Ohio Amphibian Research and Conservation Conference, March 15, 2008, Columbus, Ohio.

Keynote Presentation:
Unisexual Ambystomid Salamanders in Ohio
Jim Bogart (jbogart[at]uoguelph.ca)
Professor Emeritus, Department of Integrative Biology, University of Guelph, Ontario, Canada

It has been more than seven decades from the time that Clanton first observed populations of unisexual salamanders in southern Michigan. He interpreted the unisexuals as hybrids between two geographical races of Ambystoma jeffersonianum and many people still believe that unisexuals are recent hybrids. We now know that unisexual ambystomatid salamanders are not normal hybrids. They have a reproductive system that is likely unique in vertebrates. These females all posses a mitochondrial genome that is most similar to that found in Ambystoma barbouri but their nuclear genome contains at least one A. laterale genome and one or more genomes of A. jeffersonianum, A. texanum, or A. tigrinum. They can be diploid, triploid, tetraploid or pentaploid and are the most common salamanders found in many populations. Unisexuals are considered to reproduce by kleptogenesis because they can only reproduce by stealing sperm from sympatric males. The sperm may or may not be incorporated in the developing embryos. The characterization or identification of unisexuals is based on the nuclear genomic combinations and is particularly difficult in Ohio because all four sperm donors exist in the state and a number of different genomic combinations have been found among Ohio unisexuals. Unfortunately, unisexual genotypes are not easily distinguished by morphology so molecular and cytogenetic techniques are essential to confirm the identifications of unisexual genomes. Because Ohio unisexuals have more diversity than northern and eastern populations, that use only one or two sperm donors, they provide new insights with respect to the evolution and persistence of kleptogenesis.

Ohio: The Cradle of Herpetology
Ralph Pfingsten (rap347[at]wideopenwest.com)
347 Pineview Circle, Berea, Ohio, 44017

There is probably more known about the amphibians, turtles, lizards and snakes in Ohio than any other state. The main reason for this body of knowledge began with Roger Conant and Charles Walker who produced The Reptiles of Ohio (1938) and The Frogs and Toads of Ohio (1946) respectively. These two works stimulated a number of young men to carry out county or regional surveys. Ohio is also the place where the world’s largest professional herpetological organization had its beginnings with two high school students in Columbus in 1958. In the 1970's the knowledge base continued to grow with financial support for projects from the Ohio Division of Wildlife. In 1989 the production of The Salamanders of Ohio edited by Pfingsten and Downs quickly became the authoritative work on salamanders in the northeastern and central United States. In recent years state herp atlases documenting our knowledge of distributions have been produced: Frogs and Toads in 2000 by Davis and Menze; Salamanders in 2003 by Pfingsten and Matson; and Turtles, Lizards and Snakes in 2006 by Wynn and Moody. The state currently has many active herpetologists and ongoing projects and there are some exciting plans for the future.

Predicting Population Connectivity in a Fragmented Landscape
Katy Greenwald (Greenwald.35[at]osu.edu)
Ohio State University, Dept. of EEOB, 300 Aronoff Lab, 318 W 12th Ave., Columbus, Ohio, 43210

In areas characterized by rapid habitat destruction and fragmentation, it is increasingly important to prioritize conservation action. Amphibians are a particularly complicated group of organisms to protect, due in part to their dimorphic life cycle requiring both aquatic and terrestrial habitat. They are considered important indicators of ecosystem health, but many species are facing rapid declines due to habitat loss and fragmentation, disease, and other unknown causes. We examined the effect of upland habitat composition on the genetic isolation of marbled salamander (Ambystoma opacum) populations in southeastern Ohio. We used 9 polymorphic microsatellite loci and assignment test analyses to characterize genetic isolation. We then used Geographic Information Systems (GIS) analysis to quantify landcover types surrounding each breeding pond. We show that on a small scale (300m radius), the amount of open land – grass, hay and cropland – predicts a significant amount of the genetic isolation of populations. On a larger scale (1km radius), the most important landcover predictor is the amount of deciduous forest. We describe a combined model using these data plus pond size that describes nearly three quarters of the variation in genetic isolation in our system. Straightforward landcover analyses may serve as a useful proxy for genetic isolation when allocating limited conservation resources.

Impacts on Varying Quantity and Quality of Terrestrial Habitat on Spotted Salamanders
Rebecca Homan (homanr[at]denison.edu)
Dennsion University, Biology Dept., Talbot Hall, Granville, Ohio, 43023

Pond-breeding amphibians rely on two important habitat types, the breeding pond and the surrounding terrestrial habitat. If either of these habitats is degraded or destroyed, the population may be at risk of extirpation, especially if the species demonstrates high site fidelity. Given the relative lack of regulatory protection of terrestrial habitat surrounding wetlands, my focus is on understanding the terrestrial habitat requirements of pond-breeding amphibians with high site fidelity, including the Spotted Salamander (Ambystoma maculatum). I study a population of Spotted Salamanders whose breeding pond is surrounded by forested terrestrial habitat that varies in extent, edge type, and forest age. Although each category of habitat is apparently suitable for Spotted Salamanders, I am interested in understanding which habitats support the largest number of salamanders. To that end, my students and I measured finer scale habitat variables, including soil moisture, soil temperature, leaf litter depth, standing tree density, coarse woody debris density, and mammal burrow density in each of the broader habitat types. We also captured breeding adults as they made their way to the breeding pond in the springs of 2005, 2006, and 2007 to determine from which habitat type they were coming. Principle components analysis on habitat, when compared with the non-random pattern of breeding adult migration, suggested that habitat differences in relation to litter depth, canopy cover, and soil moisture best described the non-random distribution of Spotted Salamanders. Differences in forest extent and edge types may also have contributed to the Spotted Salamander distribution pattern.

Phylogeography of the Four-toed Salamander (Hemidactylium suctatum) Across Eastern North America
Timothy A. Herman⁺ (taherman[at]gmail.com) and Juan L. Bouzat
Bowling Green State University/⁺Toledo Zoological Society, Dept. of Herpetology, 2700 Broadway St., Toledo, Ohio, 43609

Phylogeographic analysis as a tool to reconstruct the history and evolution of species improves in resolution with increasing age and decreasing vagility of the taxon in question. In addition, as the geographic scope of the study increases, the data derived becomes informative for comparison to co-distributed taxa. As such, the four-toed salamander (Hemidactylium scutatum) is well suited to track biogeographic patterns across its broad distribution in eastern North America. The range of the monotypic genus Hemidactylium is highly disjunct in its southern and western portions, and even within contiguous portions is highly localized around pockets of preferred nesting habitat. Over 330 Hemidactylium genetic samples from 79 field locations have been collected and analyzed via mtDNA sequencing of the cytochrome oxidase 1 gene (co1). Phylogenetic analysis shows deep divergences at this marker (>10%) and strong bootstrap support of regional monophyletic clades with minimal overlap. Patterns of haplotype distribution suggest major river drainages, both ancient and modern, as boundaries to dispersal. Two distinct allopatric clades account for all sampling sites within glaciated areas of North America yet show differing patterns of recolonization. High levels of haplotype diversity were detected in the southern Appalachians, with several members of widely ranging clades represented in the region as well as other unique, endemic, and highly divergent lineages. This pattern of radiation from the southern Appalachians, coupled with the basal placement of Hemidactylium in recent family-level phylogenies of the Plethodontidae, lends support to the "out of Appalachia" hypothesis.

Changes in Abundance and Demographics of Hellbender populations in Ohio: Implications for Conservation
Gregory Lipps⁺ (GregLipps[at]aol.com)and Ralph Pfingsten.
⁺Ohio Salamander Monitoring Program, 1473 County Road 5-2, Delta, Ohio, 43515

The Eastern Hellbender (Cryptobranchus alleganiensis alleganiensis) is a subspecies widely believed to be in decline. A status assessment completed for the US Fish & Wildlife Service in 2003 found that of the 16 states that historically harbored populations of this salamander, nine have designated them as critically imperiled or imperiled (S1 or S2), and six have designated them as vulnerable (S3). Due to the paucity of data for most populations, the report concluded that the need for increased monitoring and surveying dominates the research priorities. Hellbenders were found in 16 Ohio waterways during surveys of the mid-1980's, and these plus 4 streams with anecdotal accounts of occurrence were visited during 2006-2007. Over 700 person-hours of surveying resulted in Hellbenders being captured in only seven streams, all but one east of Portsmouth. Abundance estimates place the decline in Ohio at ~78%, very similar to the results for three Missouri streams (~80%). Like Missouri, there appears to be little to no recruitment in most Ohio Hellbender populations, and many extant populations may be on the verge of extirpation. The cause of declines in unknown, but is likely the result of multiple stressors. Intensive agricultural practices that began in the late 1960's (the "Green Revolution") most likely resulted in decreases in stream substrate quality across much of the Hellbender's range. Hellbender conservation efforts in Ohio are focusing on additional surveys to locate undocumented populations, protection of riparian corridors along priority Hellbender streams, and searching for novel factors that may be playing a role in the decline.

Comparison of Health Parameters in Eastern Hellbender Populations in Ohio and West Virgina
Rachel B. Weiss¹ (rweiss[at]thewilds.org), Tiffany M. Wolf¹, Gregory Lipps, Ralph Pfingsten, Joe Greathouse², and Barb A. Wolfe¹
¹the Wilds, 14000 International Rd., Cumberland, Ohio, 43732
²Good Zoo at Oglebay Resort, Rt. 88 North, Wheeling, West Virginia, 26003

During the months of June - September 2006 and 2007, health assessments were performed on hellbenders captured as part of ongoing hellbender population surveys in Ohio and West Virginia. In 2006, a total of 21 animals were captured in three different locations. In 2007, a total of 20 animals were captured in five different locations. Physical examinations were conducted on all individuals and included body weight, snout-vent length, blood collection for hematology and serum chemistry, skin swabs for chytridiomycosis (Batrachochytrium dendrobatidis) identification (PCR) and cloacal swabs for ranavirus (family iridoviridae) detection ( PCR). Whole blood was evaluated for estimated total white blood cell count, white cell differential and chemistry. Serum chemistry analytes included alkaline phosphatase (ALP (U/L)), alanine aminotransferase (ALT ,U/L), aspartate aminotransferase (AST, U/L), creatine kinase (CK,U/L), lactate dehydrogenase (LDH ,IU/L), albumin (ALB,g/dL), total protein (TP,g/dL), globulins (GLOB,g/dL), cholesterol (CHOL ,mg/dL), glucose (GLU,mg/dL), calcium (Ca,mg/dL), phosphorus (P,mg/dL), potassium (K,mEq/L), sodium (Na,mEq/L), albumin-globulin ratio (A/G) and uric acid (UA,mg/dL). We continue to monitor regional wild hellbender health and evaluate environmental variables that are affecting population health as well as potentially limiting population growth. This work was sponsored by the Nature Conservancy of Ohio, Oglebay Resort and the West Virginia Department of Natural Resouces.

Historical and Contemporary Perspectives of Ohio's Anuran Distribution
Jeffrey G. Davis (anura[at]fuse.net)
Ohio Frog and Toad Calling Survey, 625 Crescent Road, Hamilton, Ohio, 45013

Ohio has a long history of amphibian distribution documentation. Results of surveys from as early as the 1840's were compiled by Charles Walker and added to the results of his own collection efforts and published in 1946 in The Amphibians of Ohio, Part I, The Frogs and Toads. Walker's maps showed the distributions of 14 species and pointed out major gaps in their distribution. In 2000, The Ohio Frog and Toad Atlas was published. It included all of Walker's records and approximately 1,800 new ones that resulted from field collecting by Ohio herpetologists between 1946 and 1999. Data from eight additional years of collecting combined with data from the Ohio Frog and Toad Calling Survey are presented to provide a increasingly clear picture of Ohio Frog and Toad distribution. Walker's maps are compared with current maps to demonstrate apparent reduction in the range of some species and expansion in the range of others as inferred from distribution maps for each species. The most notable changes in Ohio Frog and Toad distribution include those of Blanchard's Cricket Frog, the Northern Leopard Frog and the Eastern Spadefoot.

The Status of Blanchard’s Cricket Frog: Initial Results from a Long-term Monitoring Program in Western Ohio
Rick Lehntinen (rlehtinen[at]wooster.edu)
The College of Wooster, Dept. of Biology, 931 College Mall, Wooster, Ohio, 44691

Amphibian population dynamics are often characterized by dramatic fluctuations in space and time. This can make it difficult to assess the status and distribution of these organisms and long-term data are often necessary to distinguish declines from natural fluctuations. Blanchard's cricket frog (Acris crepitans blanchardi) is a species that has recently declined over much of its range. In Ohio, this species has disappeared from most of the central part of the state, despite being formerly common there. To monitor extant populations of this species, we surveyed 315 sites in three areas of western Ohio for the presence of Blanchard's cricket frogs using chorusing surveys. Each site was visited one to three times during the peak breeding season each year from 2004 - 2007 (1,456 total site visits). In 2004, 15.7 %, 3.7 % and 8.0 % of sites were occupied in the north, central and southern monitoring areas, respectively. By 2007, occupancy had increased to 23.1 %, 8.4 % and 9.0 %. This increase was primarily due to notable increases in 2007, as occupancy estimates in 2005 and 2006 were little changed from 2004. Our studies of detection efficiency suggest that our detection probability is relatively high and it is likely that at least some of the observed increase is due to colonization of new sites rather than sampling error. These data suggest that cricket frog populations in western Ohio are not continuing to decline and may be rebounding to some extent.

Chytridiomycosis in Ohio: What We Know so Far and What We Hope to Learn
Brandon A. Sheafor (sheaforb[at]muc.edu) and Jonathan Scott
Mount Union College, Dept. of Biology, 1972 Clark Ave., Alliance, Ohio, 44601

Declines in amphibian populations have been observed worldwide since the early 1980's and have now become a global biological concern. Although factors such as habitat destruction, introduction of exotic species, UV radiation, and anthropogenic contaminants may be involved in declines, it is now apparent that disease is an important factor in many well-documented declines and extinctions. A recently discovered zoosporic fungus, Batrachochytrium dendrobatidis, is the causative agent for the disease chytridiomycosis and has been implicated in declines and extinctions of frog populations in Central America and Australia. In Ohio, B. dendrobatidis appears to have been present from at least the mid 1970's. The effects of chytridiomycosis on Ohio amphibians, both past and present, are currently being studied in order to avoid future population crashes and to better understand how amphibian populations interact with this pathogen.

Effects of Moquitofish (Gambusia affinis) on the Tadpoles of Ohio Frogs and Toads
Geoff Smith (smithg[at]denison.edu)
Denison University, Dept. of Biology, Granville, Ohio, 43203

Among the many challenges confronting amphibian populations is the introduction of nonnative species, especially fish. Using a series of experiments, I have been examining the potential effects of an introduced fish, the mosquitofish (Gambusia affinis) on the tadpoles of Ohio anurans. Mesocosm experiments have revealed that the effects of mosquitofish on tadpoles can vary from species to species, and from trait to trait. While some species showed lethal effects (e.g., Green Frogs, Rana clamitans), most effects were sublethal (e.g., changes in time to metamorphosis, tadpole mass) and could be positive or negative. Laboratory studies on behavioral responses of tadpoles to mosquitofish also show variability among species, with some species reducing activity in the presence of mosquitofish cues (e.g., Gray Treefrogs, Hyla versicolor; Green Frogs) and other species not changing their behavior in the presence of mosquitofish cues (e.g., American Toads, Bufo americanus; Bullfrogs, Rana catesbeiana). Finally, a laboratory study on the effect of mosquitofish on the hatching of American toad eggs suggests mosquitofish can influence the timing of hatching. These results suggest that mosquitofish warrant additional consideration as an introduced species that could affect Ohio anurans and tadpole communities.

Amphibians as Indicators
Mick Micacchion (mick.micacchion[at]epa.state.oh.us)
Ohio EPA, Division of Surface Water, Wetland Ecology Group, 4675 Homer Lane, Groveport, Ohio, 43215

Ohio EPA has found that amphibians can be used as meaningful indicators of wetland condition. During the past 12 years Ohio EPA has monitored the amphibian communities of approximately 200 natural wetlands and 70 wetland mitigation projects. Data from the natural wetlands has been used to develop the Amphibian Index of Biotic Integrity that is applied in Ohio's wetland protection program. At the heart of the index is a system of assigning a score of from 1 to 10, known as a coefficient of conservatism, to each amphibian species monitored. This process requiring a thorough understanding of the population ecology of each species from a statewide prospective is explained. Recently, in an effort to improve their application, several Ohio amphibian experts met and developed coefficients of conservatism for Ohio's entire amphibian taxa list. The results and applications of that collaboration are discussed. Using this new information and existing data comparisons are made between the amphibian communities of natural wetlands and mitigation wetlands constructed in an effort to compensate for wetlands lost to development. Special attention is focused on the challenges faced by wetland amphibian populations in urban settings. The data shows that we are not even close to getting back what we are losing by constructing mitigation wetlands and that common urban development patterns are extirpating amphibian populations to an alarming degree.

Evaluating the Impacts of Sublethal Contamination on Amphibian Metamorphosis
Michelle D. Boone (boonemd[at]muohio.edu)
Miami University of Ohio, Dept. of Zoology, 212 Pearson Hall, Oxford, Ohio, 45056

While amphibian declines in nature have been correlated with upwind agriculture, the effects of realistic exposure scenarios on amphibian populations are still poorly understood. Pesticides and fertilizers can directly alter individual physiology and behavior of amphibians, which may have negative consequences for populations; additionally, contaminants can affect amphibians indirectly through positive or negative changes in the food resources base and predator communities. Understanding the combined direct and indirect effects of contaminants, as well as the potential interactive effects of contaminants in the presence or absence of other natural stressors, is necessary to make accurate predictions in the field. In my studies, I have examined the effects of multiple contaminants at expected environmental concentrations on amphibians and their food web, in the presence of other stressors like fish predators or naturally occurring pathogens. Additionally, studies conducted in areas associated with chemical contamination (i.e., golf course ponds) have also been conducted to examine how effective these habitats could be in fostering amphibian communities. My research indicates that effects of combinations of chemical contaminants with different modes of action can often be predicted based on the sum of singular effects resulting from direct and indirect impacts. In many cases, the effect of the contaminant on amphibians can be predicted based on the effects on the food resources. However, the addition of other factors in the presence of contamination increases the probability of reducing or eliminating population sizes of metamorphosing amphibians. Further, my studies indicate that amphibians at higher trophic levels (i.e., salamanders) may be more vulnerable to environmental contamination, and more likely to suffer from declines resulting from contamination. My studies suggest that chemicals used in agricultural settings could contribute to declines in some amphibian species, and favor other species, indicating that contamination may result in changes in species distributions and relative abundance in nature.

Some Factors Causing Local Amphibian Declines in Northeastern Ohio
Timothy O. Matson (tmatson[at]cmnh.org)
Cleveland Museum of Natural History, 1 Wade Oval University Circle, Cleveland, Ohio, 44106-1767

You do not need to look beyond Ohio to witness amphibian declines, both in richness and evenness of distribution, that are related to factors other than overt habitat loss or degradation. In our studies of amphibians in northeastern Ohio drought, predation, potential metal toxicity, bacterial disease, parasites, and probable synergies have been implicated in local declines of the pond breeding Jefferson Salamander (Ambystoma jeffersoniaum) and Wood Frog (Rana sylvatica). Both taxa are sensitive to [Cu] and [Al] at low pH, but the effects of these metals at elevated levels of pH remain unclear. The ameliorating and/or toxic effects of high hardness at elevated pH also remain unclear. In a laboratory study conducted at higher pH (6.5 and 7.5) high water hardness was toxic to Jefferson Salamanders but not to Wood Frogs. High pH coupled with high hardness may limit breeding site suitability to Jefferson Salamanders and may negatively impact the local persistence of the taxon, community biodiversity, and the stability of the amphibian community. This presentation will address some of our recent studies of factors affecting amphibians in some northeastern counties and will discuss the need for enhanced levels of environmental and taxonomic monitoring.

Common Goal -- Conservation
Dean Allessandrini (herpconservation[at]hotmail.com)
Vice-president and Conservation Committee Chairman, The Greater Cincinnati Herptological Society, 354 Glen Oaks Drive, Cincinnati, Ohio, 45238

Over the past few years, the Greater Cincinnati Herpetological Society (GCHS) has amplified its emphasis on local conservation. During the presentation, Dean Alessandrini will describe some of the conservation methods, strategies and challenges of the GCHS amphibian conservation efforts. A brief overview of a few recent victories will be described, including-

Working with a local DuPont plant to restore a Jefferson Salamander breeding pool.
Protecting a critical Hamilton County amphibian habitat from becoming a drag-race track.
Surveying an east-side Cincinnati recreation area (Grand Valley) and providing consultation related to amphibian conservation.

In the name of "getting things done," GCHS officers and members call on internal and external resources both from both the professional and non-professional sector. Traditionally, differences in philosophies and basic prejudices between individuals within these sectors have formed a barrier to collaboration. This presentation will stress a mindset that we believe the GCHS is demonstrating: The fortitude to put aside stereotypes of "professional" and "amateur" herpetologists and naturalists in the name of collaboration generates results. Working together will enable us to tap into a wealth of knowledge, skills, expertise, and enthusiasm. Amphibian conservation in Ohio is a critical and time-sensitive issue. Any action plan that employs effective use of all available resources will maximize our chances of attaining our Common Goal -- Conservation.