Twenty-first Annual Meeting of the Tennessee Herpetological Society
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Twenty-First Annual Meeting of the Tennessee Herpetological Society
Horse Creek Wildlife Sanctuary and Animal Refuge Savannah, TN Hosted By Freed-Hardeman University Thursday PM Location: Rockhouse Lodge (use the Rockhouse Lodge entrance) 35°06’41.11”N; 88°09’38.59”W 1:00 Welcome1:15 Joshua R. Ennen Keynote : The where and why of North American turtles: anexplanation of species richness and composition patterns. Ennen, J.R., W. Matamoros, S.C. Hazzard, M. Agha, and J.E. Lovich 2:15 Break2:30 Metler, A. If you build it, will they come? Temporary backyard ponds as analternative to traditional frog metamorphosis projects 2:50 Lillard, C.E., D.L. Miller, J.L. Howard, B.S. Wilson, R.T. Jackson, J.R. Asper, R.P.Wilkes, L. Rollins-Smith, R.D. Hill, Y. Geng, W.B. Sutton, B. Reeves, S. Reinsch, D. McGinnity, and M.J. Gray. Pathogens vs. pesticides: the threat to eastern hellbenders 3:10 Carter, E.D., M.J. Gray, J.A. Spatz, and D.L. Miller. Rapid transmission ofranavirus: role of direct contact 3:30 Spatz, J.A., M.J. Gray, E.D. Carter, and D.L. Miller. Poor biosecurity could lead todisease outbreaks in amphibian populations 3:50 Kirkpatrick, S.J. and V.A. Cobb. Does digestion affect thermoregulation infreeranging timber rattlesnakes ( Crotalus horridus)?4:10 Flaherty, J.P. and C.M. Gienger. Habitat selection and movement patterns ofcopperheads ( Agkistrodon contortrix) in fire-altered landscapes4:30 Hamed, K. and K. Cecala. Proposal for a journal published by THS5:00 Business Meeting6:00 Dinner Catered7:00-8:30 Lewis Memorial Scholarship Auction8:30 Field TripFriday AM 8:30 Colvin, R. An update of Alligator Snapping Turtle, Macrochelys temminckii,surveys in West Tennessee and potential future research 8:50 Nelson, S., B. Fitzpatrick, M. Niemiller, and M. Ogle. Evidence of a new species ofmudpuppy in Tennessee 9:10 Vannatta, J. Demographic characteristics, incidence of ranavirus infection, andcorticosterone levels in the eastern box turtle, Terrapene carolina carolina, in asuburban wetlands habitat of Middle Tennessee 9:30 Break9:50 Ennen, J.R., J.M. Davenport, and K.F. Alford. Interspecific competition amongheadwater stream vertebrates 10:10 Walker, D.M., S.P. Graham, C. Kelehear, C. Camp, A.J. Hill, M. Edelbrock, J.Rheubert, and J. Wooten. A next-generation sequencing approach to assess the diversity of the skin and gut microbiota in the slimy salamander ( Plethodonglutinosus ) species complex, with an emphasis on gastrointestinal fungal pathogens10:30 Ogle, C. An update of amphibian and reptile surveys on wildlife management areaswithin TWRA’s administrative region 4. 10:50 Barton, L.J., B.P. Butterfield, and J.B. Hauge. A preliminary survey of anuransfrom Laguna Del Lagarto Lodge, Boca Tapada, Costa Rica 11:10 Butterfield, B.P. and L.J. Barton. Amphibians and reptiles of Horse Creek with anemphasis on the turtle community Poster Burton, E., K. Hudson, and T. Blanchard. Differences in habitat use amongspecies of aquatic snakes in the Reelfoot Lake area of Northwestern Tennessee Poster Walker, D.M., S.P. Graham, C. Kelehear, C. Camp, A.J. Hill, M. Edelbrock, J.Rheubert, and J. Wooten. A next-generation sequencing approach to assess the diversity of the skin and gut microbiota in the slimy salamander ( Plethodonglutinosus ) species complex, with an emphasis on gastrointestinal fungal pathogens11:30 Lunch12:30 Field Trip
Abstracts A Preliminary Survey of Anurans from Laguna Del Lagarto Lodge, Boca Tapada, Costa Rica Lee J. Barton 1, Brian P. Butterfield1, and J. Brian Hauge21 Freed-Hardeman University, 2Peninsula CollegeThe number of anurans species found in Mesoamerica is astounding. However, our understanding of the basic biology of many of these species is incomplete, even in many well-studied regions. Here we report preliminary results of an ongoing survey of the anurans of La Laguna Del Lagarto Lodge, Boca Tapada, Costa Rica. To date, we have documented 20 species at this site. Determining the geographic distribution of species is a necessary step for the successful development and implementation of conservation plans designed to protect biodiversity. ________________________________________________________________________ Differences in Habitat Use Among Species of Aquatic Snakes in the Reelfoot Lake Area of Northwestern Tennessee Erica Burton, Kelsey Hudson, and Tom Blanchard University of Tennessee at Martin Five species of water snakes occur in Tennessee and all of them are commonly observed in the Reelfoot Lake area. Although several studies of habitat use in these animals have been published, few address large-scale differences in habitat use among them in regions where they occur together. Published information on water snakes in Tennessee suggests that all five species are habitat generalists and regularly occur in cypress swamps, bayous, lakes, streams, and ditches. Personal observations of water snakes at Reelfoot Lake however, suggest a more restrictive habitat use for some. The purpose of this study was to determine if meso-scale habitat use was different among the five species of water snakes that occur at Reelfoot Lake. We collected data from May - October of 2014 and from March - April 2015. A variety of habitat types were searched by boat or on foot and snakes were captured if possible, identified, marked, and released. A location for each animal was recorded with a hand-held GPS unit and a variety of habitat variables were obtained. Our preliminary results suggest that some species of water snakes in the Reelfoot Lake area are generalist, whereas others are more restrictive in their habitat use. ________________________________________________________________________ Amphibians and Reptiles of Horse Creek with an Emphasis on the Turtle Community Brian P. Butterfield and Lee J. Barton Freed-Hardeman University We began surveying the herpetofauna of Horse Creek Wildlife Sanctuary and Animal Rescue in 2012. Our purposes for this ongoing study are to: 1) construct a species list, 2) establish a long-term turtle community study modeled after North American Freshwater Turtle Research Group (NAFTRG) studies, and 3) provide undergraduates with opportunities to conduct field research and enhance their educational experiences. We have documented 38 species that include 10 records for Hardin County. We have captured and marked 152 individual turtles representing eight species. The most abundant turtle species is T. scripta. Our data provide insight into amphibian and reptile diversitywithin a protected area and should serve as a baseline from which to assess assemblages from more disturbed systems. ________________________________________________________________________ Rapid Transmission of Ranavirus: Role of Direct Contact E. Davis Carter 1, Matthew J. Gray1, Jennifer A. Spatz1, and Debra L. Miller1,21 Center for Wildlife Health, University of Tennessee, Knoxville, TN, 2College ofVeterinary Medicine, University of Tennessee, Knoxville, TN Amphibian populations are declining globally in part due to viral pathogens from the genus Ranavirus. Ranaviruses can be transmitted through various routes includingcontact with contaminated water and soil, ingestion of infected tissue, and direct contact with infected individuals. Our objective was to determine how quickly an outbreak of ranavirus could occur through direct contact of infected individuals. We exposed five wood frog ( Lithobates sylvaticus) tadpoles to a lethal dose of ranavirus (103 PFU/mL) inwater for 24 hours. We touched each exposed individual for 1 sec to five unexposed L.sylvaticus and five unexposed Cope’s gray tree frog (Hyla chrysoscelis) tadpoles at 24,48, 72, and 96 hours. Significant transmission of ranavirus between wood frog tadpoles started at 48 hours, with the resulting mortality increasing at each consecutive time step (48 h: 36%, 72 h: 76%, 96 h: 92%). Transmission of ranavirus between wood frog and Cope’s gray tree frog tadpoles was less, with resulting mortality <36% among the postexposure contact durations. Our results indicate that outbreaks of ranavirus in wood frog populations can occur quickly through direct contact of conspecifics; however, spillover effects to other species may be dependent on other factors, such as virion concentrations increasing beyond lethal dose levels. _______________________________________________________________________ An update of Alligator Snapping Turtle, Macrochelys temminckii, Surveys in West Tennessee and Potential Future Research Rob Colvin Tennessee Wildlife Resources Agency From 2000 through 2006, the Tennessee Wildlife Resources Agency (TWRA) conducted an alligator snapping turtle ( Macrochelys temminckii) restoration project involving therelease of adult and juvenile turtles in various watersheds. Since then, there has been limited work done with this species regarding their distribution and population status in Tennessee. Between May and June 2015 we sampled two different locations in Tipton and Dyer Counties using baited hoop nets and we were able to locate a new population of alligator snapping turtles in Tipton County. Two additional records were also documented by other individuals in the Beech River and at Radnor Lake State Park. We plan to start a multi-year survey beginning in 2016 to gather distribution and population data to better understand the status of this species in Tennessee. ________________________________________________________________________ Interspecific Competition Among Headwater Stream Vertebrates Joshua R. Ennen *, Jon M. Davenport†, and Kathlina F. Alford*1 Tennessee Aquarium Conservation Institute, Chattanooga, Tennessee, United States ofAmerica. †Department of Biology, Southeast Missouri State University, Cape Girardeau,Missouri, United States of America. Species interactions, such as interspecific competition, often influence species distribution and abundance over space and time. Some stream vertebrates, such as salamanders, can reach high abundances in headwater systems and account for a disproportionate amount of total vertebrate biomass. However, the importance of stream salamanders in structuring these headwater systems is still understudied. We conducted an artificial stream experiment to evaluate interactions among three common vertebrates. We investigated the scaled mass growth of salamanders in the black-bellied dusky complex ( Desmognathus quadramaculatus/folkertsi) in the presence of two different fishspecies, common creek chub ( Semotilus atromaculatus) and Coosa darter (Etheostomacoosae ). There was no statistical significance (t = 0.71, df = 5, P = 0.51) between thecontrol and darter present treatments (mean with darters: 0.09±0.64 SE; mean with conspecifics: 0.47±0.10 SE). However, the body condition of salamanders (-0.25±0.26 SE) were significantly reduced ( t = -2.65, df = 6, P = 0.04) in the presence of the creekchubs suggesting an asymmetric competitive interaction between those two species. Our results suggest that asymmetric competition, and not solely predation, may play an important role in limiting the distribution and abundance of stream vertebrates in headwater streams. ________________________________________________________________________ The Where and Why of North American Turtles: An Explanation of Species Richness and Composition Patterns Joshua R. Ennen, Wilfredo Matamoros, Sarah C. Hazzard, Mickey Agha, and Jeffrey E. Lovich Tennessee Aquarium Conservation Institute, Tennessee Aquarium, 201 Chestnut Street, Chattanooga, TN, 37402 USA Species richness and composition is not uniformly distributed across the landscape, and these patterns are often associated with environmental gradients. In this study, we examine species richness and composition patterns within North America turtles and use 14 abiotic variables (8 climatic, 2 topographical, 2 stream variables, as well as latitude and longitude) to construct regression models fitted to spatial data that predict species richness for all turtles (n = 84), freshwater aquatic and semiaquatic turtles, turtles in the family Emydidae, and those in the Kinosternidae. Overall, species richness was positively related to maximum temperature of the warmest month (K), precipitation of the wettest month (mm), total stream length (km), and temperature seasonality. Species richness of aquatic and semiaquatic turtles was positively related with precipitation of the wettest month, total stream length, and temperature seasonality. Emydid species richness was only related (positively) to precipitation of the wettest month. Interestingly, only kinosternid richness was related (negatively) with latitude. Also, species richness within kinosternid was positively related to precipitation of the wettest month and total stream length. Given current and projected climate trends, especially alterations of temperature and precipitation patterns, species richness of North American turtles could be impacted. ________________________________________________________________________ Habitat Selection and Movement Patterns of Copperheads ( Agkistrodon contortrix)in Fire-altered Landscapes James P. Flaherty and C.M. Gienger Center of Excellence for Field Biology & Department of Biology Austin Peay State University, Clarksville, TN 37044 Fire can functionally alter habitat available to wildlife through alteration of structural and micro-climatic characteristics. Reptiles, due to their reliance on behavioral thermoregulation and crypsis, can be especially susceptible to these post-fire changes in habitat characteristics. The copperhead ( Agkistrodon contortrix) is a widely distributedsnake throughout the southeastern US, and is found in a variety of habitats. Previous studies suggest that copperheads decrease in abundance immediately after fire events, but to our knowledge there has been no work investigating this species’ habitat selection and movements in a fire-altered landscape. We tracked 11 adult male copperheads captured in burned and unburned sites at Land Between the Lakes National Recreation Area, KY using radio-telemetry. Copperheads were tracked up to three times per week during the active season (May-October) of 2014. Home ranges were calculated by minimum convex polygons (MCP) and fixed kernel density estimation (KDE). Movement patterns were assessed by measurement of straight-line distances between successive copperhead relocations. At each copperhead location, a suite of structural and environmental habitat variables were recorded. In order to measure habitat availability, each snake relocation point was paired with a randomly selected point. There was no significant difference between MCP home ranges and movement patterns of copperheads captured in burned and unburned sites. KDE home ranges were significantly larger in copperheads captured in unburned sites than those captured in burned sites. Among the available habitats, burned sites exhibited significantly greater habitat heterogeneity than unburned sites, but overall, copperheads selected habitats more similar to those available in burned sites than in unburned sites. ________________________________________________________________________ Proposal for a Journal Published by THS Kevin Hamed 1, Kristen Cecala21 Virginia Highlands Community College, 2Sewanee; University of the SouthWe propose the development of a peer-reviewed journal published by the Tennessee Herpetological Society. Many individuals are currently contributing to improving knowledge about Tennessee’s amphibians and reptiles. We recommend building an avenue to increase awareness of new discoveries and findings by and for any individual interested in Tennessee’s herpetofauna. An option successfully implemented by Virginia’s Herpetological Society is the journal Catesbeiana. They share short papers, the results of bioblitzes and other surveys, field notes, range extensions and new species localities, as well as society business. We propose that a journal published by the Tennessee Herpetological Society include the following components: 1) Survey Results – We encourage the publication of habitat types and speciesfound during herpetological surveys of private and public lands in Tennessee. 2) Field Notes – Publication of field observations of amphibians and reptiles. Thismay include observations of aspects of the natural history of a species as well as identification of new localities. We suggest conversations with Floyd Scott and William Redmond and TWRA to facilitate incorporation of these localities into state-wide databases. 3) Original Research Papers – Publication of results not published in other venues.4) Recent Study Reviews– Short descriptions of research published about thenatural history, distribution, and ecology of Tennessee’s herpetofauna in other venues. We encourage these pieces to be less than 250 words and be written for public audiences. 5) Biographies – To facilitate getting to know the diversity of people involved in theTennessee Herpetological Society, we encourage less than 500 word profiles highlighting the background and interests of our various members. 6) Minutes or summary of annual Tennessee Herpetological Society meetings.Survey results, field notes, and original research papers would all be peer reviewed and submitted electronically to an editor. We would encourage an online publication once or twice per year depending on popularity. Content could be grouped and published into a pdf file for posting on the THS website. ________________________________________________________________________ Does Digestion Affect Thermoregulation in Free-ranging Timber Rattlesnakes ( Crotalus horridus)?Sarah J. Kirkpatrick and Vincent A. Cobb Middle Tennessee State University For terrestrial ectotherms, increasing body temperature ( Tb) during digestion can facilitatelocalized biochemical reactions and consequently increase passage rate of food through the gut. Snakes benefit particularly from an increase in digestion rate; they typically undergo a relatively large increase in body mass after feeding, which can have a negative effect on mobility and defensive behavior. Therefore, a common assumption is that digestion can alter thermoregulatory behavior, i.e., digesting snakes choose warmer environmental temperatures than non-digesting snakes, and snakes actively thermoregulate to optimize digestive efficiency. However, a predominantly inactive, infrequently-feeding ambush predator may sacrifice optimal body temperature to conserve energy and/or to avoid detection. This study investigated thermoregulation and microhabitat selection before and after feeding in telemetered timber rattlesnakes ( Crotalus horridus) in central Tennessee. Snakes were offered large food items(laboratory rats weighing 30-50% of snake body mass) or were observed feeding naturally. Data from a total of eleven individuals monitored over four active seasons indicated that neither Tb nor environmental temperatures differed significantly betweenpre-fed and post-fed treatments. This lack of a difference in temperature selection is counter to what we hypothesized and what is generally assumed for snakes. A possible explanation for this result is the tendency of C. horridus to be a thermoconformer.________________________________________________________________________ Pathogens Vs. Pesticides: The Threat to Eastern Hellbenders Carson E. Lillard 1, Debra L. Miller1,2, Jennifer L. Howard2, Benjamin S. Wilson1, ReillyT. Jackson 1, Jennifer R. Asper1, Rebecca P. Wilkes3, Louise Rollins-Smith4, Rachel D.Hill 1, Yi Geng1,5, William B. Sutton1,6, Bill Reeves7, Sherri Reinsch8, Dale McGinnity8,and Matthew J. Gray 11 Center for Wildlife Health, University of Tennessee, Knoxville, TN, USA2 College of Veterinary Medicine, University of Tennessee, Knoxville, TN, USA3 Veterinary Diagnostic and Investigational Laboratory, University of Georgia, TiftonGA, USA 4 Department of Pathology, Microbiology, and Immunology, Vanderbilt UniversityMedical Center, Nashville, TN, USA 5 College of Veterinary Medicine, Sichuan Agricultural University, Ya’an, China6 Department of Agricultural and Environmental Sciences, Tennessee State University,Nashville, TN, USA 7 Tennessee Wildlife Resources Agency, Nashville, TN, USA8 Nashville Zoo at Grassmere, Nashville, TN, USAThe eastern hellbender ( Cryptobranchus alleganiensis alleganiensis) of North America isdeclining in several watersheds across its geographic range. Frequently, these watersheds are degraded due to surrounding agricultural land use. The increase in application of glyphosate in the 1990s coincided with the decline of hellbenders in some watersheds. To date, no studies have tested the toxicity of glyphosate herbicides to hellbenders. The 1990s also were a period when several pathogens emerged in amphibian populations, but their effects on hellbenders remain unclear. We tested the toxicity Cornerstone® and Roundup® formulations of glyphosate on larval hellbenders, the susceptibility of hellbenders to ranavirus and Batrachochytrium dendrobatidis (Bd), and if there was aninteraction between these pathogens and a sub-lethal exposure to Roundup®. We found that larval hellbenders could withstand high concentrations of Cornerstone® (24 ppm) and Roundup® (12 ppm), larval and juvenile hellbenders experienced 67 – 80% mortality when exposed to ranavirus, and juvenile hellbenders did not die when exposed to Bd.There was evidence of interactive effects, with 20% greater mortality when exposed to Roundup® and either of these pathogens. Our results indicate that, of the factors we tested, ranaviruses are the greatest threat to larval and juvenile hellbenders, and sub-lethal exposure to Roundup® could contribute to mortality when pathogens are present. ________________________________________________________________________ If You Build it, Will They Come? Temporary Backyard Ponds as an Alternative to Traditional Frog Metamorphosis Projects Alli Metler Homeschooled, 6 th grade, 7007 Scepter Dr., Bartlett, TN 38135For years, homeschool curricula and schools everywhere have done the traditional project of watching a tadpole metamorphose into a frog. We sought to redefine that project by setting up a wading pool in our suburban Memphis area property with assorted cover objects, both commercial and found objects. In Year 2, plants were put near the pool. Our question was whether or not frogs would do amplexus in this pool. In Year 1, there was definite use by American Toads, Bronze Frogs, and Southern Leopard Frogs, as well as Gray Treefrogs, Copes’ Gray Treefrogs, and Green Treefrogs. In Year 2, tadpoles were found from Gray Treefrogs and there was definite colonization from Bronze Frogs, Southern Leopard Frogs, American Toads, Gray Treefrogs, and Copes’ Gray Treefrogs. Other animal use includes birds as well as mammalian species such as squirrels, possums, and cats. Others around the United States have agreed to take part in this project and record results. ________________________________________________________________________ Evidence of a New Species of Mudpuppy in Tennessee Stephen Nelson 1, Ben Fitzpatrick2, Matthew Niemiller3, and Michael Ogle41 Knoxville Zoo, 2University of Tennessee (Knoxville), 3Illinois Natural History Survey,and 4Knoxville ZooThere is currently only one necturid species (family Necturidae) currently known to occur in Tennessee, the Common Mudpuppy ( Necturus maculosus), (Pasachnik and Niemiller2011). However, there is evidence from color patterns, mitochondrial DNA (mtDNA) and nuclear DNA (nDNA) that an additional species also occurring in Tennessee. The taxonomy of mudpuppies in the southeastern United States has been in dispute for several years. Current taxonomy recognizes five species and one subspecies. Our evidence suggests that there is a new species of mudpuppy that coexists with the most well-known and widely distributed of the water dogs and mudpuppies, the Common Mudpuppy ( Necturus maculosus). The putative new species co-occurs with the Common Mudpuppyin one watershed in Southeastern Tennessee, but the extent of its geographic range is unknown. ________________________________________________________________________ An Update of Amphibian and Reptile Surveys on Wildlife Management Areas Within TWRA’s Administrative Region 4. Chris Ogle Tennessee Wildlife Resources Agency Since 2004, the Tennessee Wildlife Resources Agency (TWRA) has been inventorying nongame species on numerous Wildlife Management Areas (WMA’s). More recently, this inventory has begun to focus on species of Greatest Conservation Need (GCN) in each of the following classes: Amphibia, Reptilia, Mammalia, and Aves. Within each of TWRA’s four administrative Regions, one Wildlife Manager 1 has been tasked with this responsibility. A variety of techniques have been implemented to accomplish this goal, and presented here is an update of the work and species captured within TWRA’s administrative Region IV since the 2014 meeting. ________________________________________________________________________ Poor Biosecurity Could Lead to Disease Outbreaks in Amphibian Populations Jennifer A. Spatz 1, Matthew J. Gray1, E. Davis Carter1, and Debra L. Miller1,21 Center for Wildlife Health, University of Tennessee, Knoxville, TN 379962 College of Veterinary Medicine, University of Tennessee, Knoxville, TN 37996Outbreaks of ranavirus and chytrid fungus have contributed to amphibian population declines. It has been suspected that biologists could contribute to pathogen outbreaks through poor biosecurity practices during sampling. Biologists frequently co-house captured amphibians and do not change gloves between handling different individuals. We tested whether these poor biosecurity practices could facilitate transmission of ranavirus from infected to uninfected wood frog ( Lithobates sylvaticus) tadpoles, andincrease the likelihood of mortality. Co-housing tadpoles for only 15 minutes with 10% of individuals initially infected resulted in transmission and mortality of 50% of uninfected tadpoles. Not changing gloves between individuals when 10% were initially infected resulted in transmission of ranavirus and mortality of 70% of uninfected tadpoles. More extreme mortality was observed when tadpoles were co-housed for longer durations, or when the initial infection prevalence was >10%. Our results indicate that poor biosecurity practices can cause pathogen transmission between individuals, which could lead to disease outbreaks and decrease survival in populations. Biologists should change gloves or decontaminate them between handling individuals, and not co-house animals. ________________________________________________________________________ Demographic Characteristics, Incidence of Ranavirus Infection, and Corticosterone Levels in the Eastern Box Turtle, Terrapene carolina carolina, in a SuburbanWetlands Habitat of Middle Tennessee. Jessica Vannatta Tennessee Technological University The Eastern Box Turtle, Terrapene carolina carolina, is a species of concern inTennessee because of population decline. Decline is due mainly to anthropogenic causes including, but not limited to, road mortality, disease, and habitat fragmentation. The purpose of this study was to assess the general health of the T. c. carolina population in asuburban wetlands habitat in Murfreesboro, TN, USA (Nickajack) by measuring demographic characteristics, infection status, and physiological characteristics. Demographic characteristics recorded include population density, age class, sex ratio, and several body size measurements (e.g., carapace length and carapace height). A small blood sample ( ≤0.2 ml) was drawn to measure corticosterone levels, triglycerides, uricacid, innate immunity, and to determine Ranavirus infection status. The population density was estimated to be approximately 14-15.5 turtles per hectare. The age class structure had a normal distribution with most turtles falling within the middle age class (10-14 years). The sex ratio was 1.26 male:1 female. For body size, the only variable with a significant difference between males and females was straight-line carapace length in which males were longer than females. A single turtle (LPW) was positive for Ranavirus infection out of the 102 turtles sampled and tested, accounting for 1% prevalence in the sampled population. Females had higher body condition indices than males. In 2013, corticosterone levels were significantly higher in summer than in spring. In 2014, corticosterone levels were significantly higher in fall than in both spring and summer. The difference in the seasonal timing of peak corticosterone levels between the years may be related to weather conditions or associated with opportunistic mating. Corticosterone levels were positively correlated with hemolysis titer, possibly because of an immunoenhancing effect of stress hormones. In both 2013 and 2014, triglyceride levels were significantly higher in females than in males. The higher triglyceride levels of females were most likely associated with the energetic demands of egg production, carrying eggs, and nesting. Triglycerides were positively correlated with body condition, indicating greater lipid reserves in individuals with a better body condition. All other physiological measures, including total leukocytes and heterophil counts, failed to exhibit significant seasonal or sex related differences. To our knowledge, this is the first report of baseline corticosterone values and innate immunity in free-ranging T. c. carolina. Theresults obtained will aid in the conservation and protection of this species of concern. ________________________________________________________________________ A Next-generation Sequencing Approach to Assess the Diversity of the Skin and Gut Microbiota in the Slimy Salamander ( Plethodon glutinosus) Species Complex, withan Emphasis on Gastrointestinal Fungal Pathogens 1 Donald M. Walker, 2Sean P. Graham, 3Crystal Kelehear, 4Carlos Camp, 1Aubree J. Hill,5 Michael Edelbrock, 5Justin Rheubert, 6Jessica Wooten1 Department of Biology, Tennessee Technological University, Cookeville, TN 385052 Department of Biology, Geology, and Physical Sciences, Sul Ross State University,Alpine, TX 79832 3 Smithsonian Tropical Research Institute, Apartado 0843-03092, Balboa, Ancon,Panama, Republic of Panama 4 Department of Biology, Piedmont College, Demorest, GA 305355 Department of Natural Sciences, The University of Findlay, Findlay, OH 458406 Department of Biology, Centre College, Danville, KY 40422Amphibians have become an increasing conservation concern due to documentation of widespread declines in populations throughout the world. Many of these declines are attributable to outright habitat loss and degradation; however, many are linked to an emerging infectious fungal pathogen: Batrachochytrium dendrobatidis (Bd). Someamphibians have developed an active and potent battery of cutaneous antimicrobial peptides to control potential pathogen growth. In addition, a few studies have shown that amphibians have a symbiotic gut microbial community that either assists with digestion or is entirely benign to the host organism. Therefore, studies simultaneously considering the gut and skin microbial community of amphibians may contribute to our understanding of their peculiar susceptibility to emerging infectious diseases. Skin swabs ( n=57) andfecal samples ( n=57) were collected from eight species of Plethodon (n=57 individuals)in the southeastern US. DNA was extracted, the 16S rRNA gene (bacteria) and ITS rDNA gene (fungi) were PCR amplified, and total microbial communities sequenced on the Illumina MiSeq platform. The most common bacterial phyla on the skin and gut were Bacteroidetes, Firmicutes, Proteobacteria, and Verrucomicrobia. A non-metric multidimensional scaling ordination indicated strong differences (stress 0.13) in skin and gut microbial communities across all Plethodon species. Two-way ANOSIM analyseswere performed to assess whether host species and/or skin/fecal communities were significant predictors of variability across Plethodon species. The ANOSIM confirmedskin and fecal communities were a significant predictor of variability (R=0.976, p<0.001), whereas, host species was not (R=0.149, p<0.001). Gastrointestinal fungal pathogens in the genera Basidiobolus was detected in all eight and Mariannaea in sevenof eight salamander species. This comparative analysis documents the bacterial and fungal diversity of both the skin and gut microbiome for species of slimy salamanders.
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