23 nd Annual Meeting of the Tennessee. Herpetological Society

Transcription

1 23 nd Annual Meeting of the Tennessee Herpetological Society Middle Tennessee State University 1301 East Main Street, Murfreesboro, TN September 28-29, 2017

2 Front Cover Photo: Streamside Salamander, Nicole Witzel, Tennessee State University Name Tags: Streamside Salamander, Salamander Artwork for T-shirts: Streamside Salamander, Todd Pierson, University of Tennessee

3 Middle Tennessee State University The General Assembly of Tennessee in 1909, through the General Education Bill, moved to establish three teacher-training institutions, one in each of the grand divisions of the state. Middle Tennessee State Normal School opened as a two-year school in 1911, became a four year school in 1925, leading to a Bachelor of Science degree, and was recognized as a state college in The current name, Middle Tennessee State University, became official in With the continued addition of degrees and academic programs, Middle Tennessee State University became the largest undergraduate university in Tennessee in 2012, surpassing the University of Tennessee. Over 28,000 students are enrolled at the university. A significant addition was added to the Middle Tennessee State campus in the fall of 2014 with the opening of the new science building. The $147 million dollar project provided over 250,000 square feet state of a state of the art facility allowing teaching, faculty and student research and collaborative learning. Over 13,000 students enrolled in biology, chemistry and other science courses use the facility annually. For more history and programs/degrees at Middle Tennessee State University, please visit: University Map: Raider Xpress Shuttle Map:

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7 Thursday Afternoon, September 28, 2017 KUC Theater 12:00 Registration 1:00 Opening Remarks 1:05 Welcome - Dr. Lynn Boyd, Biology Department Chair Middle Tennessee State University 1:15 Snake Envenomation: A first-hand account of being bitten by a Timber Rattlesnake Danny Bryan Student Session I Chair - Chris Ogle 1:45 The Relative Abundance and Distribution of a Turtle Assemblage in Two Adjacent Ponds on the Department of Energy Reservation in Oak Ridge, TN. Melody Guo and Olivia Milloway 2:00 Assessment of ATV Impacts on Softshell Turtle Nests Cody D. Godwin 2:15 Larval Growth, Metamorphosis, and Reproduction in the Two-Lined Salamander (Eurycea cf. aquatica) in the Upper Tennessee River Alexander Miele 2:30 Break 3:00 The Morph A problem: alternative reproductive strategies in Eurycea Todd W. Pierson 3:15 The Use of Environmental DNA and Occupancy Modeling to Evaluate the Distribution of the Eastern Hellbender (Cryptobranchus alleganiensis) in Tennessee, U.S.A. Jeronimo Da Silva Neto 3:30 Analysis of Hellbender Skin Secretions in Tennessee and Arkansas Populations Rebecca Hardman 3:45 Neutrophil:Lymphocyte Ratios As A Measure Of Chronic Stress In Populations Of The Hellbender (Crypotbranchus alleganiensis) Across A Habitat Quality Gradient 4:00 Break Allison Litmer

8 Thursday (cont.), September 28, 2017 KUC Theater 4:15 THS Business Meeting and Annual Elections Science Building Atrium, 2 nd Floor 5:30 Poster Presentations / Social 6:30 THS Banquet Science Building, Room :30 THS Auction Friday Morning, September 29, 2017 KUC Theater Student Session II Chair - Lee Barton 8:30 Detecting the Presence and Abundance of Streamside Salamander (Ambystoma barbouri ) in Middle Tennessee Using Environmental DNA 8:45 9:00 Nicole Witzel Response of Reptile and Amphibian Communities to Prescribed Fire in an Oak/Hickory forest. Steven J. Hromada Behavioral and physiological responses to simulated predator induced stress in the Eastern Box Turtle, Terrapene carolina carolina. Victoria Lay 9:15 Preliminary Results From Alligator Snapping Turtles (Macrochelys temminckii ) Surveys in Two West Tennessee River Drainages Saidee J. Hyder 9:30 Break 10:00 Distribution and Status of the Western Pygmy Rattlesnake (Sistrurus miliarius streckeri ) in Tennessee Shawn Snyder 10:15 A Comparative Study of Stress Physiology in the Common Watersnake (Nerodia sipedon ), the Diamondback Watersnake (Nerodia rhombifer ), and the Queen Snake (Regina septemvittata ) Louise McCallie 10:30 The cutaneous snake microbiome correlates with Ophidiomyces ophiodiicola, the causative agent of snake fungal disease 10:45 Break This concludes the student presentations. Jacob E. Leys

9 Friday Morning (cont.), September 29, 2017 Professional Session II Chair - Danny Bryan Presentation Awards will be issued prior to the start of this session 11:00 Common Mudpuppy (Necturus maculosus ) not common in Southern Appalachian streams. Stephen K. Nelson 11:20 What s temperature got to do with it? Climate change interferes with stream amphibian community stability 11:40 12:00 Kristen Cecala I m not your steppin stone: Recent winter finds and current distribution of the Streamside Salamander, Ambystoma barbouri, in Tennessee. Meeting Wrap-up David Withers THS Annual Field Trip Flat Rock Cedar Glades and Barrens State Natural Area 1:30 Meet at parking area located on Factory Road (directions in meeting agenda) ,

10 Tennessee Herpetological Society Business Meeting Agenda Items Thursday, September 28, 2017 Reading of the 2016 Meeting Notes Treasurer s Report Committee Reports o Conservation Committee o Chad Lewis Memorial Grant Committee o Website o Publication/Newsletter (Society Journal) New Business o Website Recommendations o Establishing a Social Media/Website Committee Elections Positions to be elected during the 2016 meeting: Vice President Treasurer East Tennessee Representative Middle Tennessee Representative Current Tennessee Herpetological Society Board President: Josh Campbell Vice President: Kevin Hamed Secretary: Stephanie Chance Treasurer: Dustin Thames West TN Representative: Lee Barton Middle Tennessee Representative: Danny Bryan East Tennessee Representative: Chris Ogle Sergeant at Arms: Scott Dykes

11 Abstracts Student Presentations Turtles The Relative Abundance and Distribution of a Turtle Assemblage in Two Adjacent Ponds on the Department of Energy Reservation in Oak Ridge, TN. Melody Guo 1, Olivia Milloway 1, and Kathy Strunk Nature Lane, Clinton, TN (creso@acs.ac) Research shows that turtle populations can vary in similar geographic locations, even where study sites are close in proximity. In this presentation, we discuss the differences in abundance and distribution of a turtle assemblage in two adjacent ponds (L and S) that are comparable in size and depth. Over an eleven-year period, 828 total captures of 144 individuals representing six species were recorded. There were significantly more Chelydra serpentina and Chrysemys picta total captures in L pond, while there were more Sternotherus odoratus total captures in S pond. Based on initial capture there were significantly more individual C. picta from L pond and S. odoratus from S pond. There were more C. picta and C. serpentina males found in L pond, whereas there were more S. odoratus males found in S pond. Data collected during each trap session included sex, carapace length/width, plastron length/width, thickness, weight, and number of leeches found on each turtle. 1 - Melody Guo and Olivia Milloway are students at Oak Ridge High School in Tennessee. Melody has been conducting research with the Clinch River Environmental Studies Organization (CRESO) for three summers, while Olivia has worked with CRESO for two summers. 2 - Kathy Strunk is a retired math teacher that has been conducting field research with middle and high school students for the past 29 years as part of the CRESO program. Assessment of ATV Impacts on Softshell Turtle Nests Cody D. Godwin Department of Biology, Tennessee Tech University, Cookeville TN, Recreational vehicle use (e.g., all-terrain vehicles or ATV s) has become increasingly popular in recent years, and is particularly prevalent in the southeastern U.S.. Numerous studies have indicated negative effects of ATV s on the environment, especially in and around wetlands, including wetland degradation, soil erosion, destruction of vegetative communities, and direct animal mortality via impact. However, the impact of ATV s on nest success of fresh water turtles has not been documented. The beaches of the Comite River in southeastern Louisiana bring into close contact ATVs and the nesting sites for two species of softshell turtles (Apalone mutica and Apalone spinifera). The present study aims to understand the impacts of the ATVs on softshell turtle nests and thus the turtle populations. A study performed in , when ATVs were absent, provides a baseline for assessing the current impacts. Two field seasons in 2015 and 2016 reveal ATV impacts are the highest forms of mortality over naturally occurring forms of nest mortality.

12 Salamanders Larval Growth, Metamorphosis, and Reproduction in the Two-Lined Salamander (Eurycea cf. aquatica) in the Upper Tennessee River Alexander Miele 1, Todd W. Pierson 1 1 Department of Ecology and Evolutionary Biology, University of Tennessee, Knoxville, TN, USA The two-lined salamander (Eurycea bislineata) species complex is one the most widely distributed and common groups of plethodontid salamanders in North America, and it shows great variation in habitat across this distribution. Because the E. bislineata species complex occupies a large geographic extent and broad ecological niche space, it is an interesting model for studying life history variation. Currently, five species are recognized in this group, including E. bislineata, E. junaluska, E. wilderae, E. cirrigera and E. aquatica. Here, we summarized > 1 year of monthly life history and demography data from Eurycea cf. aquatica in the Upper Tennessee River. Our principle discoveries are: 1) a single-season larval period; 2) hatching in May with metamorphosis the following summer; 3) evidence of biparental care of nests; and 4) a mean clutch size of 57 eggs. We supplement these data with observations of male-male aggression, communal nesting, and expedited hatching in response to predators, and we place our data and observations in a broader evolutionary context. The Morph A problem: alternative reproductive strategies in Eurycea Todd W. Pierson 1, Samantha Wesnak 2, Jacob Fetterman 2, Kelsey Young 2, Stanley Sessions 3, Jennifer Deitloff 2 1 Department of Ecology and Evolutionary Biology, University of Tennessee Knoxville, Knoxville, TN, USA; 2 Lock Haven University, Lock Haven, PA, USA; 3 Hartwick College, Oneonta, NY, USA Although males of most plethodontid salamander species exhibit stereotyped secondary sexual characters associated with courtship and reproductive behavior, Sever (1979) described an apparent polymorphism in male Blue Ridge two-lined salamanders (Eurycea wilderae). Some males possess seasonally elongate cirri, mental glands, and protruding premaxillary teeth; others deemed Morph A lack all three features and instead grow seasonally hypertrophied jaw musculature. Speculation ensued as to whether these two forms represented a polymorphism within a species or two cryptic, sympatric species across the Appalachians. Here, we present genomic, karyotypic, morphological, and behavioral data demonstrating that these two forms represent an intraspecific polymorphism and frame this discovery within a broader discussion of reproductive morphology and behavior in spelerpine salamanders.

13 The Use of Environmental DNA and Occupancy Modeling to Evaluate the Distribution of the Eastern Hellbender (Cryptobranchus alleganiensis) in Tennessee, U.S.A. Jeronimo Da Silva Neto 1, William Sutton 1, Stephen Spear 2, and Michael Freake 3 1 Tennessee State University, Nashville, Tennessee; 2 The Wilds, Cumberland, Ohio; 3 Lee University, Cleveland, Tennessee The Eastern Hellbender (Cryptobranchus alleganiensis) is a long-lived, fully-aquatic salamander that inhabits cool, well-oxygenated streams and rivers in the eastern United States. Although once abundant, C. a. alleganiensis populations have experienced major declines across the historical range due primarily to habitat degradation. Even though Tennessee provides some of the best remaining habitat for C. a. alleganiensis, standardized state-wide distribution assessments have been limited to known populations, and status of other C. a. alleganiensis populations remains unknown. We used environmental DNA (presence/absence) data to identify important conservation areas with potential remaining C. alleganiensis populations, and estimate site occupancy at 292 sites across Tennessee. Site occupancy was estimated to be 0.45 (130 sites), and the detection probability was The overarching goals of this project is to encourage efficient use of time and resources to effectively manage and conserve the few remaining secure, yet declining, hellbender populations throughout the state of Tennessee. Analysis of Hellbender Skin Secretions in Tennessee and Arkansas Populations Hardman, R.H. 1, Oziminski, K. 1, Reinert, L.K. 2, Rollins-Smith, L. 2, Irwin, K.J. 3, and D.L. Miller 1 1 Center for Wildlife Health, University of Tennessee, Knoxville, Tennessee, USA; 2 Department of Pathology, Microbiology and Immunology, Vanderbilt University School of Medicine, Nashville, Tennessee, USA; 3 Arkansas Game and Fish Commission, Arkansas, USA The Hellbender (Cryptobranchus alleganiensis), is a large aquatic salamander containing two subspecies, Ozark Hellbender (C. a. bishopi), and Eastern Hellbender (C. a. alleganiensis), from the Ozark mountains and eastern U.S., respectively. Both subspecies have seen population declines over the past 25 years, especially in C. a. bishopi which is federally endangered. Habitat degradation alongside other factors may lead to secondary infections with amphibian pathogens such as Ranavirus and chytrid fungus (Batrachochytrium dendrobatidis) or Bd. Skin health and changes in secretions are well documented to explain some observed differences between amphibian species resistance to chytrid fungus. Antimicrobial peptides (AMPs) are secreted on the skin and part of the amphibian innate immunity. In frogs, AMPs have demonstrated growth inhibition of Bd and have correlated with prevalence of disease seen in the field. We purified AMPs from hellbender skin secretions collected in Tennessee and Arkansas during summers We performed growth inhibition assays (GIAs) against Bd for serial concentrations of AMPs for each hellbender. Animals from Arkansas had markedly less inhibitory capacity than those from TN. Within TN populations, Ranavirus negative animals had superior inhibition than those that were Ranavirus negative. We will continue to perform GIAs from our 2017 sampling season including other bacterial and fungal pathogens such as the

14 emerging Salamander Chytrid, Batrachochytrium salamandrivorans. These data will be combined with ongoing analyses of skin microbiome parameters. Neutrophil:Lymphocyte Ratios As A Measure Of Chronic Stress In Populations Of The Hellbender (Crypotbranchus alleganiensis) Across A Habitat Quality Gradient Allison Litmer and Christopher Murray Department of Biology, Tennessee Tech University, Cookeville, TN Amphibians are currently facing widespread population declines, primarily due to the introduction of anthropogenic stressors. Anthropogenic stressors have the potential to alter ecosystem dynamics, as well as elicit long-term physiological responses that could ultimately hinder population viability. Current metrics of population viability rely heavily on demographics, population fluctuation, and degree of isolation. Physiological parameters, such as stress-response, offer a new scale for population assessment and provide an explanation of the mechanism behind population declines. A method currently used to assess stress in wild populations is through plasma concentration of glucocorticoids and presence of the stress-response. However, because glucocorticoids attenuate over time and return to baseline levels, plasma concentration is an unreliable proxy of chronic stress. Utilizing an alternative method, such as neutrophil:lymphocyte (N:L) ratio, as a proxy of chronic stress may provide a metric that can be rapidly assessed and related back to microhabitat characteristics for management purposes. As a long-lived, specialist facing population declines, the Hellbender (Cryptobranchus alleganiensis) is an ideal species for applying N:L ratios to assess population viability. This study utilized chronic stress in relation to water quality and microhabitat among wild Hellbender populations to assess the implication of this method. Blood samples were taken from Hellbenders across a habitat quality gradient in Tennessee and microhabitat was assessed. Neutrophil:Lymphocyte ratios will be assessed to determine the relation between quality of microhabitat and presence of chronic stress. Detecting the Presence and Abundance of Streamside Salamanders (Ambystoma barbouri) in Middle Tennessee Using Environmental DNA Nicole Witzel, Ali Taheri, and William B. Sutton Tennessee State University, 3500 John A Merritt Blvd, Nashville, TN Amphibians represent the vertebrate taxa that has experienced the greatest declines globally. Declines tend to occur in species with geographically-isolated and/or fragmented populations. This is specifically true for the Streamside Salamander (Ambystoma barbouri), an Ambystomatid salamander that occurs in Middle Tennessee. This salamander species is active during winter and spring months when it emerges to breed in low-order, ephemeral streams. As these animals are cryptic and only surface-active for several months, they can be difficult to detect using traditional survey methods. Surveys that target environmental DNA (edna) in the form of DNA sloughed into their aquatic environment could provide an effective method for detecting the presence of this species. Water samples were collected at 50 meter stretches of 17 streams across the A. barbouri range once per

15 month for 6 months (December May 2017). Stream segments were searched for all life-stages of salamanders using rock-turning and visual surveys. We used real-time PCR to quantify DNA amounts using an A. barbouri species-specific primer. We used Generalized Linear Mixed Models to evaluate relationships between A. barbouri adult, larval, and egg counts and biomass at each site. In addition, we evaluated the change in edna amounts across the active season of A. barbouri. Primary outputs from this study include a replicable edna approach to identify A. barbouri populations in Tennessee and sampling guidelines for appropriate times to collect edna survey data for A. barbouri. This information will provide a method which can be used by wildlife agencies to further the knowledge and conservation of this species. Herpetofauna and Management Response of Reptile and Amphibian Communities to Prescribed Fire in an Oak/Hickory Forest. Steven J. Hromada, C.A.F. Howey, C.M. Gienger Department of Biology, Austin Peay State University, Clarksville, TN, Prescribed fire can have important impacts on ecosystems, including direct effects in the form of injury and mortality to individuals, or indirect, in the form of changes to preferred resources available within the environment. Changes in vegetation structure (such as a decrease in canopy cover or an increase in herbaceous cover) from prescribed fire can increase availability of preferred microhabitats for some species while reducing the availability of preferred microhabitats for others. We examined the responses of herpetofaunal communities to a prescribed fire regime in an Oak/Hickory forest in western KY. Prescribed fires were applied to a 1000-ha area twice. Herpetofaunal communities were sampled using drift fences and artificial cover object arrays, and habitat attributes were sampled via transects in burned and unburned areas. Reptile and amphibian species richness did not differ between treatments, though we found higher reptile diversity in burned areas. Reptile community structure correlated with differences in vegetation structure, suggesting a connection between prescribed fire and community response. Amphibian community structure differed on yearly, indicating that sampled communities were likely influenced by factors other than treatment. We found differences in taxaspecific abundances with snakes, Coluber constrictor, Diadophis punctatus, Sceloporus undulatus, and Lithobates clamitans higher in burned areas while amphibians, Anaxyrus sp., Notophthalmus viridescens, and Plethodon glutinosus higher in control areas. Differences in amphibian taxa abundances may have been influenced by availability of aquatic breeding habitat relative to sampled areas. Our results suggest that the habitat changes caused by prescribed fire can have indirect impacts on reptile abundances and diversity, while not impacting overall amphibian communities.

16 Reptiles Behavioral and physiological responses to simulated predator induced stress in the Eastern Box Turtle, Terrapene carolina carolina. Victoria Lay and Matthew Klukowski Department of Biology, Middle Tennessee State University, Murfreesboro, TN The behavioral and physiological responses of eastern box turtles, Terrapene carolina carolina, to a simulated predatory encounter were measured. Wild box turtles were randomly assigned to a stimulus intensity group, which consisted of observation (control), a visual stimulus, a poke on the leg, or a pinch on the leg before being flipped. Behavior in response to the stimulus was observed from a distance for at least 5 minutes. Immediately after the observation period, turtles were captured and had a small blood sample taken, after which a pinch gauge was used to measure the shell s closing force. The results indicate that the stronger the simulated predatory stimulus, the stronger the behavioral responses, with the turtles taking more advantage of their fully closable shell at the more intense stimulus groups. However, stimulus intensity did not affect the plasma corticosterone or plasma lactate response; this may be because the turtles perceived even our most intense stimulus (pinch-flip) as mild or because of seasonal effects. Upon capture and during handling almost all turtles withdrew into their shells and closed their anterior plastron, although fewer actually closed their posterior plastron, and a few engaged in other behaviors such as biting, urinating, and air walking. There appeared to be a correlation between box turtle size and pinch strength; however, the relatively large size of the pinch gauge may have put smaller turtles at a disadvantage compared to larger turtles because of excessive stretching of their closing muscles. The results of this study indicate that in spite of their fully closable shell, box turtles exhibit a complex, graded response to perceived threats. This strongly suggests that full closure of the shell is likely to be costly and occurs only in response to the most intense stimuli. Preliminary Results from Alligator Snapping Turtles (Macrochelys Temminckii) Surveys in Two West Tennessee River Drainages Saidee J. Hyder 1, Joshua R. Ennen 2, Dustin F. Garig II 1, Rob L. Colvin 3, Jeremy S. Dennison 3, Andrew J. Feltmann 1, Madison A. Herrboldt 1, Caitlin M. Weible 1, Taylor R. Simmonds 1, and Jon M. Davenport 1 1 Department of Biology, Southeast Missouri State University, Cape Girardeau, MO; 2 Tennessee Aquarium Conservation Institute, Chattanooga, TN; 3 Tennessee Wildlife Resources Agency, Jackson, TN The Alligator Snapping Turtle (Macrochelys temminckii) is the largest freshwater turtle in the United States and is distributed within the Mississippi and Gulf Coast river drainages reaching as far north as Iowa. ASTs are apex predators in these drainages, but have experienced dramatic declines throughout their range due to overexploitation. Despite the type locality from West TN, very little distribution and demographic information is available from this part of their range. For conservation measures from years 1992 to 2005, Tennessee Wildlife Resources Agency (TWRA) released 444 ASTs into West and Middle Tennessee river drainages. Unfortunately, no data is available assessing the success of those introductions along with the current status of the AST. Previously, our initial work investigated the distribution and abundance of ASTs in two West TN drainages, the Wolf and Hatchie Rivers. This year

17 we began surveying two new West TN drainages, the Obion and Forked Deer Rivers. During the spring and summer of 2017, we surveyed 15 sites with baited hoop net arrays of various sizes. The various sizes allowed us to determine what age classes if any were present. During this sampling period, we did not detect any new populations of ASTs. In the future, we plan to revisit all four river drainages to verify the presence or absence of ASTs. Distribution and Status of the Western Pygmy Rattlesnake (Sistrurus miliarius streckeri) in Tennessee Shawn Snyder and William Sutton Tennessee State University, 3500 John A Merritt Blvd, Nashville, TN Globally, reptile populations are declining at a rate quicker than most other vertebrates. A host of anthropogenic and environmental pressures such as habitat destruction, climate change, emerging pathogens, and pollution have been identified as causes behind these declines. However, when a species occurs rarely across the landscape, it is often difficult to determine population status. The Western Pygmy Rattlesnake (S. miliarius streckeri) occurs in a narrow range in west-central Tennessee along the Tennessee River drainages and Western Highland Rim. Little is known about the spatial ecology or habitat requirements of this species in Tennessee where it is listed as State Threatened. Previous studies on this species have reinforced the rarity of this species in the state with as little as 30 confirmed occurrences coming in the last 30 years. Our primary research objectives are to evaluate the distribution of the Western Pygmy Rattlesnake in Tennessee by using a variety of field-based survey methods and species distribution modeling techniques. Our preliminary habitat suitability model suggests most of the suitable habitat for pygmy rattlesnakes in Tennessee occurs on the East side of the Tennessee River and is predominantly associated with riverine and stream systems. A Comparative Study of Stress Physiology in the Common Watersnake (Nerodia sipedon), the Diamondback Watersnake (Nerodia rhombifer), and the Queen Snake (Regina septemvittata) Louise McCallie Department of Biology, Middle Tennessee State University, Murfreesboro, TN This field project examined corticosterone levels in three Natricines: the common watersnake Nerodia sipedon, the diamondback watersnake Nerodia rhombifer, and the queen snake Regina septemvittata. Corticosterone is the primary stress hormone in reptiles, and chronically elevated levels have been found to inhibit immune function along with feeding, locomotion, and reproductive behavior. We drew blood from the three species immediately upon capture and after thirty minutes of acute stress to measure corticosterone and will look at the levels in relation to sex, body condition, and levels of testosterone/estradiol. We will also compare corticosterone levels among the three species to see if these closely related snakes with distinctive life history traits have discernable differences.

18 The Cutaneous Snake Microbiome Correlates with Ophidiomyces ophiodiicola, the Causative Agent of Snake Fungal Disease Jacob E. Leys 1, Danny L. Bryan 2, Vincent A. Cobb 3, Donald M. Walker 1 1 Tennessee Technological University, 1100 N. Dixie Ave, Cookeville, TN 38505; 2 Cumberland University, 1 Cumberland Square, Lebanon, TN 37087; 3 Middle State Tennessee University, 1301 E Main St, Murfreesboro, TN The causative agent of snake fungal disease (SFD), Ophidiomyces ophiodiicola, is an emerging fungal pathogen threatening North American snake populations. Previous studies have elucidated clinical presentation, but the life history of this species remains largely obfuscated. This study aims to characterize the cutaneous snake microbiome to determine host-microbiome-pathogen effects that result from the presence or absence of O. ophioodiicola. We hypothesized that the cutaneous microbiome of snakes differs by species, geographic region, and presence of O. ophiodiicola. We also hypothesized that O. ophiodiicola persists in the environment and opportunistically infects susceptible snake species. Cutaneous snake swabs (n=116), soil samples (n=40), and water filtration samples (n=20) were collected between late spring and early fall of 2015 and 2016 across seven states in the Southern U.S.; Alabama, Arizona, Arkansas, Florida, Georgia, Tennessee, and Texas. These samples were subsequently subjected to DNA extraction and quantitative PCR to determine the presence/absence and fungal load of O. ophiodiicola. Additionally, high-throughput sequencing was utilized to characterize the bacterial microbiome of cutaneous snake swabs (n=83) and environmental samples (n=10). O. ophiodiicola was detected in 21 of 116 snake individuals, in two of 40 soil samples, and zero of 20 water filtration samples. PERMANOVA analysis indicates slight statistical differences (Pseudo- FPERMANOVA = , P < 0.05) between the assemblages of cutaneous bacterial communities where O. ophiodiicola is present rather than absent, and RELATE indicates no relationship (ρ = 0.049, P > 0.05) between compared resemblance matrices of fungal load and community assemblage. Abstracts Professional Presentations Common Mudpuppy (Necturus maculosus) Not Common in Southern Appalachian Streams. Kirsten A. Hecht 1,2, Lori A. Williams 3, Stephen K. Nelson 4, Thomas Floyd 5, & Jeremiah L. Cronin 2 1 School of Natural Resources and Environment, University of Florida, Gainesville, FL 32611; 2 Florida Museum of Natural History, University of Florida, Gainesville, FL 32611; 3 North Carolina Wildlife Resources Commission, 177 Mountain Laurel Lane, Fletcher, North Carolina 28732; 4 Zoo Knoxville, 3500 Knoxville Zoo Dr, Knoxville, TN 37914; 5 Georgia Department of Natural Resources While the Common Mudpuppy (Necturus maculosus) is considered common throughout most of its North American range there is relatively little data to support the claim, especially within Southern Appalachia. From , we sampled streams in eastern Tennessee, northern Georgia, and western North Carolina using several methods to determine presence/absence of Common Mudpuppies

19 in lotic systems as well as to collect information on basic life history traits and habitat use. While locally abundant in some drainages, N. maculosus was absent from many waterways (especially in North Carolina and Georgia) and overall we had low catch per unit effort (CPUE) and trapping success. Both immature and mature Mudpuppies were found throughout the surveyed areas. We found the majority of animals under rocks in runs and pools. Overall our preliminary research demonstrates that N. maculosus is potentially not common in Southern Appalachia and warrants future research. Specific research needs include, but are not limited to, continued distributional surveys, habitat use (specifically, micro-habitat and potential use impounded waters and deeper riverine pools), diet, density, and movement. What s Temperature Got to Do with It? Climate Change Interferes With Stream Amphibian Community Stability Kristen Cecala 1, Josh Ennen 2, Jon Davenport 3 1 University of the South; 2 Tennessee Aquarium Conservation Institute; 3 Southeast Missouri State University Temperature changes due to climate change are directly altering organismal performance and distributions, but the mechanisms behind these shifts require additional attention. Because small aquatic ectotherms are proposed to perform better at future climates, it is possible that competitive interactions in size-structured communities may also shift affecting future performance and persistence. We have evaluated the interactive effects of temperature and interspecies competition in size-structured stream salamander communities. Temperature altered morphological growth patterns. Interference competition appears to occur between species of different sizes with larger species excluding smaller species from aquatic refuge. At elevated temperatures, the larger species tended to be negatively affected whereas the smaller species had neutral or even positive responses that resulted in declining strength of competitive interactions and even higher rates of co-occurrence. Disruptions in competitive interactions within these communities affect habitat use patterns and may decrease selection for character displacement among species. Determining how biotic interactions change along abiotic gradients is necessary to predict the future long-term stability of current communities. I m Not Your Steppin Stone: Recent Winter Finds and Current Distribution of the Streamside Salamander, Ambystoma barbouri, in Tennessee. David Ian Withers Natural Heritage Program, Division of Natural Areas, Tennessee Department of Environment and Conservation An environmental consultant s 2015 find of a juvenile Ambystoma barbouri from Nolensville, TN, inspired researchers to redouble efforts to find new populations in under-surveyed portions of middle Tennessee. Surveys produced 2 new county records, 2 previously undocumented drainages, 7 additional stream systems, and 20 newly identified breeding populations. The Streamside Salamander, listed by the TWRA as in Need of Management, is believed extant from 41 locations and extirpated from at least one historic and one contemporary site. Though ostensibly protected by public lands in two counties, it remains a conservation concern in the rapidly urbanizing lands in and around

20 Nashville. Many new populations have been documented only in response to development permit review or related water quality investigations. As a winter breeder, this species historically is overlooked by field surveys conducted outside of the breeding season, and easily is missed by winter surveys if not specifically targeted. Abstracts - Poster Presentations Aggressive Interspecific Interactions Within Stream Salamander Communities Increase at Warmer Temperatures Kailey E. Bissell and Kristen K. Cecala Department of Biology, University of the South, Sewanee, TN Stream salamander communities are maintained by size-structured interactions among species. Warming temperatures are predicted to improve performance by smaller species, and recent studies have demonstrated reduced competitive strengths among species within this community at warmer temperatures. Temperature is also known to influence how organisms respond to stimuli within their environment, and these behavioral differences could contribute to shifting interactions at warmer temperatures. Therefore, we tested if aggressive, defensive, or flight behaviors among individuals of three different congeners varied with temperature. Short term behavioral observations of interacting species pairs were conducted at 15, 20, and 25C. We observed 3.8 times more aggressive interactions at 25C than at 15 or 20C, which had similar frequencies. Species identity also affected the frequency of interactions with the most aggressive behaviors occurring between the largest and smallest species whereas the highest flight frequency occurred between the largest and medium sized species. Defensive behaviors were most frequent in trials with high rates of aggression. Temperature appears to increase aggressive behaviors, but size and potentially frequency of interactions among species appear to determine outcomes of interactions among species. Morphological Plasticity in Larval Hylid Tree Frogs as a Function of Abiotic Factors Cory Blackwelder Department of Biology, Tennessee Tech University, Cookeville TN, Frogs and tadpoles can accommodate large degrees of morphological plasticity. However, most studies have only been performed on changes due to predator cues. Little is known about how abiotic variables, such as flow, impact the morphology of these animals. H. cinerea were collected during amplexus and the eggs were housed into two tanks that were set up with no flow conditions, and two that were set up with flow. Multiple clutches of H. chrysoscelis were collected and were distributed among one flow and one non-flow tank. Approximately 30 individuals were placed in each tank. H. cinerea exposed to flow were significantly more laterally compressed. H. chrysoscelis exposed to flow showed significantly more elongation.

21 Long-term nest site fidelity in some male green salamanders, Aneides aeneus Paul Cupp Eastern Kentucky University, Department of Biological Sciences Previous studies indicate that male and female Aneides aeneus exhibit site fidelity from one year to the next in their use of rock crevices, including breeding crevices. The present study examines the use of breeding crevices by A. aeneus males at field sites in SE KY over a 25 year period. Crevices in specific rock outcrops were monitored for the presence or absence of males prior to or during the deposition of eggs by females during the summer breeding period. Of the visible rock crevices with A. aeneus present, only a relatively few had brooding females with eggs. These crevices were often used every year or in alternate years over several years to brood eggs and hatchlings. Long-term site fidelity was shown by males (n=6) over a range of 4-13 years during which individual males were found in the same crevices in subsequent years at about the same time of year. Nest site fidelity by males may influence the continued use of these crevices. These crevices must have properties that result in their selection by males (and females?) as breeding sites. Going Out on a Limb: Scaling of Terrestrial and Arboreal Defensive Strikes by Ratsnakes Michael C. Fulbright 1, David A. Penning 2, and Cory J. Hillard 1 1 Department of Biology, University of Louisiana at Lafayette, Lafayette, LA, 70504, ; 2 Department of Biology and Environmental Health, Missouri Southern State University, Joplin, MO, Ratsnakes are semi-arboreal snakes that must dispel predators in both terrestrial and arboreal environments. We measured defensive terrestrial and arboreal strikes to determine if snakes modulate their strike performance relative to the habitat they occupy. To determine the scaling of striking performance, snakes were measured from an ontogenetic range of body sizes. In paired analyses terrestrial strikes were significantly higher in maximum velocity, strike duration, and strike distance than perched strikes (p < 0.01 for all) while striking performance scales isometrically with body size. Performance of snakes in arboreal habitats is likely confounded by the need to use a portion of the posterior part of the body to maintain contact with limbs and prevent falling. New county records from Tennessee s Cumberland Plateau Matt Grisnik Department of Biology, Tennessee Tech University, Cookeville TN, Tennessee encompasses one of the most biodiverse regions in the United States, and is the fourth most diverse state for Amphibians. The monitoring of amphibians and reptiles plays an important role in habitat management. Amphibians and reptiles are often considered indicator species, their presence indicates the quality of the habitat due to their susceptibility to environmental degradation. Prolonged monitoring of herpetofaua within protected areas allows for early detection of changes in environmental quality, however the first step in a long term monitoring effort is to establish presence

22 of species within an area, this is often difficult due to the cryptic nature of reptiles and amphibians. Throughout February to September 2017 I have surveyed State parks and Wildlife management areas within eight counties and established 23 county records, including for two species of conservation concern. Host Taxonomy and Geography Affect the Structure of the Salamander Cutaneous Microbiome Aubree J. Hill, John M. Griffith, Gabrielle Russell, Fantasia Erdman, and Donald M. Walker Tennessee Technological University Department of Biology Batrachochytrium dendrobatidis (Bd) is a dermatophytic pathogen that has caused enigmatic declines in amphibian populations worldwide. Salamander populations in the eastern US persist in the presence of Bd with surprisingly low rates of infection, despite relatively high rates in co-occurring anuran populations. This may be due to cutaneous probiotic defenses or other innate and/or adaptive immune responses. We conducted a foundational study to better understand effects of microbial community dynamics on pathogenicity of Bd, and to identify any probiotic members of the salamander skin microbiome. Objectives were to (1) use high-throughput DNA sequencing to characterize structure of the skin microbiome, (2) isolate bacteria from skin swabs, and (3) identify bacterial isolates that inhibit Bd. During May and July 2016, salamanders from the genera Plethodon (n=26), Eurycea (n=20), and Desmognathus (n=36) were captured from nine sites in Tennessee, and skin swab samples were obtained. From these swabs, a total of 476 bacterial colonies were isolated in pure culture. Thus far, 40 isolates have been challenged against Bd on agar plates, and five formed measurable zones of inhibition. These candidate probiotics will be identified through DNA sequencing. Additionally, we used high-throughput DNA sequencing techniques and bioinformatics analyses to define the composition and structure of the resident skin microbial community across salamander taxa, season, and ecoregion. Results indicate that both geography and host taxon significantly affect the structure of the microbiome, but seasonality does not. During future research, we will identify dominant or keystone symbionts within the microbiome of each genus and ecoregion, and we will identify probiotic members of the microbiome. The isolation and challenge phases are ongoing, and we anticipate discovery of additional Bd inhibitors during future trials. These inhibitors will be identified, and their dominance in the microbiome will be evaluated to determine candidacy for in vivo treatment trials. Results may directly impact future conservation efforts for valuable North American wildlife. Assessing Biodiversity and Leech (Hirudinea) Parasitism of Semi-aquatic Turtles in an Urbanized Wetland in Nashville, Tennessee Laura Horton, Nicole Witzel, Brittany Hogan, Shawn Snyder, Jeronimo Silva, Brent C. Newman, and William Sutton Tennessee State University, Nashville, Tennessee Aquatic turtles inhabiting a palustrine emergent wetland adjacent to Tennessee State University in Nashville, Tennessee were examined for leeches as a component of a multi-year mark and recapture

23 study. Leeches can serve as bioindicators in aquatic environments and are useful for determining the biological condition of aquatic ecosystems via species richness indices between parasite and host. Aquatic funnel traps were baited and deployed weekly from June September 2016 to sample aquatic turtles. Each captured turtle was weighed, measured, sexed, marked, and examined for external leeches. All located leeches were removed and preserved for later identification. Turtle species captured during this study, included the Common Snapping Turtle (Chelydra serpentina), Stinkpot (Sternotherus odoratus), Spiny Softshell Turtle (Apalone spinifera), and the Red-eared Slider (Trachemys scripta elegans). The leeches that were collected from the turtles represented 3 different genera under phylum Hirudinea, including Batracobdella, Placobdella, and Helobdella, representing 7 different species. Collectively, these data are being be used to understand parasite-host relationships and how patterns in leech abundance and richness vary throughout the active season. If you give a homeschooler a frog pond... Alli Metler and Donna D. Metler Mount Parnassus Academy, Bartlett, TN Homeschoolers report significant struggles with science education, and often seek out outside educational experiences. This project reviews the participation demographics and comments of 96 families who chose to participate in a backyard frog habitat observation project over a 2 year period, and examines implications for involving this population in field research and outreach. A survey of ticks (Acari: Ixodida) parasitizing reptiles from managed pinehardwood forests of northwestern Alabama, USA Brent C. Newman 1, William B. Sutton 1, T.J. Haltigan 2, and Yong Wang 2 1 Tennessee State University, Department of Agricultural and Environmental Sciences, Nashville, TN 37209; 2 Alabama A&M University, Department of Natural Resources and Environmental Sciences, Normal, AL Reptiles and amphibians were trapped and examined for ticks from May through August 2017 in 18 total forest stands treated with a variety of management treatments, including two levels of prescribed burning and three levels of thinning in the William B. Bankhead National Forest located in northwestern Alabama. Two tick species (Ixodes scapularis and Amblyomma americanum) were collected from six species of lizards (Plestiodon fasciatus, Plestiodon laticeps, Plestiodon inexpectatus, Anolis carolinensis, Sceloporus undulatus, and Scincella lateralis). Lizards parasitized by ticks occurred at all treatment sites, however, lizards parasitized by ticks were most abundant in prescribed burning and thin with prescribed burning treatments. Ticks were most commonly encountered on Plestiodon fasciatus and Plestiodon laticeps with peak infestations occurring in July. Our results indicate that lizards may play an important role in the life cycle of ticks of which forest management techniques may also influence this parasite-host relationship.

24 Metabolic Response to Predation Risk in Agkistrodon contortrix Matthew Pierson and C.M. Gienger Department of Biology and Center of Excellence for Field Biology, Austin Peay State University, Clarksville, TN Direct interactions between predators and prey can have fatal consequences, typically resulting in the predator consuming the prey. Non-fatal indirect interactions also have consequences on organisms by modifying physiology and behavior. This study examines how a snake s metabolic rate (measured by oxygen consumption) responds to the chemical scent cue of a known predator to Copperheads, the coyote. This gives us insight into how the snake perceives the predator cue without physical contact giving us insight into the snakes internal response to chemical cues in the environment. Overall, the responses to the chemical stimulus (coyote urine) were very small among our study group. Among the five individuals tested only one showed a significant response to chemical stimulus. Even after adjusting for mass specific VO2, there was very little change in measurements. Other studies show that snakes respond to various chemical cues, none have demonstrated a response metabolically to predator chemical cues. With such a small sample size, our priority was to first develop a successful method of quantifying such a response among snakes. Preliminary investigation of mercury concentrations in three freshwater turtle species of West Tennessee Taylor R. Simmonds 1, Saidee J. Hyder 1, Madison A. Herrboldt 1, Caitlin M. Weible 1, Dustin F. Garig II 1, Andrew J. Feltman 1, Rob L. Colvin 3, Jeremy S. Dennison 3, Rebecca L. Brasso 1, Joshua R. Ennen 2, and Jon M. Davenport 1 1 Department of Biology, Southeast Missouri State University, Cape Girardeau, MO; 2 Tennessee Aquarium Conservation Institute, Chattanooga, TN; 3 Tennessee Wildlife Resources Agency, Jackson, TN Mercury is a heavy metal that bioaccumulates in a variety of organisms, causing physical and neurological effects. Turtles are one group of organisms that bioaccumulate mercury in their tissues and some species (e.g., Chelydra serpentina) are used as biomonitors of mercury. In general, turtles are model organisms for studying bioaccumulation of heavy metals largely because of their longevity. In this study, we focused on three species of turtles, the red-eared slider (Trachemys scripta), the common snapping turtle (Chelydra serpentina), and the alligator snapping turtle (Macrochelys temminckii), to determine mercury concentrations in West Tennessee turtles. To achieve this goal, a total of thirteen sites were sampled across West Tennessee from March to July Blood and toenails were taken from the back feet of each captured turtle and analyzed in a Nippon MA-3000 Direct Mercury (Hg) Analyzer. We found that all species sampled had a significant level of mercury in their toenails, and there was significant difference in mercury levels between species. Secondarily, we found that some turtle species have potentially higher mercury levels than the EPA recommended consumption level for freshwater fish.

25 Development Of An edna Protocol to Detect and Quantify Streamside Salamanders (Ambystoma barbouri) in Low-Order Streams of Middle Tennessee Nicole Witzel, Ali Taheri, and William B. Sutton Tennessee State University, 3500 John A Merritt Blvd, Nashville, TN The Streamside Salamander (Ambystoma barbouri) is an Ambystomatid salamander that occurs in Middle Tennessee where it is geographically isolated. This salamander species is active for a few months during winter and spring months when it emerges to breed in low-order, ephemeral streams. As these animals are cryptic and only surface-active for several months, they can be difficult to detect using traditional survey methods. Surveys that target environmental DNA (edna) in the form of sloughed skin, sperm, and eggs provide a potentially effective method for detecting the presence of this species. However, before this method can be used, it is essential to develop species-specific genetic primers that will correctly identify presence and absence of the target species. An initial objective of this study is to identify a primer that is specific to and will successfully amplify only the DNA of A. barbouri without amplifying DNA of congeners. Primers were selected by choosing A. barbouri cytochrome B mitochondrial DNA segments with base pairs that differed from A. texanum, a closely related species. These were used to amplify A. barbouri DNA and tested for specificity among other Ambystomatid congeners in Tennessee. Following initial tests of specificity, we will develop a quantitative PCR approach to evaluate the quantity of environmental DNA in stream water samples. The long-term goal of this study is to provide a replicable edna approach to identify A. barbouri populations in Tennessee. This information will provide a method which can be used to further the knowledge and conservation of the species. Tertiary wastewater treatment wetlands provide suitable habitat for anurans Emma F. Zeitler, Erin Gill, Ansley Murphy, Deborah McGrath, Kristen Cecala Department of Biology, University of the South, Sewanee, TN Constructed wetlands are becoming important pieces of environmental technology because they can serve as a tertiary wastewater treatment, removing nutrients and pollutants that remain in the water after the primary and secondary treatments. It is hypothesized that the constructed wetlands may function similar to natural wetlands in their ability to support plant and wildlife communities, but remaining pharmaceuticals have been demonstrated to cause abnormalities in anurans using wastewater treatment wetlands. Our objective was to compare the morphology and occupancy of anuran communities between a newly constructed tertiary treatment wetland (TTW) and nearby rain filled ponds. We quantified calling behavior, tadpole and adult size and body condition among 5 unimpacted ponds, a wastewater treatment lagoon, and the TTW. We found that the majority of the anuran community colonized the TTW within 12 months of its completion, and successful reproduction was observed for 50% of species observed at the TTW quantified by tadpole presence. Generally, adult frogs and tadpoles were larger as quantified by length and body condition when they were found at the TTW relative to unimpacted ponds. TTWs appear to provide habitat for successful

26 reproduction of anurans, but carryover effects on offspring have yet to be quantified to determine the long-term viability of populations using TTWs.

27 Annual Tennessee Herpetological Field Trip This year s annual field trip will be held at Flat Rock Cedar Glades and Barrens State Natural Area. This state natural area is located just a short distance from Middle Tennessee State University and will provide a variety of habitats to search for amphibians and reptiles. The field trip will be led by Dr. Vincent Cobb of Middle Tennessee State University. Dr. Cobb and his graduate students recently completed research assessing snake fungal disease and its impacts on the population of Timber Rattlesnakes (Crotalus horridus) located on the state natural area. Flat Rock Cedar Glades and Barrens is an 846 acre state natural area which supports a variety of habitats, including oak barrens, open grassland barrens, and cedar glades. The state natural area supports a diverse number of plants, several of which are federally endangered and state listed. Despite being located just a few miles from Middle Tennessee State University in the geographic center of the Central Basin, Flat Rock Cedar Glades and Barrens State Natural Area is one of Tennessee s most ecologically significant state natural areas. In recent years, Flat Rock Cedar Glades and Barrens State Natural Area has experienced an unusual form of habitat modification that has and will continue to impact the flora and fauna inhabiting the area. Removing, stacking, and altering the exposed rocks has increased significantly. This behavior displayed by visitor s changes the habitat available to reptiles and amphibians, damages and or alters habitat for plants, and can lead to the unintended loss of rare, threatened and endangered plants. We have been given the opportunity to search for amphibians and reptiles based on the confidence of the Tennessee Department of Environment and Conservation in our members to minimize our impacts flipping rocks may have on the flora and fauna of the state natural area. All members are expected to return rocks that have been flipped and moved in the search of herpetofauna in the exact or as close to the exact spot where rocks were found prior to being moved to minimize impacts to the habitat and maintain the confidence of the managing state agency in our members.

28 Head North on N. Rutherford Blvd. toward MTSU Blvd for 07. Miles. Turn right onto Halls Hill Pike. Stay on Halls Hill Pike for 3.1 miles and turn right onto Factory Road. Stay on Factory road for 0.7 miles and the entrance to the parking area for Flat Rock Cedar Glade and Barrens State Natural Area will be on your right. ( , )