project.

Sevin has repeatedly visited 124 study plots, each 32 by 2 meters, some in talus areas and others in less rocky habitat. In addition to measuring and weighing every salamander found, she and her team also note conditions such as air temperature, relative humidity, the amount of moisture on rocks and other objects and days since last rainfall. They also have collected information on each site’s dominant tree species, the amount of canopy cover, the depth of leaf litter, the distance from roads or trails, and the kind of talus present.

Photo: Researchers from the National Zoo take a census of Shenandoah salamanders in the Shenandoah National Park. Global Positioning System coordinates are used to pinpoint the rough boundaries of the salamander’s territory. (Photos by John Barrat)

Zoo scientists are using DNA to learn if Shenandoahs and redbacks are mating across species. “If these species are hybridizing, there is the potential for the redback genes to swamp the Shenandoah salamander genes,” Sevin says. That could bring down the curtain on five million years of Shenandoah salamander history.

In addition to the DNA work and pending permission from the U.S. Fish and Wildlife Service, Sevin hopes to capture approximately 30 Shenandoahs for experiments that will be conducted in climate-controlled rooms in the basement of the Zoo’s Conservation Biology Building. National Zoo biologists want to investigate how climate change might affect the species’ use of habitat, feeding success and even its competition with redback salamanders. Brian Gratwicke, a biologist who leads the National Zoo’s Amphibian Conservation Program, also will attempt to breed Shenandoah salamanders in captivity, something never done before.

Salamanders merit attention “simply for existence value,” Gratwicke insists. “They are important because they existed in this part of the world a long time before we ever got here, and they’re beautiful, amazing creatures.” Reason enough, he believes, to try and help them stick around a few more million years.

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