Northeastern Naturalist 410 L.M. Thompson, et al. 22001199 NORTHEASTERN NATURALIST 2V6(o2l). :2461,0 N–4o1. 92 Surveys for Population Persistence and Bd at the Northeastern Range Edge of the Eastern Lesser Siren Lily M. Thompson1, Benny Pugh1, Logan A. McDonald2, Angie Estrada3, Katelyn Horn1, Bronte L.C. Gilman1, Lisa K. Belden3, Joseph C. Mitchell4, and Kristine L. Grayson1,* Abstract- Sirens are enigmatic, fully aquatic salamanders found in freshwater wetland habitats. Siren intermedia intermedia (Eastern Lesser Siren) occurs along the East Coast of the United States from Alabama to Virginia. Surveys near the northeastern range edge of the subspecies at Fort A.P. Hill in Virginia from 1995 to 1999 documented 53 Eastern Lesser Sirens in 5 wetlands. In 2015, 13 individuals were found, documenting persistence at 4 of these wetlands; none were found in 4 additional wetlands with habitat that appeared appropriate. The size distribution of captured individuals was similar for the 2 survey periods. Captured individuals in 2015 were screened for Batrachochytrium dendrobatidis (Bd), a fungal pathogen ubiquitous in aquatic habitats of the southeast and mid-Atlantic. No Bd was detected on these individuals, despite the presence of Bd on other amphibians at Fort A.P. Hill. Further investigations of the Eastern Lesser Siren populations in this area would provide important information about the persistence of this species in the region and provide more insight into the biology of this elusive salamander. Introduction Siren intermedia Barnes (Lesser Siren) is a fully aquatic salamander with 2 recognized subspecies (Petranka 1998, Powell et al. 2016). Members of the genus are nocturnal and require muddy wetland habitats with thick vegetation, sediment for burrowing, and standing water for at least 6 months of the year (Leja 2005, Petranka 1998, Snodgrass et al. 1999). The subspecies S. i. intermedia Barnes (Eastern Lesser Siren) occurs along the East Coast of the United States from southern Alabama to Virginia. Data on this subspecies are mostly from the southern portion of its distribution, and fewer studies have been conducted in northeastern populations. Range-edge populations, those near the periphery of a species’ distribution, can be more susceptible to extirpation than those in the interior of a range due to smaller population sizes, physiological constraints, and reduced gene flow when isolated (Gaston 2003). Monitoring the status of these populations is particularly important because they also can be more affected by changes in climate and anthropogenic disturbance (Rehm et al. 2015). The enigmatic nature of siren species makes them challenging to monitor, but all the more important to study because early signs of population decline could otherwise go unnoticed (e.g., Tedesco et al. 2014). In the 1Department of Biology, University of Richmond, Richmond, VA 23173. 2Department of Biology, Virginia Commonwealth University, Richmond, VA 23284. 3Department of Biological Sciences, Virginia Tech, Blacksburg, VA 24061. 4Florida Museum of Natural History, University of Florida, Gainesville, FL 32611. *Corresponding author - kgrayson@richmond.edu. Manuscript Editor: Rudolf Arndt Northeastern Naturalist Vol. 26, No. 2 L.M. Thompson, et al. 2019 411 absence of data from frequent monitoring over long periods, assessing the status of previously documented populations can provide important information on persistence (Kroschel 2012, Parra-Olea et al. 1999, Witte et al. 2008). Additionally, infection by the fungal pathogen, Batrachochytrium dendrobatidis Longcore, Pessier, & D.K. Nichols (Bd), is of particular concern for amphibian species because it is linked with population declines across the globe (e.g., Skerratt et al. 2007). In the southeastern and mid-Atlantic regions of the United States, Bd is ubiquitous in aquatic habitats; however, fewer species here appear to be experiencing disease-related declines compared to in tropical regions (Rothermel et al. 2008). Bd can infect Eastern Lesser Siren (Chatfield et al. 2012), but the potential for Bd-associated declines in this species is unclear. Here we assess the population persistence of Eastern Lesser Siren at its northeastern range edge and screen the captured individuals for Bd. In the late 1990s, surveys to determine the distribution of amphibians and reptiles at Fort A.P. Hill in Caroline County, VA, provided important information on populations of Eastern Lesser Siren in this region (see checklist in Mitchell and Roble 1998). A subset of these sites was revisited in 2015 as part of a smaller amphibian and reptile monitoring effort, and here we report on the results of both survey periods. We used these combined data to determine if these populations have persisted and if Eastern Lesser Siren occurs in other suitable nearby sites. To test if shifts have occurred in age or size structure, we compared the body sizes of individuals caught during the 2 survey periods. We also assessed recently captured individuals for the presence of Bd. Evaluating the status of these populations provides important information on the geographic range of this species and the habitats where it is found as well as local natural history data to inform management recommendations. Field-site Description US Army Garrison Fort A.P. Hill is a 30,673-ha military installation located in the northwestern portion of Caroline County, VA (Fig. 1). This Coastal Plain area is comprised of pine and mixed hardwood forests, fields, and a variety of freshwater wetland habitats, such as streams, ponds, and vernal pools. The installation is located in the northernmost portion of the distribution of the Eastern Lesser Siren (Petranka 1998, Powell et al. 2016, Roble 1995), which is known to occur in semipermanent and permanent ponds and wetlands in this area. Methods Siren surveys Initial surveys for Eastern Lesser Siren occurred in 1995–1999 (Mitchell and McNulty 1999). During the spring and summer of those years, individuals were captured using D-ring aquatic dip nets in opportunistic surveys across Fort A.P. Hill. Depending on wetland size, 5–20 dip-net samples were taken at each wetland. The original goal of these surveys was to record the amphibian and reptile species that occur on the installation and to assess their general distribution. In late Northeastern Naturalist 412 L.M. Thompson, et al. 2019 Vol. 26, No. 2 1997 and 1998, Eastern Lesser Sirens were captured using galvanized steel minnow traps in a single wetland where 50 traps were set each month for 15 months. All traps were checked within 24 h of being placed. Recent surveys for Eastern Lesser Siren occurred in the summer of 2015, when we opportunistically surveyed 8 training areas (TAs: 1A, 5A, 5C, 14A, 22A, 24B, 25A, 30B; Fig. 1) across Fort A. P. Hill 1 to 3 times during June and July based on the availability to enter these areas (i.e., when not in use for military exercises). During each survey night (n = 26), we deployed 40 plastic minnow traps, 8 crayfish Figure 1. Siren. i. intermedia (Eastern Lesser Siren) distribution shown in dark gray with inset map of capture locations at Fort A.P. Hill, VA. Symbology of capture locations represents the detection of individuals during our study period. The numbered units are training area (TA) designations used for the organization of military training and other activities. Northeastern Naturalist Vol. 26, No. 2 L.M. Thompson, et al. 2019 413 traps, and 6 trashcan traps. These 3 trapping methods were used to maximize our ability to detect sirens throughout the water column. Minnow traps target the portion of the water column near the surface and are most effective in shallow water (Willson et al. 2005). Modified crayfish traps allow for a greater sampling depth and can be more effective at catching sirens than minnow traps (Johnson and Barichivich 2004). Traps constructed from large, heavy-duty trash cans using a funnel trap design can sample near the substrate and can also be more effective than minnow traps (Luhring and Jennison 2008). We checked traps within 24 h of being set. Whenever possible, we recorded snout to posterior vent length (SVL, mm) and mass (g) for each individual. Given the differences in survey design and variation in effort depending on site availability, we provide capture numbers summarized by site and survey period. Comparisons of SVL and mass between captures in the initial and recent surveys were analyzed with t-tests (α = 0.05) using R version 3.5.1 (R Core Team 2018). Bd sampling Each Eastern Lesser Siren we captured in 2015 was handled with nitrile gloves, rinsed with 50 mL of sterile water, and swabbed with a sterile rayon swab 5 times on the ventral surface near the front limbs and 5 times on the ventral side just anterior to the vent. We put swabs in sterile 1.5-mL centrifuge tubes that were immediately placed on ice and frozen until processed. Sirens captured during the initial survey efforts in the 1990s were not screened for Bd. We extracted whole genomic DNA from skin swabs using the DNeasy Blood and Tissue kit (Qiagen, Valencia CA, USA) according to manufacturer instructions. The DNA was eluted in 200 μl of sterile molecular biology water (Millipore, Burlington, MA). To quantify Bd infection intensity, we used a TaqMan real-time PCR assay, as in Boyle et al. (2004). We prepared the DNA standards by making serial dilutions of Bd strain JEL 404 (originally isolated from Lithobates catesbeianus (Shaw) [American Bullfrog] in Maine) for 1000–0.1 zoospore genome equivalents. All sample reactions were performed in duplicate, and we considered samples infected if both replicates tested positive and the values estimated by qPCR were above 0.1. Results Between 1995 and 1999, the total captures consisted of 53 Eastern Lesser Sirens from dip nets and minnow traps in 5 TAs (1B, 5A, 5C, 24B, 30B; Fig. 1). Measurements were not recorded for the 4 individuals captured in 1995 and 1996. In 2015, a total of 13 individuals was captured in minnow traps within 4 TAs (5A, 5C, 24B, 30B; Fig. 1), all of which previously had records of Eastern Lesser Siren. One individual escaped before measurements could be recorded. Bd was not detected on any of the 12 Eastern Lesser Sirens screened in 2015. The size distribution of captured individuals was consistent between survey periods for both SVL (Fig. 2A) and mass (Fig. 2B). When comparing the means, there were no statistically significant differences in either SVL (t = -0.38, df = 22.37, Northeastern Naturalist 414 L.M. Thompson, et al. 2019 Vol. 26, No. 2 P-value = 0.71) or mass (t = 0.17, df = 19.41, P-value = 0.87) between the initial and the most recent surveys. Table 1 provides a summary of the SVL, mass, and sample size of captured individuals. Discussion Overall, we found that populations of Eastern Lesser Siren have persisted at Fort A.P. Hill without significant changes in body size over a period of 16–18 y. This finding suggests that these populations have been viable over a long time period. Not all sites with Lesser Sirens present in the initial surveys were surveyed again in 2015, but sirens were captured in all areas that we Figure 2. Boxplots of SVL (A) and mass (B) of Siren. i. intermedia (Eastern Lesser Siren) by survey period. Initial surveys represented here are from 1997 to 1999, and recent surveys are from 2015. Table 1. Summary of data collected from captured S. i. intermedia (Eastern Lesser Siren). Snout-vent length (SVL, mm) and mass (g) were not recorded for the individuals captured in 1995 and 1996. The number of captured individuals that were measured for each body size metric is indicated (n). Total SVL (mm) Mass (g) Year captured Mean (SE) Min–max n Mean (SE) Min–max n 1995 1 na na na na na na 1996 3 na na na na na na 1997 16 185.05 (15.26) 69–249 15 40.64 (7.39) 1.85–79.0 14 1998 26 206.53 (8.94) 52–259 25 56.25 (3.78) 5.50–89.0 26 1999 7 196.71 (12.55) 133–230 7 46.88 (7.70) 11.81–78.0 7 1995–1999 53 198.21 (7.06) 52–259 47 50.21 (3.33) 1.85–89.0 47 2015 13 203.00 (10.39) 112–236 12 49.08 (5.66) 9.0–86.0 12 Northeastern Naturalist Vol. 26, No. 2 L.M. Thompson, et al. 2019 415 were able to resurvey. No additional populations were found after surveying in habitats 4 additional locations at Fort A.P. Hill with wetland characteristics that we visually assessed as suitable for sirens, which suggests that the known populations are persisting but have not colonized additional sites (Schalk and Luhring 2010). Another siren species, Siren lacertina L. (Greater Siren), that occupies similar habitats has been reported at and near Fort A.P. Hill (Mitchell and Roble 1998) but was not found in either of our survey periods. Changes in body size or local occurrence could have indicated competition with other species or habitat disturbances (Luhring and Holdo 2015). Despite being secretive, siren species play an important role in freshwater aquatic communities. As generalist predators, they feed on a wide variety of invertebrates, including isopods and odonates, as well as larval amphibians and conspecific eggs (Collette and Gelbach 1961, Fauth and Resetarits 1991, Petranka 1998). For example, predation on Notophthalmus viridescens Rafinesque (Eastern Newt) larvae by Eastern Lesser Sirens resulted in decreased fecundity of Eastern Newt when newt density was low, but increased fecundity when newt density was high (Fauth and Resetarits 1991). At high newt densities, the presence of Eastern Lesser Siren can also decrease survival and growth of Eastern Newt, likely because of competition between these 2 predator species (Fauth and Resetarits 1991). At Fort A.P. Hill, Eastern Newt and Eastern Lesser Sirens co-occur in many wetlands (Mitchell and Roble 1998), which suggests that this interaction could be important for these communities. Extensive surveys across military installations of the United States, including at Fort A.P. Hill, have shown widespread occurrence of Bd in wetlands that are important to amphibians (Lannoo et al. 2011). During those surveys, Bd was detected at Fort A.P. Hill in Anaxyrus fowleri (Hinckley) (Fowler’s Toad), American Bullfrog, Lithobates clamitans (Latreille in Sonnini de Manoncourt and Latreille) (Green Frog), and Acris crepitans Baird (Northern Cricket Frog), but not in Anaxyrus americanus (Holbrook) (American Toad), Lithobates palustris (LeConte) (Pickerel Frog), or Eastern Newt that were sampled (Petersen et al. 2011). During 2015, we found no detectable levels of the pathogen on any Eastern Lesser Sirens that we captured and screened. However, it is likely that Bd persists in aquatic environments at Fort A.P. Hill since a separate study during the same time period found Bd present on Northern Cricket Frogs, though not in the same wetlands where we captured Eastern Lesser Sirens (Grayson et al. 2016). Other studies examining sirens have found populations both with and without positive detections of Bd in Lesser Sirens from Louisiana and Mississippi and Greater Sirens from Florida (Chatfield et al. 2012, Rizkalla 2010). North American amphibian species vary in susceptibility to Bd infections (Gahl et al, 2012), as has been shown for some fully aquatic salamanders (Chatfield et al. 2012); however, comprehensive work is lacking on the susceptibility of siren species to Bd infection. While we are confident that Bd was not present on the Eastern Lesser Sirens we captured in 2015, our sampling occurred in midsummer. Bd infection is generally more prevalent in the late summer and fall at Fort A.P. Hill (Lannoo et al. 2011), which corresponds to trends of higher infection rates with increased temperature (Hughey et al. 2014). A more thorough Northeastern Naturalist 416 L.M. Thompson, et al. 2019 Vol. 26, No. 2 sampling scheme with a larger sample size is required to definitively conclude that Bd is not currently found on Eastern Lesser Sirens at Fort A.P. Hill and if this pathogen has the potential to negatively impact siren populations. Conclusion This study occurred on military lands operated by the United States Department of Defense; the combination of security measures and legislation requiring comprehensive natural resource management has led to military properties harboring important, and sometimes threatened, ecological communities (Lannoo et al. 2011, Petersen et al. 2016, Stein et al. 2008, Warren et al. 2007, Zentelis and Lindenmayer 2015, Zentelis et al. 2017). Eastern Lesser Siren is one example of a species at Fort A.P. Hill that benefits from the maintenance of Coastal Plain wetlands and pine savannas. The preservation of these areas is essential for the protection of critical habitats and conservation of many amphibian species. The S. intermedia complex of species is categorized as a species of least concern by the IUCN due to its wide distribution and presumed large population sizes (Parra-Olea et al. 2008). This species is believed to be secure in many locations in the United States, and our study indicates that stable populations persist at Fort A.P. Hill at the northern range edge of the Eastern Lesser Siren subspecies (Petranka 1998, Roble 1995). However, the lack of long-term studies leaves us with little understanding of current population trends (Parra-Olea et al. 2008). Continued monitoring will be important particularly in the context of climate change, which will likely result in warmer and more variable temperatures, as well as changes in precipitation patterns that may disproportionately affect individuals at range edges (e.g., Hampe and Petit 2005). Sampling methods that are more robust could be used to provide estimates of occupancy and population density that are more precise. Future work could also compare demographics and performance of Eastern Lesser Siren populations at this isolated site to those in the contiguous portion of its range. Our results tell a cautiously optimistic story of Eastern Lesser Siren populations persisting at a range edge. Acknowledgments We thank the Fort A.P. Hill Environmental and Natural Resources Division, particularly Ben Fulton, Terry Banks, Heather Mansfield, Andrew Satterwhite, and John Yowell of the Fish and Wildlife Branch. We are grateful for their assistance with base operations and logistical support, and for sharing their extensive knowledge of the wildlife at Fort A.P. Hill. We thank Todd Georgel, who was the primary field technician during 1996–1999 and John D. Kleopfer from the Virginia Department of Game and Inland Fisheries for the use of trapping equipment in 2015. James Vonesh and Tom Luhring provided consultation on our study design. Laura Blackburn provided data for the Figure 1 range map. We appreciate reviewer and editor feedback that improved this manuscript. Funding and support was provided by the United States Department of Defense through the Legacy Research Management Program (JCM) and Army Corps of Engineers Cooperative Agreement (KLG, No. W9126G-14-2-0077), the Fort A.P. Hill Department of Public Works, and the University of Richmond School of Arts and Sciences. Northeastern Naturalist Vol. 26, No. 2 L.M. Thompson, et al. 2019 417 Literature Cited Boyle, D.G., D.B. Boyle, V. Olsen, J.A.T. Morgan, and A.D. Hyatt. 2004. Rapid quantitative detection of chytridiomycosis (Batrachochytrium dendrobatidis) in amphibian samples using real-time Taqman PCR assay. Diseases of Aquatic Organisms 60:141–148. Chatfield, M.W.H., P. Moler, and C.L. Richards-Zawacki. 2012. The amphibian chytrid fungus, Batrachochytrium dendrobatidis, in fully aquatic salamanders from southeastern North America. PLoS ONE 7:1–5. Collette, B.B., and F.R. Gehlbach. 1961. The salamander Siren intermedia intermedia Leconte in North Carolina. Herpetologica 17:203–204. Fauth, J.E., and W.J. Resetarits. 1991. Interactions between the salamander Siren intermedia and the keystone predator Notophthalmus viridescens. Ecology 72:827–838. Gahl, M. K., Longcore, J. E., and Houlahan, J. E. 2012. Varying Responses of Northeastern North American Amphibians to the Chytrid Pathogen. Conservation Biology, 26:135–141. Gaston, K.J. 2003. The Structure and Dynamics of Geographic Ranges. Oxford University Press, New York, NY. Grayson, K.L., J. Vonesh, L. Bullock, and C. Viverette. 2016. Baseline surveys for amphibians and Prothonotary Warblers at Fort A.P. Hill in Virginia. Internal Report for Cooperative Agreement No. W9126G-14-2-0077. Fort A.P. Hill, VA. 153 pp. Hampe, A., and R.J. Petit. 2005. Conserving biodiversity under climate change: The rear edge matters. Ecology Letters 8:461–467. Hughey, M.C., M.H. Becker, J.B. Walke, M.C. Swartwout, and L.K. Belden. 2014. Batrachochytrium dendrobatidis in Virginia amphibians: Within- and among-site variation in infection. Herpetological Review 45:428–438. Johnson, S.A., and W.J. Barichivich. 2004. A simple technique for trapping Siren lacertina, Amphiuma means, and other aquatic vertebrates. Journal of Freshwater Ecology 19:263–269. Kroschel, W.A. 2012. Revisiting the ecological status of the Cheat Mountain Salamander (Plethodon nettingi) after 32 years. MS.Thesis. Marshall University, Huntington, WV. 118 pp. Lannoo, M.J., C. Petersen, R.E. Lovich, P. Nanjappa, C. Phillips, J.C. Mitchell, and I. Macallister. 2011. Do frogs get their kicks on route 66? Continental US transect reveals spatial and temporal patterns of Batrachochytrium dendrobatidis infection. PLoS ONE 6. Leja, W.T. 2005. Siren intermedia Barnes 1826, Lesser Siren. Pp. 910–912, In M.J. Lannoo (Ed.). Amphibian Declines: The Conservation Status of United States Species. University of California Press, Berkeley, CA. Luhring, T.M., and R.M. Holdo. 2015. Trade-offs between growth and maturation: The cost of reproduction for surviving environmental extremes. Oecologia 178:723–732. Luhring, T.M., and C.A. Jennison. 2008. A new stratified aquatic sampling technique for aquatic vertebrates. Journal of Freshwater Ecology 23:445–450. Mitchell, J.C., and S. McNulty. 1999. Distribution and habitat affinities of amphibians and reptiles on Fort A.P. Hill, Virginia: A gap analysis study. Final Report to Environmental and Natural Resources Division, Fort A.P. Hill, Bowling Green, VA. Mitchell, J.C., and S.M. Roble. 1998. Annotated checklist of the amphibians and reptiles of Fort A.P. Hill, Virginia, and vicinity. Banisteria 11:19–32. Parra-Olea, G., M. García-París, and D.B. Wake. 1999. Status of some populations of Mexican salamanders (Amphibia: Plethodontidae). Revista de Biología Tropical 47:217–223. Northeastern Naturalist 418 L.M. Thompson, et al. 2019 Vol. 26, No. 2 Parra-Olea, G., D.B. Wake, and G.A. Hammerson. 2008. Siren intermedia. The IUCN red list of threatened species. Available online at http://dx.doi.org/10.2305/IUCN.UK.2008. RLTS.T59491A11936674.en. Accessed October 1, 2018. Petersen, C., R.E. Lovich, M.J. Lannoo, and P. Nanjappa. 2011. Do frogs still get their kicks on Route 66? A transcontinental transect for amphibian chytrid fungus (Batrachochytrium dendrobatidis ) infection on US Department of Defense installations. Final Report for Legacy Resource Management Program Project No. 09-426. Naval Facilities Engineering Command Atlantic, Norfolk, VA. 50 pp. Petersen, C.E., R.E. Lovich, C.A. Phillips, M.J. Dreslik, and M.J. Lannoo. 2016. Prevalence and seasonality of the amphibian chytrid fungus Batrachochytrium dendrobatidis along widely separated longitudes across the United States. EcoHealth 13:368–382. Petranka, J.W. 1998. Salamanders of the United States and Canada. Smithsonian Institution Press, Washington, DC. Powell, R., R. Conant, and J.T. Collins. 2016. Peterson Field Guide to Reptiles and Amphibians of Eastern and Central North America. Fourth Edition. Houghton Mifflin Harcourt Publishing Company, New York, NY. R Core Team. 2018. R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. Rehm, E.M., P. Olivas, J. Stroud, and K.J. Feeley. 2015. Losing your edge: Climate change and the conservation value of range-edge populations. Ecology and Evolution 5:4315–4326. Rizkalla, C.E. 2010. Increasing detections of Bactrachochytrium dendrobatidis in Central Florida, USA. Herpetological Review 41:180–181. Roble, S.M. 1995. Geographic Distribution: Siren intermedia intermedia. Herpetological Review 26:150–151. Rothermel, B.B., S.C. Walls, J.C. Mitchell, C.K. Dodd, L.K. Irwin, D.E. Green, V.M. Vazquez, J.W. Petranka, and D.J. Stevenson. 2008. Widespread occurrence of the amphibian chytrid fungus Batrachochytrium dendrobatidis in the southeastern USA. Diseases of Aquatic Organisms 82:3–18. Schalk, C.M., and T.M. Luhring. 2010. Vagility of aquatic salamanders: Implications for wetland connectivity. Journal of Herpetology 44:104–109. Skerratt, L.F., L. Berger, R. Speare, S. Cashins, K.R. McDonald, A.D. Phillott, H.B. Hines, and N. Kenyon. 2007. Spread of chytridiomycosis has caused the rapid global decline and extinction of frogs. EcoHealth 4:125–134. Snodgrass, J.W., J.W. Ackerman, A.L. Bryan Jr., and J. Burger. 1999. Influence of hydroperiod, isolation, and heterospecifics on the distribution of aquatic salamanders (Siren and Amphiuma) among depression wetlands. Copeia 1999:107–113. Stein, B.A., C. Scott, and N. Benton. 2008. Federal lands and endangered species: The role of military and other federal lands in sustaining biodiversity. BioScience 58:339. Tedesco, P.A., R. Bigorne, A.E. Bogan, X. Giam, C. Jézéquel, and B. Hugueny. 2014. Estimating how many undescribed species have gone extinct. Conservation Biology 28:1360–1370. Warren, S.D., S.W. Holbrook, D.A. Dale, N.L. Whelan, M. Elyn, W. Grimm, and A. Jentsch. 2007. Biodiversity and the heterogeneous disturbance regime on military training lands. Restoration Ecology 15:606–612. Willson, J.D., C.T. Winne, and L.A. Fedewa. 2005. Unveiling escape and capture rates of aquatic snakes and salamanders (Siren spp. and Amphiuma means) in commercial funnel traps. Journal of Freshwater Ecology 20:397–403. Northeastern Naturalist Vol. 26, No. 2 L.M. Thompson, et al. 2019 419 Witte, C.L., M.J. Sredl, A.S. Kane, and L.L. Hungerford. 2008. Epidemiologic analysis of factors associated with local disappearances of native ranid frogs in Arizona. Conservation Biology 22:375–383. Zentelis, R., and D. Lindenmayer. 2015. Bombing for biodiversity: Enhancing conservation values of military training areas. Conservation Letters 8:299–305. Zentelis, R., D. Lindenmayer, J.D. Roberts, and S. Dovers. 2017. Principles for integrated environmental management of military training areas. Land Use Policy 63:186–195.