Evaluating translocation success of wild eastern hellbenders (Cryptobranchus alleganiensis alleganiensis) in Blue Ridge Ecoregion streams using pre- and post-translocation home range sizes and movement metrics.

Translocations of freshwater species have become a widespread conservation strategy to mitigate the impacts of habitat fragmentation, yet they are not often rigorously monitored using animal movement data to determine their success. We demonstrate the value of monitoring pre- and post-translocation movements and home-range sizes of a fully-aquatic, benthic stream salamander, the eastern hellbender (Cryptobranchus a. alleganiensis) to determine translocation success. We studied the home range sizes, movements, and habitat use of individuals (n = 27) in two self-sustaining populations (S1 & S2) for one year, and then subsequently collected similar data from a subset of these individuals (n = 17) that were translocated into two nearby streams (T1 & T2) with dam-isolated, declining populations in the Blue Ridge Ecoregion of Tennessee. We collected 1,571 location data points (869 pre-translocation and 715 post-translocation) from four study sites, and evaluated effects of mass, sex, and pre-translocation home range size/sedentariness, as well as habitat covariates on home range size and movements. Hellbender home range sizes increased from pre-translocation estimates at both sites, but response depended primarily on physical characteristics of release sites. Home range and fine-scale movement metrics indicated that hellbenders translocated from S1 to T1 settled in more quickly, had greater site fidelity, and smaller home ranges than hellbenders translocated from S2 to T2. Hellbender movements were influenced by cover rock size and density rather than individual characteristics. Study-long survival rates of translocated hellbenders increased from S1 to T1 (80% to 100%) and decreased from S2 to T2 (76% to 33%). Monitoring pre- and post-translocation movements was a valuable method for evaluating short-term translocation success in a freshwater environment. For future hellbender translocations, managers should prioritize selecting suitable release sites with contiguous boulder-dense areas (1-2 per m2), adequate prey (crayfish) densities (>1/m2), and habitats with low risk of predation.

Prevalence and Molecular Analysis of Hellbender (Cryptobranchus alleganiensis) Trypanosomes in Tennessee.

Populations of eastern hellbenders (Cryptobranchus alleganiensis alleganiensis) have been declining for multiple decades because of a variety of stressors associated with anthropogenic habitat disturbance followed by riparian sedimentation. The influence of parasite-associated morbidity and mortality on wild hellbender populations is poorly understood. Research has detected widespread trypanosome infection in hellbenders in Virginia, US, with no other reported detections within the eastern hellbender's extensive range. In our study, trypanosomes mostly closely resembling Trypanosoma cryptobranchi were observed in a blood smear of a hellbender from Tennessee during a population survey. Banked whole blood from this hellbender along with 51 other hellbenders was molecularly tested for the 18s rRNA gene of amphibian trypanosomes using newly designed PCR primers. In total, 3/52 (5.8%) hellbenders were PCR- and sequence-positive for trypanosomes. This is the first report of the partial 18s rRNA sequence from hellbender trypanosomes from North America. Further research into trypanosome epidemiology and hellbender health implications is warranted.

Translocation does not influence amphibian chytrid fungus prevalence among wild eastern hellbenders Cryptobranchus alleganiensis.

Disease monitoring is an essential step in translocation projects, specifically in amphibians where emerging pathogens such as the chytrid fungus Batrachochytrium dendrobatidis (Bd) are linked to population declines. The eastern hellbender Cryptobranchus alleganiensis is a large, fully aquatic salamander experiencing precipitous range-wide population declines; however, the role Bd plays in these declines is unclear. To augment declining hellbender populations and determine effects of translocation on Bd prevalence, we conducted a translocation study of wild adult hellbenders from 2 source streams with abundant hellbender populations to 2 streams with declining populations in east Tennessee, USA. In 2018, we implanted radio transmitters into 30 hellbenders and sampled them periodically for Bd until 17 of the 30 hellbenders were translocated in 2019. We attempted to recapture translocated hellbenders approximately every 45 d for 3 mo to determine Bd prevalence post-release. We used qPCR to detect Bd and quantify zoospore loads on positive samples. Hellbenders had a pre-translocation Bd prevalence of 50% (15 of 30), which decreased to 10% (1 of 10) post-translocation. The average zoospore load for positive samples was 73.63 ± 30.82, and no hellbenders showed signs of chytridiomycosis throughout the study. Although we detected no significant effect of translocation on Bd prevalence, we observed a reduction in Bd prevalence post-release. Our results indicate that translocation did not lead to an increase in pathogen prevalence in translocated wild adult hellbenders, suggesting that chytrid did not impact the success of short-term translocations of eastern hellbenders in the Blue Ridge ecoregion.

GEOGRAPHIC AND INDIVIDUAL DETERMINANTS OF IMPORTANT AMPHIBIAN PATHOGENS IN HELLBENDERS (CRYPTOBRANCHUS ALLEGANIENSIS) IN TENNESSEE AND ARKANSAS, USA.

Wildlife diseases are a major threat for species conservation and there is a growing need to implement disease surveillance programs to protect species of concern. Globally, amphibian populations have suffered considerable losses from disease, particularly from chytrid fungi (Batrachochytrium dendrobatidis [Bd] and Batrachochytrium salamandrivorans [Bsal]) and ranavirus. Hellbenders (Cryptobranchus alleganiensis) are large riverine salamanders historically found throughout several watersheds of the eastern and midwestern US. Populations of both subspecies (Ozark hellbender, Cryptobranchus alleganiensis bishopi; eastern hellbender, Cryptobranchus alleganiensis alleganiensis) have experienced precipitous declines over at least the past five decades, and emerging pathogens are hypothesized to play a role. We surveyed Ozark hellbender populations in Arkansas (AR) and eastern hellbender populations in Middle Tennessee (MTN) and East Tennessee (ETN) for both chytrid fungi and ranavirus from swabs and tail tissue, respectively, from 2011 to 2017. Overall, we detected Bd on hellbenders from nine out of 15 rivers, with total prevalence of 26.7% (54/ 202) that varied regionally (AR: 33%, 28/86; MTN: 11%, 4/36; ETN: 28%, 22/80). Ranavirus prevalence (9.0%, 18/200) was comparatively lower than Bd, with less regional variation in prevalence (AR: 6%, 5/ 85; MTN: 11%, 4/36; ETN: 10%, 8/79). We did not detect Bsal in any hellbender populations. We detected a significant negative correlation between body condition score and probability of ranavirus infection (ß=-0.13, SE=0.06, 95% confidence interval: -0.24, -0.02). Evaluation of infection load of positive individuals revealed different trends than prevalence alone for both ranavirus and Bd, with MTN having a significantly greater average ranaviral load than both other regions. We documented a variety of lesions that likely have multiple etiologies on hellbenders located within all geographic regions. Our data represent a multiyear pathogen dataset across several regions of C. alleganiensis, and we emphasize the need for continued pathogen surveillance.

Importance of demographic surveys and public lands for the conservation of eastern hellbenders Cryptobranchus alleganiensis alleganiensis in southeast USA.

Comparisons of recent and historic population demographic studies of eastern hellbenders Cryptobranchus alleganiensis alleganiensis have identified significant population declines and extirpations associated with habitat degradation, poor water quality and disease, leading to nomination as a candidate for listing under the Endangered Species Act. However, populations in the southern Appalachian region of the range have received less attention despite relatively high levels of watershed protection due to the establishment of federally protected National Forest and National Park public lands. These watersheds likely represent some of the best remaining available habitat, yet the lack of published studies make assessment of population stability and viability very difficult. Our objectives were to (1) conduct a capture-mark-recapture (CMR) demographic study and a point transect survey on the Hiwassee River in Tennessee which is designated a National Scenic River, and is largely contained within the Cherokee National Forest, (2) quantify the size structure of the population, (3) compare abundance, survival and recruitment with historic and contemporary hellbender populations across the range, (4) assess the importance of this population and the significance of National Forest and National Park lands in the context of hellbender population conservation in the southeastern United States. We detected all age classes present, with larval hellbenders comprising 21.5% of captures. Using a combination of static life table and CMR methods, we determined that survival rates during the first year were low (~10%), but were high (68-94%) for taggable sized hellbenders. Density of hellbenders at the study site was very high (84 taggable sized hellbenders per 100m of river) compared to recent demographic studies conducted in other regions of the range. We detected hellbenders over ~28 km of river, with a mean density of 23 taggable sized hellbenders per 100m of river, and a total population estimate of 6440 taggable hellbenders. National Forest and National Park lands are likely to continue to play a particularly important role in providing suitable habitat for hellbenders in the southern Appalachians. In fact, only six of 21 known hellbender locations in Tennessee appear to show consistent larval recruitment, all of which are located within or adjacent to National Forest or National Park land.

Predicting the potential distribution of the amphibian pathogen Batrachochytrium dendrobatidis in East and Southeast Asia.

Batrachochytrium dendrobatidis (Bd) is the pathogen responsible for chytridiomycosis, a disease that is associated with a worldwide amphibian population decline. In this study, we predicted the potential distribution of Bd in East and Southeast Asia based on limited occurrence data. Our goal was to design an effective survey area where efforts to detect the pathogen can be focused. We generated ecological niche models using the maximum-entropy approach, with alleviation of multicollinearity and spatial autocorrelation. We applied eigenvector-based spatial filters as independent variables, in addition to environmental variables, to resolve spatial autocorrelation, and compared the model's accuracy and the degree of spatial autocorrelation with those of a model estimated using only environmental variables. We were able to identify areas of high suitability for Bd with accuracy. Among the environmental variables, factors related to temperature and precipitation were more effective in predicting the potential distribution of Bd than factors related to land use and cover type. Our study successfully predicted the potential distribution of Bd in East and Southeast Asia. This information should now be used to prioritize survey areas and generate a surveillance program to detect the pathogen.

Batrachochytrium dendrobatidis haplotypes on the hellbender Cryptobranchus alleganiensis are identical to global strains.

To determine whether the chytrid fungus Batrachochytrium dendrobatidis (Bd) found on the hellbender Cryptobranchus alleganiensis in the southern US is endemic or exotic, we identified the genetic type of this fungus using partial DNA sequences of the internal transcribed spacer (ITS) region. We identified 3 genetic types, which are found on Japanese amphibians other than the Japanese giant salamander Andrias japonicus, a species that belongs to the same family (Cryptobranchidae) as hellbenders. The fungus collected from hellbenders exhibited low genetic diversity and matched the common Bd genetic types which have been detected from around the world. These results support that the chytrid fungus on the hellbender is a novel pathogen, as proposed by previous studies. Although we have not observed disease symptoms directly linked to this fungus on this endangered salamander, further evaluation of the influence of this exotic fungus on this species is warranted.

Magnetic maps in animals: a theory comes of age?

The magnetic map hypothesis proposes that animals can use spatial gradients in the Earth's magnetic field to help determine geographic location. This ability would permit true navigation--reaching a goal from an entirely unfamiliar site with no goal-emanating cues to assist. It is a highly contentious hypothesis since the geomagnetic field fluctuates in time and spatial gradients may be disturbed by geological anomalies. Nevertheless, a substantial body of evidence offers support for the hypothesis. Much of the evidence has been indirect in nature, such as the identification of avian magnetoreceptor mechanisms with functional properties that are consistent with those of a putative map detector or the patterns of orientation of animals exposed to temporal and/or spatial geomagnetic anomalies. However; the most important advances have been made in conducting direct tests of the magnetic map hypothesis by exposing experienced migrants to specific geomagnetic values representing simulated displacements. Appropriate shifts in the direction of orientation, which compensate for the simulated displacements, have been observed in newts, birds, sea turtles, and lobsters, and provide the strongest evidence to date for magnetic map navigation. Careful experimental design and interpretation of orientation data will be essential in the future to determine which components of the magnetic field are used to derive geographic position.

'Fixed-axis' magnetic orientation by an amphibian: non-shoreward-directed compass orientation, misdirected homing or positioning a magnetite-based map detector in a consistent alignment relative to the magnetic field?

Experiments were carried out to investigate the earlier prediction that prolonged exposure to long-wavelength (>500 nm) light would eliminate homing orientation by male Eastern red-spotted newts Notophthalmus viridescens. As in previous experiments, controls held in outdoor tanks under natural lighting conditions and tested in a visually uniform indoor arena under full-spectrum light were homeward oriented. As predicted, however, newts held under long-wavelength light and tested under either full-spectrum or long-wavelength light (>500 nm) failed to show consistent homeward orientation. The newts also did not orient with respect to the shore directions in the outdoor tanks in which they were held prior to testing. Unexpectedly, however, the newts exhibited bimodal orientation along a more-or-less 'fixed' north-northeast-south-southwest magnetic axis. The orientation exhibited by newts tested under full-spectrum light was indistinguishable from that of newts tested under long-wavelength light, although these two wavelength conditions have previously been shown to differentially affect both shoreward compass orientation and homing orientation. To investigate the possibility that the 'fixed-axis' response of the newts was mediated by a magnetoreception mechanism involving single-domain particles of magnetite, natural remanent magnetism (NRM) was measured from a subset of the newts. The distribution of NRM alignments with respect to the head-body axis of the newts was indistinguishable from random. Furthermore, there was no consistent relationship between the NRM of individual newts and their directional response in the overall sample. However, under full-spectrum, but not long-wavelength, light, the alignment of the NRM when the newts reached the 20 cm radius criterion circle in the indoor testing arena (estimated by adding the NRM alignment measured from each newt to its magnetic bearing) was non-randomly distributed. These findings are consistent with the earlier suggestion that homing newts use the light-dependent magnetic compass to align a magnetite-based 'map detector' when obtaining the precise measurements necessary to derive map information from the magnetic field. However, aligning the putative map detector does not explain the fixed-axis response of newts tested under long-wavelength light. Preliminary evidence suggests that, in the absence of reliable directional information from the magnetic compass (caused by the 90 degrees rotation of the response of the magnetic compass under long-wavelength light), newts may resort to a systematic sampling strategy to identify alignment(s) of the map detector that yields reliable magnetic field measurements.

Behavioral titration of a magnetic map coordinate.

Spatial variation in the inclination of the geomagnetic field has been implicated in the map component of homing by eastern red-spotted newts Notophthalmus viridescens. Here we show that when newts are exposed to small changes in magnetic inclination, the most dramatic effects on homing orientation occur at values close to the 'home value', as predicted by the magnetic map hypothesis (Phillips 1996). Newts reverse the direction of homing orientation over a range of inclination of 0.5 degrees spanning the home value, providing further evidence that magnetic inclination or one of its components (i.e., vertical or horizontal intensity) is used to derive map information.