Bufo margaritifera by John Wilkinson ISSN 1026-0269 F ROGLOG Newsletter of the Declining Populations Task Force

February 2003, Number 55.

The Himalayan ( swamps, rain puddles and artificial -To plan interventions of pond fishing ponds, but the majority of the restorations. verrucosus) , an Endangered breeding sites are temporary ponds -To collect data in the regions where Species of India with very low depth. presence of this species is argued or Despite the relative abundance anecdotally reported (Sikkim, of T. verrucosus in the area (as a Arunachal Pradesh, Meghalaya, matter of fact all the suitable sites Manipur). Reliable data on the extent By Daniele Seglie & examined are inhabited by the of occurrence and area of occupancy Prof. Debjani Roy, species), the habitats are of the Himalayan newt will allow DAPTF Seed Grant Holders 2002 disappearing at a dangerously rapid assessment of the risk of extinction in rate: about 30% of the sites found the whole Indian territory. Tylototriton verrucosus is the only from 1998 to 2000 were destroyed -To collect data on life history, biology species of the order Caudata in the before June 2002. The most common and population structure in the Indian Indian subcontinent. It is reported from cause of destruction is the draining by Himalayan region to better plan the the mountainous region of southwest local people for utilizing the land for long-term conservation of T. China, North Vietnam, Thailand, cultivation. Furthermore, 50% of the verrucosus. Burma, Bhutan,east Nepal and remaining sites are under threat of northeast India. Acknowledgements. destruction as a consequence of the We thank Prof. C. Giacoma for Although the Himalayan newt development of tourist and economic is listed under the endangered valuable suggestions and opinions. activities in the area. Our best thanks to Mr. Bijoy Tamang, category of the India Wildlife The India Wildlife (Protection) (Protection) Act of 1972, the actual for field assistance, and to his family Act of 1972 only protects the for hearty hospitality. conservation status of this species has against wildlife trafficking and does not Contact: D. Seglie, Dipartimento di not been systematically investigated. prevent habitat loss. The Social The lack of precise knowledge on life Forestry Department of the Darjeeling Biologia Animale e dell’Uomo, history, morphology and distribution Gorkha Hill Council has undertaken Università di Torino. Via Accademia prevents the formulation of any long- some initiatives to preserve the Albertina, 17, 10123 Torino, Italy. [email protected] term conservation action plan. To fill habitats of the Himalayan newt (like this information gap, Professor D. Roy periodical surveys of some of the Prof. D. Roy, Institute of Self and M. Mushahidunnabi conducted a breeding ponds) but, due to the lack of Organising Systems and Biophysics, survey in the Darjeeling District from funds, these actions aren’t sufficient to North Eastern Hill University, Shillong 1998 to 2000 with assistance from grant a long-term protection of the – 793 022, India. [email protected] WWF for Nature, India. During this sites. period 23 habitats of T. verrucosus From the observations M. Mushahiddunnabi, c/o Sukbir were identified. collected during the survey evidencing Tamang, Forest Range Office, Sukhia In June 2002, under the a rapid population decline, we suggest Pokhrii, Darjeeling – 734221, West permission of the Chief Wildlife to insert T. verrucosus in the Red List Bengal, India. Warden of the state of West Bengal, of IUCN. Furthermore, the focusing of Pathogens and another intensive survey was international attention on the conducted to examine the status of conservation problems of this species Amphibian breeding sites and to evaluate the will help the local conservation Declines decline of this species, in order to organisations and authorities to get formulate further action plans for the funds for the protection of the By James P. Collins conservation of the animals. The study breeding sites. area covered during the survey of Emerging infectious diseases, June 2002 was approximatively 800 Recommendations for future especially those caused by fungi km2 in the Darjeeling District, West conservation plans include: (chytrids) and viruses (ranaviruses), Bengal, India. The area is located on -To insert T. verrucosus in the IUCN are among the suspected causes of the Himalayan flanks in one of the Red List to focus the attention of amphibian declines. In 1998 an biodiversity hotspots and wilderness National and International international team developed a areas identified by Conservation organisations on the conservation of research program that integrated International. this species. disciplines from molecular biology to T. verrucosus occurs in all the -To guide the local authorities in the global change to address the types of aquatic lentic habitats protection of the threatened sites in question: Why are pathogens causing investigated -permanent ponds, the Darjeeling District. some amphibian population to decline,

The World Conservation Union (IUCN)/Species Survival Commission (SSC) The Open University ● The World Congress of Herpetology ● Harvard University even to extinction? Two grants from infection? Are there environmental crevices in the rocks. Due to the U.S. National Science conditions that increase the likelihood excessive grazing pressure these Foundation’s (NSF) Integrated that animals become ill? An sponges are rapidly being destroyed Research Challenges in expanded international team will and less and less seepage is Environmental Biology (IRCEB) continue using experiments and occurring. Trees growing in cracks in program have funded the collaborative observations in the field and the rocks are being cut down for research projects of 26 principal laboratory to answer these and related firewood which, again, alters the investigators plus their graduate questions. Modeling is a central rainfall infiltration and increases runoff. students and postdoctoral associates. element of the new award that will Due to aggressive new settlers, Alliances have also been forged with enhance our ability to integrate this site is no longer available to me other investigators funded through a diverse biological sub-disciplines. but I would imagine that the situation joint NSF/National Institutes of Health We welcome collaborations is now worse than on my last visit. program for the Ecology of Infectious with other investigators studying All in all, this is a man-made Diseases and another NSF-funded pathogens and . Our disaster that will have a widespread program, Research and Analysis experience is that researchers and and long-term impact on Zimbabwe's Network for Neotropical Amphibians students derive great benefits from fragile environment and its (RANA). being part of a larger set of amphibians. In many populations, hosts and interactions. It also seems clear that, pathogens coexist and each shows in addition to the efforts of individual Adaptive Cluster regular increases and decreases in investigators, we need interdisciplinary Sampling for population size. At times, however, a research and training strategies for Amphibians pathogen nearly or completely analyzing the declining amphibian decimates its host population. Our problem in an integrated way. Our first grant (“Host-pathogen biology and webpage address is: ByLee-Ann C. Hayek & Ron Heyer, the global decline of amphibians,” http://lifesciences.asu.edu/irceb/am Smithsonian Institution 1999-2002) tested the basic phibians/ Although amphibian fieldwork mechanisms underlying each of these sampling needs to be done according patterns using amphibians as a model. Zimbabwe's Fast Track Land to what we know of statistical The leading questions were: How do Re-Settlement and its Impact on principles, we often wish our random pathogens influence host population Amphibians selection had been in a better, more dynamics? Are these newly productive area. Field samples are introduced pathogens, or has the From a correspondent in Zimbabwe often pitiably small in number or non- virulence of historically benign Zimbabwe's fast track land re- informative, yet we know the amphibian associates changed? settlement programme has received amphibians are out there, somewhere Have recent environmental changes considerable world attention, mainly beyond the boundaries of our altered amphibian-pathogen because of its impact on the human samples. Use of the techniques of interactions? The team of molecular population involved. There has been, Adaptive Cluster Sampling (ACS) is biologists, immunologists, patholo- however, a considerable modification one way to enhance our catch or gists, population ecologists and to the environment which has observations for certain types of epidemiologists is using experiments impacted all life forms, including sampling techniques. Below we and observations in the field and amphibians. Indigenous forest is describe the basics of this approach laboratory to answer these and related being destroyed at an alarming rate. and give a reference for further questions. Partially because of the Uncontrolled fires have burnt out a information. different roles they apparently play in high percentage of the former DEFINITION Adaptive Cluster declines, we initially hypothesized that commercial farming areas. Where ranaviruses have an older relationship Sampling (ACS) designs are statistical ploughing is taking place, little strategies for the selection of initial with amphibians, but that chytrids consideration is being taken of soil have recently evolved pathogenicity or random (unrestricted or restricted) conservation practices. Stream bank samples of plots, areas, transects, or are newly introduced. However, cultivation, formerly illegal, is now ranavirus and chytrid phylogeographic traps that allow for inclusion of all widespread. As most grazing has relevant observations (animals, calls, analyses both suggest recent and been destroyed by fire, serious possibly anthropogenic spread. Thus, signs) in the vicinity of the initial overgrazing of what grassland sample. we consider both ranaviruses and the remains is commonplace. chytrid fungus Batrachochytrium All of these factors will PURPOSE ACS increases sampling dendrobatidis emergent amphibian inevitably lead to soil moisture deficits, effectiveness by taking advantage of pathogens. The complete genetic excess run-off of rain water, siltation of amphibian population characteristics, sequence of Ambystoma tigrinum rivers and dams, diminished water which you observe in your field virus is now completed, opening the tables and progressive desertification. sampling effort. ACS allows for a possibility of searching the genome for This is bound to adversely affect more accurate statistical estimate of pathogenesis genes. amphibian populations. the population parameters for animals A second research award For a number of years, a that are patchily distributed in the (“Emerging wildlife diseases: Threats population of Bufo fenhouloti (a toad habitat being sampled. to amphibian biodiversity,” 2002 – with a specialised habitat requirement) ADVANTAGES 2005) was made recently. It builds on has been observed. This particular 1. In conventional amphibian field projects in the first grant to test how population occurred on an isolated, sampling, once the random selection extinction, disease, and environmental large granite outcrop on a former has been made, the amphibian change are linked. Do novel, highly commercial farm. They breed in fieldworker is not allowed to look virulent pathogens increase the seepage water from "sponges" of beyond the pre-determined chance of extinction? Does the health vegetation growing in hollows and boundaries of the samples or final of the host change the risk of

2 population estimates will be biased more amphibians and reptiles are The adaptive cluster (see Heyeretal.,1994,especially seen. sampling data yield anestimateof a Chapters 2 and 6). 3. When this predefined condition is total of 40 specimens. 2. In cases where the amphibians are not met we continue to our next If neighborhood overlaps are clearly more abundant outside of the sample. taken into statistical consideration, the sample area boundaries, any final 4. If this condition obtains, say we adjusted adaptive cluster sampling estimate will be an underestimate. found a number of in our estimate is 176 specimens. ACS allows boundaries to be plot, we lay the box as close as RECOMMENDATIONS As can be extended when high density patches possible to or original placement spot, seen from the above example, for are discovered during the fieldwork. in four adjacent areas - above, below, patchily distributed amphibians, the ACS thus provides for better estimates and each of the sides in some ordered adaptive cluster sampling technique of population numbers. manner. performs better than the simple 3. ACS provides for more efficient, 5. We continue laying the plots about random sampling method, but only the smaller variance, unbiased statistical the four sides of each and every neighborhood overlap adjustment of estimates of population richness or sample plot that meets our condition the adaptive cluster sampling method mean density. (say that we find at least 1 comes close to estimating the true DISADVANTAGE ), until we find each of the number of individuals in the area of Because additional samples are four adjacent plots to be without any interest. added next to sampling units that have salamanders. We then continue with If the patches can be the target organisms, this technique our original sampling plan, stopping to recognized before sampling takes will be reliable only for non-invasive take additional samples whenever our place, then the best technique to use techniques where organisms are not condition is met. is the patch sampling technique disturbed by the sampling procedure 6. In this way we have not only located (Heyer et al. 1994:107-109). involved. Thus, large ground litter a patch of observation but we have If the patches cannot be quadrat sampling would not be included them into the sample and recognized before sampling takes appropriate, as the clearing of leaf made an attempt to avoid a severe place, then the adaptive cluster litter outside the randomly located underestimate. sampling technique will do a better job quadrat perimeter would disturb the 7. The added plots are included in an than simple random sampling. amphibians in that area. The ordered manner and thus become part However, the user must bear in mind technique could be adapted for large of the statistical sample. that even the adaptive cluster ground litter quadrat sampling by 8. The final parameter estimates are sampling technique will seriously determining what the disturbance unbiased and more accurate with underestimate the population size of zone would be between two quadrats lower variance than if the observed very patchily distributed organisms and then using that distance to patchiness had been ignored. unless neighborhood overlap is taken separate “adjacent” quadrats. In such EXAMPLE For illustrative purposes, into statistical consideration. cases, it is absolutely critical to we set up a 20x30 grid in which Standard random quadrat associate these modifications with the numbers representing individuals were sampling is appropriate when the data so that other workers can distributed in three clusters. We distribution of amphibians is not highly replicate studies or compare results summarize the results of this example clumped in the area of interest. accordingly. The technique should here and explain the example in detail, References work best for methodssuch as the including all steps in sampling and Heyer, W.R., Donnelly, M.A., McDiarmid, small quadrat sampling technique for analysis of the data, on the DAPTF R.W., Hayek, L.C. & Foster, M.S. (eds). such species as terrestrial web site ( http://www.open.ac.uk/da (1994) Measuring and Monitoring salamanders. ptf/index.htm ). Biological Diversity: Standard Methods for Our example data set Amphibians. Smithsonian Institution Press: GENERAL DESIGN In the field, Washington, DC. contained a total of 126 specimens. whenever an inventory object, or an Thompson, S.K. (1991) Adaptive Cluster unusually large number of such We first randomly sampled 10 quadrats (= grid cells). Only one of Sampling: Designs with Primary and objects, is located within a sample Secondary Units. Biometrics 47(1103- plot, areas adjacent to the plot are our quadrats had two individuals. The 1115). searched. These new areas define estimate under simple random sampling for these data gives an USGS Press Release, neighborhoods, which may contain the rd target objects that can then be added estimated total of six specimens. December 3 2002 to the initial sample and increase the We next used the adaptive Shedding Light on Amphibian accuracy (and decrease the variance) cluster sampling technique for these Declines: New Research Finds That of population estimates. same ten quadrats. One of the Ultraviolet Radiation May quadrats was in one of the three METHOD Not be a Factor in Amphibian clusters of specimens. The adaptive 1. A statistical sample size and Population Declines cluster sampling estimate for these method is determined, which uses data gives an estimated total of 27 Two reports published in a leading unrestricted or restricted (cluster, specimens. science journal cast doubt on the stratification, systematic) sampling. We then increased our sample importance of ultraviolet-b radiation For example, determining transects, size to 20 random quadrats. The 20 (UV-B) as a factor driving amphibian placing box plots etc. For random quadrats found one of the population declines. Scientists with the convenience, let us discuss the other clusters. Only two quadrats had U.S. Geological Survey (USGS), the placing of box plots along a gradient a total of six specimens from the 20 University of Washington, and the (looking for specimens in leaf litter). random quadrats. The estimate under U.S. Environmental Protection Agency 2. A predefined condition is defined, simple random sampling for these just released their research findings in for example (a) in any given sample, if data gives an estimated total of 15 the journal Ecology . one salamander is observed or found; specimens. Because UV-B has been or (b) in a given sample, if three or shown in field and laboratory

3 experiments to cause deformities and years and that the UV-B exposure De Queiroz, Arthur C. Echternacht, Sandra increased mortality in amphibian could be a factor in the species’ Essbauer, Evan C. Evans, Robert embryos, some scientists have decline. Gunderman, Julian R. Harrison, Jane D. contended that increases in UV-B “The results of our study Hey, Ronald Humbert, Richard Hunnewell, Robert Inger, Jerry Johnson,Elizabeth from thinning of atmospheric ozone suggest that the timing of breeding Kaeding, Fred Kraus, Nancy Karraker, Beth have contributed to declines of frog must also be taken into account, and E. Leuck, Michael Lodato, John McGrath, populations worldwide. However, one that the earlier breeding after dry Karen Menczer, Richard Montanucci, John of the shortcomings of this earlier winters may alleviate some of the UV- C. Murphy, Neiko Murphy, Richard Nord, research has been a lack of B exposure resulting from shallower Kelly O’Neill, Stephane Ostrowski, Thomas knowledge about the actual exposure water,” Corn said. K. Pauley, F. Harvey Pough, A. Stanley of amphibians to UV-B in their natural Biologists from the USGS are Rand, Terri Roth, Rudolfo Ruibal, Anthony habitats. The research presented in helping determine why amphibians are Russell, Richard D. Sage, David A. Saugey, Danna Schock, David M. Sever, the journal Ecology sheds light on UV- disappearing in the United States and Max Sparreboom, Albert Spencer, B as a factor in amphibian declines. across the globe. Research by these Raymond J. Stein, Glenn Stewart, Edward According to USGS research scientists and others has identified Styskel, Mills Tandy, Heather Brooke ecologist Michael Adams, “This is only many deadly viral infections as well as Taylor, Rodger Waldman, Peter R. Warmy, the second study to look at how the the chytrid fungus as factors in some Richard Zweifel. Institutions: Center for distribution of amphibians relates to amphibian die-offs and population North American Herpetology, Chicago potential UV-B exposure. Most declines. Scientists are actively Herpetological Society, Conservation previous studies only addressed investigating a suit of hypotheses that International, Charles D. Sullivan Co. Inc., JETT, San Antonio Zoo. physiological effects of UV-B but did could help explain these worldwide not provide evidence that any negative declines, including global change, Call for papers and registration for the Seventh Meeting of the DAPTF, effects translated into population contamination from pesticides and Southwestern U.S. Working Group: March losses.” other chemicals, increased exposure 10, 2003, Arizona-Sonora Desert Museum. Research by Adams and his to ultraviolet radiation due to ozone Would all persons wishing to present colleagues showed that dissolved thinning, and the spread of non-native papers relating to amphibian research in organic matter in the water absorbs predators. Many biologists suspect Arizona or New Mexico and/or wishing to UV-B in amphibian habitats and that a combination of factors may be attend the meeting, please e-mail protects 85 percent of the amphibian responsible. [email protected] for details. In habitats the researchers sampled. Die-offs are of great concern addition, Partners in Amphibian and Reptile Conservation (PARC) will be holding an This study sampled 136 because amphibians may be good organizational meeting for the Arizona potential amphibianbreeding sites in barometers of significant working group on March 11th , also at the the Olympic Mountains of Washington environmental changes that may go Desert Museum. and the Cascade Mountains of initially undetected by humans. Oregon and measured how well UV-B Amphibians, unlike people, breathe at could penetrate the water. The levels least partly through their skin making of dissolved organic matter found in them much more sensitive to this study were high enough to protect environmental disturbances. the majority of amphibian populations In 2000, the USGS initiated the from the levels of UV-B that are known Amphibian Research and Monitoring to be harmful to amphibians. Initiative, a national effort to detect The second study, which trends in amphibian populations and began in 1986, discussed the conduct research into causes of breeding behavior of boreal chorus declines. This week scientists and RANA and the US National Science frogs at a pond in the Front Range of collaborators are meeting at the San Foundation grant DEB-0130273 the Rocky Mountains west of Fort Diego Zoo to exchange information helped support the publication of Collins, Colorado. USGS researcher and plan activities for 2003. this issue. Stephen Corn and his colleagues Other contributors to the observed that the timing of breeding research published in Ecology include FROGLOG is the bi-monthly depended on snow. In years with USGS scientists Erin Muths, newsletter of the Declining Amphibian below average snow frogs bred in Christopher Pearl and Bruce Bury; Populations TaskForce. Articles on mid-May because the snow melted Wendy Palen and Daniel Schindler of any subject relevant to the earlier, and in years with heavy snow the University of Washington; and understanding of amphibian declines accumulation breeding was delayed Stephen Diamond of the U.S. should be sent to: until mid to lateJune. These Environmental Protection Agency. John W. Wilkinson, Editor, observations were combined with Contacts: Ruth Jacobs 541-750-1047 Department of Biological Sciences, satellite-based estimates of UV-B. The [email protected] The Open University, Walton Hall, scientists found that frogs breeding in Danielle Jarkowsky 541-758-8801 Milton Keynes, MK7 6AA, U.K. May are exposed to less UV-B than [email protected] Tel: +44 (0) 1908 - 652274. frogs that breed in June. Fax: +44 (0) 1908 - 654167 Another study by scientists at Froglog Shorts E-mail: [email protected] Oregon State University had shown that boreal toad eggs developed in DONATIONS We gratefully acknowledge Funding for FROGLOG is shallower water in years with low receipt of these donations, received prior to underwritten by the Detroit snow accumulation. Because January 17. Individuals: Kraig Adler, Zoological Institute, P.O. Box 39, penetration of UV-B in water David P. Badger, Craig Bienz, Marilyn & Royal Oak, MI 48068-0039, USA diminishes with increased water Randy Blasus, Malcolm R. Braid, Bayard depth, scientists in that study had Brattstrom, Robert T. Brooks, Alan Byboth, Helene Cooper, Andrew J. Crawford, suggested that toad embryos received Elizabeth Davidson, Ted M. Davis, Kevin greater UV-B exposure in low water

4