LeafThe Magazine of TreeLitter Walkers International and Conservation EDITOR Ron Skylstad Leaf Litter VOLUME 4 | ISSUE 1 ASSISTANT EDITOR Ed Kowalski 1 FROM THE EDITOR 2 AMPHIBIAN HEALTH AND NUTRITION PHOTO EDITOR 5 FOLIUM Tim Paine

FEATURES • 11 The Darwin’s Conservation Initiative TWI DIRECTOR Ron Skylstad 20 Innovation in Conservation: The Case of the Strawberry TWI PROGRAMS DIRECTOR Brent L. Brock 26 Islands in the Sky

40 Developing a Captive Breeding Protocol for Georgia’s Blind Salamander (Haideotriton wallacei) at the Atlanta Botanical Garden MISSION STATEMENT Tree Walkers International supports the protection, conservation, and restoration of wild amphibian populations through hands- on action both locally and internationally.

We foster personal relationships between people and nature by providing opportunities for citizens of all ages to become directly involved in global amphibian conservation.

Through this involvement, our volunteers become part of a growing and passionate advocacy for the protection and restoration of wild amphibian populations and the environments on which they, and ultimately COVER Rhinoderma darwinni we, depend. Photo by Danté Fenolio

the magazine of tree walkers international and amphibian conservation from the director

The hellbender (Cryptobranchus alleganiensis) is a species of giant salamander endemic to eastern North America. Larger than any other salamander in its range, it is a specialist, filling a very specific niche within a very specific environment. To survive the hellbender requires a consistently high level of dissolved oxygen found in chronically cold, swift flowing riffles of water. However, due to a variety of factors, its numbers are in dramatic decline throughout much of its range.

It is that last fact that makes such a unique creature so common among other .

You have no doubt heard the warnings over the last couple of years: we are in the midst of the 6th Great Extinction. Amphibian populations all over the globe are in decline due to habitat loss and modification, pollution, climate change and the spread of an amphibian- killing fungus, Chytridiomycosis.

At one time, illustrations like the one above were the only glimpse people had of creatures like the hellbender. We hope, that through awareness and novel approaches to conservation, that it does not become the only way people will be able to view them in the future.

from the editor 1 amphibian health & nutrition

The back and head are generally solid black, although in some individuals there is an interrupted yellow median dorsal stripe. A wide gold, red-orange or orange dorsolateral stripe runs from each side of the snout over the eyes and back, down to the base of the thigh. A single white line projects from the shoulder, at the insertion of the upper arm, and runs along the lip to just under the eye. The dorsal surfaces of the limbs show dense blue-green speckling on a black background, while the venter and ventral surfaces of the limbs are The Quality of a Frog

by ed kowalski

Every now and then in various discussions there are It has been well established in the literature that size references to “poor quality” , usually referring to at directly affects adult size. smaller adult size or less colorful as compared growth is directly affected by light, water quality, to either the initial wild caught imports or frogs that temperature and to a lesser extent by the (protein) were directly observed in the wild. There are usually composition of their diet. High protein diets combined two main theories put forth as to the reason for with optimal rearing conditions will not only encourage the disparity between the frogs: nutritional and/or an optimal rate of growth, but should also maximize genetic. In actuality, the difference is probably due size at metamorphosis. Slower growth rates and/or to a convergence of more than one factor, and a basic reduced size at metamorphosis can usually be resolved review and discussion of these issues may help provide when conditions for the tadpole are improved. With a little clarity as well as direction on this topic. This respect to dendrobatids, there have been fads over the article should in no way be considered definitive and years for different tadpole diets (excluding obligate probably should be considered speculative at this time feeders) ranging from primarily plant-based diets due to the speed at which the available information is (nettle, spirulina, algae based flake foods) to diets changing. high in matter (Tadpole Bites©, black worms,

2 LEAF LITTER • VOL 3 ISSUE 2 fish-based flake foods). The diets utilizing higher in conditions which optimize activity and breeding protein levels should result in maximizing size at periods with little to no seasonal variations to separate metamorphosis, yet adult size of the frogs in captivity breeding from non-breeding periods, this is an area does not appear to have varied significantly. where changes in not only time to maturation but reproductive frequency can significantly impact the Temperatures for rearing are often subject growth of the animals in question. In the wild, active to day/night drops as the tadpoles are often reared foraging for food items, food items that are less calorie in ambient room temperatures. These temperatures dense (ants versus crickets, and fruit ), as well as anecdotally appear to be within the range of what is seasonal limits on reproduction would reduce not only known to occur in the wild, but the fairly constant the rate of reproduction, but reduced fat stores allowing temperatures in captivity should lead to standardized for greater periods of growth. (also see thoughts below conditions for growth and consequently should not on Phenotypic Plasticity.) be a primary cause in the differences seen between wild and captive populations (for a number of One of the potential differences in the size of the anuran species in the literature, lower temperatures frogs may be due to phenotypic plasticity (which is increases size at metamorphosis by increasing time to where an animal demonstrates different phenotypes metamorphosis). in response to environmental conditions) as many of the dendrobatids are housed in conditions that Light can have a significant effect not only on the vary significantly from their natal environment. tadpoles but post metamorphosis. Tadpoles that For example, leaf litter dwelling species are often are deficient in melatonin (such as being kept under maintained in extremely heavily planted enclosures red lights and/or mainly in the dark), then time to which while aesthetically pleasing to the eye provide metamorphosis is reduced with a subsequent loss different environmental conditions than those in the in size at metamorphosis. As a further complication wild. The density of the plantings in these enclosures photoperiods that outside of normal day light (10:14 may actively inhibit the movement of the frogs which day/night to 14:10 day/night) can change not only along with the conditions which optimize reproduction growth but can significantly affect feeding behaviors. may result in differences in size (as larger frogs may In some species, as the ratio of day to night decreased, be at a disadvantage for example in foraging, and feeding decreased as well. This lowered intake will have courtship behaviors. Phenotypic plasticity can affect a direct impact on the size at metamorphosis. a population very quickly with significant difference appearing as quickly as one generation. If phenotypic It is also well known in the literature that the adult plasticity is the component (or one of the components) size of the frog also depends on the size at which of the differences seen between captive and wild reproduction begins to occur as resources are diverted populations then the captive population(s) may be at from growth to reproduction. A number of different risk of losing alleles (genetic variation) via relaxed or factors can affect the size at reproduction of the frog, passive selection for smaller frogs. including but not limited to photoperiod, nutrition, as well as genetics. The rest of the article will touch upon Simple genetic effects may also be a cause in the these topics and their potential effects. differences between the populations of wild and captive frogs. Unless the captive populations are managed in Photoperiod can also change the rate at which the frog a manner that encourages maximal genetic diversity, reaches sexual maturity as well as modifying growth the genetics of the population will shift over time rates of the frog. Day lengths that have a light period potentially fixing one or more undesirable traits like longer than 14 hours can cause frogs to reach sexual smaller size at maturity. Passive and relaxed selections maturity at an earlier age and smaller size. In addition are two more potential reasons for the differences seen differing photoperiods can either increase the rate at in the populations of the frogs. Both of these methods which a frog grows or decrease the rate of growth. It is of genetic selection are similar in their effects and hard to generalize in these cases as there appears to be depending on the size of the population in question a lot of variation not only between species but within may have a very rapid effect on the morphology. As species. a further complication, these changes can eventually result in the captive population being rendered non- It has been established in other animals that the viable (this may be best exemplified by problems time to first reproduction is also partly controlled by seen in larger scale aquaculture requiring a continual nutrition. Given that many frogs in captivity are fed to reacquisition of brood stock) excess resulting abundant fat reserves, are maintained

amphibian health & nutrition 3 In the end, there are potentially multiple causes that way to changing the metabolic usage of the animal. may or may not be acting in a manner which magnifies Combinations of a reduction in total calories offered the result, but it should be noted that most of these in combination with seasonal reproductive periods can can be dealt with via relatively basic changes to the allow the frogs to utilize nutrients for growth instead management of the frogs. Enclosures which more of reproduction. Changes in the allele frequency can be closely mimic the correct biotope of the frog in question largely controlled by monitoring the level of relatedness (for example, an enclosure for D. auratus may include of the frogs and utilizing one of the populations plans less dense plantings on the floor resulting in more open put forth by TWI’s Amphibian Steward Network. In spaces and patches of leaf litter) and include seasonal short poor quality frogs don’t have to be inevitable. and daily variations in activity periods can go a long

References

Alvarez, D., Nicieza, A.G. 2002. Effects of Modzelewski, Edward H.; Culley, Dudley D.; 1974. temperature and food quality on anuran larval Growth responses of the bullfrog, Rana catesbeiana growth and metamorphosis. Functional Ecology fed various live foods. Herpetologica 30(4):396-405 16:640-648 Olvera-Novoa; Miquel A.; Ontiveros-Escutia, Brown, Lauren E.; Rosati, Ronald R.; 1997. Effects Victor M.; Flores-Nava, Alejandro; 2007. Optimum of three different diets on survival and growth of protein levels for growth in juvenile bullfrog (Rana larva of the African clawed frog Xenopus laevis. The catesbeiana Shaw, 1802). Aquaculture 266 (1): 191- Progressive Fish-Culturist 59:54-58 199

Brown, R. K.; Pomering, M.; Hamer. A. J.; 2003. Pahkala, M.; Merila. J.; Ots, I.; Laurila, A.; 2003. High density effects on the growth, development and Effects of ultraviolet-B radiation on metamorphic survival of Litoria aurea tadpoles. Aquaculture 215 traits in the common frog Rana temporia. Journal of (1): 109-121 Zoology 259 (1): 57-62

Jasienski, Michal; 2008. The potential for recovery Pancharatna, Katti; Patil, Mahantesh Mudigouda; growth in stunted larva of Rana sylvatica and its 1997. Role of temperature and photoperiod in the decline with developmental states in R. temporaria. onset of sexual maturity in female frogs, Rana Amphibia-Reptilia 29(3):399-404 cyanophlyctis. Journal of Herpetology 31(1):111-114

Joshi, Bhaskar N.; Mohinuddin, Khaja; 2003. Rich, C. Nelson; Talent, Larry G. The effects of prey Red light accelerates and melatonin retards species on food conversion efficiency and growth of metamorphosis of frog tadpoles. BMC Physiology an insectivorous lizard. Zoo Biology 27:181-187 3(9) Richards, Christina M.; Lehman, Grace C.; Li, H.; Vaughan, M. J.; Browne, R. K. 2009. A 1980. Photoperiodic stimulation of growth in complete enrichment diet improves growth and postmetamorphic Rana pipiens. Copeia 1:147-149 health in the endangered Wyoming Toad (Bufo baxteri). Redford, Kent H.; Dorea, Jose G. The nutritional value of invertebrates with an emphasis on ants and Lovern, Matthew B.; Adams, Amber L. 2008. The termites as food for mammals. Journal of Zoology effects of diet on plasma and yolk steroids in lizards 203(3):385-395 (Anolis carolinensis). Symposium Consequences of Maternally-Derived Yolk Hormones for Offspring: Van der Have; Rom M.; 2008. Slaves to the Eyring Current Status, Challenges, and Opportunities. San Equation: Temperature dependence of life-history Antonio Texas characters in developing ectotherms. Thesis, Wageningen University, Netherlands

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CR-047 08

INinsp iraSITUtion PHotos by Wayne Kelterer

“The jungle closed around us, green on green on green. The foliage was so dense that the eye had no point of reference. It was not until we stopped for a breather that I could actually see the forest in its compenent parts rather than in its baffling entirety. On the branches, in the crevices of trees, on the ground - wherever I looked there was hardly a foot of space that was not occupied. Every available niche was filled. Life here overlapped itself, layer on layer.”

Don Moser Central American Jungles

folium 5 6 LEAF LITTER • VOL 4 ISSUE 1 folium 7 8 LEAF LITTER • VOL 4 ISSUE 1 microgramma sp.

folium 9 10 LEAF LITTER • VOL 4 ISSUE 1 The Darwin’s Frog Conservation Initiative

By Danté Fenolio, Mauricio Fabry, Andres Charrier, Marcela Tirado, Martha Crump, and Bill Lamar

Photographs By Danté Fenolio

darwin’s frog 11 Within the humid leaf litter of Chile’s temperate southern forests is a frog so unique that it captured the attention of Charles Darwin when he visited the region.

It was known as sapito vaquero or “cowboy However, the courtship behavior of the two frog” by the Spaniards (Cei, 1962; Torres species can include direct kicks to the male’s and Castillo, 1973). Stranger than the head and body by the female (Busse, 1989, fictional animals reported in the day, 1991, 2002). Females deposit a clutch of these amphibians have a bizarre physical on the damp forest floor and the male appearance to match their colorful name, fertilizes them. At that point, the female sporting a long, fleshy, nose-like appendage. leaves and her parental duties are done. Collectively referred to as “Darwin’s Frogs,” The male remains in the area of the clutch the two species are in the genus Rhinoderma. as it develops. As the tadpoles hatch, the The Darwin’s Frog, Rhinoderma darwinii, male takes the developing larvae into his was formally described in 1841 (Duméril mouth and manipulates them through an and Bibron, 1841) and Chile’s Darwin Frog, opening (the vocal slit) below his tongue R. rufum, was described in 1902 (Philippi, and into his vocal sac. The tadpoles develop 1902). The former is found in Chile and inside the structure that is normally used Argentina while the latter is a Chilean for vocalizing! After a developmental endemic. period which can last from 45 to 60 days and is temperature dependent, the male Darwin’s Frogs have a reproductive mode “spits out” fully formed froglets. Male that makes them noteworthy. Like many Rhinoderma rufum spit out tadpoles in a anurans, male Rhinoderma darwinii water source rather than accommodating call to attract females on the forest floor. the larvae all the way through development.

12 LEAF LITTER • VOL 4 ISSUE 1 The unique reproductive strategy of these frogs (vocal found nowhere else; there are minimally 37 anurans sac brooding) has long been recognized and new unique to these forests in Chile, constituting no less details about the behavior were added by 19th and than 85% of the regional amphibian fauna including 20th century investigators (e.g. De la Espada, 1872; three endemic genera and an endemic family to Chile Harting, 1879; Howes, 1888; Barros, 1918; Wilhelm, (, Insuetophrynus, , and 1927, 1932; Pflaumer, 1935; Busse, 1970). the , respectively) (Díaz-Páez et al. 2008; Rabanal and Nuñez, 2008; Vidal et al. 2008; Unfortunately the Chile’s Darwin Frog, R. rufum, has Fenolio et al., 2011). Many of them are now threatened. not been seen since the 1970s (Busse and Werning, Conservation actions will be required to ensure the 2002; Crump, 2003). The decline of the Darwin’s Frog, survival of these species, and useful activities include R. darwinii, is also documented and looks to be most the implementation of captive breeding and assurance apparent in the northern portion of its limited range colonies. (Busse, 2002; Busse and Werning, 2002; Crump, 2003; Crump and Veloso, 2005). No single cause Darwin’s Frogs have long been bred in captivity. The accounts for all amphibian declines (e.g., Blaustein, earliest recorded success that we have been able to find 1994; Pounds et al., 2006; Mendelson et al., 2006; in the literature was by Pflaumer (1935). Also in the Collins and Crump, 2009). Likely contributing to the early 1930s, Wilhelm removed tadpoles from a male Darwin’s Frog declines are problems of deforestation, R. darwinii and artificially reared them (Wilhelm, replacement of native forests with mono-cultured exotic 1932). Wilhelm was also the first to witness and tree species, and non-sustainable land management report a male R. darwinii taking larvae into his mouth practices (Armesto et al., 1996; Aravena et al., 2002; (Wilhelm, 1932). More recently, Klaus Busse bred Cuevas and Cifuentes, 2009). Many of the frog a captive group in Germany multiple times (Busse, species that inhabit the dwindling southern temperate 1970, 1989, 1991, 2002). One of us (DBF) had a humid forests of southwestern South America are captive group in the 1980s and 1990s and they bred for

A monocultured pine plantation in the middle of R. darwinii habitat.

Darwin’s Frog 13 several years. Recently, a captive breeding program public regarding problems that Chilean amphibians at the Universidad de Concepción captively produced face. Ultimately, we aim to provide information about R. darwinii and the program we detail here, at the declining Chilean amphibians (including the spread of National Zoo of Chile, has groups of frogs breeding. emergent infectious amphibian disease) and to provide This is all great news. Having multiple programs in conservation options. Another goal is to establish the Chile working with this endangered amphibian helps Darwin’s Frog as a flagship species, thus bringing to bolster the likelihood that they will survive into the attention to the plight of all of south Chile’s endangered future. Collaboration between these groups is in the amphibians. best interest of these endangered amphibians. One of our first projects was to develop a captive The Darwin’s Frog Conservation Initiative is a breeding facility within Chile. Many projects involving collaboration between the Atlanta Botanical Garden, amphibian conservation have removed individuals of The National Zoo of Chile in Santiago and personnel the in question from the range from the Center for Advanced Studies of Ecology and country for captive reproduction efforts elsewhere. Biodiversity at the Catholic University of Chile, from We feel that by aiding local conservation efforts, Northern Arizona University at Flagstaff, and from complete with trained personnel and a modern GreenTracks, Inc. The goals of the initiative focus on captive reproduction facility, the chances of long elucidating the reasons behind the declines of Darwin’s term conservation success are greater. We hope the Frogs and other amphibians endemic to Chile’s program functions in perpetuity as long as Darwin’s temperate humid forests. Further, goals include Frogs and other endemic amphibians of Chile require attempts to curb the declines using techniques such conservation assistance. A grass roots approach as: (1) development of captive assurance colonies with harnessing local enthusiasm and pride is an important ex-situ breeding of endangered amphibians within part of the equation. Additionally, by removing Chile, (2) monitoring of diseases in wild populations individuals of an endangered species from the range to better inform policymakers and conservation country, the frogs might be exposed to new pathogens authorities in Chile, and (3) education of the general in the host country. This risk complicates the

14 LEAF LITTER • VOL 4 ISSUE 1 prospects of returning captive bred individuals to the We have also emphasized education for the personnel range country for reintroduction should that become a running the facility. The lead veterinarian for the zoo, necessity. This is not a condemnation of past captive Marcela Tirado, visited the Atlanta Botanical Garden reproduction efforts outside of host countries. Rather, for captive amphibian husbandry training. She also we feel that as the collective amphibian conservation attended the Association of Zoos and Aquarium’s community has learned through its experiences, captive amphibian management program. range country breeding facilities, when possible, are preferable. Cooperative projects involving the Atlanta The captive breeding facility at the National Zoo of Chile Botanical Garden have led to the same conclusion. is modeled after the amphibian breeding facilities at the Atlanta Botanical Garden. Front opening terrariums Our captive breeding facility is located on the grounds streamline maintenance and upkeep. ZooMed of the National Zoo of Chile in Santiago (Parque Laboratories supported our project with discounted Metropolitano de Santiago, Chile). The building was rates on equipment and in serving as a staging ground designed with one of its walls made of glass so that for a host of equipment purchased in the United States the visiting public could see into the laboratory. We before it was shipped to Chile. There are 32 18x18x18 wanted to use the facility for education and outreach inch (roughly 46x46x46cm) terraria in the facility to with the general public. To draw visitors to the facility, accommodate the same number of breeding groups. we commissioned a large sculpture of a Darwin’s Frog The glass terraria were drilled and fitted with bulkhead made by a Chilean artist and placed it to one side fittings to implement drainage systems. Each unit has of the lab. Additionally, signage with information a false bottom made of “egg crating” to keep fouled about Darwin’s Frogs and other threatened Chilean water away from the inhabitants and to provide ample amphibians educates visitors. To emphasize education drainage of the media. The terraria are supported for the general public, we developed a bilingual website on lab racks. The lab racks are arranged so that the featuring our project: www.savedarwinsfrogs.org. visiting public can see them through the wall of glass.

Darwin’s Frog 15 Automated misting systems keep the humidity in Our project has combined three key components: (1) an each enclosure at appropriate levels and dampen the ex-situ captive breeding facility within Chile, complete terraria a number of times daily (the number of misting with trained personnel, (2) a field program to help us events is altered to replicate field conditions depending better understand the causes for the declines of Darwin’s on the season). Fluorescent lighting illuminates Frogs, and (3) an education and outreach program to the enclosures. The building has an environmental help us communicate our message to the public. We control system strong enough to keep the ambient feel that projects with all of these components have a temperatures cool during the hottest of Santiago’s better chance at producing viable conservation options summer days. Floor drains facilitate cleaning. Should for declining amphibian species. We are currently the electricity fail, a self-starting generator has been looking to expand our capacity at the National Zoo of provided that can power the building for an extended Chile to accommodate six to twelve additional species period through potential blackouts. A spare water of amphibians that have experienced marked declines storage unit was also implemented on a hillside above in recent years. Sadly, there is no shortage of candidate the breeding facility, should the water supply to the species. Building from the platform of an educated zoo be interrupted. The 350-gallon (1325L) reserve husbandry staff in Santiago, the experiences we have tank has enough water to run the facility for more than had with Darwin’s Frogs, and the relationships we have four weeks. The water will gravity-feed to the breeding developed with wildlife authorities in the government facility in the absence of electricity. The bottom floor and throughout Chile, we are optimistic about our of the captive breeding facility is an insect culturing chances. Nevertheless, the issues threatening the facility. Crickets, collembolans, and flour are amphibians of Chile’s southern humid forests are not already in production. We are working to add bean going to be solved through these measures alone. Non- beetles, aphids, and wingless fruit flies. Additional sustainable forestry practices are eliminating critical good news is that in 2010 our facility survived the fifth habitat. Alteration of habitat in other ways is also strongest earthquake in recorded history. In particular, contributing to the problems. Invasive species, like the self-starting generator and backup water supply trout, could be impacting local anurans by feeding on worked flawlessly. aquatic larvae. Captive assurance colonies, field work, and educational outreach programs offer steps in the Our first live frogs were added in 2010. We are pleased right direction, but there is still much work to be done. to report that they are already breeding. The goal is to maintain the collection of wild collected frogs by Charles Darwin referenced the origin of species as locality. We would like to have as many localities the “mystery of mysteries” in the opening paragraph represented in our facility as space permits. Breeding of On the Origin of Species (Darwin, 1859). Over 150 groups are three to five frogs in size. Captive offspring years after his landmark publication, the mystery of are accommodated individually in deli cups when mysteries might not be the evolutionary origins of young, and then in plastic tubs on shelves. Darwin’s Frogs, but rather their fate at the hands of humans. Field work has involved visiting historic localities where Darwin’s Frogs have been reported. It also includes checking forests without reports of Rhinoderma that have appropriate habitat. Since 2008, we have visited dozens of localities looking for the frogs. When we find populations, skin swabs are collected. The swabs are Acknowledgements: We are grateful to the following tested for the presence of amphibian chytrid fungus for financial support: The Association of Zoos and (Bd) in the laboratory of M. Levy at North Carolina Aquariums and their Conservation Endowment Fund State University, School of Veterinary Medicine. At the (grant No. 08-809), The Shared Earth Foundation, end of our study, we will publish a report documenting The Sophie Danforth Conservation Biology Fund, where we observed frogs and where we have detected The Chicago Board of Trade Endangered Species amphibian diseases. Fund, Cisco Systems, and the George and Mary Rabb Foundation. We would like to thank Klaus Busse for Visits to field sites have involved a few localities his help throughout our project and with this article. accessible via a 2-wheel-drive vehicle. However, Klaus has more experience in keeping and breeding 4-wheel-drive vehicles are usually required. Hiking in Rhinoderma than anyone else so we are grateful for his from the ends of logging roads and other non-developed input. We would like to thank ZooMed Laboratories access ways is also routine. For a few remote localities, for their generous assistance with this project. We renting horses and mules has been necessary. The pack thank Robert Hill and Mark Mandica for their time animals allow for access to distant forest fragments and suggestions as we developed the breeding center. with camping equipment and a field crew for several Osvaldo Cabezas, our amphibian keeper at the National days. The trips to these remote localities have been Zoo of Chile, has done a tremendous job in keeping our memorable. Uncut forest fragments offer a glimpse animals happy and healthy. into what southern Chile was once like.

16 LEAF LITTER • VOL 4 ISSUE 1 Literature Cited

Barros, R. 1918. Notas sobre el sapito de vaquero (Rhinoderma darwinii). Revista Chilena de Historia Natural 22: 71–75.

Blaustein, A.R. 1994. Chicken little or Nero’s fiddle? A perspective on declining amphibian populations. Goicoechea, O., O. Garrido, and B. Jorquera. 1986. Herpetologica 50: 85–97. Evidence for a trophic paternal-relationship in the Busse, K. 1970. Care of the young by male frog Rhinoderma darwinii. Journal of Herpetology Rhinoderma darwinii. Copeia 2: 395. 20(2): 168–178.

Busse, K.1989: Zum Brutpflegeverhalten des Harting. 1879. Rhinoderma darwini- Album der Nasenfrosches Rhinoderma darwinii (Anura: Natur. Haarlem (Venesch Bijland) 78. ). Tier u. Museum 1 (3): 59 63. Howes, B.G. 1888. Notes on the gular brood-pouch Busse, K.1991: Bemerkungen zum of Rhinoderma darwini. Proc Zool Soc 231-237. Fortpflanzungsverhalten und zur Zucht von Mendelson, J.R. III, E.D. Brodie Jr., J.H. Malone, Rhinoderma darwinii: Balz bis Eiablage. M.E. Acevedo, M.A. Baker, N.J. Smatresk, and Herpetofauna 13 (71): 11 21. J.A. Campbell. 2004. Factors associated with the Busse, K. 2002 (2003): Fortpflanzungsbiologie von catastrophic decline of a cloudforest frog fauna Rhinoderma darwinii (Anura: Rhinodermatidae) in Guatemala. Revista de Biologia Tropical 52(4): und die stammesgeschichtliche und funktionelle 991–1000. Verkettung der einzelnen Verhaltensabläufe. Bonn. Ottmar, W.G. 1927. La Rhinoderma darwinii D. y B. zool. Beit.r 51 (1); 3-34. Boletin de la sociedad de Biologia de Concepción Año Busse, K. and H. Werning. 2002. Another extinct 1, Nos. 1 and 2: 4–31. amphibian species? The fate of the Darwin Frogs. Ottmar, W.G. 1932. Nuevas onservaciones acerca de ZGAP Mitteilungen 18(2) 2: 16–18. la neomelia de la Rhinoderma darwinii. Extraxto de Cei, J.M. 1962. Batracios de Chile. Ediciones de la la Revista Chilena de Historia Natural XXXVI (1932): Universidad de Chile. Santiago de Chile. 128pp. 166–170.

Cei, J.M. 1980. Amphibians of Argentina. Italian Pflaumer, K. 1935. Observaciones biológicas acerca Journal of Zoology Monograph 2: 1–609. de la Rhinoderma darwini D & B. Revista Chilena de Historia Natural 39: 28-30. Collins, J.P. and M.L. Crump. 2009. Extinction in Our Times: Global Amphibian Decline. Oxford Philippi, R.A. 1902. Suplemento a los Batraquios University Press. Chilenos Descritos en la Historia Fisica I Pilítica de Chile de don Claudio Gay. Santiago de Chile: Librería Crump, M. 2003. Vocal sac-brooding frogs. From Alemana de Jose Ivens. – Grzimek’s Animal Life Encyclopedia, second ed. Hutchins, M. ed. Gale Group. Pounds, J. A., A.C. Carnaval, R. Puschendorf, C.F.B. Haddad, and K.L. Masters. 2006. Responding Darwin, C. 1859. On the Origin of Species by Means to amphibian loss. Science 314 (8 December of Natural Selection, or the Preservation of Favoured 2006):1541–1542. Races in the Struggle for Life. John Murray, London. Rabanal F.E. and J. Nuñez. 2008. Anfibios de De La Espada, J. 1872. Sobre la reprodución de los Bosque Templados de Chile. First Edition. Rhinoderma darwinii. Annales de la Sociedad de Universidad Austral de Chile, Valdivia , Chile. 206 pp. Historia Natural de Madrid I: 139-151. Torres, D. and H. Castillo. 1973. Notas sobre Duméril, A.M.C. and G. Bibron. 1841. Erpétologie la distribución geográfica del sapito vaquero Genérale ou Histoire Naturelle Compléte des Rhinoderma darwini Dum y Bibr 1841 (Amphibia, Reptiles. Volume 8. Paris: Librarie Enclyclopedique Anura, Dendrobatidae). Boletín del Museo Nacional de Roret. de Historia Natural (Chile) 203-204: 7-9.

Darwin’s Frog 17 Other Chilean Amphibians of Note Text and Photos By Danté Fenolio

The primary goal of the Darwin’s Frog Conservation Initiative (DFCI) is the protection and long-term survival of Chile’s endangered amphibian fauna. Darwin’s Frogs are not the only species in peril. The Chilean endemic family Calyptocephalellidae includes the “False Toads” of the genus Telmatobufo and the Chilean Wide Mouth Frog, Calyptocephalella gayi. Based on genetics studies, it has been hypothesized that the closest living relatives to these frogs are Australian, supporting the notion that all contemporary land masses were once one part of supercontinents, one of which was known as “Gondwana.” If this hypothesis is true, the Calyptocephalellidae are an ancient lineage and true relics of an era long since past. In modern times, the Calyptocephalellidae find themselves running out of suitable habitat and living in areas with increased agricultural and forestry activities. Add to these problems introduced predators, such as trout, and a proposed series of hydroelectric dams... the future for many of Chile’s endemic amphibians is in question.

Support in the United States for amphibian conservation efforts has dropped off owing to the economy. We need your help to continue our protection of these ancient and magnificent species. Please donate through our website. Donations go to one of the key partners in the DFCI, the Atlanta Botanical Garden. Because the Garden is a registered 501c3 non-profit, donations from American citizens are tax deductible.

18 LEAF LITTER • VOL 4 ISSUE 1 antartandica The Marbled Wood Frog, Batrachyla antartandica, is not yet considered threatened. An arboreal species with a stunning pattern, it is always a great encounter on night hikes.

Two endangered “False Toads” are and T. bullocki. Both have suffered considerable habitat loss whereby T. bullocki is now listed by the UK’s “EDGE” program as the 5th most endangered amphibian on Earth. Some in Chile had considered T. bullocki extinct until a recent rediscovery of a small population in a forest fragment. The DFCI has intervened and is establishing captive assurance colonies of Telmatobufo at the National Zoo of Chile in Santiago. More information on the development of captive assurance colonies at the National Zoo of Chile can be found at www.savedarwinsfrogs.org. Telmatobufo venustus

The author with two of the several T. bullocki that were collected as part of an assurance population to be housed at the National Zoo of Chile. Darwin’s Frog 19 Innovation in Conservation The Case of the Strawberry Poison Dart Frog

Text & Photographs by J.P. Lawrence

We live in an era that is perhaps the most biodiverse that the Earth has ever seen. Unfortunately, we also live in the midst of what is likely the largest extinction event the world has experienced in millennia. There have been five major extinction events throughout history, all seemingly caused by stochastic events, the most famous of which was the asteroid that served as a death-knell for the dinosaurs. However, unlike these previous extinction events, the one we are in today is being caused by a single species: Homo sapiens.

As we gain awareness of this crisis, conservation efforts are spawned from the ashes, as it were. We live in an era where people are most aware of the biodiversity crisis and conservation efforts are being pushed all over the world to protect and preserve species. Amphibians are likely the most threatened group of animals with at least a third of the species being threatened or implicated in declines and extinctions in a number of declining, and another third are unknown. With so amphibian species around the world. Global climate many species threatened with immanent extinction, change can affect the precipitation patterns around the aggressive conservation efforts are being enacted each globe which, for animals whose lives are so intimately year to prevent this loss. tied to water, can spell disaster. Global climate change can also have synergistic effects with the spread of Amphibians are threatened by a variety of factors chytrid and facilitate its expansion. Many species of including habitat loss, disease, global climate amphibians are highly sought after in the pet trade— change and overexploitation. Amphibian species as recently as this past year we have seen the damage predominantly found in tropical areas often have overexploitation by the pet trade can have on a species. relatively limited population ranges, and with Kaiser’s Spotted Newt (Neurergus kaiseri) of Iran was increasing human encroachment, these animals often elevated earlier this year to the Convention for the lose out. Habitat is commonly destroyed for a variety International Trade of Endangered Species (CITES) of reasons including agriculture, human settlement Appendix I, the convention’s strictest regulation. This and habitat conversion. To make matters worse, some was done because of unregulated online trading of this governments consider some agricultural systems species. Although the future appears bleak for many such as banana plantations to be forested land, even species, efforts are being made to solve some of these though these areas cannot support natural life due to problems. the intensive pesticide and fungicides commonly used. The hot topic in amphibian conservation right now Current Initiatives (and what receives quite a bit of press and funding) is Chytridiomycosis, a fungal disease that has been These issues have driven zoos and conservationists

20 LEAF LITTER • VOL 4 ISSUE 1 external conservation efforts and scientific investigation from happening. But out of bad, there can be some good. Along with this initiative, El Valle Amphibian Conservation Center (EVACC) was established in order to perform in situ conservation and breeding of threatened species rather than ex situ conservation done by zoos.

These initiatives, while helpful, do not provide a solution for the majority of declining or threatened amphibian species across the globe. Many ambitious conservationists often suggest that zoos or even private individuals can be the solution— and there is no denying that these people can greatly aid conservation efforts—but in order to have the truly large scale effects necessary to slow, stop or even reverse the global amphibian crisis, we need to think outside of the box and look for large-scale solutions to these very large-scale problems. Ex situ conservation has its place, especially for species that, like the Panamanian Golden Frog (Atelopus zeteki) or Kihansi Spray Toad (Nectophrynoides asperginis), are in immediate threat of extinction. The reality of effective conservation for the majority of amphibian species, however, is in situ efforts. These efforts need to be carefully examined in order to balance effort versus reward. to come up with intensive management strategies Strawberry Poison Dart to help slow, stop and reverse this crisis with mixed results. Successes and failures can be found all over Frog Research the board. The Kihansi Spray Toad (Nectophrynoides asperginis) is among the most recent success Enter my research on the Strawberry Poison Dart stories from conservation initiatives. This critically Frog (Oophaga pumilio). This species is a common endangered species was found in only one area of Dendrobatid throughout the Caribbean lowlands of Africa and threatened by habitat loss and invasive Nicaragua, Costa Rica, and Panama. It is a species species. American zoos opted to collect the remaining that is actually doing relatively well, even though the individuals of the population (which was not many) overall trend is a decline through most of its range. and breed them in captivity. This was so successful So you may ask why I am talking about this species in that this year the first toads were returned to Africa in reference to conservation of threatened species. Low an attempt to reestablish the population. concern species such as O. pumilio act as great test subjects in new conservation methods because they But with success there is also failure. Several years can withstand the typical trial and error involved in ago there was a massive import by American zoos such studies. What attracted me to O. pumilio are its out of Panama to try and save these species from phenotypically distinct populations which consist of the wave of chytrid sweeping through the country. varying degrees of density, some being very low while Unfortunately, the situation was not handled with others are quite high. These individual populations care and the Panamanian government felt poorly are good candidates for in situ conservation efforts, treated by the deal, so they closed future exports for especially given the popularity of this species in several years, which also thereby prevented other the pet trade. Without conservation efforts these

innovation in conservation 21 populations could likely be extirpated due to a variety The rearing sites posed a particular problem. Oophaga of factors. pumlio uses bromeliads, tree holes and other small sources of water for tadpole deposition sites. Given My research is centered on gaining a better the nature of these sites (many of which are 100 feet understanding of what types of resources serve to limit or more in the trees) I could not realistically remove populations of O. pumilio on Isla Colon in the Bocas rearing sites. In order to observe effects of rearing del Toro archipelago of Panama. I have hypothesized site addition I had to drastically increase their that there are two likely factors that influence the availability. Adding 200 bromeliads to each of nine population density of these frogs: rearing sites and different RSA quadrats was simply out of the question, food (using leaf litter as a surrogate for food). so I started looking for alternatives and, in this case, I drew on knowledge from the pet trade. For these In order to test my hypotheses, I set up three replicates small amphibians, keepers often use film canisters as of six 10m x 10m quadrats for a total of eighteen. Each deposition sites. This, however, posed a problem as set of six quadrats consisted of a different treatment: my original plan called for 4,500 deposition sites (500 (1) Leaf Litter Added (LLA), (2) Leaf Litter Removed per RSA quadrat) and good luck to anyone trying to (LLR), (3) Rearing Sites Added (RSA), (4) Rearing obtain such a large volume of canisters in this era of Sites Added and Leaf Litter Added (RSA + LLA), (5) digital photography. Another option was test tube Rearing Sites Added and Leaf Litter Removed (RSA + caps, which I could acquire the proper number of, but LLR), and (6) a Control. By using this combination would cost well over $1,000 to obtain. So that option of treatments I can weed out what resource is most was out. important to the frogs. In order to add leaf litter to plots, I took all of the leaf litter from the removed plots I ended up gambling on a novel and innovative method and spread it evenly over the added plots. for rearing sites: drinking straws. Specifically, I used large diameter (0.5”) bubble tea straws which cost a grand total of $70 for 4,500 straws. I cut the tips off each end to mimic the axil of a bromeliad, then folded them in half and attached them to trees and vines using zip-ties or fence staples. I was extremely nervous about this aspect of the project as it was a completely new method and could quite possibly not work at all.

But work it did…and incredibly well. Within two months of my setting up these straws, frogs were using them to deposit tadpoles. At that point, in 2009, I at least knew that the frogs would use my straws and that the experiment was not a complete flop. I returned a year later in 2010 to find the straws were still being used. During this second trip I started my data collection before the wet season had gotten into full swing. As a result, my observations of larvae deposition in the straws were low, between 5% and 15% of all of the straws being used at that time. During the wet season, however, I would assume the usage to go up substantially. After completing my research in June 2010, I had a mountain of data to analyze, out of which a few key results emerged.

First, leaf litter was not important…at least in the density it was looked at in this study. Leaf litter houses many of the food resources that these frogs depend on, so density of leaf litter almost certainly affects the frogs, but I would likely need to have more drastic manipulations to see effects

22 LEAF LITTER • VOL 4 ISSUE 1 of the leaf litter. The second result was that the straws worked and incredibly well. In 2009 my plots averaged approximately 4.5 frogs per 10 m2 plot, both for the straws added plots and the no-straw plots. In 2010 the plots averaged 4.5 frogs per no-straws plot, and in the straws added plots I found an average of 9.5 frogs per plot, doubling (or tripling in some individual quadrats) the population of frogs. This indicates that these frogs are strongly limited by rearing site availability.

The implications of these results are twofold. First, this species of frog is severely rearing site limited, at least on Isla Colon, but likely throughout much of the rest of its range (similar research done several decades ago suggests the same results1). The second implication is that straws can be used, both effectively and cheaply, to help a threatened population rebound from perturbations. This research opens up all sorts of questions regarding other species. In Dendrobatids, many species will utilize these small water sites for tadpole deposition, and the same or similar methods, depending on the species, could be utilized. Even further, this method could potentially allow for frogs to be sustainably harvested by increasing the yield a particular patch of habitat can produce, thereby reducing the stress on the population as a operate at the microhabitat level rather than the whole. This provides an excellent example of the need huge spatial scales typically engaged through habitat for innovation when approaching conservation issues. protection. Managers and conservationists need to develop methods to improve microhabitat quality Innovation in Conservation for these small animals if we are to conserve them at large spatial scales. Many may argue that ex situ The “same old, same old” conservation methods conservation is the key in this predicament, especially help, but when 2,000+ species of amphibians are when we involve the pet trade community. In some hurting, we need large scale solutions that allow for cases that is true, but I would argue that in most cases in situ conservation. Habitat protection is certainly it is not. Only in rare instances is ex situ conservation important, but for small animals like amphibians, better than in situ conservation, but on the large scale we need something more since most of these animals of things we need to be focusing on large scale—we

1 Donnelly, M. A. 1989. Demographic Effects of Reproductive Resource Supplementation in a Territorial Frog, Dendrobates pumilio. Ecological Monographs, 59:207-221.

innovation in conservation 23 Ten transects were run 100 m into the forest from a forest edge for nine different populations of Oophaga pumilio. Frogs were counted along the way and noted how far along the transect in order to determine how frogs related to forest edges, as well as to determine population density for each population.

24 LEAF LITTER • VOL 4 ISSUE 1 simply cannot conserve 2,000+ species using ex situ conservation methods. We need to focus on methods that can improve microhabitat quality on a large scale so that we can conceivably conserve populations or species in situ.

Many reading this article are involved in the pet trade and likely have amphibians of their own. I would even venture a guess that many reading this article have not been so fortunate as I to be able to observe and study these animals in their natural . I see, over and over again, frustration expressed by people in the pet trade interested in conservation because they are not the managers out there making these decisions. I see, over and over again, frustration expressed by these people over not being able to be involved in zoo conservation programs because of protocol. Many seem to think that their voice is meaningless in the effort to conserve these species. However, if I have learned anything from this study, it’s that this could not be further from the truth. My idea and subsequent success was based upon common practices used in the pet trade for rearing these frogs. Innovation is the true key to conservation, and if experience has taught me anything, it is that those working with amphibians in captivity are nothing if not innovative.

innovation in conservation 25 Islands in the Sky Cool and moist mountaintops surrounded by a sea of warmer valleys create environmental “islands in the sky” for Appalachian salamanders. A recently released online film attempts to spread awareness about these crucial habitats and the amphibians that depend on them for survival. text and photOs by Joe Milmoe and Peyton Hale

The Appalachian region of the United States is home to 14% of the world’s 535 salamander species, making it an extraordinary salamander biodiversity hotspot, and a priority region for salamander conservation.i A perfect storm of environmental stressors such as residential and commercial development, energy production and mining, invasive species, transportation corridors and a warming climate pose serious threats to these vulnerable species. The Hidden Jewels of Appalachia is a short film that utilizes compelling still photography and HDdSLR video that communicates the beauty, importance and needs of Appalachian salamanders. This film works to fulfill a conservation outreach objective of “educating local residents to highlight how important and unique salamanders are and identifying ways in which they can change their own behavior to help salamanders” as identified by the IUCN/CBSG Appalachian Salamander Conservation working group, formed in 2008 at the Smithsonian Conservation Biology Institute in Front Royal, VA. Filming for the project took place from May 2009 to April 2010 throughout much of the Appalachian range (New York to Georgia).

The film can be viewed at: http://vimeo.com/26202702 i Gratwicke, B (ed). 2008. Proceedings of the Appalachian Salamander Conservation Workshop. IUCN/SSC Conservation Breeding Specialist Group: Apple Valley, MN.

26 LEAF LITTER • VOL 3 ISSUE 2

28 LEAF LITTER • VOL 4 ISSUE 1 Previous Shenandoah National Park in Virigina is home to three isolated populations of the federally endangered Shenandoah salamander (Plethodon shenandoah). This image helps visualize the “islands in the sky” where terrestrial salamanders occupy a unique habitat niche. These mountaintop habitats are particularly sensitive to a changing climate with warmer and drier conditions. left Peyton Hale photographing the Eastern hellbender (Cryptobranchus alleganiensis alleganiensis) in western North Carolina. Hellbenders have strict habitat requirements consisting of cool and clean stream riffles with varied gravel substrates and large rocks for cover.

islands in the sky 29 Rivers and streams in Great Smoky Mountains National Park provide quality habitat for salamanders such as hellbenders and common mudpuppies. Both species are permanently aquatic, completing their entire life cycles within river and stream habitats. These pristine waterways are still sensitive to the spread of disease and contaminants such as ranavirus, chytrid fungus and fecal coliform.

30 LEAF LITTER • VOL 4 ISSUE 1 islands in the sky 31 32 LEAF LITTER • VOL 4 ISSUE 1 The Red-legged salamander (Plethodon shermani) is a medium to large Plethodontid native to the Unicoi and Nantahala Mountains of North Carolina. The IUCN classifies P. shermani as a vulnerable species, with a dense abundance in a limited range. When threatened this species releases a noxious, sticky skin secretion in order to deter predators.

islands in the sky 33 Mid to high elevation mixed hardwood forests in western North Carolina provide excellent habitat for terrestrial salamanders. Thick canopies shade a dense and moist ground cover consisting of plants, mosses, decaying leaves, logs and bark, supporting the world’s greatest diversity of terrestrial salamanders. Nantahala National Forest

34 LEAF LITTER • VOL 4 ISSUE 1 islands in the sky 35 36 LEAF LITTER • VOL 4 ISSUE 1 Blue Ridge Two-lined Salamander (Eurycea wilderae) found on a stream bank in the Nantahala Mountains of North Carolina.

islands in the sky 37 The Pygmy Salamander (Desmognathus wrighti) is endemic to the high elevation spruce and fir tree forests of the Appalachians. The smallest species in its genus, D. wrighti undergoes direct development and does not have a free-living larval stage of development. Photo by Brian Gratwicke

38 LEAF LITTER • VOL 4 ISSUE 1 islands in the sky 39 Developing a Captive Breeding Protocol for Georgia’s Blind Salamander (Haideotriton wallacei) at the Atlanta Botanical Garden An Update on TWI Grant No. 01

by DantÉ fenolio, ronald bonett and matthew niemiller photographs by DantÉ fenolio

40 LEAF LITTER • VOL 4 ISSUE 1 Haideotriton wallacei. Almost nothing is known of the reproductive biology of this species nor has any study clarified population ecology, behavior, genetic similarities/differences between known populations, etc. In fact, the limits of its range are not even clearly defined owing to the inaccessibility of the habitat within which the salamander lives (the Floridan Aquifer). Because these salamanders inhabit groundwater, any study aiming to clarify their biology and ecology will be a challenge.

We may lose some of the biodiversity inhabiting groundwater before we have a chance to understand the biology and ecology of these fascinating fauna. Human activities on the surface and within recharge zones of aquifers can degrade groundwater quality, threatening future human use of the resource as well as subsurface ecosystems that exist there (Crunkilton, 1982, 1984; Tercafs, 1992; Simon and Buikema, 1997; Wood et al., 2002; Graening and Brown, 2003). Agricultural pollution and industrial runoff have been documented to pose serious risks to groundwater quality (Crunkilton, 1982, 1984; Gunn et al., 2000; Culver et al., 2000; Wood et al., 2002). In fact, the risk of groundwater contamination in the U.S. is greatest in agricultural areas where, ironically, an estimated 95% of local residents rely directly on the resource for their freshwater needs (USGS, 2005). Organisms inhabiting extreme environments Groundwater communities often include amphibians can teach us about the flexibility of life through and invertebrates that are sensitive to contaminants in adaptations in morphology, ecology, and behavior that the environment, particularly fertilizers and pesticides often demonstrate unique and peculiar phenotypes. (e.g., Cole and Casida, 1983; Crunkilton, 1982, 1984; Salamanders have successfully exploited the extreme Hecnar, 1995; Sparling et al., 2001; de Wijer et al., environments of subterranean waters that lack light 2004; Relyea 2005a, 2005b, 2005c). From a wildlife and often have very low food and nutrient resources. management perspective, groundwater contamination Scientists are just beginning to unravel the complexities is a serious problem; because so many groundwater in biodiversity, adaptation, and population ecology species are found in a single aquifer system, one of groundwater inhabiting salamanders (termed contamination event can pose an extinction threat. stygobitic salamanders). Intensified cave exploration For example, in 1981 a liquid ammonia nitrate and coupled with genetic analyses has produced a rapidly urea fertilizer spill and subsequent contamination of expanding list of caudates that are restricted to, and Ozark groundwater (in Missouri), demonstrated the adapted to, subterranean habitats; at least 11 North seriousness of such events. Roughly 21 km from the American species from four different lineages exist spill site, living and dead groundwater fauna ranging (Chippindale et al., 2000; Wiens et al., 2003; Bonett from blind cavefish to groundwater crayfish to cave and Chippindale, 2004; Niemiller et al., 2008) and salamander larvae washed out of a spring fed by the more are on the way (Bonett et al., in prep.). One aquifer (Crunkilton, 1982, 1984). Because an aquifer is convergent characteristic in all but one or two of the the entire available habitat for groundwater fauna, any North American species is paedomorphosis, whereby change in water quality, and especially contamination salamanders maintain a larval morphology and achieve of it, poisons the environment without anywhere for reproductive maturity in this condition, all the while fauna to seek refuge. living in persistent bodies of water (i.e., groundwater). This strategy may be advantageous when aquatic Over-harvest of groundwater also threatens the environments are more productive, or terrestrial communities of organisms living in the habitat. habitats are inhospitable or nonexistent (Wilbur and There are some points to consider that demonstrate Collins, 1973; Sprules, 1974; Bruce, 1976; Bonett and groundwater fauna as exceptionally vulnerable to Chippindale, 2006; Niemiller et al., 2008). However, human activities, including excessive water extraction. very little is known about the life history strategies or For example, subsurface habitats typically display population ecology of any subterranean salamander. decreased diversity in community complexity and reduced species abundance relative to above ground Perhaps the most enigmatic of the stygobitic ecosystems; this means there are fewer species in salamanders is the Georgia Blind Salamander, subterranean habitats and fewer individuals than in

Haideotriton wallacei 41 Haideotriton wallacei

Cambarus cryptodytes

42 LEAF LITTER • VOL 4 ISSUE 1 related species living in surface habitats (Holsinger, 1988). Processes that isolate subterranean populations of organisms, and evolutionary adaptation of those species to their environments, can produce extreme patterns of endemism (Barr and Holsinger, 1985; Culver et al., 2000). For example, the combined ranges of over 50% of the described species and subspecies of groundwater dwelling fauna in the continental United States are estimated to constitute less than 1% of the total surface area of that region (Culver et al., 2000). Within the United States, subterranean fauna constitute more than 50% of the imperiled (G1–G2) species recorded in the Natural Heritage Program; however, less than 4% have received federal protection (Culver et al, 2000). These conditions in subterranean species render them using local limestone so that any trace minerals that vulnerable to human activities. For example, it is may be present in the limestone are also present in believed that no fewer than 10 species of troglobites and the water of the system. A chiller maintains the water stygobites have gone extinct owing to anthropogenic temperature at 68°F, the groundwater temperature in activities (Elliott, 2000). the aquifer where these species are found. Capacity for 24 groups of salamanders and crayfish (12 of each) The situation for the Georgia Blind Salamander and are present; although much smaller groups have been other species that inhabit the Floridan Aquifer is no collected to start the project. The system is maintained different. Despite the fact that the Floridan Aquifer in complete darkness. Live specimens of both the has been designated as an at risk aquifer for fertilizer Georgia Blind Salamander and the Dougherty Plain contamination by the United States Geological Survey Cave Crayfish have been collected from Florida and (Nolan et al., 1998), no regular activities are in place by Georgia. These animals are settling into their captive any agency or institution to monitor the populations of conditions nicely. Importantly, this project would not endemic and imperiled wildlife within the aquifer. be possible without the assistance of a number of cave To begin to understand and monitor the populations divers, four of which have participated so far (Kelly of groundwater organisms inhabiting the Floridan Jessop, Ben Martinez, Mike Stine and Bonnie Stine). Aquifer, a group of biologists has teamed up to study The Georgia Blind Salamander, and all the inhabitants the biology and of the Georgia Blind of the Floridan Aquifer, face daunting environmental Salamander and one of its fellow aquifer inhabitants, challenges. As water quality deteriorates in this aquifer, the Dougherty Plain Cave Crayfish (Cambarus the species that call it home are likely to disappear. cryptodytes). Both in-situ and ex-situ studies are Taking steps now to develop captive breeding protocols planned. Further, Tree Walker International and for some of these imperiled species will enable the a private donor have supported the work through a rapid establishment of assurance colonies in the research grant to help establish a basic lab wherein future, should they be necessary. Further, learning live specimens are being maintained in captivity at the about the reproductive biology of the salamander Atlanta Botanical Garden. and the crayfish will expand upon what little we know Using the TWI grant, a laboratory has been developed at of these “extremophiles.” Immediate goals of this the Atlanta Botanical Garden to study the reproductive project include developing a protocol for the captive biology and initiate a breeding colony for both the management of the crayfish and the salamander as Georgia Blind Salamander and Dougherty Plain Cave well as learning more about the reproductive output Crayfish. The design of the systems has been modeled and behavior of both species. Future goals include after the systems used by the United States Fish and learning more about the range of both aquifer Wildlife Service to breed the Texas Blind Salamander inhabitants, learning how different populations are (Eurycea rathbuni) and the Austin Blind Salamander similar or different based on genetic analysis, studying (E. waterlooensis), which is bred in captivity by the the population ecology of these unique creatures, City of Austin. Our system is composed of a series and determining if the salamander is suffering from of aquaria that are plumbed into a central filtration emergent infectious amphibian diseases. The long- unit. The central filtration unit is filled with limestone term conservation of imperiled groundwater fauna, from one of the localities where the salamanders and starting with the Georgia Blind Salamander and the crayfish have been collected. The approach includes Dougherty Plain Cave Crayfish, is our key goal.

haideotriton wallacei 43 Acknowledgements: We would like to thank Amphibian decline and aquatic pollution: Effects of TWI and Lee Moran for funding assistance. We are nitrogenous fertilizer on survival and development indebted to several cave divers who have assisted us in of larvae of the frog Rana temporaria. Applied acquisition of live animals (Kelly Kessop, Ben Martinez, Herpetology 1(1-2): 3–12. Mike Stine, Bonnie Stine). We appreciate the time and assistance of the Georgia Department of Natural Elliott, W. 2000. Conservation of the North American Resources (permit No. 29-WBH-11-79) and the Florida cave and karst biota. From: Culver, D.C., W. Humphreys Fish and Wildlife Conservation Commission (permit (eds.). Subterranean ecosystems. Elsevier. No. LSSC-09-0288). Valuable time and assistance has been provided from Atlanta Botanical Garden’s Graening, G. O. and Brown, A. V. 2003. Ecosystem amphibian specialist, Robert Hill, and amphibian dynamics and pollution effects in an Ozark cave conservation coordinator, Mark Mandica. We are stream. Journal of the American Water Resources also thankful for the valuable time and suggestions Association 39(6): 1497-1507. regarding this project from W.W. Lamar and Dr. S. Gunn, J. 1991. Water tracing experiments in the Opsahl. Castleton karst, 1950-1990. Cave Science 18: 43-46. Hecnar, J.S. 1995. Acute and chronic toxicity of ammonium nitrate fertilizer to amphibians from Literature Cited southern Ontario. Environmental Toxicology and Chemistry 14(12): 2131–2137. Bonett, R.M. and P.T. Chippindale. 2004. Speciation, phylogeography and evolution of life history and Holsinger, J.R. 1988. Troglobites: the evolution of morphology in plethodontid salamanders of the cave dwelling organisms. American Scientist 76: Eurycea multiplicata complex. Molecular Ecology 147–153. 13, 1189–1203. Niemiller, M.L., B.M. Fitzpatrick, and B.T. Miller. 2008. Recent divergence-with-gene-flow in Tennessee Bonett, R.M., and P.T. Chippindale. 2006. Streambed cave salamanders (Plethodontidae: Gyrinophilus) microstructure predicts evolution of life history in inferred from gene genealogies. Molecular Ecology plethodontid salamanders. BMC Biology 2006 4:6 (12 17: 2258–2275. pp). Nolan B.T., B.C. Ruddy, K.J. Hitt, and D.R. Helsel. Bruce RC. 1979. Evolution of paedomorphosis in 1998. A national look at nitrate contamination of salamanders of the genus Gyrinophilus. Evolution 33: ground water. Water Conditioning and Purification 998-1000. 39(12): 76–79. Chippindale, P.T., A.H. Price, J.J. Wiens, and D.M. Relyea, R.A. 2005a. The lethal impacts of Roundup Hillis. 2000. Phylogenetic relationships and systematic and predatory stress on six species of North American revision of central Texas hemidactyliine plethodontid tadpoles. Archives of Environmental Contamination salamanders. Herpetological Monographs 14: 1–80. and Toxicology 48(3): 351–357. Cole, L.M., and J.E. Casida. 1983. Pyrethroid toxicology Relyea, R.A. 2005b. The impact of insecticides and in the frog. Pesticide Biochemistry and Physiology herbicides on the biodiversity and productivity of 20: 217–224. aquatic communities. Ecological Applications 15(2): 618–627. Crunkilton, R. 1982. Bitter Harvest. Missouri Conservationist Nov.: 4–7. Relyea, R.A. 2005c. The lethal impact of Roundup on aquatic and terrestrial amphibians. Ecological Crunkilton, R. 1984. Subterranean contamination Applications 15(4): 1118–1124. of Meramec Spring by ammonium nitrate and urea fertilizer and its implication on rare cave biota, p.p. Simon, K.S. and A.L. Buikema. 1997. Effects of 151–158. Proceedings of the 1984 National Cave organic pollution on an Appalachian cave: Changes Management Symposium – Journal of the Missouri in macroinvertebrate populations and food supplies. Speleological Society (25)1–4. American Midland Naturalist 138: 387-401. Culver, D. C., L. L. Master, M. C. Christman, and H. Sparling, D.W., G.M. Fellers, and L.L. McConnell. H. Hobbs III. 2000. Obligate cave fauna of the 48 2001. Pesticides and Amphibian population declines contiguous United States. Conservation Biology in California, USA. Environmental Toxicology and 14(2): 386–401. Chemistry 20(7): 1591–1595. de Wijer P., P.J. Watt, and R.S. Oldham. 2004. Sprules WG. 1974. Environmental factors and the incidence of neoteny in Ambystoma gracile (Baird)

44 LEAF LITTER • VOL 4 ISSUE 1 (Amphibia: Caudata). Canadian Journal of Zoology: Wiens J.J., Chippindale P.T., Hillis D.M. 2003. When 52:1545–1552. are phylogenetic analyses misled by convergence? A case study in Texas cave salamanders. Systematic Tercafs, R. 1992. The protection of the subterranean Biolology 52: 501-514. environment. Conservation principles and management tools. From Camacho, A. I. (ed.) The Wilbur, H. M. and J. P. Collins. 1973. Ecological aspects natural history of biospeleology. Monografias del of amphibian metamorphosis. Science 182: 1305-1314. Museu Nacional de Ciencias Naturales Madrid. pp. 479-524. Wood, P. J., J. Gunn, and J. Perkins. 2002. The impact of pollution on aquatic invertebrates within USGS (United States Geological Survey). 2005. a subterranean ecosystem – out of sight out of mind. Pesticides in Groundwater. http://ga.water.usgs.gov/ Archives in Hydrobiology 155(2): 223-237. edu/pesticidesgw.html

The Haideotriton captive breeding project is modeled after the USFWS program in San Marcos, Texas, for breeding the Texas Blind Salamander, Eurycea rathbuni, and the program run by the city of Austin, Texas, for the Austin Blind Salamander, E. waterlooensis.

Eurycea rathbuni

Eurycea waterlooensis

haideotriton wallacei 45 Amphibian Related Papers & Publications

“the repellents emanating from these frogs are believed to be volatile...”

Weldon, Paul J., Kramer, Matthew; Gordon, Scott; Spande, Thomas F.; Daly, John W. (2006). A common pumiliotoxin from poison frogs exhibits enantioselective toxiciy against . Proceedings of the National Academy of Science of the United States of America http://www.pnas.org/content/103/47/17818

ABSTRACT: Neotropical poison frogs (Dendrobatidae) contain a variety of lipophilic alkaloids in their diffusely distributed cutaneous glands, including a major class of compounds known as pumiliotoxins. Pumiliotoxins are highly toxic and are believed to protect frogs against predators. Their potential activity against ectoparasites, however, has not been investigated. We tested female yellow fever mosquitoes (Aedes aegypti) for responses to 8-hydroxy-8-methyl-6-(2′-methylhexylidene)-1-azabicyclo[4.3.0]nonane, designated pumiliotoxin 251D [PTX (+)-251D], a skin alkaloid present in all genera of dendrobatids and in other anurans, and to its unnatural enantiomer, PTX (−)-251D.

Brown, Jason L. et al. (2011) A taxonomic revision of the Neotropical poison frog genus Ranitomeya (Amphibia: Dendrobatidae). Zootaxa 3083: 1–120 http://www.mapress.com/zootaxa/2011/f/z03083p120f.pdf

ABSTRACT: The Neotropical poison frog genus Ranitomeya is revised, resulting in one new genus, one new species, five synonymies and one species classified as nomen dubium. We present an expanded molecular phylogeny that contains 235 terminals, 104 of which are new to this study. Notable additions to this phylogeny include seven of the 12 species in the minuta group, 15 Ranitomeya amazonica, 20 R. lamasi, two R. sirensis, 30 R. ventrimaculata and seven R. uakarii.

Andreone, Franco; Mercurio, Vincenzo; Mattioli, Fabio (2006). Between environmental degredation and international pet trade: conservation strategies for the threatened amphibians of Madagascar. Natura 95 (2):81-86 http://www.nandoperettifound.org/documents/2003_28_Strategy.pdf

ABSTRACT: We present and discuss data on the conservation status and threats regarding some “” and “endangered” frogs of Madagascar (Mantella cowani, M. bernhardi, M. viridis, M. expectata, gottlebei), and one species currently classified as “near threatened” and included in CITES I (Dyscophus antongilii). All these frogs suffer from a combination of habitat degradation and – to some extent – collection for the international pet trade. Updated data indicate however that some of these species are locally abundant and could possibly be harvested (sometimes at varying quotas) for the terrarium market. We also discuss the significant differences between collecting for commercial and scientific purposes, together with modalities for establishing commercial export quotas. Furthermore, we comment and present possible conservation strategies, showing the main results based on an understanding of the sensitivities and needs of amphibian conservation in Madagascar. Our Sincere Thanks

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American Frog Day Anna Diaz Royce Gouge Nicholas Smeenk Sally Rothfuss Michael D’Aquino JP Lawrence Brett Cope Zachary Hill Shae Pridemore Timothy Steward Christopher Park Christian Langner Mitchell Heller Jason Descamps Timothy Paine Mitchell Kaliner Field Smith Lori Minch Erik Sweet Joshua Wells Adam Rees Devin Edmonds Richard Terrell Colin Clark W. Ruprecht Wiedemeyer John Sabol Michael Davis Andrew Romano Justin Pierce Stephen Stock Jeffe Mette Dendrobatidae Nederlands Jason Etgen Steven Wilfong Antonina Katayama James Florian Don Fleming Diane Remmer Joshua Mappes Christopher Miller Rhonda Terry Matthew Runo Michael Pantliano Ryan Lawrence Matthew Lubinn Seabird McKeon Tor & Tiffany Linbo Sean Higgins Timo Paasikunnas Kenda Deford Alex Shepack Greg Allen Jesse Kooiman Luke Keeton Justin McColley Taylor Kennedy Jeanne Marioni Volume 4, Issue 1 www.treewalkers.org