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Environmental DNA improves Eastern Hellbender (Cryptobranchus alleganiensis alleganiensis) detection over conventional sampling methods
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Effective conservation planning relies on accurate species detection. However, conventional sampling methods used for detecting rare and cryptic aquatic species suffer from low probabilities of detection. Environmental DNA (eDNA) has emerged as an innovative and powerful sampling tool for detecting aquatic species, with previous studies suggesting a detection advantage over conventional sampling. However, comparative studies often fail to consider the appropriate sampling frameworks to adequately compare sampling methodologies and account for the influence of environmental variables on eDNA detection probabilities. In this study, we paired two detection methods (eDNA and physical sampling) at 22 sites in West Virginia, USA, to compare the probability of detecting a cryptic, elusive, and imperiled species of giant salamander, the Eastern Hellbender (Cryptobranchus alleganiensis alleganiensis). We used a multimethod occupancy modeling framework to compare method‐specific detection probabilities using a suite of predictor variables based on environmental conditions thought to influence hellbender detection. We detected hellbenders at 19/22 sites using eDNA and at 13/22 sites using physical sampling methods. The best supported model indicated that detection probability for eDNA (0.84 ± 0.06) was three times higher than conventional methods (0.28 ± 0.07). Water turbidity was the best predictor of hellbender detection and negatively impacted our ability to detect eDNA. We failed to detect an association between eDNA concentration and hellbender catch per unit effort. Our study supports previous findings that suggest eDNA sampling methods greatly increase the probability of detecting aquatic species. However, with little known about the influence of environmental variables on eDNA detection, our results highlight the negative influence turbidity and other physiochemical factors have on eDNA detection and suggest that further research on eDNA detection in turbid environments is needed.
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Noninvasive Method for a Statewide Survey of Eastern Hellbenders Cryptobranchus alleganiensis Using Environmental DNA
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Traditional survey methods of aquatic organisms may be difficult, lengthy, and destructive to the habitat. Some methods are invasive and can be harmful to the target species. The use of environmental DNA (eDNA) has proven to be effective at detecting low population density aquatic macroorganisms. This study refined the technique to support statewide surveys. Hellbender presence was identified by using hellbender specific primers (cytochrome b gene) to detect eDNA in water samples collected at rivers, streams and creeks in Ohio and Kentucky with historical accounts of the imperiled eastern hellbender (Cryptobranchus a. alleganiensis). Two sampling protocols are described; both significantly reduced the amount of water required for collection from the previously described 6 L collection. Two-liter samples were adequate to detect hellbender presence in natural waterways where hellbenders have been previously surveyed in both Ohio and Kentucky—1 L samples were not reliable. DNA extracted from 3 L of water collected onto multiple filters (1 L/filter) could be combined and concentrated through ethanol precipitation, supporting amplification of hellbender DNA and dramatically reducing the filtration time. This method improves the efficiency and welfare implications of sampling methods for reclusive aquatic species of low population density for statewide surveys that involve collecting from multiple watersheds.
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Prevalence of Batrachochytrium dendrobatidis in Eastern Hellbender (Cryptobranchus alleganiensis) Populations in West Virginia, USA
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The eastern hellbender (Cryptobranchus alleganiensis alleganiensis) is a North American salamander species in decline throughout its range. Efforts to identify the causes of decline have included surveillance for the chytrid fungus Batrachochytrium dendrobatidis (Bd), which has been associated with global amphibian population losses. We evaluated the prevalence of Bd in 42 hellbenders at four sites in West Virginia, US, from June to September 2013, using standard swab protocols and real-time PCR. Overall prevalence of Bd was 52% (22/42; 37.7–66.6%; 95% confidence interval). Prevalence was highest in individuals with body weight ≥695 g (χ2=7.2487, df=1, P=0.007), and was higher in montane sampling sites than lowland sites (t=−2.4599, df=44, P=0.02). While increased prevalence in montane sampling sites was expected, increased prevalence in larger hellbenders was unexpected and hypothesized to be associated with greater surface area for infection or prolonged periods of exposure in older, larger hellbenders. Wild hellbenders have not been reported to display clinical disease associated with Bd; however, prevalence in the population is important information for evaluating reservoir status and risk to other species, and as a baseline for investigation in the face of an outbreak of clinical disease.
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Improving the Utility of Artificial Shelters for Monitoring Eastern Hellbender Salamanders (Cryptobranchus alleganienses alleganiensis)
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Artificial shelters show great promise as novel, non-invasive tools for studying
hellbenders, but their use thus far has faced several challenges. During initial trials in multiple
river networks, artificial shelters routinely became blocked by sediment and dislodged during
high stream discharge events, and were rarely used by hellbenders. We sought to determine
whether these complications could be overcome via alternative shelter design, placement, and
maintenance. Between 2013 and 2018, we deployed 438 artificial shelters of two different
designs across ten stream reaches and three rivers in the upper Tennessee River Basin. We
assessed evidence for several hypotheses, postulating broadly that the availability, stability, and
use of artificial shelters by hellbenders would depend on how shelters were constructed,
deployed, and/or maintained. We found that maintaining shelters at least once every 40 days
limited sediment blockage, and building ~ 40 kg shelters with 3-4 cm thick walls and recessed
lids improved their stability during high discharge events. Additionally, we found that
hellbenders most frequently occupied and nested in artificial shelters when they were deployed
in deeper (~50+ cm) portions of reaches with high adult hellbender densities. Our results suggest
that artificial shelters can serve as effective tools for studying hellbenders when designed,
deployed, and maintained with these advancements, but also highlight some limitations of their
use.
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Artificial Nest Box Research
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Evaluating artificial shelter arrays as a minimally invasive monitoring tool for the hellbender (Cryptobranchus alleganiensis)
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Hellbenders Cryptobranchus alleganiensis are critically imperiled amphibians throughout the eastern USA. Rock-lifting is widely used to monitor hellbenders but can severely disturb habitat. We asked whether artificial shelter occupancy (the proportion of occupied shelters in an array) would function as a proxy for hellbender abundance and there by serve as a viable alternative to rock-lifting. We hypothesized that shelter occupancy would vary spatially in response to hellbender density, natural shelter density, or both, and would vary temporally with hellbender
seasonal activity patterns and time since shelter deployment. We established shelter arrays (n = 30 shelters each) in 6 stream reaches and monitored them monthly for up to 2 yr. We used Bayesian mixed logistic regression and model ranking criteria to assess support for hypotheses concerning drivers of shelter occupancy. In all reaches, shelter occupancy was highest from June–August each year and was higher in Year 2 relative to Year 1. Our best-supported model indicated that the extent of boulder and bedrock (hereafter, natural shelter) in a reach mediated the relationship between hellbender abundance and shelter occupancy. More explicitly, shelter occupancy was positively correlated with abundance when natural shelter covered <20% of a reach, but uncorrelated with abundance when natural shelter was more abundant. While shelter occupancy should not be used to infer variation in hellbender relative abundance when substrate composition varies among reaches, we showed that artificial shelters can function as valuable monitoring tools when reaches meet certain criteria, though regular shelter maintenance is critical.
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Artificial Nest Box Research
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The use of nest boxes by the hellbender salamander in Western North Carolina
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The hellbender salamander (Cryptobranchus alleganiensis) is a unique, large-bodied amphibian that serves as an excellent water quality indicator species in Western North Carolina. This animal has suffered substantial population declines over the past four decades throughout its range. Increased stream siltation largely attributed to human development fills the concave undersides of large rocks, consequently destroying hellbender breeding habitat. Habitat degradation has contributed to reductions in North Carolinian populations to such a degree that the species is now considered of Special Concern in the state. In order to restore hellbender population sizes under current land use conditions, researchers have recently begun developing artificial nest boxes that exclude sediment and promote increased reproduction. To identify the short-term efficacy of these shelters as substitutes for natural hellbender habitat in Western North Carolina, I constructed and placed 54 boxes across five river sites throughout the region. Following summer nest box installment, I examined each shelter through the breeding season for hellbender in habitation and to determine the quality of water passing through the structures. Additionally, I created a maximum entropy species distribution model and conducted a spatial connectivity analysis for the hellbenders of Western North Carolina to identify ideal locations for nest boxes installation in the future. Although no hellbenders have yet been detected in the artificial shelters, additional structural improvements and time may reveal nest boxes to be useful conservation tools for this iconic species of Special Concern.
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Artificial Nest Box Research
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Movement and habitat use of Eastern hellbenders (Cryptobranchus alleganiensis alleganiensis) following population augmentation
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With amphibian declines at crisis levels, translocations, including population augmentations, are commonly used for amphibian conservation. Eastern Hellbenders (Cryptobranchus alleganiensis alleganiensis) have declined to low densities in many areas of their range, making them ideal candidates for population augmentation. Both wild adults and captive-reared juveniles have been used for augmentations, but their suitability has never been directly compared. Herein, we use radio telemetry with Eastern Hellbenders to examine patterns of site fidelity, movement, and habitat use over a 2-yr period for adult residents, wild adult translocates, and captive-reared juvenile translocates. We used generalized linear models and generalized linear mixed models to identify temporal trends and explore the effects of residential status (resident vs. translocate) and origin/age (captive-reared juveniles vs. wild adults) on various ecological and behavioral traits relating to habitat. Site fidelity was high in adult residents and wild adult translocates, but lower in captive-reared juvenile translocates. Both adult and juvenile translocates had greater mean movement distances than residents, leading to larger home range sizes, but these differences decreased over time. Wild adult translocates had a higher probability of using artificial nest rocks than adult residents or captive-reared juvenile translocates. This pattern was most prevalent early in the study, indicating these shelters are particularly useful during the transition to release sites. Captive-reared juvenile translocates had lower site fidelity and utilized suboptimal habitat (smaller and fewer shelter rocks) compared to wild adults. Compared to previous studies, translocations had fewer negative effects on site residents or wild translocates and might be effective at promoting growth of Hellbender populations. However, translocations of captive-reared juveniles were less successful. As we are uncertain whether captive-rearing or ontogeny led to these differences, both longer head-starting times and conditioning should be explored to improve outcomes in captive-reared juvenile cohorts.
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Artificial Nest Box Research
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Theory and practice of the hydrodynamic redesign of artifical hellbender habitat
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The success of nest boxes in Missouri led researchers to test whether similar management tools could increase C. a. alleganiensis populations in the streams of western North Carolina, where these salamanders are listed as a Species of Special Concern (Messerman 2014). Fifty-four nest boxes were constructed following the boot-shaped design of Briggler and Ackerson (2012) in May 2013, and were installed across five known C. alleganiensis stream sites between late June and early August 2013. Messerman (2014) then monitored each nest box every three to four weeks through November 2013, and the boxes were revisited in August 2014 and July 2015 to observe structural condition and occupancy (Messerman, pers. obs.). Of the 54 nest boxes, only two structures at a single site were confirmed as inhabited in 2014 and 2015, and no breeding events were detected (Messerman, pers. obs.). Moreover, many of these ~50 lb concrete boxes moved in flood events or accumulated sediment at the downstream tunnel entrance (Messerman 2014). The low success of the boot-shaped nest box design in North Carolina may be attributed to the sites generally being narrower and shallower than those in Missouri, with much of the substrate consisting of bedrock slabs covered by relatively thin layers of rock, gravel and silt. Here we address the observed shortcomings of the original North Carolina design through the lens of engineering, and present a new and easily implemented nest box model for use in streams like those found in western North Carolina.
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Artificial Nest Box Research
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Partners in Amphibian and Reptile Conservation
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Partners in Amphibian and Reptile Conservation (PARC) is an inclusive partnership dedicated to the conservation of the herpetofauna--reptiles and amphibians--and their habitats. Our membership comes from all walks of life and includes individuals from state and federal agencies, conservation organizations, museums, pet trade industry, nature centers, zoos, energy industry, universities, herpetological organizations, research laboratories, forest industries, and environmental consultants. The diversity of our membership makes PARC the most comprehensive conservation effort ever undertaken for amphibians and reptiles.
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Southeastern Hellbender Conservation Initiative
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The Southeastern Hellbender Conservation Initiative (SEHCI), a collaboration between Defenders of Wildlife, NRCS and other conservation partners to support farmers using conservation practices on their lands that help restore hellbender habitat.
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