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    Poster Session 2

    Room: Ballroom 100

    2022-07-30   16:00 - 18:00



    1.    Contributed Ichthyology  Otoliths through time: a multidecadal characterization of fish otoliths in a changing climate. Jordan Waldron*, University of North Florida; Brittany Bottom, FWC Fish and Wildlife Research Institute; Jessica Carroll, FWC Fish and Wildlife Research Institute; Lexi Lindsley, University of North Florida; Brian Wingender, Materials Science and Engineering Research Facility, University of North Florida; M. Laura Habegger, University of North Florida   jordanwaldron82@gmail.com

    Otoliths are biomineralized calcium carbonate stones in the inner ear of bony fishes important for auditory perception and vestibular function. These structures have been used to answer a variety of questions in numerous fields of research because of their unique development and growth patterns. Though otoliths have been researched extensively over the past few decades, there is still limited data that describes how widespread environmental changes in marine ecosystems affect wild teleost fish otoliths. This knowledge gap limits our ability to understand the potential impacts that alterations to otoliths can have on fish physiology and ecology. In this study, potential morphological and chemical changes in wild adult fish otoliths will be traced over a large timescale among species common in Florida’s coastal habitats – red drum (Sciaenops ocellatus) and common snook (Centropomus undecimalis). Morphometrics, materials science techniques, and biomechanical methods will be used to characterize otolith morphology, microstructure, mechanical properties, and composition. Changes in all variables are hypothesized to be present in correlation with environmental fluctuations over time. Although the physiological and ecological consequences of changes in otoliths are not fully understood, some studies suggest that they can impact otolith function, which could potentially have ecological implications.


    2.    Contributed Ichthyology  Investment in Otolith Collection Capacity at the UW Burke Museum. Katie Bennett*, University of Washington; Divinity Paterson*, University of Washington; Katherine Maslenikov, University of Washington   pearsonk@uw.edu

    Founded in 1919, the University of Washington Fish Collection is home to over 13 million preserved specimens, making it the largest fish collection in North America. The otolith collection alone hosts over 2.5 million pairs across a myriad of families with a primary geographic focus on the eastern North Pacific Ocean. With a growing collection of approximately 50,000 to 70,000 new otolith vials annually, spatial constraints have been a significant concern, inhibiting expansion while posing security and fire safety risks. Our goal was to expand our otolith collection into new storage facilities that would provide more security and additional space necessary for the future expansion of the collection. We applied for an IMLS grant to obtain shelf compactors, expand the otolith collection into a new room, and employ undergraduate students to assist with the transition. The result has been the expansion of the collection between two secured rooms, with the addition of compactor shelves to allow for the continued growth of the collection and the necessary inclusion of space above the shelves to allow for code-compliant sprinkler operation. Otoliths are critical to resolving pressing questions in contemporary marine science addressing changes in ocean chemistry, evolutionary relationships, and population dynamics throughout history and into the future. The otolith collection at the Burke Museum is an ever-expanding public resource that is freely available for anyone to utilize and explore.


    3.    Contributed Ichthyology  Environmental Justice for Marshall Island Communities: Confronting our Fish Collection’s Colonial History. Kalena Kattil-deBrum*, University of Washington; Samara Tobey, University of Washington; Luke Tornabene, University of Washington; Holly Barker, University of Washington   luke.tornabene@gmail.com

    Natural history collections are catalogs of life on Earth. The specimens housed in collections around the world are rich with stories of how our planet has changed through human history and how life evolved in the hundreds of millions of years prior. However, there are other stories to be told through the specimens in natural history collections, including stories about the exploitative, extractive, and or violent ways in which specimens are sometimes collected. One such example can be found at the University of Washington Fish Collection (UWFC), which houses more than 15,000 specimens from the Republic of the Marshall Islands (RMI). These specimens were collected during three decades of nuclear weapons testing on the RMI, during which the US detonated 67 nuclear weapons that caused unprecedented environmental contamination, the loss of land and livelihoods, and long-term adverse impacts to human health. These specimens serve as a constant reminder to the Marshallese community of a period in history where federally funded scientific research took priority over the quality of life of the Marshallese people. Here we present a brief review the history of the UWFC during these times and a summary of similar collections from the RMI around the US. We also present examples of potential research projects that could use these specimens to support Marshallese communities today, representing the first of many necessary steps towards confronting the injustice done to the RMI. We hope that this report serves as a primer for future decolonization of collections.


    4.    Contributed Ichthyology  Conservation Efforts and Preserving Imperiled Ichthyofauna of the American Southwest. Emily DeArmon*, University of New Mexico; Thomas Turner, University of New Mexico   esdearmon@unm.edu

    The Museum of Southwestern Biology (MSB) Division of Fishes houses specimens from rivers and tributaries of the American Southwest. Surveys and collections have been ongoing at the same sites for over two decades to monitor native and nonnative fish communities. Due to increased population and climate change, habitats are declining at a rapid rate making it essential to preserve these species. Conservation efforts are implemented throughout these desert river basins with the help of hatcheries and federal and state agencies. Project-based collections monitor vulnerable species native to the southwest. These collections and monitoring sites allow us to understand how fishes are interacting with hatchery bred and if natural spawning events are occurring. Over the last few years, we have expanded our genetic collection of fishes by more than 10,000 samples. Many of these samples are now housed in the MSB’s Division of Genomic Resources for long-term storage. Our near future goal is to collect tissue samples of fishes throughout the American Southwest and to expand on our genetic collection and thus enable development of genomic resources for conservation of an imperiled ichthyofauna.


    6.    Contributed Ichthyology  Interspecies and Individual Variation in Sculpin Intracranial Pressures During Ventilation. Jeremiah Pouncy*, North Carolina Agricultural and Technical State University; Sarah Handy, Arizona State University; Stacy Farina*, Howard University   stacy.farina@howard.edu

    Intracranial pressures of the buccal and gill chambers are often used to compare the mechanics of pumping across fish species, particularly in ventilation and feeding behaviors. While it is generally expected that patterns of intracranial pressures will vary widely across large taxonomic groups, little is known about variation in pressures among closely related species and among different behaviors of the same individual. We compared the dominance of the buccal pump relative to the gill chamber pump during ventilation, in a multispecies study of sculpins and relatives (Cottoidea). Most benthic species are known to have a dominant gill chamber pump, in terms of ventilatory pressures. Because the studied species were benthic, we predicted that they would primarily use gill chambers as the dominant pump. We compared the amount of pressure generated by each pump during each phase of the ventilatory cycle using surgically implanted pressure transducers. We then quantified the ratio of buccal to gill chamber pressure for 50 breaths per individual. We also looked at morphological correlates of the ratio among species using measurements from CT scans. We found convergent evolution in pump ratio across species. We also found that individuals of a species exhibited a range of buccal and gill chamber pumping, with the ability to switch between the two behaviors.


    7.    Contributed Ichthyology  Critical Thermal Maxima of Arkansas Stream Fishes Determined Using Field Acclimatization Protocols. Reid Adams*, University of Central Arkansas; Ginny Adams, University of Central Arkansas; Willow Newman, University of Central Arkansas; Robert Remy, University of Central Arkansas; Chance Garrett, University of Central Arkansas; Cade Richesin, University of Central Arkansas; Matthew Gifford, University of Central Arkansas   gadams@uca.edu

    Upland fishes in the southeastern United States may be especially vulnerable to increased water temperatures due to climate change, yet thermal tolerances of these faunas have not been extensively quantified. Furthermore, existing data have been mostly collected using lab acclimation procedures that may not accurately reflect thermal tolerances in the field. To address these data gaps, we quantified Critical Thermal Maxima (CTMax) of 23 fishes in an Ozark river and 16 species from the Arkansas River Valley spanning seven families (Cyprinidae, Catostomidae, Fundulidae, Poeciliidae, Cottidae, Centrarchidae, and Percidae). Fishes were collected with seines and held for 24-48 hours prior to testing. CTMax was measured using Precision CIR 19 heated water baths that were increased approximately 1.0℃ per minute until loss of equilibrium. Measures of CTMax in our study are among the highest recorded for several species (e.g., Fundulus olivaceus, Lepomis megalotis, and Campostoma spadiceum). Variation in CTMax was observed due to ecoregion, stream macrohabitat, range extent, and taxonomic groupings. On average Cottidae (34.9℃) and Cyprinidae (36.8℃) had the lowest CTMax, with Fundulidae (40.8℃), and Poeciliidae (41.8℃) tending to have the highest mean values. Lowest CTMax tended to be observed in Ozark fishes, species occupying run habitats, and fishes with restricted ranges. Summer thermal safety margins averaged near 8 to 9℃ for fishes in the Arkansas River Valley but were 3 to 5℃ for several Ozark fishes. By understanding thermal tolerances of fishes, we can better predict how community assemblages may change in the future as climate change continues.


    8.    Contributed Ichthyology  Distribution and community structure of deep-sea demersal fishes in the central Pacific. Gina Selig*, University of Hawaii at Manoa; Jeffrey Drazen, Univeristy of Hawaii at Manoa; Peter Auster, University of Connecticut at Avery Point   ginams@hawaii.edu

    Due to the remoteness of deep-sea habitats and their distance offshore, deep-sea fishes have been poorly sampled globally resulting in much of the descriptions of demersal fish communities to occur along temperate continental margins. Demersal fish communities from islands and seamounts, are poorly described even in the Hawaiian archipelago past 500 meters. Knowledge across all depths, in similar settings, is even more sparse for other archipelagoes in the central Pacific. However recent remotely operated vehicle (ROV) explorations of the central Pacific provide an opportunity to explore these communities. Here we describe demersal fish assemblages of archipelagoes across the Pacific, including in multiple Marine National Monuments, and examine the relationship of the assemblages to depth and environmental conditions. Over 200 dives were conducted by ROVs and 318 taxa (189 identified to species and 129 to genera) belonging to 94 families were recorded across the regions at depths ranging from 250-4000m. The assemblages were numerically dominated by Halisauridae, Macrouridae, Ophiidae, and Synaphobranchidae. Preliminary results indicate strong changes in the fish assemblages with depth and that assemblages become more similar between regions with greater depths. Overall, understanding the distribution patterns of demersal fishes across archipelagoes in this region will increase knowledge of mesophotic and deep sea-fish biogeography. Further, given the region’s oligotrophic conditions it is susceptible to global change so characterizing these communities can aid in the development of conservation and management strategies.


    10.    Contributed Ichthyology  Aerobic Scope in Stream Fishes as a Fitness Indicator of Climate Change Impacts. Richard D. Durtsche*, Northern Kentucky University; Samuel P. Bauer, Northern Kentucky University; Mackenzie Danker, Northern Kentucky University   durtsche@nku.edu

    Predicted environmental temperatures will increase up to 3˚C or more over the next 100 years, which could drastically change streams conditions and aquatic ecosystems. As many areas of the world are finding these thermal changes occurring faster than anticipated, testing of how stream fishes respond to varying temperatures can provide insight into their current and future capacity to respond to these changes. This study addresses how adult stream fish respond metabolically to different temperatures. We chose three fish species to test that represent bottom dwelling (rainbow darters, Etheostoma caeruleum), midwater occurring (bluntnose minnows, Pimephales notatus), and top water (mosquitofish, Gambusia affinis) fishes. A static-flow respirometry system was used to measure metabolism in these fish. Metabolic responses as an indicator of fitness were measured as the minimum metabolic rate (or standard metabolic rate, SMR) needed for fish to survive, as their maximum metabolic rate (MMR) used in predator escape behavior or foraging, and a calculation of their overall metabolic capacity or their aerobic scope (AS) which is the difference between MMR and SMR. This study further investigated the plasticity of metabolic response in these fish after they were raised through embryogenesis and into feeding juvenile stages at temperatures that mimic current stream temperatures (cold) for one cohort, and at future climate change temperatures +3˚C above current stream temperatures (warm) for another cohort. Juvenile fish of both cohorts were then tested for their aerobic scope at a common temperature to determine how they differ depending on their development temperatures.


    11.    Contributed Ichthyology  Body shape evolution in cryptobenthic reef fishes found in the Tropical Eastern Pacific. Cindy Fajardo*, Natural History Museum of Los Angeles County; Michael Burns, Cornell University; William Ludt, Natural History Museum of Los Angeles County   cfajardo@nhm.org

    Body shape plays a key role in how a species interacts with its ecosystem, and is often influenced by the ecology of a species and is indicative of their function within a community. In tropical reef fish communities, both shape and size vary greatly across lineages and ecological gradients. On the smaller end of the size spectrum, cryptobenthic reef fishes (CRFs) are an influential sub-community that are essential for the overall health of coral reefs. CRFs are generally <50 mm in size and comprise nearly 50% of biomass consumed in reefs. The Tropical Eastern Pacific (TEP) is a biogeographic region that spans from southern Baja California to northern Peru and comprises three marine provinces: the Cortez, Panamic, and offshore islands. Throughout these provinces are 245 species of CRFs from 12 families, yet little is known about the community composition and evolutionary processes that have shaped these CRF communities. In this study, we quantified body shape variation of CRF communities throughout the TEP using geometric morphometrics and assessed morphological disparity between biogeographic provinces, among families, and across different habitats. Furthermore, we assessed whether CRFs that have adapted to the same habitat exhibit parallel phenotypes, indicating shared selective pressures across biogeographic regions, or if lineages in each biogeographic region evolved unique morphologies, indicative of differential selection. Collectively, these results shed light on the evolutionary history and formation of CRF communities in the TEP, and provide critical insight into the functional dynamics of these CRF communities throughout this unique biogeographic region.


    12.    Contributed Ichthyology  Fishes of Oklahoma App. Sara Cartwright*, Sam Noble Museum; Dahiana Arcila, Sam Noble Museum   isisrayn@gmail.com

    The Ichthyology Collection of the Sam Noble Museum (OMNH) is the largest biorepository of freshwater fish biodiversity in the state of Oklahoma. Although more than 90% of the specimens deposited in the OMNH collection are catalogued and georeferenced in different biological databases, most of the date remain largely inaccessible to the public and fragmented across different websites. The mobile app can be downloaded by anyone with a smartphone to assist with the identification of fishes from streams and lakes throughout Oklahoma. Individuals can also learn about the habitat, biology, morphology, and geographic range distributions, as well as view photos of each species.


    13.    Contributed Ichthyology  Morphological Study of Bregmaceros atlanticus and B. japonicus (Bregmacerotidae: Teleostei), with Recognition of a New Species from the Eastern Pacific. Antony S. Harold*, Grice Marine Laboratory, College of Charleston; D. Andrew Baltzegar, Genomic Sciences Laboratory, North Carolina State University   gobyharold@gmail.com

    Bregmaceros species (Codlets) are miniature, epi- to mesopelagic gadiform fishes occurring circumglobally in tropical to warm temperate coastal regions. They have a moderately elongate body shape, a well-defined caudal fin, and a subdivided dorsal fin including a single occipital ray followed by a long-based second dorsal fin typically of about 40 to 65 rays. There have been two major reviews of the genus but although many of the species are distinct there remains a number of areas of confusion. There has been progress in recent years, however, including descriptions of several new species. In our study we re-examine two darkly pigmented species, B. atlanticus and B. japonicus, and a complex of similar forms. In studying specimens of nominal B. japonicus from the eastern Pacific we reach the conclusion that there is a morphologically distinct, undescribed species present. Meristic and morphometric character distributions and a multivariate shape analysis, are used in combination with pigmentation features to demonstrate the existence of this form. The new species bears considerable similarity to other species with relatively high dorsal- and anal-fin ray counts, but is distinguished from both B. atlanticus and B. japonicus, based on its high precaudal vertebral counts (typically 15 or 16) and dark pigmentation concentrated dorsally and with the ventral portions of the head largely unpigmented. The geographic distribution of the new species includes parts of the eastern tropical Pacific, from near the Hawaiian and Line Islands eastward to the California Current region.


    14.    Contributed Ichthyology  The Guam Biorepository: a Path Towards Rescuing and Expanding a Fish Collection. Terry Donaldson*, University of Guam Marine Laboratory/EPSCoR; David Burdick, University of Guam Marine Laboratory/EPSCoR   terryjdonaldson@gmail.com

    The University of Guam’s (UOG) Biorepository, housed at the UOG Marine Laboratory, was established in 2015 with funds from the U.S. National Science Foundation’s Established Program to Stimulate Competitive Research (EPSCoR) under the Guam Ecosystems Collaboratorium (GEC). A second larger award from EPSCoR for the Guam Ecosystems Collaboratorium for Corals and Oceans (GECCO) was received in in 2020. Both awards have enabled the Biorepository to apply state-of-the-art methods towards the curation of significant existing, new, and planned fish collections from Micronesia and the western Pacific region. The quickly growing Biorepository collection includes physical and voucher specimens, cryo-preserved tissues, microscope slides of histological (gonad) tissues and otoliths, photographs and other imagery, and metadata. Collections data and specimen images are curated using the Specify data curation database, and are made available online to the global scientific community and general public following the standards and protocols of the Integrated Digitized Biocollections (iDigBio) project. New instrumentation being utilized include a micro-computer assisted tomography (m-CT) scanner, a next generation sequencer, a scanning electron microscope, a microscope slide scanner, and various other imaging systems for use underwater and in the laboratory. These acquisitions increase analytical capabilities of institutional and visiting researchers alike. The Biorepository collections will lay the foundation towards enhancing our understanding of the region’s marine biodiversity and biogeography, promote new collaborative research initiatives at local, regional and global scales, and promote the development and implementation of effective plans for the conservation of threatened species, communities and their habitats in changing environments.


    15.    Contributed Ichthyology  Morphometric and meristic disparity within the zeiform genus Parazen correlated with its geographic distribution. Randy Singer, University of Michigan Museum of Zoology; Terry Grande*, Loyola University Chicago; Mark Wilson, Loyola University Chicago   tgrande@luc.edu

    The zeiform genus Parazen Kamohara, 1935, is known from one species, P. pacificus, the Slender Dory, distributed in the tropical western North Atlantic (Caribbean region), western and southwestern Pacific (near Japan, Taiwan, and Australia), and Indian oceans, at depths ranging from 145 to 500 meters. The family Parazenidae traditionally has included also the genera Cyttopsis and Stethopristes. Recent phylogenetic studies based on both morphological and molecular data strongly support a sister-group relationship between Cyttopsis and Stethopristes, but only weakly recover Parazen as sister to those two genera. This tenuous relationship suggests that the family Parazenidae might not be monophyletic. Morphometric analyses further suggest that the shallower body shape, relatively larger eye placed more laterally, and shorter jaws of Parazen evolved differently than comparable traits of Cyttopsis and Stethopristes, both of which resemble more the primitive body shape of zeiforms. Within P. pacificus, morphometric and meristic data further suggest morphological disparity that is correlated with geographic distribution. For example, in addition to morphometric differences, samples from the Atlantic and Indo-Pacific Oceans (Cuba and the Indian Ocean, respectively), exhibit considerably different dorsal fin-ray and lateral-line scale counts. This poster explores morphological variation within Parazen via morphometric and meristic data as well as morphometric analysis, comparing these to the distribution of known Parazen populations in different ocean basins. Results to date suggest that Parazen could contain at least two distinct species.


    16.    Contributed Ichthyology  The Caudal Bioluminescent Organ of the Pelican Eel, Eurypharynx pelecanoides, and Its Evolution (Anguilliformes). Michael Ghedotti*, Regis University; Kandice Agudo, Regis University; Flor Gonzalez, Regis University; Matthew Davis, St. Cloud State University; W. Leo Smith, University of Kansas; Benjamin Frable, Scripps Institution of Oceanography, UCSD   mghedott@regis.edu

    The Pelican Eels (Eurypharynx pelecanoides) are deep-water fishes with extremely enlarged jaws producing enormous oral cavities and a caudal light organ that has been suggested to be used in prey luring. In this study we seek to provide a first description of the histologic structure of the bioluminescent organ in Eurypharynx and to place its structure into an evolutionary context. We used histological sectioning and gross examination to explore the anatomy of the bioluminescent organ. We generated and used a DNA-sequence based phylogeny for the anguilloid eels (a clade including the eels previously placed in Saccopharyngoidei and other families). The bioluminescent organs in Eurypharynx are intrinsic and use partial screening via ultra-black melanosome anatomy within the bioluminescent organ. The anatomy in the context of the phylogeny suggests that a dermal anatomy associated with ultra-black coloration evolved in the ancestor of the deep-ocean anguilloids and the that intrinsic caudal bioluminescent organs evolved once in the most recent common ancestor of Eurypharynx and Saccopharyx (the Swallower Eels). We also discuss the evolution of other anatomical characteristics likely associated with a deep-sea habitat in this group.


    17.    Contributed Ichthyology  Global Macrogenetic Analysis of Fish Diversity and Demographic History. Justin Bagley, Jacksonville State University; Rachel Bonner, Jacksonville State University; Christian Braswell, Jacksonville State University; Diego Elias*, Louisiana State University; Macee Glick, Jacksonville State University; William Hanson-Regan, University of Tennessee at Chattanooga; Kayla Morrow, Jacksonville State University; Kevin O'Malley, University of Tennessee at Chattanooga; Elyse Parker, Yale University; Mackenzie Smith, University of Tennessee at Chattanooga; Fernando Alda, University of Tennessee at Chattanooga    fernando-alda@utc.edu

    The increased capacity for generating DNA sequences and open data initiatives have made thousands of records of molecular, geographical, natural history or environmental data publicly accessible. Shifts in data accessibility now permit leveraging these data for macrogenetic studies to analyze thousands of species, over large spatial extents or long time series. In this study, we examine patterns of intraspecific genetic variation (IGV) across the fish tree of life from a macrogenetic perspective to understand the ecological and evolutionary processes that generate, maintain and erode fish IGV. Specifically, we are interested in testing the role of geographical and natural history factors (e.g. dispersal capacity) in driving patterns of genetic diversity and species richness, genetic diversity gradients, and demographic responses to historical climate change. We repurposed legacy mtDNA sequences from the phylogatR database. phylogatR is a free web database that aggregates phylogeographic data (sequences and geographic occurrences) from online repositories and produces DNA alignments that are taxonomically organized and ready to use. We downloaded an initial dataset of cytochrome c oxidase I (COI) sequences from 8065 species of actinopterygian and elasmobranch fishes. We pruned this dataset to only keep species with a minimum of n = 5 COI sequences, resulting in a dataset of 4879 fish species from 425 families. All data (sequence identification, alignments, and maps of sampling localities corresponding to sequences) were reviewed manually. The final, curated dataset was analyzed to elucidate geographical patterns of IGV and demography, which we compared between freshwater and marine fishes.




    18.    Contributed Ichthyology  Phylogeography of Stonerollers (Campostoma: Cyprinidae) across North America. Fernando Alda*, University of Tennessee at Chattanooga; Taya de Blonk, University of Tennessee at Chattanooga; Juliet Wimberly, University of Tennessee at Chattanooga; Susana Schönhuth, St. Louis University; David Neely, Tennessee Aquarium Conservation Institute; Richard Mayden, St. Louis University; Michael Blum, University of Tennessee-Knoxville   fernando-alda@utc.edu

    Stonerollers from the genus Campostoma are widespread and abundant across North America. This genus of herbivorous minnows inhabits streams along a broad range between southern Canada to central Mexico and span across central and eastern North America. The dynamic geographic history of North America has hindered the proposal of highly resolved hypotheses about the evolutionary relationships of widespread taxa like Campostoma. This is because previous methods have relied heavily on geographically variable morphological characteristics. A comprehensive systematic analysis involving sufficiently informative markers and thorough taxon sampling has yet to be conducted. A potential issue is seen in the widespread taxon C. anomalum where preliminary mitochondrial molecular evidence suggests it is not a monophyletic group. The purpose of this study is to identify inconsistencies across previously constructed phylogenies and provide a robust hypothesis by using extensive DNA sequence data from the mitochondrial cytochrome b gene. We uncover hidden genetic diversity that may warrant taxonomic revision and/or species recognition taxa. This study represents the most comprehensive analysis to date of wide-range patterns of genetic variation and evolutionary relationships exhibited by the genus Campostoma. These findings will ultimately contribute to existing knowledge on the species diversity of freshwater fish and their distributions in hyper-diverse regions.


    19.    Contributed Ichthyology  Systematics of the Pirate Perches,Aphredoderus. Tyler Muller*, Conservation Sciences Graduate Program, University of Minnesota; Andrew Simons, University of Minnesota   mulle651@umn.edu

    Pirate Perch, Aphredoderus, are a lowland species of freshwater fish with a U-shaped distribution present in the Great Lakes, Mississippi River valley, Gulf of Mexico and Atlantic slope. Peripheral populations of this species show species level differences in pigmentation, scale counts and fin spine counts. Populations in the Gulf Coast however, have been considered intergrades leading to a subspecific status following a systematic evaluation by Boltz and Stauffer in 1993. April et al.’s 2011 genetic analysis disagrees with this, and suggests Pirate Perch contain COI divergences to a nearly family-level status. My preliminary morphologic data suggest that Atlantic slope and Mississippi valley Pirate Perch are diagnosable, and that the Eastern Gulf Coast and Florida contains sympatric populations of Eastern and Western forms. Rare intermediate forms are also present in Florida and Georgia which may be hybrids or additional species. My preliminary genomic data suggests 14 million years divergence between Atlantic and Mississippian forms, and significantly diverged lineages within the Gulf of Mexico.


    20.    Contributed Ichthyology  The Phylogeny and Taxonomy of the Stomiiform Bristlemouths and Portholefishes. Leo Smith*, University of Kansas; H.J. Walker, Scripps Institution of Oceanography; Matthew Girard, National Museum of Natural History; Matthew Davis, St. Cloud State University   leosmith@ku.edu

    For the last 50 years, the limits and relationships of the Gonostomatidae have been variously examined with little consensus beyond support for a "core" gonostomatid clade composed of Bonapartia, Cyclothone, Gonostoma, Margrethia, and Sigmops that is potentially allied with or separated from other putative gonostomatids. The inclusion or exclusion of Diplophos, Manducus, and Triplophos and the relationships of all of these genera relative to Polymetme, Sternoptychidae, and Photichthyidae + Stomiidae has varied. In this study, we combined genetic and morphological data to assess the relationships among stomiiform families with a focus on species traditionally included or allied with the gonostomatids. Preliminary analyses separate Polymetme and Triplophos and ally these genera with Photichthyidae + Stomiidae. Further, these analyses suggest that the core gonostomatid clade is sister to a clade composed of Diplophos and Manducus. In light of these relationships and their morphological support and anatomical diagnoses, we will present revisions to the typical classification of these abundant deep-sea fishes.


    22.    Contributed Ichthyology  Species delimitation in four putative species pairs of North Pacific flatfishes. Calder Atta*, School of Aquatic and Fishery Sciences, University of Washington; Duane Stevenson, Alaska Fisheries Science Center, NOAA National Marine Fisheries Service; James Orr, Alaska Fisheries Science Center, NOAA National Marine Fisheries Service; Alison Deary, Alaska Fisheries Science Center, NOAA National Marine Fisheries Service; Luke Tornabene, School of Aquatic and Fishery Sciences, University of Washington   calderatta@gmail.com

    Flatfishes of the family Pleuronectidae are widely abundant in the North Pacific and contribute heavily to commercial fisheries. Management of these species is more likely to succeed if species definitions reflect lineages that are naturally separated by reproductive boundaries, however, most economically important species were described prior to the rise of molecular taxonomy and have not yet been evaluated using genetic data. Four species pairs from Alaska, one each from the genera Atheresthes, Hippoglossoides, Lepidopsetta, and Limanda, are examined in this study due to their morphological similarity and overlapping distributions. The pairs were tested for being distinct species using several delimitation analyses (phylogenetic species, multispecies coalescence, genetic distance, and admixture coefficients) based on exon-capture data. For each pair, there were variable levels of support across methods. Overall, species of Lepidopsetta and Limanda are well-supported evolutionary lineages with reliable diagnostic morphologies, and thus are good management species. Species of Hippoglossoides were also supported as distinct lineages, but there was evidence of high rates of genetic exchange across lineages, possibly caused by recent or incomplete speciation. Species of Atheresthes failed to resolve as distinct lineages in all analyses, but admixture analysis found potential for the existence of two populations that were poorly correlated with species identifications.


    23.    Contributed Ichthyology  A New Lineage of Deep-reef Gobies from the Caribbean, Including Two New Species and One New Genus (Teleostei: Gobiidae: Gobiosomatini). Luke Tornabene*, University of Washington; Rachel Manning, University of Washington; Ross Robertson, Smithsonian Tropical Research Institute; James Van Tassell, American Museum of Natural History; Carole Baldwin, National Museum of Natural History: Smithsonian Institution   luke.tornabene@gmail.com

    Fish communities on tropical deep reefs are dominated by species that belong to families primarily composed of shallow-water species. Collections of deep-reef fishes via submersibles have allowed us to include these deep-reef species in molecular phylogenies, providing insights into the timing and frequency of invasions from shallow to deep reefs. Here we provide evidence of a new deep-reef invasion in the tribe Gobiosomatini in the family Gobiidae (gobies). We describe two new species, one of which belongs to a new genus, and incorporate these taxa into a time-calibrated molecular phylogeny of Gobiosomatini to show that collectively these two genera represent a previously unreported independent invasion onto deep reefs that occurred approximately 20-30 million years ago. These new taxa are readily distinguished from related genera and species by a combination of live coloration, pelvic-fin morphology, meristic characters, head-pore patterns, and other osteological characters. We discuss the relevance of these two new species to the systematics of the tribe Gobiosmatini and include a comparison to all known genera in the tribe.


    24.    Contributed Ichthyology  Phylogeny of Damselfishes (Teleostei: Pomacentridae). Kevin Tang*, University of Michigan-Flint   kltang@umflint.edu

    The family Pomacentridae is a species-rich and widespread group of primarily marine fishes, forming a major component of the world’s reef communities. Currently, there are over 400 recognized damselfish species divided among 29 genera and four subfamilies. Sequence data from mitochondrial (12S, 16S, ATPase 8/6, cytochrome b, cytochrome c oxidase I) and nuclear (histone 3, recombination activating gene 1, and tmo-4c4) genes were analyzed for over 300 damselfish species, representing all 29 pomacentrid genera. The results of these analyses corroborate the recent revision of the classification, finding support for the monophyly of the family, the four subfamilies, and the revised genera. These relationships will be discussed as they relate to the updated classification and taxonomy of Pomacentridae.


    25.    Contributed Ichthyology  Unexpectedly high diversity in a small basin again: the genusPareiorhaphis(Siluriformes: Loricariidae). Roberto Reis*, PUCRS; Edson Pereira, PUCRS   reis@pucrs.br

    We recently demonstrated that the sucker-mouth catfish genus Eurycheilichthysis highly diverse, with eight species in the Jacuí-Taquari basin and one species in the contiguous upper Uruguay River. We now discovered that Pareiorhaphisis equaly highly diverse, but shows the opposite distribution. Pareiorhaphisincludes 27 species distributed on the highlands of the Brazilian Shield, in coastal rivers from Bahia to Rio Grande do Sul and in the upper Paraná and São Francisco basins. Three of these species are known to occur in the upper Uruguay River basin in south Brazil: P. vestigipinnis described in 1992 and P. eurycephalus and P. hystrix described simultaneously in 2002, the latter also occurring in the Taquari basin. During the past 20 years additional collecting efforts in the upper Uruguay River basin revealed the existence of four additional, undescribed species of Pareiorhaphis, elevating to seven the number of species in that basin. Accordingly, while Eurycheilichthyshas one species in the upper Uruguay and eight in the Jacuí-Taquari basin, Pareiorhaphishas seven species in the upper Uruguay and one in the Jacui-Taquari basin. The fact that the members of these two genera share the same microhabitat, living among rocks in swift portions of the headwater creeks, may suggest that they are outcompeting each other in the contiguous basins. The four new species inhabit the upper portions of creeks tributaries to the Uruguay River basin, one of them in the major tributary Ijuí River. These four species are clearly diagnosed and will be described in this study.


    26.    Contributed Ichthyology  Population Structure of Coral Reef Fishes in the Line Islands. Mackenzie Woods, University of California, San Diego   mfwoods@ucsd.edu

    The Line Islands are a remote archipelago in the Central Pacific that offer unique insight into marine genetic dynamics of this type as a result of their geography and the numerous climatological gradients that they encapsulate. The archipelago spans different major currents, varying levels of human exploitation, and differing susceptibility to major climate events, offering the ability to examine the impact of these variables on genetic structure within a single chain of islands. To this end, we examine the population structure of Stegastes aureus, a highly data deficient damselfish, using 275 COI sequences from individuals across the Line Islands with the intent of elucidating any present population structure. Our findings indicate that there is a statistically significant population divide between the two southernmost islands and the rest of the chain. This result is mainly driven by a genetically distinct population that is almost exclusively found in the southern group, and rarely found in the northern group. This finding holds significance not just for S. aureus, but likely also has implications for the population structure of the many small-bodied, monogamous, demersal-spawning reef fishes that also occur in the Line Islands. This study is ongoing, and will include a similar COI analysis for Pseudanthias bartlettorum, a data deficient basslet, as well as a ddRAD-seq analysis of Lutjanus bohar, a significant fisheries species.


    27.    Contributed Ichthyology  Conservation genomic assessment of imperiled freshwater fishes endemic to the Pecos and Devils Rivers. Kayla Dye*, Texas A&M University - Corpus Christi; Kevin Conway, Texas A&M University - College Station; Chris Hollenbeck, Texas A&M University - Corpus Christi; Megan Bean, Texas Parks and Wildlife Department; Sarah Robertson, Texas Parks and Wildlife Department; David Portnoy, Texas A&M University - Corpus Christi   kdye1@islander.tamucc.edu

    Many freshwater fishes endemic to spring-fed tributaries of the lower Rio Grande within Texas exhibit small, highly fragmented distributions and are a priority for conservation. The five target species (Dionda diaboli, D. argentosa, Cyprinella proserpina, Etheostoma grahami, and Notropis megalops) in this study have distributions that lie almost entirely within two native fish conservation areas, deemed important for maintaining the state’s native fish diversity. Previous research reported low levels of haplotype and microsatellite diversity of D. diaboli and D. argentosa as well as a sympatric species of Notropis. Due to the extreme vulnerability of these species, assessing levels of standing genetic diversity at the genome level is essential for conservation. Each target species will be collected across multiple sites in the Pecos and/or Devils Rivers. Double digest restriction-site associated DNA (ddRAD) sequencing will be used to characterize single nucleotide polymorphisms (SNPs) for each species across the genome, and patterns of genetic variation within and among sampling localities will be assessed. Additional museum samples (>30 for each species) will be used to facilitate comparisons of contemporary diversity with that of the recent past. This project will provide information on standing genetic diversity (neutral and putatively adaptive) and connectivity among sampling localities within species, allowing managers to prioritize actions on areas harboring vulnerable populations.


    28.    Contributed Ichthyology  Population Genomics of Red Grouper (Epinephelus morio) in the Western North Atlantic and Gulf of Mexico. Nick Weber*, Texas A&M University-Corpus Christi; Berverly Barnett, NOAA Fisheries; Christopher Hollenbeck, Texas A&M University-Corpus Christi; David Portnoy, Texas A&M University-Corpus Christi   dweber@islander.tamucc.edu

    Understanding how genetic variation is partitioned within and among populations is critical to fisheries management because not accounting for independent populations can lead to overexploitation. In the western North Atlantic and Gulf of Mexico, the red grouper (Epinephelus morio) is an economically important reef-associated species targeted extensively by commercial and recreational fisheries. Due to this importance, a rigorous assessment of genetic population structure is warranted. Therefore, patterns of population structure and genetic diversity within populations will be assessed using a reduced representation genomic sequencing approach. Preliminary results obtained from 198 individuals genotyped at 7,005 loci containing single nucleotide polymorphisms (SNPs) indicated significant heterogeneity among geographic samples (FST = 0.0013, p < 0.0001), though pairwise comparisons were only significant when they involved Cuba. Estimates of genetic diversity were significantly lower in the eastern Gulf of Mexico when compared to all other regions besides Cuba (i.e., Carolinas – Atlantic, Florida – Atlantic, northeastern Gulf of Mexico, and southern Gulf of Mexico; p < 0.001). Future research will involve the use of landscape genomic approaches (e.g., redundancy analysis) to disentangle the potential influence of geographic distance and environmental variables and assess their interaction on observed patterns of genomic variation among samples.


    29.    Contributed Ichthyology  Genetic and morphological identification of formalin fixed larval fishes; how long is too long? Tony Miskiewicz*, UNSW; Sharon Appleyard, CSIRO; Safia Maher, CSIRO; Ana Lara-Lopez, University of Tasmania; Paloma Matis, UNSW; Stewart Fielder, NSW DPI; Iain Suthers, UNSW   tonymisk@yahoo.com

    Identification of larvae of fish is usually based on assembling a developmental series of wild collected larvae, usually formalin preserved, using pigmentation patterns, morphology and fin meristics. However, for many species, larvae are still undescribed or there are only limited descriptions of larvae development. Formalin fixation of larval fish was previously thought to prevent genetic sequencing compared to ethanol preserved larvae. In this poster we detail the results of an integrative taxonomic approach based on morphology, imaging and DNA barcoding of the mitochondrial (mtDNA) cytochrome c oxidase subunit (COI) gene. We used this approach in both cultured yellow tail kingfish, Seriola lalandi and wild sourced fish larvae fixed in 5% formalin. DNA barcoding and genetic species identification was 100% successful in S. lalandi fixed in formalin for up to 6 months, while barcoding of wild caught fish larvae enabled species identification of 93% for up to 8-weeks formalin fixed specimens. While COI genetic identifications from the in-field experiments were patchier than the controlled experiments, our study highlights the possibility of recovering suitable DNA from formalin fixed larvae for up to six months. This was achieved by applying DNA extraction methods that use de-cross-linking steps and species identification based on both full-length reference and mini-barcodes. Our study provides a practical framework for undertaking both morphological and genetic identification to document the larval development of previously undescribed species from historic and current formalin fixed samples collected around southern Australia.


    30.    Contributed Ichthyology  Hybridization and Introgression of Two Imperiled Headwater Fishes, Leon Springs Pupfish, Cyprinodon bovinus, and Pecos Gambusia, Gambusia nobilis. Robert Bretzing*, Texas A&M University-Corpus Christi; Kevin Conway, Texas A&M University-College Station; Chris Hollenbeck, Texas A&M University-Corpus Christi; Megan Bean, Texas Parks and Wildlife; Sarah Robertson, Texas Parks and Wildlife; David Portnoy, Texas A&M University-Corpus Christi   roberttungate12@gmail.com

    Two freshwater fishes, Leon Springs Pupfish, Cyprinodon bovinus, and Pecos Gambusia, Gambusia nobilis, are federally endangered and constrained to extremely narrow habitat ranges in the Southwestern United States. Furthermore, habitat degradation and invasive congers have accelerated population declines in both species. Therefore, next-generation sequencing will be an assessment of standing genetic diversity for both species, as well as historical levels of hybridization and introgression between Leon Springs Pupfish and Pecos Gambusia with invasive congeners. Results of this genomic assessment will provide information for management plans that can be used for conserving wild populations and/or managing refuge populations.


    31.    Contributed Ichthyology  Estimating Atlantic red snapper population size with genetic close-kin mark recapture. Alison Monroe*, Texas A&M University Corpus Christi; Katherine Lanoue, Texas A&M University Corpus Christi; William Patterson, University of Florida; Paul Rudershausen, North Carolina State University; Jeffrey Buckel, North Carolina State University; Eric Anderson, NOAA; Christopher Hollenbeck, Texas A&M University Corpus Christi; David Portnoy, Texas A&M University Corpus Christi   monroea22@gmail.com

    Red snapper, Lutjanus campechanus, is an ecologically and economically important reef fish in the Atlantic Ocean off the southeastern US. The stock is currently estimated to be overfished and undergoing overfishing, but stakeholders have questioned the veracity of population abundance estimates derived from the stock assessment. Using genetic close-kin mark recapture (CKMR) analysis we aim to provide an independent assessment of the population size of red snapper to inform future fisheries management decisions. With existing double digest restriction-site associated DNA sequencing data we developed a panel of 500 single nucleotide polymorphism (SNP) containing loci to be used in genotyping in thousands by sequencing (GT-seq). These SNP containing loci were chosen for high observed heterozygosity, the presence of multiple alleles, and location in the genome. The loci, also known as microhaplotypes, provide greater power than simple SNP loci for identifying kin relationships, while GT-seq provides an efficient and reproducible method for genotyping thousands of organisms. Primers designed to amplify the selected loci will be used to genotype over 7,000 fin clips from Atlantic red snapper caught by both fishermen and researchers throughout 2021. A CKMR model will then be able to estimate population size from the identified half siblings. Beyond an independent estimate of stock size, this study will provide further information on genetic stock structure and can help estimate the effective number of breeders in the population. It will greatly benefit both reef fish assessment and management in the future.


    32.    Contributed Ichthyology  Significant Female Skew in Six Species of EtheostomaDarters in the Subgenus Simoperca. Bruce Stallsmith*, University of Alabama in Huntsville   stallsb@uah.edu

    Sex ratios are important to evolution and ecology. A balanced 1:1 ratio is seen as an evolutionarily stable strategy (Fisher's principle). Yet skew can be found in nature and can vary within a reproductive season. Skew can affect mate choice and sexual selection since the rarer sex should be more attractive. Investment in gametes can be affected by skew. From my experience making monthly collections of two species of subgenus Simoperca darters, Etheostoma duryiand E. simoterum,I found a strong pattern of female skew in both species. I examined museum collections of these two species for more data, and also examined collections of four other Simopercaspecies: E. blennioides, E. coosae, E. flavumand E. ramseyi. For each species I examined 12-26 collections of at least 20 adults each collected between early February and late May, the spawning season for these species. At least 88% of the collections per species exhibited female skew. To test the strength of this pattern I did a chi-square test for each species, in a 2Xn format where nwas the number of collections and the null assumption was an equal number of both sexes per collection. All test results yielded significant rejections of the null with p<0.001 (with Bonferroni correction). It is likely that all of these species have a 1:1 sex ratio at conception. What shapes the strong female skew of breeding adults is still unclear. A likely outcome of this strong skew is stronger male-male competition than is often assumed for these species.


    33.    Contributed Ichthyology  Where is ASIH headed? An open discussion and listening space. Prosanta Chakrabarty*, LSU Museum of Natural Science; Emily Taylor, California Polytechnic State University   prosanta@lsu.edu

    This poster is meant to invite an open discussion about the role of ASIH leadership in working towards meaningful goals for change and strengthening areas that we already do well. We can also discuss the future of ichthyology and herpetology generally (genomics, MRI/CT scanning), diversity and student issues, or anything you might want to bring up with the President, President-Elect, and other members of ASIH leadership.


    34.    Contributed Ichthyology  Female participation and representativeness in scientific publications in the journals of the North American and Brazilian Societies of Ichthyology. Barbara Calegari*, University of Bern; Maria Laura Delapieve, PUCRS; Marina Mendonça, Universidade federal do Pará; Rafaela Priscila Ota, UNESP; Renata Rubia Ota, Nupelia; Karla Soares, UFRJ; Andrea Thomaz, Universidad del Rosario   barbara.calegari@gmail.com

    Scientific publications consist of the main products and metrics by which scientists are evaluated. This study aimed to quantify the female representation in ichthyological publications. For this purpose, data regarding authorship from publications in the journals Neotropical Ichthyology (NI) and Ichthyology & Herpetology (IH) were collected between 2003 and 2020. The total number of females and males authors per publication was evaluated and the frequency of females in first and last authorship positions. In total, 1925 publications were analyzed (1141 from NI and 784 from IH) and, of all, 54.7% had female authors. Among the single-authored publications, females represented only 3% of the records. Considering the publications including at least one female, 55.2% had females as the first author, and 39.5% as of last. Of all publications from NI, females represented 26% of the total number of authors, with 64.8% of the publications including at least one female among the authors of the study. Among these, 57.8% had a female as the first author, and 41.5% as of last. In comparison, the IH had less female representativeness, with only 19% of all authors being females and 38.8% of publications having at least one female author. Among these, 47.9% had a female as the first author and 33.8% as the last. Temporal analyses indicate an increase in the female representativeness, but not in first or last authorship positions. A considerable portion of all publications had only males in the authorship, demonstrating the necessity of implementing gender equality strategies in ichthyology.


    35.    Contributed Ichthyology  Environmental DNA (eDNA) Assessment of Freshwater Fishes from the Guayas River Basin Threatened by Coastal Incursion. Olivia Schweikart*, DePaul University   olivia.schweikart@gmail.com

    By 2030, the National Aeronautics and Space Administration (NASA) estimates coastal incursion events to significantly increase due to rising sea levels associated with climate change, coupled with tidal shifting.Whereas increased tidal size is a natural phenomenon associated with a change in lunar cycle, this process becomes drastically amplified as sea volume increases, leading to instances of coastal flooding. Specifically in the Guayas River Basin in Western Ecuador, sea-level is expected to rise 0.47 m, increasing flooding by a magnitude of 14.1. With my project, I have selected five representative species from different taxa to assess species abundance and habitat quality along three rivers in the Guayas River Basin. Some species are endemic, some not, but all are either economically or ecologically important to the Guayas. Certain populations of chosen taxa are thought to have been isolated for large durations of time, so I will also assess population genetic structure between the lowest and highest elevations within species. Using environmental DNA as an indicator, the results of my study will couple with any existing data on species abundance and geographic distribution, providing a clearer picture of the current threat levels the study species face. By understanding the current endangerment of species, we are better able to make inferences on future threat levels of the species as climatic patterns continue to shift and the ecology of the habitats of these organisms shift in quality as well. Additionally, comprehensive understanding of the future threats to a species allows for better adaptive management models.


    36.    ASIH Storer Ichthyology  Evolution of the Long Wavelength-Sensitive Opsin (LWS) Gene in South American Electric Fishes (Gymnotiformes). Esme Macpherson*, Department of Biological Scineces; University of Toronto Scarborough; Frances Hauser, Department of Biological Scineces; University of Toronto Scarborough; Ahmed Elbassiouny, Department of Cell & Systems Biology; University of Toronto; William Crampton, Department of Biology; University of Central Florida; Belinda Chang, Department of Cell & Systems Biology; University of Toronto; Nathan Lovejoy, Department of Biological Scineces; University of Toronto Scarborough   esmesbmacpherson@gmail.com

    Vision is essential for survival in many animals as it facilitates rapid relay of sensory information regarding the environment. In addition to their vision, South American weakly electric fishes (Gymnotiformes) navigate and communicate using an electrosensory system. All Gymnotiformes have a reduced number of opsin genes, which encode the first step of visual transduction, and several species have poorly developed eyes. Additionally, these fishes inhabit rivers varying in clarity, colour, and amount of light, suggesting that the relative importance of vision may vary among species. We investigated the evolution of the long wavelength-sensitive opsin (LWS) in Gymnotiformes, hypothesizing that this gene would experience relaxed selection relative to non-electric fishes due to reduced reliance on vision. We also tested whether light environment influenced the evolution of this gene within Gymnotiformes. We obtained LWS sequences from 90 gymnotiform species, and used codon-based likelihood models to investigate patterns of molecular evolution in this gene. Contrary to our expectations, we found no evidence for relaxed selection in gymnotiform LWS relative to other fishes, and instead found positive selection across the gene. Additionally, LWS in Gymnotiformes living in clearwater and blackwater habitats showed significantly elevated dN/dS relative to those living in whitewater (turbid) waters. We further identified mutations in LWS that may promote spectral tuning of the opsin protein. While our findings do not exclude the possibility of evolutionary trade-offs between vision and electroreception, detection of positive selection in the LWS gene suggests long-wavelength photopic vision remains an important component of gymnotiform sensory ecology.


    37.    Contributed Herpetology  Characterization of Major Histocompatibility Complex gene diversity in invasive populations of American Bullfrogs (Rana [Lithobates] catesbeiana) in California. Luisa Segovia*, Cal Poly Humboldt; Angel Klawiter, Cal Poly Humboldt; Karen Kiemnec-Tyburczy, Cal Poly Humboldt   lns193@humboldt.edu

    The American Bullfrog (Rana [Lithobates] catesbeiana) is a highly invasive species that has successfully colonized habitats around the world. The level of immune gene genetic variability in an invasive population may contribute to how resistant a population is to pathogens and how persistent a population may be over time. Our project’s objective is to characterize the genetic diversity of a rapidly evolving immune gene in four invasive bullfrog populations in California. Our hypothesis is that our focal population harbors high genetic diversity, consistent with its presence in California for decades and complex introduction history. To test this hypothesis, we isolated a Major Histocompatibility Complex (MHC) gene from 15-20 individuals per population. We then assessed individual heterozygosity, number of alleles in population and genetic diversity of those alleles using standard population genetic metrics. Finally, we compared the genetic diversity within these bullfrog populations to that found in other frog populations, including both native and non-native populations. We found that the levels of diversity in our populations were similar to the levels found in native populations of other anurans but higher than in native species that have undergone population declines.


    38.    Contributed Herpetology  Genetic Variation and Demographic History of the Santa Cruz Black Salamander (Aneides niger). Brandon Kong*, U.C. Santa Cruz; Sean Reilly, U.C. Santa Cruz   brandonrkong@gmail.com

    The Santa Cruz black salamander (Aneides niger) is endemic to the Santa Cruz Mountains of California and is particularly vulnerable to decline due to its restricted range size. Anecdotal observations suggest that the species was relatively abundant in the early 1900’s and has become more difficult to find in the past few decades. However, little work has been done to understand the basic biology of this species including a lack of intraspecific genetic analysis. We re-analyzed mitochondrial and nuclear sequence data from a previous study to examine levels of genetic variation, phylogeographic structure, and test for signatures of population size change within A. niger. Our analyses detected low levels of genetic variation and a general lack of genetic structuring across the range. While neutrality tests of individual loci were non-significant, extended Bayesian skyline plot analysis detected signals of population size fluctuation, and isolation-with-migration analyses estimate a relatively recent population decline. These results suggest that Aneides niger may lack the adaptive genetic variation needed for long-term persistence and that their population size may be unstable. This study affirms the need for a follow-up genetic study with greater geographic sampling, population-level surveys to estimate abundance, and a conservation management plan for this small-range California endemic salamander.


    39.    Contributed Herpetology  Genetic Connectivity of the Western Toad (Anaxyrus boreas) Along Interstate-90 in the Snoqualmie Pass Area of Washington State (USA). Anneliese Myers*, Yakima-Klickitat Fisheries Project, Yakama Nation; Jason Irwin*, Central Washington University   irwinj@cwu.edu

    Single Nucleotide Polymorphism (SNP) analysis was used to assess the genetic connectivity of Western Toad (Anaxyrus boreas) breeding populations along I-90 near Snoqualmie Pass, WA. Sites north and south of the freeway were sampled during the breeding season of 2019. SNP loci were subsequently generated using the proprietary DArTseqTM (Canberra, ACT, Australia) method. A total of 15,468 SNPs were used to calculate pairwise FST values and three distinct breeding populations were identified, two north and one south of I-90. All pairwise FST values between these sites were low (>0.02) but significantly different from 0. The lowest pairwise FST was between the two northern sites, representing the longest distance (11.6 km) and indicating higher levels of connectivity along vs. across the freeway. A de novo Discriminant Analysis of Principal Components (DAPC) confirmed this division between sites on either side of I-90. Although I-90 is the most prominent potential barrier on the landscape, the Yakima River and/or Keechelus Lake may also be contributing to this division. An a priori DAPC was able to distinguish between all populations with enough confidence to assign toads that were encountered away from thre three main breeding sites to their most likely population of origin and will be a useful tool in future studies.


    41.    Contributed Herpetology  Peptide phage display for high-throughput, unbiased profiles of snake venom function. Matthew Holding*, University of Michigan; Laura Haynes, University of Michigan; David Siemieniak, University of Michigan; Colin Kretz, McMaster University; David Ginsburg, University of Michigan   venomatt@umich.edu

    In 2017, snakebite envenoming was recognized by the World Health Organization as a Category A Neglected Tropical Disease, reflecting that snakebite is both far-reaching (5.4 million bites per year) and that further research efforts are expected to produce measures for mitigation of the disease. Venom has also served as a rich source of bioactive proteins developed for both therapeutics and biomedical research. Discoveries in both snakebite treatment and venom bioprospecting have come through more than a century of significant work on individual protein isoforms, while most venom proteins and their complex functions remain poorly characterized. Venom variation is the uniting theme between the biomedical relevance and biological intrigue of venoms, as venom composition can vary among snake life stages, populations, and species. Diverse libraries of peptides displayed on bacteriophage may provide a tool to rapidly characterize the realized functional space of snake venom proteases in high throughput. We have conducted phage-assisted screens of specificity for three purified venom serine proteases from vipers to evaluate the usefulness of phage display. We infer unique substrate specificity profiles for each venom protein, confirming phage display can discriminate the substrate preferences of venom serine proteases. We next relate cut sit preferences to features of the venom protease active site, and use them to predict cut sites in natural human and mouse protein targets. We discuss the potential of phage-display for increasing our knowledge of snakebite physiology, facilitate venom bioprospecting, and linking venom variation to venom function in natural systems.


    42.    Contributed Herpetology  NEW DISTRIBUTION RECORDS OF AMPHIBIANS AND REPTILES OF THE SIERRA DEL CARMEN, COAHUILA, MEXICO. Fabiola Baeza-Tarin, Archbold Biological Station; Tomas Hernandez, Midland College; Mark Herr, University of Kansas; Laine Giovanetto*, New Jersey City University; David Lazcano, Univseridad Autonoma de Nuevo Leon; Sean Graham, Sul Ross State University   lgiovanetto@ymail.com

    We surveyed the Área de Protección de Flora y Fauna Maderas del Carmen (APFFMC) in Coahuila, Mexico for amphibians and reptiles in 2018 and 2019. This is a remote and understudied region of northern Mexico. Survey methods included active visual encounter surveys and nightly road surveys. During our surveys we documented 15 new distribution records including one new state record for Coahuila (Aspidoscelis exanguis) one significant range extension (Sceloporus parvus), and several new municipality records. Our results and previous collections indicate the presence of 67 native amphibian and reptile species and one introduced gecko (Hemidactylus turcicus) within the APFFMC.


    43.    Contributed Herpetology  Investigating Mitonuclear Co-Evolution and Introgression in the North American Cornsnakes (Pantherophis guttatus complex). Thomas Marshall*, University of Texas at Austin; Justin Havird, University of Texas at Austin; David Hillis, University of Texas at Austin   thomm80@utexas.edu

    The Mitonuclear Species Concept recognizes species as lineages with uniquely co-evolved mitochondrial and nuclear genomes. Under this framework, hybridization can disrupt these mitonuclear pairings and produce low-fitness offspring, generating reproductive barriers that maintain species boundaries. However, mitochondrial introgression has been documented in a wide range of organisms, leading to a question of how mitochondrial genomes can cross species boundaries while also maintaining them. One possibility is that hybrid breakdown of fitness is avoided through the selective co-introgression of nuclear alleles essential for maintaining mitochondrial function. In this study, we investigated a likely mitochondrial introgression event in the North American cornsnakes (Pantherophis guttatus complex). We resequenced whole genomes of 34 individuals (4 at high coverage; 30 at low coverage). First, we used a comparative genomic framework to detect evidence of mitochondrial introgression as well as introgression of nuclear genes known to be involved with mitochondrial function (nuclear mitochondrial, or N-mt, genes). Second, we investigated individuals from a contact zone of two divergent mitochondrial haplotypes and asked whether N-mt alleles segregated according to mitochondrial haplotype. The results of this study will shed light on the importance of mitonuclear interactions in shaping the evolutionary dynamics within and among species.


    44.    Contributed Herpetology  Determining Best Thermal Mimic. Andrea Colton*, University of Illinois; Michael Dreslik, University of Illinois   acolton@illinois.edu

    Microhabitat variation creates diversity in thermal niches across a landscape which ectotherms use to effectively thermoregulate. Determining how turtles use available thermal niches allows for a greater understanding of their ecology in light of predicted temperature shifts associated with global climate change. In instances where real animals or materials are not feasible to identify thermal niches across a landscape, researchers employ models mimicking thermal properties of the target organism. Although copper models are considered effective, plastic materials have not been thoroughly tested. Using a real Ornate Box Turtle (Terrapene ornata) shell, we made models of 3D printed plastic and resin casts. We affixed iButtons to each and placed them across various natural thermal gradients to determine which material best thermally mimicked real turtle shells.


    46.    Contributed Herpetology  Phylogenetic Systematics of Ground Agamas (Agama) in Southern Africa. Elizabeth Patton*, Villanova University; Stuart Nielsen, Louisiana State University Shreveport; Todd Jackman, Villanova University; Aaron Bauer, Villanova University   epatton1@villanova.edu

    The ground agamas of Southern Africa are conspicuous, diurnal lizards ubiquitous throughout the subcontinent. Morphological conservatism, incomplete taxon sampling and inadequate geographic coverage has resulted in uncertainties with respect to species boundaries and the validity of certain taxa. Previous data has established deep divergence within more widespread taxa including the A. atra/A. knobeli complex in the southwest and A. aculeata and A. anchiete to the North, but whether such substructure is consistent with species-level divergence remains an open question. We utilized increased gene sampling of all available taxa (notably highly increased coverage of A. atra and A. aculeata) including novel specimens from historically understudied regions (especially Angola) to construct fine-scale, time-calibrated molecular phylogenies in maximum likelihood and Bayesian frameworks using a combination of two mitochondrial (ND4, 16s) and two rapidly evolving nuclear (Rad1, PDC) genes. We were able to resolve the phylogenetic relationships and divergence times between more recently diverged clades and substantiate findings of genetically distinct populations withinA. atra but failed to find support for the elevation of A. knobeli to independent species status.


    47.    Contributed Herpetology  Diversity of the poorly known Neotropical genus Synapturanus (Microhylidae: Otophryninae). Mariela Osorno*, Instituto SINCHI; João Tonini, University of Richmond; Rafael de Sá, University of Richmond   rdesa@richmond.edu

    The family Microhylidae includes three subfamilies of Neotropical frogs: the monotypic Adelastinae, the large Gastrophryninae (83 species) and the Otophryninae. Otophryninae consists of two genera Otophryne Boulenger 1900, with three recognized species, and Synapturanus Carvalho, 1954 with seven species. Species of Synapturanus are terrestrial, small body size, and recently have been reported to lay eggs underground. Until recently, the genus Synapturanus consisted of only three species; however, four new species were described, three of them related to S. salseri or S. mirandariberoi, and one of the S. rabus species complex. The Synapturanus rabus species complex consists of the smaller species in the genus. Our fieldwork in the Amazonia of Colombia collected additional specimens Synapturanus. Morphological and molecular analyses, coupled with phylogenetic analyses, identify samples that could represent new species. Our phylogenetic analyses recovered these samples as three new species associated with the S. rabus species complex, increasing the total diversity of the genus to ten species.


    48.    Contributed Herpetology  Access for all: 3D digitization of wet collections using photogrammetry. Zach Randall*, Florida Museum of Natural History   zach150@ufl.edu

    Photogrammetry is a cost-effective and flexible solution in producing 3D models of biological collection objects. This imaging modality excels at capturing external morphology while retaining realistic colors and textures in a wide range of collection object sizes. Photogrammetry uses photography and computer software to create high resolution and scalable 3D models by texturing a polygonal mesh with a series of overlapped photographs. The Florida Museum has been utilizing photogrammetry to create 3D models of its fluid-preserved specimens to increase digital access to ichthyology and herpetology collection objects for research, education, and outreach. Workflows for imaging, model reconstruction, and data management/access will be discussed.


    49.    Contributed Herpetology  Community-Level Detection of Amphibian Host-Pathogen Distributions Using Museum Frozen Tissue Collections. Daniele Wiley*, Museum of Southwestern Biology, University of New Mexico; Jonathan Duran, Museum of Southwestern Biology, University of New Mexico; Naija Cuzmar, Museum of Southwestern Biology, University of New Mexico; Nalanie Miranda, Museum of Southwestern Biology, University of New Mexico; Matthew Atkinson, University of Central Florida; Anna Savage, University of Central Florida; Lisa Barrow, Museum of Southwestern Biology, University of New Mexico   dlfwiley@gmail.com

    Global wildlife populations are exposed to a variety of emerging pathogens. Anurans are especially vulnerable to introductions of widespread disease-causing organisms and viruses in their environments, such as the fungus Batrachochytrium dendrobatidis (Bd), the virus Ranavirus (Rv), and the protozoan known as amphibian Perkinsea (Pk). These pathogens cause varying effects in amphibians as extreme as community-level mass mortality events. To better understand where and to what degree different species have the potential to interact with these pathogens, we conducted surveys and analyzed frozen tissues archived at the Museum of Southwestern Biology, using established quantitative (q)PCR protocols. We screened 277 individuals from multiple species sampled across the Central and Eastern U.S. for all three pathogens. We also examined environmental and geographic variables for each sampled locality to investigate the potential association between climate and elevation on infection. We found that the most common pathogen detected was Bd with 64 of 277 (23.1%) individuals infected, followed by Rv with 11 of 277 infected (4.0%), and Pk with 2 of 277 infected (0.72%). There were four accounts of co-infection equally split between Bd and Pk, and Bd and Rv. We also found that infection dynamics in Bd did not vary across geographic space or environmental conditions, but there were large differences in prevalence among species. Our study highlights the critical role of frozen tissue collections for uncovering pathogen prevalence and host range, as well as the importance of sampling throughout species’ distributions to better capture insights of host-pathogen evolution and ecology.


    50.    Contributed Herpetology  Arctos: A Collaborative Collection Management Solution For Amphibian, Reptile and Fish Collections. Carol Spencer*, Museum of Vertebrate Zoology, University of California, Berkeley; Emily Braker, University of Colorado Museum of Natural History; Mariel Campbell, Museum of Southwestern Biology, University of New Mexico; Carla Cicero, Museum of Vertebrate Zoology, University of California, Berkeley; Emily DeArmon, Museum of Southwestern Biology, University of New Mexico; Andrew Doll, Denver Museum of Nature and Science; Lindsey Frederick, New Mexico Museum of Natural History and Science; J. Tom Giermakowski, Museum of Southwestern Biology, University of New Mexico; Kyndall Hildebrandt, University of Alaska Museum of the North; Michelle Koo, Museum of Vertebrate Zoology, University of California, Berkeley; Angela Linn, University of Alaska Museum of the North; Teresa Mayfield-Meyer, Museum of Southwestern Biology, University of New Mexico; Christopher Witt, Museum of Southwestern Biology, University of New Mexico; Elizabeth Wommack, University of Wyoming Museum of Vertebrates   atrox@berkeley.edu

    Arctos (arctos.database.museum) is a web-based collaborative collection management system and data portal serving global data on 4.3 million biodiversity and cultural records from more than 247 collections, including 54 fish, amphibian and reptile collections, and nearly 800,000 media objects. Arctos is a leader in providing museums with community-driven solutions to manage and improve data and develop workflows for data cleaning and publication. Pioneered in 1999 and securely hosted since 2012 at the Texas Advanced Computing Center, the portal provides numerous tools and services to manage museum data and make them instantly publicly available. A web interface supports data entry and editing, with integrated tools for transaction and permit management, mapping, citations, object tracking of tissues and specimens, and automated IPT publishing. Arctos strives for superior data quality through its highly-normalized model, controlled vocabularies, and authorities. Shared standardized data has led to innovative ways of relating objects within or between collections, promoting data exploration and interdisciplinary research. Arctos also leverages external services to extend capabilities and generate reciprocal links with collaborators, including Barcode of Life, GBIF, GenBank, Global Biotic Interactions, Global Genome Biodiversity Network, Global Names, iDigBio, iNaturalist, MorphoSource, World Register of Marine Species, and VertNet. Arctos is a community of museum professionals who collaborate on best practices and work together to improve data richness and expand functionality via the Arctos Working Group. By connecting cultural and natural history collections and promoting high quality data, Arctos enables easy data access to ultimately empower research, education, and conservation.


    51.    Contributed Herpetology  Metamorphosis of the Tiger Salamander, Ambystoma tigrinum. Grace Davis*, HALO   grace.e.davis@colostate.edu

    Ambystoma tigrinum, known as the “Tiger Salamander”, remains one of the most well-known amphibians in the nation. Comparatively, very little is known about the details of their metamorphosis, specifically the rate of these processes. To investigate and record data on this topic, 10-20 larval-stage A. tigrinum individuals will be separated and externally influenced for a period of 5 months. Group A water levels will be gradually reduced to mimic the droughts that occur in their natural habitats, whilst Group B’s water level will be maintained at a constant depth. Data will be collected by means of photographs, measurements of individuals, and written behavioral/morphological observations. All will receive the same type and amount of food to ensure accurate results. The research team will include members from the organization, “HALO” as a method of inspiring youth to conserve and research herpetofauna. Results will be presented at the 2023 JMIH.


    52.    Contributed Herpetology  Investigating the Temperature-Size Rule among Eurycea cirrigera. Charlie Holguin*, Virginia Tech; Nicholas Caruso, Virginia Tech; Holly kindsvater, Virginia Tech   holguinc@vt.edu

    An amphibian’s life history includes patterns such as growth, development, and reproduction, which vary due to different environmental conditions. At high latitudes and altitudes, some amphibians take a longer time to reach a minimum size for sexual maturity and the age of first reproduction. This slower life history could have important effects on population ecology, such as age structure, but has only been documented in a few species, so far. This pattern can follow the Temperature-Size Rule (TSR), which describes the phenotypically plastic response of size at different temperatures. However, resources (food) are another source of variation on body size, and it is unclear how it might interact with temperature. In August of 2020, we collected a total of 120 southern two-lined salamanders (E. cirrigera) and we exposed salamanders to a factorial design containing two food treatments (1 and 2 black worms per week) crossed with two temperature treatments (Warm and Cool). Once a month, the salamanders were measured for total length (TL), and mass (g). The data were analyzed in a linear mixed modeling framework with fixed effects of temperature and food treatments and a random effect for individual identity. We found that body size was higher in the high food and high-temperature treatments after 5 months; however, after 10 months the other three treatment groups achieved a similar size. By addressing the TSR through laboratory experiments, we will be able to determine how environmental conditions at early life stages influence the growth of the southern two-lined salamander.


    53.    Contributed Herpetology  THERE’S BABY SNAKES OUT THERE THIS BIG?!?! Dustin Siegel, Southeast Missouri State University; Christopher Murray*, Southeast Missouri State University   dsiegel2@gmail.com

    In the hit movie Anaconda, rapper Ice Cube (as Danny Rich) emphatically asked John Voight (as Paul Serone), “There’s snakes out there this big?” This question was a result of John Voight unravelling a monstrous (approximately 6 meters) shed from an anaconda, most likelyEunectes murinus. Outside of popular fictional movies, the answer to Ice Cube’s question is no, as the largestE. murinusare slightly over 5 meters in length, and few records exist of extant snakes over 6 meters, with the longest records coming from captive individuals (e.g., Medusa the Reticulated Python, 7.67 meters); however, 58-60 million years ago, the answer was yes. During this time, a snake roamed the neotropics of South America with a length of up to 13 meters. The authors who described this extinct species named itTitanoboa cerrejoensisbecause of its gigantic size and discovery in the Cerrejón Formation of northeast Columbia.T. cerrejoensiswas diagnosed as a member of the boid subfamilly Boinae and, thus, it is assumed thatT. cerrejoensiswas viviparous.


    54.    Contributed Herpetology  External nasal gland morphology of Eurycea bislineata (Amphibia, Urodela, Plethodontidae). Samantha Trame, Southeast Missouri State University; Jacob Denova, Southeast Missouri State University; Kenzie Pereira, Duquesne University; Sara McClelland, Moravian University; Kevin Gribbins, University of Indianapolis; Justin Rheubert, University of Findlay; Dustin Siegel*, Southeast Missouri State University   dsiegel2@gmail.com

    Plethodontid salamanders possess numerous courtship glands. Previous studies have shown that the glands are more prominent in male individuals than females, and often experience periods of atrophy and hypertrophy throughout the year that correlate to the non-mating and mating seasons, respectively. We sampled male and female Eurycea bislineatathroughout the year to test the hypothesis that external nasal glands are courtship glands. External nasal glands are paired, branched tubular glands that extend from excretory ducts dorsal to the nares to terminal secretory units posterior to the eyes. We found that the glands hypertrophy and stain/react more intensely with histochemical procedures during the mating season. Hypertrophy of the glands is more pronounced in males, and seasonal variation in epithelial height of external nasal glands of males is significantly correlated to that of seasonal variation in mental gland epithelial height, a known courtship gland found in males, when compared throughout the year. This correlation was not as strong in females, confirming sexual dimorphism of external nasal glands in terms of seasonal variation. We found no ultrastructural differences between male and female external nasal glands. In all specimens, the glandular tubules were lined by a simple, columnar epithelium that was packed with secretory granules that often obscured other cytoplasmic contents.


    55.    Contributed Herpetology  Environmental Enrichment During Development: Neurodevelopmental Impacts of Rearing Tadpoles in Either Empty Tanks or an Enriched Tank Environment. Sara McClelland*, Moravian University   mcclellands@moravian.edu

    Enriched environments have been shown to positively impact neurodevelopment, learning, and memory across vertebrates, with environments that are devoid of stimuli causing increased stress and depression. Amphibians are facing global declines with multiple controlled studies dedicated to better understanding how different variables are impacting amphibian development and survival. These studies often employ standard methods rearing tadpoles in empty tanks devoid of any stimuli or shelter. This environment is not ecologically realistic. Yet, little work has been done analyzing how environments with more stimuli impact amphibians, with even fewer analyzing the impacts on tadpole development. The aim of this research was to determine how developing in a sparse environment (an empty tank) or an enriched environment (a tank with rocks and shelter) would impact tadpole neurodevelopment. To do this, Xenopus laevis tadpoles were raised in either a sparse or enriched environment. Tadpole brains were then extracted, imaged, and linear dimensions measured using ImageJ. By understanding the impacts of environmental conditions on tadpole neurodevelopment, we can better understand how our standard rearing techniques are impacting amphibians. Further, this work could have implications for amphibian conservation by helping us understand how environmental enrichment can contribute to amphibian wellbeing.


    56.    Contributed Herpetology  The Effect of Drought on Alligators’ Eggshell Morphology. Aaron Kaplan*, University of North Florida; Albina Mikhaylova, University of North Florida; Adam Rosenblatt, University of North Florida; M. Laura Habegger, University of North Florida   arkaplan24@gmail.com

    The eggshell is one of the most crucial adaptations that has enabled vertebrate’s transition to land. Among the different components and materials that make up this structure there is a network of pores that facilitate the exchange of vital resources between the developing embryo and the external environment. The size, number and distribution of these pores are variable and dynamic, as they are partially shaped by microbes; environmental changes could also affect these pores leading to embryo mortality. Although eggshell research is abundant, most work has been centered on birds compared to other vertebrates, and even less is known on the effect environmental changes (i.e., climate change) may have on eggshell morphology and embryo survival. Therefore, the goal of this research is to determine if changes in nest humidity may impact eggshell morphology in the American alligator (Alligator mississippiensis). Alligator eggs collected in the wild were incubated in the lab at different humidities and eggshell pore morphology was characterized with advanced microscopy techniques. Pore numbers, size, and distribution, among other variables, were assessed along the eggshells of over 200 eggs. Patterns in pore distribution and size will be analyzed in the context of varying humidities and embryo viability to correlate any possible findings. Our results will serve as the foundation for future work focusing on how climate change may affect alligator’s embryo viability.


    57.    Contributed Herpetology  Limbs, Shoulders, Necks, and Trunks: An Investigation of the Neck-Trunk Boundary in Limbless Squamates. Krista Koeller*, Unaffiliated   kkoeller3115@gmail.com

    At least 26 squamate lineages have independently evolved an elongate, limbless body plan. This transition always involves body elongation, but which body regions contribute most to elongation has been difficult to discern, largely due to a breakdown in the boundary between the neck and trunk regions. In limbed species, this boundary, defined by the location of the sternum, is associated with distinctive features (e.g. limbs, vertebral characters). However, many elongate species lack sterna, and other neck-trunk boundary characteristics are displaced relative to each other. This has caused confusion about where the neck-trunk boundary is located in elongate groups. To bring clarity to this issue, CT data from limbed and limbless taxa across Squamata were examined to determine the axial positions of neck-trunk boundary markers such as the pectoral girdle, the posterior extent of cervical vertebral characters, and overall vertebra shape. Results suggest that generally, the position of the pectoral girdle is conserved in species that retain it and cervical characters are the most posteriorly displaced, compared to the plesiomorphic body plan. In elongate species with highly reduced pectoral girdles, the cervical characters usually associated with the girdle are not, meaning vertebral characters may not be a reliable indicator of where the traditionally defined neck-trunk boundary would be. Interestingly, few characters or clades show evidence of neck elongation, but in some clades, there is a clear tendency towards a reduction in neck length. This means that in these groups, presacral elongation has occurred solely in the trunk region.


    58.    Contributed Herpetology  The Skull of the Semi-Aquatic Neotropical Lizard Genus Echinosaura (Gymnopthalmidae: Cercosaurinae) With Comments on Important Characters Within Gymnophthalmoidea. Cristian Hernandez-Morales*, University of Texas at Arlington; Alison Ngo, University of Texas at Arlington; Leila Abdelhadi, University of Texas at Arlington; Hannah Falks, University of Texas at Arlington; Walter E. Schargel, University of Texas at Arlington; Eric N. Smith, University of Texas at Arlington; Juan D. Daza, Sam Houston State University   nandezsendo@gmail.com

    The Neotropical genus Echinosaura is composed of eight semi-aquatic species of lizards distributed from northern Panama to Ecuador. Most of these species are distributed throughout the Biogeographical Choco region in Colombia and Ecuador, except E. panamensis from Panama and E. centralisfrom the Inter-Andean Valley of the Magdalena River. The current hypothesis of relationships within Echinosaura relies mainly on genotypic and integumentary characters, while osteological data have been historically overlooked. Here we provide a detailed qualitative analysis of the skull of five species of Echinosaura (Viz. E. brachycephala, E. fischerorum, E. horrida, E. keyi, and E. palmeri) and one species used as an outgroup (Andinosaura hyposticta). We found some putative synapomorphies for Echinosaura including (1) a well-developed posterior process of the jugal, (2) dermal sculpturing of the dorsal surface of the parietal and frontal, and (3) jaw adductor musculature originating on the ventral side of the parietal. Also, we found some character states shared by Andinosaura and Echinosaura but variable in the more comprehensive context of Gymnophthalmoidea (Alopoglossidae, Teiidae, Gymnophthamidae), such as (1) ethmoidal foramina piercing the ascending process of the premaxilla, (2) postorbital excluded from the orbit in lateral view, and (3) lateral process of the orbitosphenoid reduced or absent. This pilot study reaffirms osteology as a rich source of data to corroborate and resolve classifications based on genotypic and external morphology.


    59.    Contributed Herpetology  Comparative Skull Morphology and Body Size Evolution of Mainland African Plated Lizards (Squamata: Gerrhosauridae). Kelsey Fenner*, Villanova University; Aaron Bauer, Villanova University   kfenner@villanova.edu

    Mainland African gerrhosaurs have a clear bimodal body size distribution that is unique among lizards. Three genera, Broadleysaurus, Matobosaurus and Gerrhosaurus, exhibit large to very large body size (up to 300mm SVL) and two genera, Tetradactylus and Cordylosaurus, exhibit small to very small body size (up to 80mm SVL). We hypothesize that the two small genera have undergone a heterochronic shift and evolved a miniaturized body plan. Gerrhosaurs are heavily armored lizards encased by adherent osteoderms which have previously made detailed observations of their osteology difficult. Previous studies on the skull morphology of gerrhosaurs have mainly focused on features of the intact skull, and only looked at a few representative species that do not fully capture the diversity of this group. We obtained scans for 17/18 extant species and used micro-CT-scanning to remove many of the osteoderms exposing the underlying structures. This allowed for in depth analysis of both the skull as a whole and each individual skull bone across all five genera. All mainland gerrhosaurs were found to have more cancellous skull bones in comparison to most other lizards. The volume of the skull of the largest species (295 mm3) was found to be 74 times that of the smallest (4 mm3). The two smaller genera exhibit many traits that have been identified as good indicators of miniaturization in other squamate groups including an increase in the size of the neurocranium in relation to the dermatocranium, greater fusion in the jaw bones, and a closure of the post-temporal fenestrae.


    60.    Contributed Herpetology  Comparative osteology of Western Australian toadlets (Myobatrachidae; Uperoleia). Ryan Cook*, Villanova University; Aaron Bauer, Villanova University   ryanacook2@gmail.com

    Australian toadlets (genus Uperoleia) are the most species-rich frogs in the Family Myobatrachidae, itself the most diverse anuran family in Australia. The genus includes 29 currently recognized species (approximately half of which occur in Western Australia), and relatively little is known about phenotypic variation across this genus. External morphology of Uperoleia is largely conservative, and most species are not easily identifiable based on traditional characters. Other sources of phenotypic variation, however, may be more informative. My study uses micro-computed tomography (micro-CT) scans, bone-by-bone analyses, geometric morphometrics, and phylogenetic comparative methods to analyze variation in the osteology of Western Australian Uperoleia frogs. I predict that (1) miniaturization of the genus may have imposed a superficial similarity across congeners that has previously obscured functional and phylogenetic differences; (2) bone-by-bone analyses will reveal diagnostic features consistent with the molecular phylogenetics of Uperoleia; and (3) variation in bone shape, reflected through morphometrics, will reveal functional differences among Uperoleia species, as has been demonstrated in family-wide studies. Based on the results from this combination of analyses, I show that careful examination of Uperoleia osteology highlights a significant phylogenetic relationship across the genus and provides some clarity on the uncertain identity of the type species, U. marmorata. By employing multiple phenotypic traits in an integrative taxonomic context, my study provides a model for studying the phenotype of other traditionally “difficult” taxonomic groups).


    61.    Contributed Herpetology  An Evaluation of Sexual Dimorphism in Head Size and Shape in Pseudotriton ruber (Red Salamander). Kenneth Wang*, Milton High School; Kelly-Ann McDonald, Kennesaw State University; Tyshiona Brandon, Kennesaw State University; Todd Pierson, Kennesaw State University   haotzuw@gmail.com

    For many secretive species of amphibians and reptiles, observations of reproductive behavior are limited to sparse anecdotes from the field or to animals held in captivity. However, a careful examination of morphological differences between the sexes can help shape hypotheses about behaviors with which they may be correlated. For example, sexual dimorphism in head size and shape are correlated with courtship, territoriality, and mate-defense behaviors in some salamanders. One widespread species with a poorly described reproductive natural history is the Red Salamander (Pseudotriton ruber). Here, we measure head size and employ geometric morphometric methods to evaluate head shape in P. ruber, with the goals of: 1) quantifying and visualizing sexual variation; 2) forming hypotheses about reproductive behavior. We found preliminary evidence for differences in head size and shape that are consistent with mate-guarding behavior, and we remark upon directions for future research.


    62.    Contributed Herpetology  Morphology of the olfactory organs of the California Slender Salamander (Batrachoseps attenuatus). Emily, E. Gremling*, Cal Poly Humboldt; Karen, M. Kiemnec-Tyburczy, Cal Poly Humboldt; John, O. Reiss, Cal Poly Humboldt   eeg36@humboldt.edu

    The amphibian olfactory system typically functions both for aerial and aquatic olfaction. In salamanders, the peripheral olfactory system is composed of paired, sac-like structures, which are the main olfactory cavities (MOC). Each cavity has a lateral diverticulum, the vomeronasal organ (VNO). Research on olfactory morphology has been conducted on several model amphibians, but no previous studies of the most species-rich family of salamanders, the Plethodontidae, have looked at cellular morphology at the level of electron microscopy. Plethodontid salamanders are documented using olfaction for a variety of social behaviors including courtship and territory recognition. Nose-tapping plays a large role in plethodontid behavior. We examined the morphology of the MOC and VNO of adult Batrachoseps attenuatus (Urodela: Plethodontidae: Hemidactyliinae) using light and electron microscopy, as well as microCT scanning. We found that olfactory epithelium is thicker in the anterior regions of both the MOC and VNO. A fold extends ventrally from the roof of the organ, separating the MOC and VNO. Two types of cilia are present in different regions of the MOC and VNO. One type is long and spreads parallel to the surface of the epithelium; these are presumably olfactory cilia. The other is shorter and extends perpendicular to the epithelium; these are likely motile cilia of respiratory epithelium. The nasolacrimal duct extends anteriorly from the eye before turning and entering the VNO dorsally. Thinning of anterior olfactory epithelium and its replacement by posterior nonsensory epithelium resembles the condition observed by Dawley (2017) in other plethodontid genera (Plethodon and Desmognathus).


    63.    Contributed Herpetology  Reproductive Behaviors of the Sidewinder Rattlesnake (Crotalus cerastes). Jacob Apjohn*, Castleton University; Brad Coupe, Castleton University   jakeapjohn@gmail.com

    Rattlesnakes are reclusive, so their social behaviors are rarely seen. Sidewinder rattlesnakes employ a list of intricate behaviors when they come together to mate. We watched footage of courtship and copulation of sidewinders to generate an ethogram, and then recorded the individual behavioral events in the behavior-logging software BORIS. Courtship behaviors were only observed in one pair of snakes. The behaviors included head jerking, where the male flicks his tongue and jerks his head along the female’s dorsum, and tail search copulatory attempts. The courted female only displayed the occasional rattle, and rapidly moved away from the male multiple times. The male repeatedly performed head-jerks along the female’s dorsum and would then immediately follow it by a tail search copulatory attempt. This was repeated over the next few hours. The duration and frequency of these behaviors have been documented in other rattlesnakes, but there is little information on their copulatory behaviors. The female displayed a traveling behavior, where she would crawl during copulation and drag the male behind her, cloacal contractions, and tail-lifting. These behaviors were observed in five pairs of snakes. In four of the mating pairs, the males would perform a periodic sequence of tail contractions that often followed female cloacal contractions. The function of these tail contractions is unknown, and we do not know of this behavior in any other species of snake.


    64.    Contributed Herpetology  Investigating Personality in Venomous Snakes: Individual Rattlesnakes Exhibit Consistent Behavioral Responses in Defensive and Exploratory Contexts. Ricardo Gibert, San Diego State University; Dylan Maag*, San Diego State University; Rulon Clark, San Diego State University   dmaag3229@sdsu.edu

    A burgeoning literature in animal behavior has demonstrated that most animals exhibit consistent individual variation in core behavioral traits, or personality. However, the taxonomic spread of animal personality studies is uneven, with some ecologically important and diverse taxa still unstudied. Some of these understudied groups, such as venomous snakes, are also frequent targets of mitigation due to human/wildlife conflict, and conservation researchers have been increasingly focused on developing a general understanding of how individual personality or temperament mediates wildlife responses to management or mitigation actions. In this study, we used twenty captive Southern Pacific rattlesnakes (Crotalus oreganus) in standardized assays to test for repeatable behaviors (i.e., personalities) between individuals and examine possible relationships in personality traits across contexts (i.e., behavioral syndromes). Repeatability of behaviors was assessed over five repeated trials consisting of a handling assay, an open field test, and a threat assay. We found five of our scored behaviors related to exploration/avoidance, activity level, and boldness/shyness showed significant repeatability. However, we found no evidence for behavioral syndromes in these behaviors. Our analysis shows that, similar to many other species and taxonomic groups, rattlesnakes also display individual personality traits when tested under standardized conditions, and we discuss the implications of this finding for mitigation, conservation, and comparative analyses across broadly similar species groups.


    65.    Contributed Herpetology  Social Behavior Deficiencies in Captive American Alligators (Alligator mississippiensis). Zane Cullinane Walsh*, New College of Florida; Hannah Olson*, New College of Florida; Miranda Clendening, New College of Florida; Athena Rycyk, New College of Florida   zane.cullinanewals19@ncf.edu

    Complex social behaviors are observed in wild alligator congregations, but it is uncertain if similar behaviors are observed as frequently in captive alligator congregations. Social behaviors can be indicative of captive animal wellbeing and by comparing the behavior of captive American alligator (Alligator mississippiensis) congregations to wild alligator congregations, we can create a baseline to measure the welfare of captive alligators. In January 2021, we studied the behavior of captive and wild American alligator congregations in Florida, USA. Our study indicated that the wild alligator congregation, on average, displayed a 827% higher frequency of social behaviors than those observed in captivity. Types of social behaviors performed were more diverse in the wild congregation than those observed in the captive congregation. The most common social behaviors observed in the wild congregation were growling and HOTA (head oblique tail arched) posturing while the most common social behavior in the captive congregation was fighting. In the captive congregation, high walking occupied 94% of general behaviors (not including feeding) and presented as a potential stereotypy. Behavioral differences identified in this study were likely a result of clear differences in the surrounding environment of the captive and wild alligator congregations. Possible contributing factors include the presence of visitors, available space, the availability of resources, and overall stocking density. The striking differences in social and general behavior between the captive and wild congregations can serve as a basis for future investigations of other indicators of alligator welfare in captivity.


    66.    Contributed Herpetology  Skin Lipids Alone Enable Scent Trail Discrimination in an Invasive Reptile, the Argentine Black and White Tegu. Lauren A. Nazarian, James Madison University; Eric A. Tillman, USDA-APHIS NWRC; Bryan M. Kluever, USDA-APHIS NWRC; M. Rockwell Parker*, James Madison University   mrockwellparker@gmail.com

    Mate searching is facilitated by a variety of cues provided by conspecifics, and chemical cues are the most ubiquitous, especially in squamate reptiles (snakes and lizards). In the majority of squamate taxa, skin lipids serve as the primary source of valuable chemosensory information that guides both mate searching and discrimination. Invasive reptiles are adept at finding mates in novel habitats, and our research focuses on assessing the influence of chemical cues on mate searching. Previously, we found that both male and female Argentine black and white tegus (Salvator merianae) could follow conspecific-laid scent trails in a Y-maze and that females were more discriminatory than males. We therefore isolated skin lipids from male and female tegu shed skins via large-scale solvent extraction to determine if lipids were the source of the information used for discrimination. We used a fully randomized, repeated measures design and tested male (n=7) and female (n=7) tegus in each of the following Y-maze conditions using isolated skin lipids to create scent trails: male lipids vs. control lipids (peanut oil), female lipids vs. control, and male vs. female lipids. Tegus (both sexes) showed a strong preference for male scent trails; they also preferred opposite-sex skin lipid trails when presented with both (male vs. female). Surprisingly, there was no variation in any of the behaviors we quantified based on either sex or trial type. Isolated skin lipids are therefore potent and consistent cues motivating scent trailing behavior in this invasive reptile.


    67.    Contributed Herpetology  The boys in blue: information content of color signals in the eastern fence lizard. Parker Woodward*, Marian University; Jacob Pressler, Marian University; Siera Stuart, Marian University; Alison Ossip-Drahos, Marian University; Julio Rivera, Arizona State University; Emilia Martins, Arizona State University   parker61402@icloud.com

    Color patches frequently convey useful information during aggressive male-male encounters, with the patch size often signaling dominance or aggression. Most male lizards in the genus Sceloporus have paired blue belly patches rimmed with black borders that are actively displayed during territorial disputes. Territorial disputes will often escalate to fighting, where lizards will bite one another, potentially resulting in injuries. Though males display these two colors simultaneously, previous behavioral work indicates that they may act as separate signals and are interpreted differently by receivers. Here, using territorial male Sceloporus undulatus lizards in the field, we examine the relationship between color patch size and bite force, in order to tease apart the information content of these two color signals. We find that blue belly patches do not predict bite force, but the combination of black throat and black belly borders is predictive of bite force. This indicates that in this species, multiple color traits are used to convey different types of information.


    68.    Contributed Herpetology  Calling Phenology of Amphibian Assemblages in the Hudson Highlands. Patrick Baker*, United States Military Academy; Paris Williams, United States Military Academy; Rachel Hughes, Oregon State University; Patrick Wolff, Engineer Research and Development Center; Aaron Rice, Cornell Lab of Ornithology, Cornell University   dr.pjbaker@gmail.com

    Passive acoustic monitoring (PAM) of advertisement calls allows for non-invasive assessment of species diversity and reproductive activity of amphibian communities. The PAM approach is useful when surveying assemblages of amphibians in training areas on military lands that would be otherwise difficult to access for traditional surveys. We deployed PAM recorders at three locations on the United States Military Academy Reservation from 2020 to 2022 to determine species composition, phenology of breeding based on calling activity, and to relate the timing of activity to environmental conditions. Although our locations are separated by less than 15 km they span > 350 m of elevation from Constitution Island (sea level) to Bear Swamp, near the summit of Bull Hill. The date of first chorus varied by year and location; however, the onset of calling activity typically began in March (Julian Day 62-84) with Spring Peepers (Pseudacris crucifer) and Wood Frogs (Lithobates sylvaticus) followed by American Toads (Anaxyrus americanus) and Pickerel Frogs (Lithobates palustris). Choruses of Grey Tree Frogs (Dryophytes versicolor), Green Frogs (Lithobates clamitans), and Bullfrogs (Lithobates catesbeianus) began in early to late May (Julian Day 124-139). Fowler's Toads (Anaxyrus fowleri) were only found at one location (Morgan Farms) and were the last species to initiate chorus activity (Julian Day 180). The Date of First Call for Spring Peepers and Wood Frogs was correlated with Growing Degree Days (Base temperature 3C).


    69.    Contributed Herpetology  Do Varying Levels of Predation Risk Affect Territorial Behavior by Male and Female Ozark Zigzag Salamanders? Alicia Mathis*, Missouri State University; Sarah Heimbach, Missouri State University   aliciamathis@missouristate.edu

    Territorial defense incurs costs, including increased visibility to predators. Salamanders that can accurately assess the level of predation threat might be able to adjust their territorial behavior to maximize the difference between costs and benefits. For example, when predation risk is high, territorial individuals might shift to lower-visibility displays. In this study, adult male and female Ozark Zigzag salamanders were exposed to different degrees of predation stress immediately prior to territorial contests: blank control (none), predator chemical cues only (low), physical attacks only (medium), and a combination of chemical cues and physical attacks (high). In comparison to the control, females decreased their time in the high–visibility posture and decreased their time in the low visibility posture across all treatments. Although males also decreased time in the high-visibility posture across all risk treatments they only increased time in the low-visibility treatment in response to the highest level of threat. The differential response by males and females may indicate that the sexes differ in the costs and benefits associated with territorial defense and antipredator behavior.


    70.    Contributed Herpetology  Pond community structure and climate change resilience: restoration and management applications in the Willamette Valley, OR. Courtney Hendrickson*, Oregon State University; Tiffany Garcia, Oregon State University; Andrew Blaustein, Oregon State University   hendrcou@oregonstate.edu

    Understanding how abiotic conditions differentially impact community organization is crucial for management and conservation of species in complex systems. Predictable habitat variability plays an important role in shaping community composition. However, community response to unpredictable variability, such as stochastic temperature shocks characteristic of climate change, remains unclear. Ponds are composed of complex biotic and abiotic interactions that vary with seasonal shifts in precipitation and snowpack. These freshwater habitats are critical targets for restoration efforts as they provide important ecosystem services and are hotspots for biodiversity. Communities structured around, and adapted to, temporary vs. permanent water availability likely experience differential responses when challenged with unpredictably high temperature spikes. Community-based management, rather than focusing on single-taxa, might enhance our ability to build resilience to climate change related disturbances. The goals of this project are to understand the role of community structure in resilience to climate change related disturbance and inform pond restoration and management efforts in the Willamette Valley, OR. We have characterized macroinvertebrate and amphibian assemblages in ponds across the Willamette Valley with varying levels of water permanence. Our findings are consistent with theory that predicts more diversity and less population variability in ponds with permanent hydroperiods, with the inverse in temporary ponds. These observations will allow us to design experiments for examining community-level climate change resiliency thresholds. This study supports management goals by identifying restoration priorities and developing scenario plans for climate change mitigation.


    71.    Contributed Herpetology  Below the Undergrowth: Quantifying Soil in the Range of Two Microendemic Plethodontids Compared to the Widespread Plethodon cinereus. Catherine Renton, University of Scranton; Janice Healey, University of Scranton; Vincent Farallo*, University of Scranton   vfarallo@gmail.com

    The role of micro-scale environments in species ecology has received more attention in recent years. Species that individually exist at micro-scale, such as salamanders, often have substantially different geographic species distributions, including the widespread Plethodon cinereus (>1 million km2) and microendemics P. hubrichti and P. sherando (<100 km2). These differences in geographic scale can make understanding their ecology more challenging. Furthermore, the micro-scale environment that salamanders occupy includes a significant amount of time spent underground, despite little understanding of their interactions with soil and other components of the underground structure (e.g., rocks). Here we compare and quantify soil properties within the range of these three species. Bulk density, porosity, and pH significantly differed between the two microendemic salamander habitats. However, when comparing the soil within the range of each microendemic species to locations directly adjacent to their range where P. cinereus is present, only pH was significantly different with a more acidic pH in soil from the range of P. sherando compared to an adjacent P. cineresus site (the trend was similar when comparing P. hubrichti to nearby P. cinereus but the differences were not significant). We also observed notably large rocks (>10 cm) within random samples of both microendemic locations and none in the range of P. cinereus. Our study suggests that more data is needed on the interaction of salamanders and their belowground environment. This may be pivotal in understanding speciation and providing further information on their ability to deal with a changing environment.


    72.    Contributed Herpetology  Road ecology through the lens of snakes in the Columbia Basin: Patterns of mortality, occurrence, and activity. Adrian Slade*, Central Washington University; Tyler Larsen*, Central Washington University   slade.oreganus@gmail.com

    Snakes bear an array of life history characteristics making them particularly vulnerable to the impact of roads. From May through October 2017, we examined the crepuscular and nocturnal road-crossing activity patterns of four sympatric Columbia basin snakes: Crotalus oreganus oreganus (Northern Pacific rattlesnake), Pituophis catenifer deserticola(Great Basin gopher snake),Charina bottae bottae (Northern rubber boa), andHypsiglena chlorophaea deserticola (Northern desert nightsnake). To do this, we conducted nightly transect surveys on three roads in central Washington State. We collected data on 924 individual snakes and analyzed patterns of occurrence, movement, and mortality in the context of anthropogenic factors, natural history, season, and habitat. Gopher snakes and rattlesnakes comprised the majority (90%) of the observed species. Occurrence of both species on road transects peaked in June. Gopher snakes were encountered more frequently than rattlesnakes in spring and fall, while rattlesnakes dominated the hottest part of the summer. Our data reveal distribution patterns for cryptic species (C. b. bottae and H. c. deserticola) and identify areas of high snake density and road-crossing activity. Our data also show that snakes traversing the road with highest traffic volumes have about a 30% chance of surviving the crossing.


    73.    Contributed Herpetology  Patterns Affecting Snake Species Occupancy within a Chronosequence of Tallgrass Prairie Restorations. Grace Wu*, University of Illinois Urbana-Champaign; Michael J. Dreslik, Illinois Natural History Survey   gwu21@illinois.edu

    A primary objective of restoration is to increase the extent and diversity of tallgrass prairie ecosystems often only use plant diversity metrics as delineators of success. Although the diversity and occupancy of fauna can be readily measured, they are typically not considered measures of success. The impact of restored habitat on snake species is particularly difficult to assess due to their secretive nature, but their role as predators and prey are important ecosystem functions. We investigated snake species occupancy of a chronosequence of 13 restored sites within the Midewin National Tallgrass Prairie in Wilmington, IL using wood and tin artificial cover objects from 2018 through 2019. Of the seven species detected during the study, occupancy could be modeled for the five most abundant. Results showed differing occupancy estimates between species with no positive correlation between occupancy and the age of restored prairie habitat suggesting snake species occupancy of restored prairies is affected by something other than time since initial restoration.


    74.    Contributed Herpetology  Daily and Seasonal Activity of Northern Watersnakes (Nerodia sipedon sipedon) in a Central Michigan Wetland. John Rowe*, Alma College; Tyler Goerge, Ohio University; Chelsea . Martin, Loma Linda University; William Mulligan, Grand Valley State University   microlophus@gmail.com

    Understanding spatial ecology of organisms can lead to insights into the influences of habitat heterogeneity and seasonal variations on resource use. Over a three-year period, we used radiotelemetry to study patterns of space use in Northern Watersnakes (Nerodia sipedon sipedon) in a northern Michigan wetland. Multiple daily radiolocations (n = 3) throughout the summer months revealed that snakes spent a clear majority of their time during the daylight hours concealed in submerged and emergent vegetation along the littoral shelf of the lake. At other times, snakes were observed in aquatic vegetation at the water’s surface or basking on, or concealed in, vegetation of the Sphagnum mat. Snakes spent the night largely stationary while suspended in aquatic vegetation. Total daily distances moved (based on three radiolocations) were typically less than 5 m although longer distances (> 30 m) were sometimes observed during shifts among core activity areas. Seasonally, snakes remained along the lake edge during summer although some pregnant females traveled to surrounding wetlands, presumably for the purpose parturition. Snakes overwintered near the lake in mostly terrestrial habitats. We did not observe differences in spatial ecology between the sexes but our sample size for male snakes was small.


    75.    Contributed Herpetology  Spatial distribution and population density of Plethodon cinereus using mark-capture-recapture methods. Shayla Graham, Lock Haven University; Jillian Fleming, USGS Eastern Ecological Science Center; Jennifer Deitloff*, Lock Haven University   jenneymd@gmail.com

    Plethodon cinereus (Eastern Red-backed Salamander) can be found throughout the Eastern United States. Individuals of this species establish and defend territories, and typically remain near these territories when foraging for prey. Territories can be beneficial for finding prey, attracting mates, and sheltering from harsh weather conditions during dry or warm weather. In this study, we observed the movement, density, and spatial distribution of Plethodon cinereus using capture-mark-recapture methods. We used six study plots, and each plot contained fifty artificial coverboards. When a salamander was captured, we tagged them with a unique color code using an elastic polymer in multiple colors. For animals that were recaptured, we recorded the color code and marked the salamander as a recapture. We recorded sex and snout vent length (SVL) of all the adult salamanders (>30mm) captured. To assign sex, we used three categories: male (vas deferens present), gravid females (no vas deferens but eggs present), and non-gravid females (no vas deferens or eggs present). In general, we found more gravid females than non-gravid females, and more females than males. We also found that the salamanders tended to stay within a specific area and, often, were recaptured under the same coverboards. This demonstrates site fidelity in our population of salamanders. The number of salamanders captured and recaptured between plots differed. Possibly, the different plots contain variability in territorial quality or prey availability. Recording site fidelity and movement patterns could help us understand the intraspecific and territorial behaviors of Plethodon cinereus.


    76.    Contributed Herpetology  Testing a Collaborative CURE Model Using a Large-scale Salamander Research Network. Louise Mead*, Michigan State University; Kristine Grayson, University of Richmond; Caitlin Fisher-Reid, Bridgewater State University; Raisa Hernandez-Pacheco, California State University; Alexa Warwick, Michigan State University; Tanya Matlaga, Susquehanna University; Sean Sterrett, Monmouth University; Jill Fleming, USGS Patuxent Wildlife Research Center; Chris Sutherland, University of St. Andrews; David Miller, Pennsylvania State University; Evan Grant, USGS Patuxent Wildlife Research Center   lsmead@msu.edu

    Course-based Undergraduate Research Experiences (CUREs) engage students in the practice of science and can enhance undergraduate education by including a large number of students in authentic research. We developed a CURE framework that integrates students into an intellectually diverse and established research network. Our course-based research modules leverage the Salamander Population Adaptation Research Collaboration network (SPARCnet), founded in 2013 by researchers at Pennsylvania State University and USGS Northeast Amphibian and Research Monitoring Initiative. Network research questions examine individual and population level responses of Plethodon cinereus to environmental variation and changes expected under human disturbance. Applying the same cover-board plot study design and sampling methods, participants mark salamanders, and thus have long-term spatial-capture-recapture data and basic population demographic data. Our modules engage students in collecting and analyzing field data from a wide geographic range, emphasizing ecology and evolutionary processes, while simultaneously helping students understand both the collaborative nature of science and the quantitative skills required for working with large data sets. We implemented modules in a variety of undergraduate biology courses engaging students in original and publishable data collection, leading to analysis and interpretation of large, multi-year data sets. We paired classrooms during the research experience to develop cross-institution student collaborations. We measured student self-efficacy, science identity, and quantitative and scientific literacy in classrooms using our modules, comparing paired and unpaired implementations. The results of the project will advance knowledge of how this pedagogical approach impacts student outcomes using a cross-institution collaborative CURE framework.


    77.    Contributed Herpetology  Microbiome Characterization and Comparative Diversity Among Biotypes in the Blue-spotted Salamander (Ambystome laterale) Complex in Central Michigan. Zachary Laughlin*, Central Michigan University; Kirsten Nicholson, Central Michigan University   laugh1zt@cmich.edu

    An explosion of recent studies has demonstrated broad diversity and unexpected patterns in the microbiomes of many organisms. Published evidence suggests microbes may play important roles in immune defense and chemical cues for identification of species and or sex. This study sought to extend that knowledge by investigating the interaction of genetics and environment on microbiome diversity. The objective of this study was to describe and compare the microbial communities of four genetic biotypes (LL, JL, JLL, JLLL) within the Blue-spotted Salamander complex (Ambystoma laterale) from two pond communities in Central Michigan. We hypothesized that (a) some microbes would be shared among the biotypes and pond communities, (b) unique microbiota would significantly distinguish all biotypes and (c) pond communities. Ninety salamanders were swabbed to examine their microbiomes. Preliminary data collected at the time of this abstract submission indicates support for our hypotheses; banding patterns from initial DGGE gels indicates some microbes are shared among ponds and biotypes, but variation is observed and unique bands appear to distinguish ponds and biotypes. Final analyses and interpretation will be presented at the presentation.


    78.    Contributed Herpetology  The interactions of Batrachochytrium dendrobatidis infection levels and frog cutaneous microbiomes in Cheney, WA, USA. Krista Dodd*, Eastern Washington University; Philip Campos, Eastern Washington University; Autumn Holley, Eastern Washington University; Angela Denton, Eastern Washington University; Jonah Piovia-Scott, Washington State University; Kimberly Cook, Washington State University; Jenifer Walke, Eastern Washington University    kdodd2@ewu.edu

    The fungal pathogen, Batrachochytrium dendrobatids (Bd), has led to the decline and extinction of many amphibian populations, but some bacteria in the skin microbiome can inhibit its growth. In Turnbull National Wildlife Refuge (TNWR) in eastern Washington, Bd is highly prevalent, but the role of the skin microbiome in Bd infection dynamics have not been examined in this region. We hypothesized that frogs with lower Bd infection intensities would have higher skin bacterial diversity and more abundant anti-Bd bacteria, indicative of a more protective function. Our study combined culture-independent and culture-dependent methods to assess the relationship between Bd and the microbiome of the Columbia Spotted Frog (Rana luteiventris, N=46) and the Pacific Chorus Frog (Pseudacris regilla, N=72) in TNWR. We characterized skin bacterial diversity with 16S rRNA gene amplicon sequencing on Illumina MiSeq, and quantified Bd infection intensity with qPCR. P. regilla had significantly higher Bd infection intensities (14,480 zoospore equivalents) and prevalence (91.43%) compared to R. luteiventris (intensity: 1,647.36 zoospore equivalents, prevalence: 67.74%). To evaluate whether these infection differences correlate with the skin microbiome, a culture-dependent method was used to determine which bacterial isolates produce anti-Bd metabolites in in vitroco-culture assays, followed by a comparison of culture and culture-independent DNA sequences to determine relative abundance of anti-Bd bacteria on wild frogs. We found that highly infected P. regilla had different skin microbiomes than those with low/no Bd infection (Mantel test, p=0.02, rs=0.43). If our hypotheses are supported, an anti-Bd probiotic could be developed to protect threatened amphibians.


    79.    Contributed Herpetology  New Insights from Old Specimens: The Autecology of Angolan Snakes. Calum Devaney*, Villanova University   calumdevaney@comcast.net

    There are 126 species of alethinophidian snakes recorded from Angola, of which five are endemic. Most of the remaining taxa are broadly distributed in either Central or Southern Africa, although some are shared only with immediately contiguous countries. Data on the autecology of Angolan alethinophidians are extremely limited. Although the parity mode for nearly all has been established, information about the seasonality of reproduction and clutch/litter size is available for only six species based on Angolan material. Likewise, dietary data are available for only 17 species. By revisiting historical material as well as newly collected specimens I have been able to provide some autecological data for additional species, including Causus maculatus, Boaedon variegatus, and Lycophidion namibianum. Clutch sizes from Angolan specimens were found to be generally smaller than those of the same species in other countries. Most species were reproductively active in the wet season. Dietary habits of Angolan snakes are consistent with those of conspecifics in more well-studied Central and Southern African countries. Overall, new data collected from Angolan snake specimens, especially from the Vernay and Pulitzer expeditions, reveals first accounts of diet, clutch size, and seasonality of reproduction for several poorly studied species as well as some recently described species in Angola; and this data confirms; and this data supports previous findings for intraspecific parallels in the ecology of widespread species in other parts of Africa.


    80.    Contributed Herpetology  Preliminary Observation on the Interaction Dynamics of Three Frogs’ Genera (Amphibia:Ranidae) Inhabiting Torrential Stream Habitats in Sumatra, Indonesia. Umilaela Arifin*, Leibniz Institute for the Analysis of Biodiversity Change   u.arifin@leibniz-lib.de

    The formation of the Bukit Barisan Mountain range that stretches along Sumatra’s longitudinal axis had profoundly shaped the topology of the island. As consequence, various types of forest ecosystems are available across elevational gradient, providing plentiful habitats and microhabitats for each Sumatran species, including the cascade frogs’ species. Odorrana, Wijayarana, and Sumaterana are the three genera of ranid frogs that commonly observed to inhabit torrential stream habitats in Sumatra. However, the interaction among those species have never been recorded to date. During fieldwork between 2014–2015, we conducted Visual Encounter Surveys in several torrential streams in Sumatra and recorded various ecological parameters (e.g., distance from water and microhabitat) for each encountered individual of the three frogs’ genera. Standard statistical analysis was performed to investigate interaction dynamics among the three genera. Our findings indicate the possibility of interspecific competition between Odorrana-Wijayarana, Odorrana-Sumaterana and Sumaterana-Wijayarana. Further studies are required to confirm our hypothesis. Additionally, this study shows that in biodiversity hotspot areas like Sumatra, ecological study is as important as study on species discovery.


    81.    Contributed Herpetology  Exploring Evolution Along the Vertical Dimension: Ground-to-Canopy Microhabitat Selection and Physiological Evolution in Neotropical Tree Frogs (Anura: Hylidae). Julia Laterza Barbosa*, Yale University   julia.laterzabarbosa@yale.edu

    Arboreality unlocks a third spatial dimension for terrestrial life, creating ecological opportunities and evolutionary challenges for arboreal organisms. Yet, verticality is a severely understudied axis of the niche. For wet-skinned ectotherms like amphibians, arboreal habitats can exert selective pressures on two important physiological fronts: thermal physiology and maintenance of water balance. For my dissertation, I will investigate how environmental gradients in the vertical niche axis interact with organismal behavior (specifically, microhabitat selection) to sculpt patterns and rates of physiological evolution in anurans in the family Hylidae, a diverse group comprising more than 1,000 species. I will sample multiple localities along a geographical gradient in the two major neotropical rainforests, Amazonia and the Atlantic Forest, performing nocturnal time-constrained frog surveys from ground to canopy using the single-rope climbing technique, measuring relevant microhabitat and environmental variables. In addition, I will collect individuals of multiple species and measure the preferred temperature, critical thermal limits, and desiccation and rehydration rates. Using the field and laboratory data, I will use PGLS to explore how ecological, behavioral, and physiological variation is phylogenetically structured. The proposed research will help elucidate the coupled evolution of behavior and physiology, and how phylogenetic relationships might help shape those relationships in Neotropical frogs.


    82.    Contributed Herpetology  Ecology of Gekkota: Testing island effects on body size and habitat evolution in geckos. Yuanxing Liao*, University of Kentucky; Phillip Skipwith, University of Kentucky   yli271@uky.edu

    Understanding the patterns of diversity and the processes by which the diversity has evolved remains a major theme in evolutionary biology. Islands have long been known to be interesting testing grounds for evolution. However, the potential effects of islands on the evolution of geckos are generally unknown. Geckos are an ideal study system given that they reached a pan-tropical distribution and have dispersed to many oceanic islands. Here, we used phylogenetic comparative methods to test how insular endemism affects body size and habitat evolution in geckos. We generated a supertree and gathered body size and habitat data for 750 species from literatures. We found that the overall rate of body size evolution on islands is higher when compared to mainland lineages. The speciation rate, on the other hand, did not correlate with island colonizations. We also found that there were transitions of habitat usage amongst island species and that these taxa are disproportionately more arboreal when compared to mainland taxa. We recognize that different island systems have unique geological histories which possibly impact resident lineages. Thus, we are cautious in generalizing to all islands


    83.    Contributed Herpetology  Are Mediterranean Geckos (Hemidactylus turcicus) Choosing to Hunt Near Lights? Year-Round Light Surveys at the University of Central Oklahoma. Cass Condray*, University of Central Oklahoma; Lauren Branham, University of Central Oklahoma; Maryem Assaoui, University of Central Oklahoma; Danielle Bright, University of Central Oklahoma; Allyson Fenwick, University of Central Oklahoma   afenwick@uco.edu

    Mediterranean geckos (Hemidactylus turcicus) are small nocturnal lizards that live on building walls in their invaded range worldwide. We expect them at the edge of light sources, balancing both insect capture and predator avoidance. During a year-round survey of temperature conditions at the University of Central Oklahoma (April 2018-December 2021) we also measured light. Once a week we surveyed both the original site of gecko introduction and a building near the outer range of the local colonization. For each gecko sighting we measured light at the individual’s perch, one meter to each side, and one meter away from the wall. Both buildings had geckos with similar responses to light. Most were found within 5 lux of their surroundings. Many were found at 0 lux with 0 lux nearby. Of individuals with differences between light at the perch and light away from the wall, more were in brighter locations than darker ones. Fewer differences were found between light at the perch and on the surrounding wall, again with more individuals in brighter locations than darker. Lack of behavioral response to light agrees with the few studies we have found that evaluate light conditions of this species, and agrees with our and others’ findings of a lack of behavioral responses to temperature.


    84.    Contributed Herpetology  Evaluating Evidence for Karyotypic Speciation in Spiny Lizards, Genus Sceloporus. Erin Westeen*, UC Berkeley; Isaac Krone*, UC Berkeley   ewesteen@berkeley.edu

    What drives some clades to diversify greatly while other, related groups fail to do so? Many diversification mechanisms contribute to postzygotic reproductive isolation and thus speciation in animals, but few are as contested as chromosomal speciation: structural changes in chromosome arrangements that lead to reproductive isolation. Observations that many species differ in chromosomal arrangements and that these rearrangements reduce hybrid viability are compelling. However, it is difficult to disentangle chromosomal rearrangements that have arisen after speciation from those that contributed to speciation itself, especially given that novel rearrangements arise as underdominant heterozygotes that require very small population sizes to rise in frequency. Most models of chromosomal speciation assume allopatry or otherwise geographic isolation to deal with the fixation of underdominant rearrangements (but see statispatric model). Recent work in a species-rich and chromosomally diverse clade of lizards (genus Sceloporus) found coincident bursts of speciation and chromosome number (karyotype), suggesting a role for chromosomal speciation. If karyotypic rearrangements contribute to the creation or maintenance of new species in Sceloporus, then sympatric sister taxa should be more karyotypically diverged than allopatric sister taxa and allopatric taxa should accumulate differences more gradually. We evaluate evidence for chromosomal speciation in this group by examining the relationships between divergence time, geographic range, and karyotype, while also testing explicitly for trait-dependent diversification. We find little evidence for geographic or temporal signatures of chromosomal speciation, suggesting that chromosomal rearrangements have not influenced the diversification of this widespread and speciose clade.


    85.    Contributed Herpetology  Deep Homology of Follicular Glands in Divergent Lizard Clades. Joseph Rangel*, University of Texas at Arlington; André Carvalho, University of Washington; Adam Leaché, University of Washington; Matthew Fujita, University of Texas at Arlington   joseph.rangel@mavs.uta.edu

    Intra- and interspecies communication is vitally important among animals, with the oldest form of communication involving chemical signaling. Lizards accomplish this by secreting chemicals via their skin, feces, and glands. Generation glands are a conglomeration of novel cell types that sit within an undifferentiated layered epithelium and do not have an external pore. Follicular glands, however, do excrete chemicals via a pore and function morphologically and chronologically independent of the surrounding epidermis. This research will focus on using follicular glands to understand their evolution and to infer the deep homology through the lizard tree of life. We conducted field work and collected follicular glands from several species of lizards from divergent families. We then sequenced and de novo assembled the transcriptomes of the follicular glands before using tools to determine gene orthology and ontology. We used a comparative approach to identify the gene composition of the transcriptomes to identify whether divergent lizards share similar or distinct genetic circuity that can inform the deep homology of follicular glands. This research will allow for a deeper understanding of the evolution of epidermal gland types through their genetic pathways, contributing to our perception of how these mechanisms have shaped communication in diverse lizard clades over time.


    86.    Contributed Herpetology  Epidermal Gland Evolution in Whiptail Lizards (Aspidoscelis). Joshua Rivera*, University of Texas at Arlington; André Carvalho, University of Washington; Adam Leaché, University of Washington; Matthew Fujita, University of Texas at Arlington   jorlandoriver@gmail.com

    The secretion and recognition of chemical stimuli in squamate reptiles has been a burgeoning field of research. These chemical stimuli (i.e., pheromones) serve a role in various intraspecific and interspecific behaviors, including kin identification, mate choice, and heterospecific aggression. Furthermore, studies have shown that some morphologically conservative lizard radiations exhibit divergence in pheromone composition. Whiptail lizards of the genus Aspidoscelis possess femoral glands, though neither the composition of the pheromones, nor the underlying genetic mechanisms regulating expression of the glandular content, are understood. In this study, we compare pheromone expression between parthenogenetic and sexually reproducing whiptails, which offers an exciting system to explore the diversity of these glands. Due to the presence of pseudocopulation in the all-female parthenogens, we expect there to be minimal differences in expressed genes, though there may be some expressed genes that are absent in asexuals that are specific for male-male aggression. Lizards caught in the field were immediately processed, which included extraction of the gland chemicals and excision of both the femoral gland tissue and forelimb tissue (control). Transcriptomic data, acquired through gland RNA extraction and library preparation, will be used to perform gene ontology analyses to investigate the diversity of proteins associated with glandular tissue. The results of this study will be important in understanding the genetic machinery responsible for maintaining these evolutionarily ubiquitous structures necessary for chemical communication.


    87.    Contributed Herpetology  Evaluation of insect inclusion in gel diets for juvenile and adult freshwater turtles (Trachemys spp.) under captivity conditions. Ana Camila Beltran Urrego*, Santacruz Zoo   acbeltranu@unal.edu.co

    Insects are considered as highly nutritious and high-quality, and sustainable dietary protein source (Lange et al., 2021), for which are currently used as an alternative ingredient for feeding a wide variety of animals, including reptiles such as freshwater turtles, which are commonly kept in captivity (Rawski et al., 2018). However, their nutrition is challenging, and the diseases associated with this field are common (NSHP, Ca and Vit. A deficiency) (Mans & Braun, 2014). For the proposed project, it is sought to measure the response in consumption, body condition, health status and digestibility in a group of Trachemys spp. kept in captivity at the Santacruz zoo (Colombia), of 4 nutritionally adequate gelatin-diets with replacement rates of the fish resource of 25, 50, 75 and 100% by gut loaded (Ca and vitamins) insects (Hermetia illucens y Acheta domesticus). Turtles will be exposed to each diet for 1 month and will be feed 4 times per week at an energy rate of 32 kcal/ (BW0.75) per day at same environmental conditions. To determine digestibility all feces will be collected during the experimental periods in water balloons for analysis. Also remaining food will be weighed at the end of feeding days. Physical examinations and morphometrics, including weight, length, and body condition score will be measure at the beginning and end of each month period. At the end of the experiment, it is expected to implement the level of insect inclusion with the best consumption, digestibility ratio and general health status reflected in the animals.


    89.    Contributed Herpetology  Assessing Kirtland's Snake (Clonophis kirtlandii) detection probabilities. Tyler Stewart*, Illinois Natural History Survey; Michael Dreslik, Illinois Natural History Survey; Andrew Kuhns, Illinois Natural History Survey; John Crawford, National Great Rivers Research and Education Center; Christopher Phillips, Illinois Natural History Survey   tyler11@illinois.edu

    Population declines threaten many reptile species worldwide, creating a need to establish monitoring programs. Researchers often rely on presence/absence surveys to estimate population status and distribution, species-level trends, and identify areas for conservation. However, an inherent problem with presence/absence surveys is estimating true absence. Repeated surveys can address this issue, but repeated site visits can quickly drain resources (i.e., time, labor, money). This issue can be alleviated by first estimating detection probabilities, which give insight into the effort needed to assess species' absence. Here, we monitored three Kirtland's Snake populations to determine detection probabilities for this elusive snake. We built detection models to determine the environmental and temporal scenarios under which Kirtland's Snake detection can be maximized and make recommendations for future presence/absence surveys.


    90.    Contributed Herpetology  Influence of Nearby Basking Opportunities on Eastern Massasauga Hibernacula Selection. Emily Sunnucks*, Illinois Natural History Survey, University of Illinois Urbana-Champaign; Seth LaGrange, Illinois Natural History Survey, University of Illinois Urbana-Champaign; Michael Dreslik, Illinois Natural History Survey, University of Illinois Urbana-Champaign   sunnucks@illinois.edu

    Identifying and designating critical habitat is vital to the long-term survival of threatened and endangered species. However, evaluating habitat requirements can be challenging for species whose habitat choices are seasonally or ontogenically influenced, such as snakes. In particular, data regarding hibernacula selection remains limited for many snake species. Using 21 years of spring emergence data, we studied the influence of environmental characteristics on hibernacula selection in the Eastern Massasauga. Specifically, we tested the significance of nearby basking opportunities by calculating vegetation height, vegetation density, and solar radiation using high-resolution lidar data. Values for emergence sites were compared to those from randomly generated points within the study extent. We found emergence locations significantly differed from the random points for all variables; however, only vegetation density demonstrated a significant effect. Vegetation density was higher in areas where hibernacula occurred, suggesting available cover may be more important than basking during ingress and egress. Based on our study, we would suggest continuing habitat management to include areas of dense vegetation. However, future research should consider incorporating other sites with more terrain and canopy variation to properly assess the effect of solar radiation and vegetation height on hibernacula selection.


    91.    Contributed Herpetology  Unraveling the Relationships Between Immunity, Reproduction, and Stress in Four Species of Watersnake. Emily Field*, Arkansas State University; Jennifer Terry, Arkansas State University; Lorin Neuman-Lee, Arkansas State University   emilyfield479@gmail.com

    All physiological processes require energy, a finite resource, which must be allocated amongst essential activities to ensure fitness and survival. Maintenance processes such as sustaining a competent immune system, while critical to survival, is energetically costly. During the active season, energy allocation is expected to shift to prioritize reproductive activities such as mating, offspring development, and oviposition or birth. Further, reproduction is likely more energetically demanding for females compared to their male counterparts, thus, physiological tradeoffs may differ between the sexes. When encountering a stressor, the balance between reproductive and maintenance processes may be disrupted to instead fuel the stress response, a process crucial to survival. Reptile physiology is poorly studied, but reptiles provide a unique study organism as ectotherms with a wide variety of reproductive strategies. We captured four water snake species throughout Northeast Arkansas throughout May 2020 and from April–October 2021. We subjected animals to an acute stressor by restraint. We collected blood samples prior to the stressor and then at 30, 60, and 120 minutes. We performed bacterial killing assays to assess functional innate immunity. We measured estradiol, progesterone, testosterone, and corticosterone concentrations using radioimmunoassays to evaluate reproductive investment and stress-responsiveness. Energetic tradeoffs between the immune system, reproductive system, and the stress response are complex. Our study adds to the growing body of literature allowing us to understand the energetic shifts in wild organisms in the context of stress.


    92.    Contributed Herpetology  Reproductive biology and population demographics of Eurycea chamberlaini. Dustin Siegel, Southeast Missouri State University; David Beamer, Nash Community College; Jon Davenport*, Appalachian State University   dsiegel2@gmail.com

    Eurycea chamberlaini (Chamberlain’s Dwarf Salamander) is a recently described small spelerpine salamander with scant reproductive and life history data available since the original species description. Therefore, the objectives of our study were to examine the reproductive life history and population structure of E. chamberlaini from a North Carolina population. From February 2008 to February 2009, monthly collections were made in Craven County, North Carolina. All specimens (n=392) were histologically examined for reproductive life history and population characteristics by month. Overall, male and female E. chamberlaini follow a reproductive cycle similar to other spelerpine species with sperm in the Wolffian ducts of specimens from September to February; however, spermatidogenesis was delayed until August and September with the presence of mature sperm in testes from August through November. We captured 201 female salamanders of which 36 were considered immature. We caught 191 males of which 27 were considered immature. The mean SVL (+1 SD) of mature females (27.13 mm +2.33) and males (25.84 mm +2.03) was statistically significant. Females tended to be larger than males from our monthly samples with a greater proportion of the largest specimens being female. Overall, E. chamblerlaini population structure appears similar to other coastal plain species. We document some variability in reproductive timing and efforts between E. chamaberlaini and other dwarf salamander species.


    93.    Contributed Herpetology  State of the Amphibia 2020: Five Years of Amphibian Research, Diversity, and Resources. Molly C. Womack, Department of Biology, Utah State University; Emma Steigerwald, Museum of Vertebrate Zoology, University of California, Berkeley; David C. Blackburn, Department of Natural History, Florida Museum of Natural History, University of Florida; David C. Cannatella, Department of Integrative Biology and Biodiversity Center, University of Texas at Austin; Alessandro Catenazzi, Centro de Ornitología y Biodiversidad; Jing Che, State Key Laboratory of Genetic Resource and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences; Michelle S. Koo, Museum of Vertebrate Zoology, University of California, Berkeley; Jimmy A. McGuire, Museum of Vertebrate Zoology, University of California, Berkeley; Santiago Ron, Museo de Zoología, Escuela de Biología, Pontificia Universidad Católica del Ecuador; Carol Spencer, Museum of Vertebrate Zoology, University of California, Berkeley; Vance T. Vredenburg, Museum of Vertebrate Zoology, University of California, Berkeley; Rebecca D. Tarvin*, Museum of Vertebrate Zoology, University of California, Berkeley   rdtarvin@gmail.com

    Amphibians are a clade of over 8,400 species that provide unique research opportunities and challenges. With amphibians undergoing severe global declines, taking stock of our current understanding of amphibians is imperative. Focusing on the past five years (2016–2020), we assessed trends in amphibian research, data, and systematics. New species of amphibians continue to be described at a pace of ~150 per year. Phylogenomic studies are increasing, fueling a growing consensus in the amphibian tree of life. Over 3000 species of amphibians are now represented by expert-curated accounts or data in AmphibiaWeb, AmphibiaChina, BioWeb, or the Amphibian Disease Portal. Nevertheless, many species lack basic natural history data (e.g. diet records, microCT scans, call recordings) and major gaps exist for entire amphibian clades. Genomic resources appear on the cusp of a rapid expansion, but large, repetitive amphibian genomes still pose significant challenges. Conservation continues to be a major focus for amphibian research and cataloged threats from AmphibiaWeb on 1261 species highlight the need to address land use change and disease using adaptive management strategies. To further promote amphibian research and conservation, we underscore the importance of database integration and suggest that other understudied or imperiled clades would benefit from similar assessments of existing data.


    94.    Contributed Herpetology  Using Erythrocyte Cell Area or Nuclear Length for Rapid Assessment of Ploidy Level for the Ambystoma laterale Complex. Tess Santoro, Central Michigan University; Zachary Laughlin, Central Michigan University; Kirsten Nicholson*, Central Michigan University   norops@gmail.com

    The Jefferson/Blue-spotted Salamander complex consists of several species (Ambystoma jeffersonianum, A. laterale, A. tigrinum, A. texanum, and A. barbouri) and unique unisexual lineages formed by ancient hybridization events between the species. The unisexual lineages are all females that are usually polyploid, although some hybrid diploids occur in low frequency. Unisexuals cannot interbreed and require sperm from males within the complex in order to produce offspring. Advanced molecular techniques may be used to distinguish the various ploidy combinations, termed biotypes, but those take time and money and some studies would benefit from a faster and cheaper method. Three previous publications reported the use of erythrocyte cell area or nuclear length as a potential alternative method, however the efficacy of this method was questioned by another publication, and all previous publications focused on A. jeffersonianum and assumed results would be the same for A. laterale. Our study investigated the efficacy of cell size and nuclear length for distinguishing polyploid from diploid individuals of A. laterale in Central Michigan. We found significant differences in cell size and nuclear length between diploids and polyploids and report on the relative ease of use for the method with some caveats to consider for future use. It is important to note that while this method can distinguish diploid from polyploid, it cannot determine the specific biotype, thus will not be useful for studies requiring that information.


    95.    Contributed Herpetology  Characterization of Steroid Hormone Receptor Expression During a Major Life-History Transition in Thamnophis sirtalis parietalis. Rysa Thomas*, James Madison University; M. Rockwell Parker, James Madison University   thoma6rl@dukes.jmu.edu

    Vertebrates exhibit a diversity of life-histories that also involve dynamic transition phases requiring tight regulation of energy stores. Across annual patterns of life-history transitions, glucocorticoids (GCs) promote energy mobilization via activation of two receptors, glucocorticoid (GR) and mineralocorticoid (MR) receptors. Species with annual and dynamic life-history transitions, such as those that exhibit migratory behavior, balance expression of these two receptors in target tissues to manage energy allocation. Annually, red-sided garter snakes emerge from hibernacula after a protracted dormancy and engage in a brief-but-explosive mating phase followed by a relatively long-distance migration, and these events occur while snakes are aphagic/fasting. A physiological mechanism driving this transitional shift from mating to migrating must exist. We tested the hypothesis that GR and MR work reciprocally in liver, skeletal muscles, and cardiac muscles to regulate energy budgets in red-sided garter snakes along the transition from mating to feeding. By creating tissue-specific cDNAs and using qPCR, we determined relative receptor expression in each tissue. We expected to observe sex differences in expression of both receptors and elevated GR expression during mating when circulating GCs are high. We also expected decreased MR expression in spring, thus the two receptors should behave oppositely with MR expression increasing during migration. Among the target tissues, we predicted the associated receptor patterns would be most pronounced in liver due to its substantial glycogen stores and its role as the primary site for gluconeogenesis. Results of this study may illuminate the relationship between ectothermic energy budgeting and the physiological response.


    96.    Contributed Herpetology  Examining the Physiological Stress Response of Prairie Lizard, Sceloporus consobrinus, following Extirpation of Keystone Predator, Crotaphytus collaris. Michael Deutsch*, University of Central Arkansas; David Adams, University of Central Arkansas; Matthew Gifford, University of Central Arkansas   mdeutsch@cub.uca.edu

    Keystone predators, defined by their pivotal role in food webs, exert top-down control via predation and competition, which promotes biodiversity by regulating population abundances at lower trophic levels. Under the risk of predation, prey organisms must trade off opportunities for foraging, mating, growth, with other physiological requirements for more immediate energetic mobilization. Consequently, these physiological and behavioral trade-offs further alter community dynamics and nutrient cycling, through modified energy allocation and fitness. In the Ozark Highlands of Arkansas and Missouri, regionally-imperiled glade habitat is home to state-threatened keystone predator the Eastern Collared Lizard, Crotaphytus collaris. Habitat degradation has led to local extinctions which enables comparative studies of keystone predator-prey interactions between glades with and without C. collaris. While keystone predator studies often focus on resulting diversity and abundances of prey in response to presence/absence of keystone species, here I investigate the physiological stress responses and life history tradeoffs of collared lizard prey, Sceloporus consobrinus. Specifically, I assess metabolic and reproductive metrics, through corticosterone, glucose, and lactate concentrations and maternal investment, between S. consobrinus populations which are sympatric and allopatric to C. collaris. Results will provide insight to both the ecological and physiological consequences of keystone species loss.


    98.    Contributed Herpetology  Sizzling Salamanders: Thermal Physiology of Plethodon cinerus from Burned and Unburned Habitats. Cloe Capalongo*, The University of Scranton; Vincent Farallo, The University of Scranton; Nicole Freeman, The University of Scranton   cloe.capalongo@scranton.edu

    Natural wildfires are predicted to become more frequent due to climate change. Burned habitats typically result in warmer microhabitat conditions with a reduction of shade. Plethodontid salamanders are especially sensitive to environmental change, especially warmer temperatures, as they breathe via cutaneous respiration which requires cool and moist conditions. The objective of this study is to assess the thermal physiology of Plethodon cinereus in burned and unburned habitats to determine their ability to adapt to environmental changes. I predict that salamanders from burned sites will have a higher thermal preference, critical thermal minimum, and critical thermal maximum compared to salamanders from unburned sites. Salamanders will be collected from populations in Pennsylvania State Game Lands that have undergone prescribed burns and from populations where no burns have occurred. To measure the critical thermal minimum and maximum, salamanders will be placed into a water bath, which will be cooled or heated by approximately 1°C per minute. The salamander will be flipped onto its back and given the chance to turn itself upright. The water temperature at the point when the salamander loses its ability to turn itself will determine the critical thermal minimum and maximum. To measure thermal preference, salamanders will be placed in a thermal gradient box ranging from 0°C to 40°C. Location and temperature will be measured every 10 minutes for a 4-hour period. I will then compare individuals from burned and unburned populations to determine if there are differences in the salamander’s thermal physiology.


    99.    Contributed Herpetology  The Effect of Burned Habitat on the Metabolism of Plethodon cinereus Salamanders: Can They Stand the Heat? Nicole Freeman*, University of Scranton; Vincent Farallo, University of Scranton; Cloe Capalongo, University of Scranton   nafreeman01@gmail.com

    Animals need energy, obtained by metabolizing food, to perform tasks needed for survival and reproduction. This energy use can be measured by quantifying an organism’s respiration to determine their standard metabolic rate (SMR). As ectotherms, salamanders’ internal body temperature and metabolism are dependent on environmental temperature. Increased temperatures cause an increase in the metabolic rate of salamanders and therefore increase total energy consumption. Plethodon cinereus, commonly known as the eastern red-backed salamander, use their skin for gas exchange in a process called cutaneous respiration which is most efficient in cool, moist environments. Due to ectotherms having higher SMRs in higher temperatures, global warming could impact their survival, especially those that rely on cutaneous respiration. Studies have predicted that the increased warming and drying of the planet will increase forest fires. Forest fires can cause increases in overall temperature with a reduction in shade provided by vegetation. Prescribed burns in specific sections of forests are performed to maintain forest health and can be used to study consequences of global warming as their effects mimic those of wildfires. In this project, we will use respirometry to measure and compare the SMRs of Plethodon cinereus collected from burned and unburned environments. Many organisms have the capability to respond to environmental cues and make physiological adjustments. If salamanders can reduce their metabolic rate when exposed to warmer temperatures it would provide a mechanism to combat habitat alterations such as climate change and forest fires that cause temperature increases.


    100.    Contributed Herpetology  Student-lead Coverboard Monitoring following Ecological Restoration. Alistair Dobson*, UC Santa Barbara; Lisa Stratton, Cheadle Center for Biodiversity and Ecological Restoration; Kyra Sullivan, UC Santa Barbara   dobsonalistair@gmail.com

    In 2017, UC Santa Barbara’s North Campus Open Space (NCOS) Ecological Restoration Project began major excavation on a golf course to convert it back into the upper arms of a slough and coastal mesa which historically existed in the area. Before restoration began, twenty-six coverboards were monitored between 2012 and 2014 to assess terrestrial populations of reptiles, amphibians, and invertebrates. In 2020, forty-four plywood coverboards with a uniform area (1526 in2) were deployed across various planting and restoration regimes on these 300 acres of recently restored open space. Boards are monitored weekly by a student research intern, who records vertebrate species abundance and invertebrate species presence at each site. So far, species richness and composition of herpetological inhabitants remained the same before and after restoration (six reptile and two amphibian species). However, encounter rates (our proxy for abundance), have changed for some species following restoration. Most notably, we recorded an increase of garter and gopher snakes, especially in adjacent salt marsh habitats, and a decrease of slender salamanders, which have not been found in areas of high soil movement. Our results demonstrate that the processes and outcomes of ecological restoration affect populations of reptiles and amphibians, and that these species should be considered in monitoring efforts that determine the “success” of a project. We hope to continue this monitoring for many years, both to examine the impacts of the restoration at a longer timescale and provide undergraduates with opportunities to conduct scientific research related to herpetology near their campus.


    101.    Contributed Herpetology  Invasive Bullfrogs in Western Colorado: Dietary Profile and Batrachochytrium dendrobatidisStatus of Lithobates catesbeianus Removed From Local Wetlands. Alexander Burdick*, Colorado Mesa University; Bella Barker, Colorado Mesa University; Denita Weeks, Colorado Mesa University   dweeks@coloradomesa.edu

    The American Bullfrog (Lithobates catesbeianus) is an invasive species in southwestern Colorado that occasionally preys on native amphibians. In a recent survey, stomach contents of one L. catesbeianus included a vertebral column putatively identified as a Canyon Tree Frog (Dryophytes arenicolor), a state species of concern. The bullfrog is also a reservoir for Batrachochytrium dendrobatidis (Bd), a fungal pathogen that causes chytridiomycosis in amphibians. This emerging infectious disease has played a significant role in amphibian declines in Colorado including the decline of the Northern Leopard Frog (Lithobates pipiens) which shares similar habitat with L. catesbeianus. Predation on native species and spread of disease by L. catesbeianus is a concern for native species conservation in Western Colorado. As part of a Bullfrog removal effort, a citizen science program trained volunteers to capture and submit L. catesbeianus from local wetlands. The Bullfrogs collected were assessed for Bd and the contents of their stomachs were surveyed for vertebrates and invertebrates. Bullfrogs tested positive for Bd in 2019 from these wetlands but tested negative for Bd in 2021. A dietary profile of L. catesbeianus that confirms native amphibians are being consumed would be an important contribution to state agency management decisions.


    102.    Contributed Herpetology  Boreal Toad (Anaxyrus boreas) Microbiome Response to Itraconazole Treatment for Batrachochytrium dendrobatidisInfection. Bella Barker*, Colorado Mesa University; Alexander Burdick, Colorado Mesa University; Amanda Schrager-Lavelle, Colorado Mesa University; Denita Weeks, Colorado Mesa University   dweeks@coloradomesa.edu

    The Boreal Toad (Anaxyrus boreas) is threatened in its native region of the southern Rocky Mountains. The fungal pathogen Batrachochytrium dendrobatidis (Bd), which infects the skin, has been implicated in population declines. Captive breeding programs have used the anti-fungal itraconazole to successfully treat other Bd-infected amphibians, but little is known about the impacts itraconazole has on commensal fungi and bacteria in A. boreas. The microbiome of amphibians is important for immunocompetence and pathogen defense with disturbances potentially shifting or removing that protective role. Bd-infected A. boreas individuals from a captive breeding program were treated with itraconazole. Skin swabs were collected before and after treatment to assess any changes in the microbiome with Illumina Sequencing. This research contributes an important gap in knowledge toward conservation efforts of A. boreas.


    103.    Contributed Herpetology  Short-Term Responses of Terrestrial Salamanders to the Restoration of Fire in the Southern Appalachian Mountains. John Maerz*, University of Georgia; Lauren Head, University of Georgia; Meghan Bierden, University of Georgia; Cynthia Carter, University of Georgia; Brian Crawford, University of Georgia   jcmaerz@uga.edu

    Fire is an important natural disturbance that can shape plant and associated animal communities. After a long history of fire suppression in the U.S., there is increasing use of prescribed fire to manage southern Appalachian forests. Southern Appalachia is an area of global importance for salamanders, particularly within the family Plethodontidae, and there is interest in understanding whether and how fire may affect salamander populations. Using a paired watershed manipulation over four years, we monitored salamander microhabitat use and age-specific abundance before and after a sequence of two prescribed burns within the Coweeta Hydrologic Laboratory in Macon County, North Carolina. As expected, fire reduced shrub cover and increased the amount of bare ground compared to the reference watershed. We estimated a relatively small decline in abundance of all age classes following the first burn. We are currently estimating abundance changes following the second burn. Following burns, we observed a significant reduction in salamander surface activity and greater nighttime use of burrows compared to salamanders in the reference watershed. However, salamander surface activity appeared to return to pre-burn levels the year following a burn. Early results suggest prescribed fire has limited direct impacts on salamander abundance; however, effects on behavior may indirectly influence abundance through more chronic effects on fitness, depending on fire frequency. Evolutionary impacts on salamander phenotypes will also be discussed.


    104.    Contributed Herpetology  Prioritizing Conservation Area In Species Management Strategy For The Edible Bornean Giant River FrogLimnonectes leporinus. Ramlah Zainudin*, Universiti Malaysia Sarawak; Elvy Quatrin Deka, Universiti Malaysia Sarawak; Julius Georgy, Universiti Malaysia Sarawak   ramlahzainudin29@gmail.com

    The endemic giant river frog, Limnonectes leporinus , is a completely riparian (stream dwelling) species and lives along streams with moderate to steep gradients. Deforestation by severe clear cutting that leads to fragmentation of its distribution, and also overhunting for local consumption, are the greatest threats to the species. Excessive landscape modification leads to habitat modification and thus is crucial for an organism to maintain heterozygosity so that the population has strong fitness to adapt to a changing environment. This study aimed to project suitable habitats and predicts the potential for habitat connectivity to allow gene flow across the Sarawak landscape. ArcGIS was used to convert the presence data file and environmental layers into ascii format and were then implemented in MaxEnt modelling to generate the map of suitable habitats. The potential habitat model and genetic attributes of haplotype data were computerized in Circuitscape software, to perform connectivity model. The results showed suitable habitats corresponded to the species distribution in lowland areas with sustainable stream networks as breeding sites, whereas higher elevations were recognized as unsuitable habitats. On the other hand, gene flow data had shown connectivity circuits fall largely in unprotected forest which include of development areas and private lands. Hence, prioritizing conservation area should include the local authorities and landowners to proper management of the landscape as well as species management strategy. This indirectly sustains and protects the forests, flora and fauna of Borneo.


    105.    Contributed Herpetology  Long-term Trends in Occurrence of a Population of Narrow-headed Gartersnakes. Erika Nowak, Northern Arizona University, Colorado Plateau Research Station; Brian Halstead*, U.S. Geological Survey, Western Ecological Research Center   bhalstead@usgs.gov

    Northern Arizona University (NAU)’s Gartersnake Program has been conducting annual surveys for Narrow-headed Gartersnakes (Thamnophis rufipunctatus) in Oak Creek, Arizona, USA, since 1999, with standardized monitoring transects developed at five sites in 2003. Results of the surveys showing a decline in detection rates compared to historical data (as estimated by catch per unit effort during surveys) were used, in part, as support for listing T. rufipunctatus as federally threatened in 2014. Recapture rates are very low under the typical monitoring strategy, so we used occupancy modeling and analysis of count data to evaluate hypotheses developed over the course of this long-term research, including: an apparent decline in detection rates indicative of an actual population decline; detection rates varying among monitoring transects; and detections are negatively correlated with increases in human disturbance over time and among sites. Initial dynamic occupancy models indicate that survey-specific detection probabilities decreased from 1999–2021, despite recent increases in survey effort. The decline in detection probability is likely related to decreased abundance, but other mechanisms cannot be ruled out. Most monitoring transects remain occupied, however, with high persistence probabilities and low site turnover. We describe further caveats of our results and discuss implications for persistence of this population, as well as improving long-term monitoring under a limited-budget scenario.


    107.    Contributed Herpetology  eDNA Concentration Positively Correlates to Abundance and Biomass in Amphibians. Padraic Robinson*, University of Georgia SREL; Jason Jason O'Bryhim, University of Georgia SREL; David Scott, University of Georgia SREL; Adam McFall, University of Georgia SREL; Gabriela Rodriquez, University of Georgia SREL; Stacey Lance, University of Georgia SREL   padraic.robinson@uga.edu

    The decline of biodiversity worldwide has been well-documented, and amphibians are among the most threatened of all taxa. Because of these declines, there is a great need to understand both the distribution and population size of these animals. Environmental DNA (eDNA) has emerged as a powerful tool in the detection of a wide range of species. eDNA methods are most commonly used for obtaining presence/absence data either from a single species or an entire community through metabarcoding. Elevating eDNA studies from presence/absence data to estimating abundance and ultimately population size would greatly increase its utility. Recent studies have shown positive relationships between eDNA concentrations and abundance in laboratory, mesocosm, and natural experiments for several species of fish and amphibians. In this study, we examined the relationship between eDNA concentration and abundance as well as eDNA concentration and biomass using controlled aquaria experiments. Our focal species were two common southeastern amphibians: the Southern Toad (Anaxyrus terrestris) and the Southern Leopard Frog (Rana sphenocephalus). Individuals were placed in tanks of varying density (ranging from 0–20 tadpoles/20 L) and eDNA samples were taken one- and four-days following introduction into the experimental tanks. We found eDNA concentrations were positively correlated to abundance and biomass in both species. We also compared how catch per unit effort estimates of abundance compare to eDNA concentrations from samples collected in a constructed wetland. We discuss these results and their implications for using eDNA to monitor amphibian populations.


    108.    Contributed Herpetology  Efficacy of Sampling Techniques for Estimating Population Sizes of Ambystomatid Salamanders. Brock Lorenzen*, Illinois Natural History Survey; Andrew Kuhns, Illinois Natural History Survey; Michael Dreslik, Illinois Natural History Survey; John Crawford, National Great Rivers Research and Education Center; Christopher Phillips, Illinois Natural History Survey   brockl@illinois.edu

    Amphibians are vital components of food webs, facilitating nutrient cycling between terrestrial and aquatic ecosystems. As the most threatened species group worldwide, high quality monitoring is essential to amphibian conservation. When monitoring ephemeral wetland breeding amphibians, biologists must choose from numerous traditional and emerging techniques, including raw abundance counts from drift fence sampling, capture-mark-recapture estimates, and N-mixture abundance modeling. To find the most balanced method in terms of cost, logistics, and data quality, it is essential to understand the strengths, weaknesses, and differences between monitoring techniques and evaluate the utility of new methods. We evaluated an N-mixture modeling approach by comparing abundance estimates to those using drift fences and capture-mark-recapture techniques for Jefferson Salamander, Ambystoma jeffersonianum. A comparatively accurate N-mixture model could become a standard tool for monitoring pond breeding salamanders and potentially other amphibians.


    109.    Contributed Herpetology  Non-Profit use of Social Media and Citizen Science as tools for the conservation and research of turtles in Texas. Sal Scibetta*, Texas Turtles; Carl Franklin, Texas Turtles; Viviana Ricardez, Texas Turtles   sscibetta@yahoo.com

    Texas Turtles is a 501(c)(3) non profit organization focused on discovering and studying the ecology and natural history of the chelonian diversity in Texas. Social media connections with volunteers and land owners across the state has permitted a wide range of field research. Our research has greatly benefited from a core group of skilled and trained volunteers. The organization has four board members and a team of over 40 volunteers from diverse backgrounds including: university faculty, students, veterinarians, biologists, zookeepers, military personnel and interested citizens. Access to 6 of our major study sites was achieved via social media contacts from land owners. During the pandemic, outreach was accomplished through Animal Planet and Youtube with Coyote Peterson resulting in more than 1.2 million views. Currently Texas Turtles operates a website, has a presence on Facebook, Instagram, Twitter, YouTube, Researchgate and a project on iNaturalist. Since 2021, Texas Turtles has produced a total of 28 publications (25 of which are peer reviewed) pertaining to the conservation, ecology and natural history of Texas turtles. These collective efforts are helping us fill a “Texas-sized hole” in our knowledge of chelonians from the Lone Star State.


    110.    Contributed Herpetology  After the Invasion: Monitoring Long-term Ecological Trends in Nonnative Cuban Treefrogs (Osteopilus septentrionalis) in Louisiana. BRAD GLORIOSO, U.S. Geological Survey; Hardin Waddle*, U.S. Geological Survey; Matthew Atkinson, University of Central Florida; Anna Savage, University of Central Florida; Robert Mendyk, Audubon Zoo   gloriosob@usgs.gov

    Introductions of invasive species can have long-term consequences for ecosystems. In late 2017, we confirmed the presence of an established population of the invasive Cuban Treefrog, Osteopilus septentrionalis, in Audubon Park in New Orleans, Louisiana. Since then, we have confirmed establishment of this species in a second location and suspected establishment in several other locations, all within the Greater New Orleans metropolitan area. To determine the long-term impact of this invasion, we sought to determine the specific interactions Cuban Treefrogs had with the native anuran community across multiple gradients. To accomplish this, we increased our removal efforts for these populations, adding artificial refugia at the Audubon Park invasion site, which ultimately led to the removal of over 2,000 individuals. We then dissected a subset of these individuals, removed toe/tail tissue for pathogen analysis, examined the entire digestive tract for food items, and evaluated reproductive condition. Additionally, we are characterizing genetic lineages and comparing them to other native and invasive populations to determine if the New Orleans area populations result from single or multiple introductions. We confirmed infections of Cuban Treefrogs with Ranavirus and Amphibian Perkinsea in the Audubon Park population as well as in co-occurring native anurans and are examining if the presence of Cuban Treefrogs alters disease dynamics of native host communities. Ultimately, this work provides a better understanding of the impacts of this invasion in Louisiana and can potentially help mitigate negative effects on native species.




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