Desmognathus Ochrophaeus Research Articles

An Evaluation of Immersive and Handling Methods for Collecting Salamander Skin Peptides

Amphibian skin secretions are a rich source of antimicrobial peptides that have long interested both the biomedical and herpetological research communities. While anuran skin peptides are well studied, little is known about skin peptides in salamanders, in part because methods for collecting them are poorly developed. In anurans, pharmacological agents are commonly used to stimulate skin peptide responses, thus increasing the amount of peptide collected. It was unclear whether similar methods would work for salamanders. We conducted a series of experiments to test the effects of immersion in pharmaceutical agents (norepinephrine hydrochloride [NE]; acetylcholine chloride [Ach]), handling, and the interaction of both on peptide collection from the skin of Red-Spotted Newts (Notophthalmus viridescens) and Allegheny Mountain Dusky Salamanders (Desmognathus ochrophaeus). Our results suggest that vigorous handling, but not immersion in NE or Ach, is a better method for collecting salamander skin peptides. Despite the widespread use of pharmaceuticals to collect anuran skin peptides, we found little evidence that NE or Ach induces peptide release in salamanders. In fact, we show that inclusion of NE in collection solutions interferes with peptide quantification in protein assays.

Combined Effects of Experimentally Elevated CORT and Predation Threat on Exploratory and Foraging Behavior of Desmognathus ochrophaeus

Amphibian responses to stress are generally mediated through glucocorticoids produced by the hypothalamic–pituitary–interrenal axis. Chronic elevation of glucocorticoids can result in delayed wound healing and growth, but less is known about its influence on behavioral responses to predators. We examined the effect of acute and chronic CORT elevation on exploratory and foraging behaviors of Allegheny Mountain Dusky Salamanders (Desmognathus ochrophaeus) exposed to kairomones from a predatory snake. We established ‘No CORT,' ‘Acute CORT,' and ‘Chronic CORT’ conditions during an 18-d preparation phase by varying salamander exposure to an exogenous source of CORT (0.51 mg/mL) delivered via cutaneous patch. During the trial phase, we conducted assays to evaluate exploratory and foraging behavior of salamanders from the different CORT groups when exposed to snake kairomones or control. Exploratory behavior was examined within a circular arena with a series of surmountable concentric barriers, and foraging behavior was observed within Petri dishes containing Drosophila prey. Salamanders experiencing both Chronic CORT and snake kairomones exhibited the least exploratory behavior, escaped the array with the lowest frequency, climbed the fewest barriers, and delayed their movement. However, foraging behavior (latency to strike at prey, number of strikes, number of prey captured) was not affected by exposure to CORT or snake kairomone. Our findings suggest that artificially elevated CORT can modify some behavioral responses to predator kairomones, but only when individuals experience multiple CORT applications in the period preceding the trials.

Salamander survey of Jakey Hollow Natural Area, Columbia County, Pennsylvania

AbstractWith current declines in amphibian and reptile populations, herpetological surveys of protected natural areas are crucial to management of these organisms, especially as natural areas become more insular due to surrounding land use. Jakey Hollow Natural Area (JHNA) is a locally significant biological site in Columbia County, PA, consisting of wooded slopes bisected by a small-order stream. This report presents the results of a salamander survey at JHNA during the spring of 2020. Cover object surveys revealed a total of 190 salamanders belonging to seven species. The most common salamanders observed at JHNA were Plethodon cinereus and Desmognathus ochrophaeus, which typify other surveys of similar habitats in Pennsylvania. Other species found in lower abundance were D. fuscus, Eurycea bislineata, Gyrinophilus porphyriticus, P. glutinosus, and Pseudotriton ruber. Future surveys utilizing other techniques may reveal the presence of additional salamander species at this site.

Salamander stress and duress: the relationship between CORT, autotomy and regeneration, and exploratory behaviour

Responses to stress are generally mediated through the production of glucocorticoids by the hypothalamic-pituitary-adrenal (or -interrenal) axis. The prolonged production of stress hormones can contribute to delayed wound healing and growth, but little is known about their influence on regeneration following tail autotomy, or exploratory behaviour in autotomized individuals. Here we examined the relationship between stress, regeneration, and exploratory behaviour in Allegheny Mountain dusky salamanders (Desmognathus ochrophaeus) by manipulating corticosterone (CORT) levels via cutaneous patch. First, we measured tail regeneration in salamanders with elevated CORT for 13 weeks after the induction of tail autotomy. Test subjects received a weekly patch to wear for one hour that was saturated with either a low CORT (0.25 mg/ml) or high CORT (0.50 mg/ml) solution. Individuals receiving CORT patches regenerated significantly less of their tail length and volume (versus control), but without exhibiting dose-dependent effects. Second, we used a factorial design to evaluate the effects of autotomy and elevated CORT on exploration within a test arena consisting of low barriers arrayed in concentric rings. Individuals experiencing tail autotomy exhibited significantly less exploratory behaviour indicated by an increased latency to cross first barrier and a decreased number of barriers crossed. Neither elevated CORT (0.50 mg/ml), nor the interaction between elevated CORT and tail autotomy significantly affected salamander activity within the array. Although CORT did not have a direct effect on explorative behaviour, a delay in regeneration attributed to CORT could lead to changes in patterns of movement in autotomized individuals.

A Survey of Terrestrially Active Salamanders from Two Different Woodlands at the Powdermill Nature Reserve, Pennsylvania

Variation in habitat and microhabitat can affect salamander populations. We systematically surveyed terrestrially active salamanders in an old forest tract and a young forest tract at the Powdermill Nature Reserve in southwestern Pennsylvania during 2009–2011. We examined the effect forest age had on the 2 salamander assemblage structures. While species compositions were similar, assemblage structures were uneven in both the old and young forest sites. Differential use of survey coverboards across species was detected at both sites. This difference in use may be best explained by ground-moisture levels as a function of coverboard distance to water. Counterintuitive findings of low numbers of Desmognathus ochrophaeus (Mountain Dusky Salamander) and Plethodon cinereus (Red-backed Salamander) compared to past studies at these 2 sites appears most likely related to differences in trapping methodology. Patterns of differential use of coverboards and inter-site differences in seasonal activity patterns point to P. glutinosus (Northern Slimy Salamander) as the least restricted species in terms of microhabitat conditions and most seasonally variable in activity.

The effect of predator kairomones on caudal regeneration by Allegheny Mountain Dusky Salamanders (Desmognathus ochrophaeus)

Many prey use autotomy as an antipredator mechanism. Rapid regeneration of autotomized appendages is beneficial because forfeited tissues may serve as organs for energy storage, accessories for locomotion, or indicators of social status. We monitored levels of caudal regeneration by Allegheny Mountain Dusky Salamanders (Desmognathus ochrophaeus Cope, 1859) exposed to kairomones from predatory Eastern Garter Snakes (Thamnophis sirtalis (Linnaeus, 1758)). After the induction of autotomy, salamanders were exposed to one of three treatment regimens: blank (water), or acute (30 min per week) or chronic (constant) exposure to predator kairomones during a 12-week study period. Overall, the mean volume of regenerated tissue, as a percentage of the original tail volume, was highest for individuals exposed to the blank versus predator kairomones. When the combined effects of time elapsed since the induction of caudal autotomy and the different treatment regimens were considered, we found that the mean volume of regenerated tissue was significantly greater for control salamanders beginning 8 weeks after autotomy. The mechanism contributing to the differential rates of regeneration among individuals in our treatment groups is unknown, but previous work suggests that elevated stress related to predation threat can have detrimental effects on wound healing and growth in amphibians.

The Hip-twist Jump: A Unique Mechanism for Jumping in Lungless Salamanders

Abstract Many terrestrial and aquatic animals jump to escape predators. Most terrestrial animals use robust hindlimbs to produce the power needed to lift their bodies off the ground. This paper explores an alternative jump mechanism used by the lungless salamander, Desmognathus ochrophaeus. Previous work on this jump mechanism suggested that lungless salamanders invoke rapid torso flexion to hurl themselves into the air with minimal hindlimb contribution. The current study was designed to investigate how salamanders pushed off the ground to jump and to determine whether elastic structures contribute to jump power amplification. Specifically, we focused on trunk action, hip rotation, limb placement, and jump power production. We used high speed cameras to film salamander jumps and used these videos to conduct a subsequent motion analysis. Our results demonstrate that prior to jumping, the trunk flexes laterally into a C-shape while the hindlimb (inside of the bend) is planted in front of the hips. Next, th...

Effects of size, caudal autotomy, and predator kairomones on the foraging behavior of Allegheny Mountain dusky salamanders (Desmognathus ochrophaeus)

Prey must balance the conflicting demands of foraging and defensive behavior. Foraging under the threat of predation may be further complicated among species that engage in caudal autotomy, the loss of a portion of the tail at preformed breakage planes, because the tail may serve as an important energy storage organ and contribute to motility, culminating in a trade-off between foraging and predator avoidance. As a result of the advantages conferred by the presence of a tail, individuals that have recently undergone autotomy may be more motivated to forage despite elevated levels of threat indicated by predator kairomones. We used a full factorial design to evaluate the combined effects of body size, exposure to predator kairomones, and experience with autotomy on the latency to strike at Drosophila prey, number of strikes, and prey captured per strike by Allegheny Mountain dusky salamanders (Desmognathus ochrophaeus). In our study, caudal autotomy was the only significant main effect and influenced both the latency to attack prey and the number of strikes attempted. In terms of latency to attack prey, there was a significant interaction between body size and autotomy such that “small” salamanders (≤3.2 cm SVL) without tails delayed their foraging behavior. In terms of the number of strikes toward prey, there was a significant interaction between autotomy and exposure to predator kairomones such that individuals with intact tails exhibited a greater number of strikes, with the exception of the “large” (>3.2 cm SVL) salamanders, which performed fewer strikes when exposed to the snake kairomones. There was no significant effect on foraging efficiency, although the trend in the data suggests that autotomized individuals forage more efficiently. This study was designed to evaluate the confluence of factors related to size, caudal autotomy, and exposure to stimuli from predators and hints at the magnitude of caudal autotomy on antipredator decision-making. Our data suggest that despite the importance of tail tissue for energy storage, locomotion, and mating, salamanders without tails are cautious when foraging under elevated risk.

Woodland salamander responses to a shelterwood harvest-prescribed burn silvicultural treatment within Appalachian mixed-oak forests

Forest management practices that mimic natural canopy disturbances, including prescribed fire and timber harvests, may reduce competition and facilitate establishment of favorable vegetative species within various ecosystems. Fire suppression in the central Appalachian region for almost a century has contributed to a transition from oak-dominated to more mesophytic, fire-intolerant forest communities. Prescribed fire coupled with timber removal is currently implemented to aid in oak regeneration and establishment but responses of woodland salamanders to this complex silvicultural system is poorly documented. The purpose of our research was to determine how woodland salamanders respond to shelterwood harvests following successive burns in a central Appalachian mixed-oak forest. Woodland salamanders were surveyed using coverboard arrays in May, July, and August–September 2011 and 2012. Surveys were conducted within fenced shelterwood-burn (prescribed fires, shelterwood harvest, and fencing to prevent white-tailed deer [Odocoileus virginianus] herbivory), shelterwood-burn (prescribed fires and shelterwood harvest), and control plots. Relative abundance was modeled in relation to habitat variables measured within treatments for mountain dusky salamanders (Desmognathus ochrophaeus), slimy salamanders (Plethodon glutinosus), and eastern red-backed salamanders (Plethodon cinereus). Mountain dusky salamander relative abundance was positively associated with canopy cover and there were significantly more individuals within controls than either shelterwood-burn or fenced shelterwood-burn treatments. Conversely, habitat variables associated with slimy salamanders and eastern red-backed salamanders did not differ among treatments. Salamander age-class structure within controls did not differ from shelterwood-burn or fenced shelterwood-burn treatments for any species. Overall, the woodland salamander assemblage remained relatively intact throughout the shelterwood-burn silvicultural treatment compared to previous research within the same study area that examined pre-harvest fire effects. However, because of the multi-faceted complexities of this specific silvicultural system, continued research is warranted that evaluates long-term, additive impacts on woodland salamanders within managed central Appalachian deciduous forests.

Amphibians in the Canaan Valley Drainage

Abstract I have been studying amphibians in Canaan Valley and its surrounding region since 1976. Most of my studies inventoried populations of Plethodon nettingi (Cheat Mountain Salamander). I sampled 29 upland sites on the mountains around Canaan Valley: (1) twelve sites on Canaan Mountain, which forms Canaan Valley's western rim, were distributed from Chimney Rock, at the southern end of Canaan Mountain, northwards to the Blackwater River at Camp 70; (2) on the eastern rim of Canaan Valley, 17 sites were on Cabin Mountain, from Snowy Point in the north to Weiss Knob in the south. At the montane sites, I observed 11 salamander species and 1 toad species. Salamanders found included Ambystoma maculatum (Spotted Salamander), Notophthalmus v. viridescens (Red-spotted Newt), Desmognathus monticola (Seal Salamander), Desmognathus ochrophaeus (Allegheny Mountain Dusky Salamander); Gyrinophilus p. porphyriticus (Northern Spring Salamander), Hemidactylium scutatum (Four-toed Salamander), Eurycea bislineata (North...

Modeling cross-scale relationships between climate, hydrology, and individual animals: generating scenarios for stream salamanders

Hybrid modelling provides a unique opportunity to study cross-scale relationships in environmental systems by linking together models of global, regional, landscape, and local-scale processes, yet the approach is rarely applied to address conservation and management questions. Here, we demonstrate how a hybrid modelling approach can be used to assess the effect of cross-scale interactions on the survival of the Allegheny Mountain Dusky Salamander (Desmognathus ochrophaeus) in response to changes in temperature and water availability induced by climate change at the northern limits of its distribution. To do so, we combine regional climate modelling with a landscape-scale integrated surface-groundwater flow model and an individual-based model of stream salamanders. On average, climate scenarios depict a warmer and wetter environment for the 2050 horizon. The increase in average annual temperature and extended hydrological activity time series in the future, combined with a better synchronization with the salamanders’ reproduction period, result in a significant increase in the long-term population viability of the salamanders. This indicates that climate change may not necessarily limit the survivability of small, stream-dwelling animals in headwater basins located in cold and humid regions. This new knowledge suggests that habitat conservation initiatives for amphibians with large latitudinal distributions in Eastern North America should be prioritized at the northern limits of their ranges to facilitate species migration and persistence in the face of climate change. This example demonstrates how hybrid models can serve as powerful tools for informing management and conservation decisions.

Treatment with corticosterone delays cutaneous wound healing in male and female salamanders

In vertebrates, exposure to stressors and stress hormones has a number of physiological effects including modulation of immune function. These effects on immune function have been well studied in mammals, but less is known in other groups, in particular amphibians. To analyze the effects of exposure to stressors and the stress hormone corticosterone, we monitored cutaneous wound healing as a measure of integrated immunity in male and female semi-terrestrial salamanders (Desmognathus ochrophaeus) that were chased to induce endogenous release of corticosterone or were treated with physiologically relevant doses of corticosterone. As predicted, subjects treated daily with corticosterone healed more slowly than did controls. In contrast, subjects that had been chased daily healed at the same rate as controls. Surprisingly, repeated chasing did not elevate plasma corticosterone despite causing drops in body mass and survival. Additionally, females healed more slowly than males, possibly due to energetic constraints.

Testing the predation stress hypothesis: behavioural and hormonal responses to predator cues in Allegheny Mountain dusky salamanders

The predation stress hypothesis posits that exposure to predators and/or predator cues causes release of glucocorticoid hormones which coordinate behavioural responses that facilitate predator avoidance. We measured responses to short-term and repeated exposure to predator-derived kairomones in Allegheny Mountain dusky salamanders (Desmognathus ochrophaeus). Salamanders expressed predator avoidance behaviours (reduced locomotion, reduced mating behaviour) in the presence of predator kairomones. However, plasma glucocorticoids after short-term exposure to predator kairomones were similar to levels after exposure to controls. After repeated exposure to predator-derived kairomones, locomotory activity and plasma glucocorticoids were similar compared to controls. There was no evidence of habituation to predator kairomones. Overall, results did not support the predation stress hypothesis in Allegheny Mountain dusky salamanders in either an acute or chronic context. Use of glucocorticoids to mediate antipredator responses may occur when predation pressure is unpredictable, and when energetic and opportunity costs of linking glucocorticoids to anti-predator responses are low.

Effects of temperature and surface water availability on spatiotemporal dynamics of stream salamanders using pattern-oriented modelling

Ecological models are powerful tools for generating predictions about the viability of populations of endangered species, especially in landscapes where they may be subject to complex, cross-scale disturbances. Spatially explicit, individual-based approaches are particularly promising due to their ability to simulate the effect of landscape level changes in habitat on individual-level behaviour, thus predicting possible emergent responses from the bottom up. We apply this approach to modelling the movement behaviour and the complex life cycles of two species of stream-dwelling salamanders (the Allegheny Mountain Dusky Salamander, Desmognathus ochrophaeus, and the Northern Spring Salamander, Gyrinophilus porphyriticus) in response to a spatially and temporally varying environment. Despite the poor state of ecological knowledge about these species, our model provides reasonable predictions about life cycle, as well as the density and distribution of salamanders. When tested with a dynamic, drought prone environment, the model predicts viability levels that are biologically plausible. By simulating the cross-scale interactions between organisms and their environment, individual-based models such as we have developed here provide a new tool for in silico investigations of the expected impacts of varying landscape scenarios and environmental changes.

Decline of the Cheat Mountain Salamander over a 32-Year Period and the Potential Influence of Competition from a Sympatric Species

We evaluated trends in occupancy of the Cheat Mountain Salamander (Plethodon nettingi) over a 32-yr period and examined the potential influence of competition by sympatric salamander species on these changes. We conducted surveys at 36 locations along four transects on an elevational gradient in the Appalachian Mountains geographic province of West Virginia, USA. We used occupancy modeling to examine patterns in species distributions for three focal species: P. nettingi, a federally threatened species, the Eastern Red-Backed Salamander (Plethodon cinereus), and the Allegheny Mountain Dusky Salamander (Desmognathus ochrophaeus). The probability of occupancy for P. nettingi was considerably lower in 2011 compared to 1978–79 at medium and high elevations (1,169–1,378 m). Additionally, occupancy of P. nettingi was associated negatively with P. cinereus at the highest elevations. These data suggest that these P. nettingi populations have declined and P. cinereus have possibly expanded their vertical distribution. Thus, P. cinereus may be negatively affecting mid- and high-elevation populations of P. nettingi. Alternatively, environmental changes (e.g., habitat disturbance and altered weather patterns) may have contributed to declines of P. nettingi at mid- and high-elevation sites, facilitated by colonization by P. cinereus. Due to the endemic and federally threatened status of P. nettingi, conservation efforts to avoid fragmentation of P. nettingi habitat should be maintained and possibly enhanced, as ecological impacts of environmental changes can be exacerbated in high-elevation habitats.

Modeling low-flow bedrock springs providing ecological habitats with climate change scenarios

Groundwater discharge areas, including low-flow bedrock aquifer springs, are ecologically important and can be impacted by climate change. The development of and results from a groundwater modeling study simulating fractured bedrock spring flow are presented. This was conducted to produce hydrological data for an ecohydrological study of an endangered species, Allegheny Mountain Dusky Salamanders (Desmognathus ochrophaeus), in southern Quebec, Canada. The groundwater modeling approach in terms of scale and complexity was strongly driven by the need to produce hydrological data for the related ecohydrological modeling. Flows at four springs at different elevations were simulated for recent past conditions (2006–2010) and for reference (1971–2000) and future (2041–2070) periods using precipitation and temperature data from ten climate scenarios. Statistical analyses of spring flow parameters including activity periods and duration of flow were conducted. Flow rates for the four simulated springs, located at different elevations, are predicted to increase between 2% and 46% and will be active (flowing) 1–2% longer in the future. A significant change (predominantly an increase) looking at the seasonality of the number of active days occurs in the winter (2–4.9%) and spring seasons (−0.6–6.5%). Greatest flow rates were produced from springs at elevations where sub-horizontal fractures intersect the ground surface. These results suggest an intensification of the spring activity at the study site in context of climate change by 2050, which provides a positive habitat outlook for the endangered salamanders residing in the springs for the future.

Antipredator Behavior in Desmognathus ochrophaeus: Threat‐Specific Responses to Chemical Stimuli in a Foraging Context

AbstractPrey species may reduce the likelihood of injury or death by engaging in defensive behavior but often incur costs related to decreased foraging success or efficiency. To lessen these costs, prey may adjust the intensity or type of antipredator behavior according to the nature of the perceived threat. We evaluated the potential for threat‐sensitive responses by Allegheny Mountain dusky salamanders (Desmognathus ochrophaeus) exposed to chemical stimuli associated with predation by asking three questions: (1) Do individual D. ochrophaeus respond to chemical cues in a threat‐sensitive manner? (2) Do salamanders exhibit the same pattern of behavioral response while foraging? and (3) Is foraging efficiency reduced when focal individuals are exposed to stimuli from predators or predation events? In our first experiment, we evaluated salamander chemosensory movements (nose‐taps), locomotor activity (steps), and edge behavior in response to chemical stimuli from disturbed and injured conspecifics as well as predatory Gyrinophilus porphyriticus and found that individual D. ochrophaeus show a significant graded increase in nose‐taps when exposed to cues from conspecifics and a reduction in activity when exposed to the predator. In our second experiment, we again observed salamander responses to the same chemical stimuli but in this instance added five Drosophila prey to the test dishes. We found that salamanders exhibited a similar pattern of response to the chemical stimuli in the presence of prey, showing a graded increase in nose‐taps to cues from conspecifics and a reduction in activity when exposed to the predator. However, foraging efficiency (i.e. the proportion of successful strikes) did not vary significantly among treatments. Our data show that individual D. ochrophaeus detect and differentially respond to chemical stimuli associated with predation, but do not significantly reduce foraging efficiency. Overall, the type and relative intensity of these responses is largely unaffected by the presence of potential prey.

Environmental acidification is not associated with altered plasma corticosterone levels in the stream-side salamander, Desmognathus ochrophaeus

As environments become increasingly altered due to anthropogenic factors, interest is growing in how endocrine systems respond to pollution and environmental degradation. Glucocorticoid hormones (GCs) are a type of stress hormones that are released upon activation of the hypothalamic–pituitary–adrenal axis and have widespread effects throughout the body. We tested the hypothesis that exposure to environmental acidification is associated with altered levels of plasma GCs in adult, stream-side Allegheny Mountain dusky salamanders (Desmognathus ochrophaeus). We compared plasma corticosterone (CORT) in salamanders living in 9 streams that differed in pH. Although capture and handling induced a robust increase in plasma CORT in all populations of salamanders, we discerned no significant effect of environmental pH on baseline CORT or handling-induced CORT levels. In a laboratory study, low pH decreased salamander locomotory activity compared to acid-neutral controls, but there was no effect of pH on plasma CORT. Decreased locomotory activity is a common amphibian response to stress, indicating that low pH has adverse effects on Allegheny Mountain dusky salamanders. Overall, we conclude that the effects of environmental pH on salamander behavior and other potential responses are not mediated by changes in plasma CORT levels. We discuss alternative explanations for our results and describe difficulties involved in searching for relationships between plasma GCs and environmental degradation.

Changes in faunal and vegetation communities along a soil calcium gradient in northern hardwood forests

Depletion of Ca from forest soils due to acidic deposition has had potentially pervasive effects on forest communities, but these impacts remain largely unknown. Because snails, salamanders, and plants play essential roles in the Ca cycle of northern hardwood forests, we hypothesized that their community diversity, abundance, and structure would vary with differences in biotic Ca availability. To test this hypothesis, we sampled 12 upland hardwood forests representing a soil Ca gradient in the Adirondack Mountains, New York (USA), where chronic deposition has resulted in acidified soils but where areas of well-buffered soils remain Ca rich due to parent materials. Along the gradient of increasing soil [Ca2+], we observed increasing trends in snail community richness and abundance, live biomass of redback salamanders ( Plethodon cinereus (Green, 1818)), and canopy tree basal area. Salamander communities were dominated by mountain dusky salamanders ( Desmognathus ochrophaeus Cope, 1859) at Ca-poor sites and changed continuously along the Ca gradient to become dominated by redback salamanders at the Ca-rich sites. Several known calciphilic species of snails and plants were found only at the highest-Ca sites. Our results indicated that Ca availability, which is shaped by geology and acidic deposition inputs, influences northern hardwood forest ecosystems at multiple trophic levels, although the underlying mechanisms require further study.

An Assessment of Distribution Patterns of Terrestrial Salamanders in the Central Appalachians Using Two Landscape Models

Abstract Conservation of terrestrial salamanders requires consideration for landscape-level features and processes; and, therefore, tools are needed to support planning at large spatial scales. We assessed terrestrial salamander distribution patterns using an existing salamander data set and two previously developed landscape-level spatial tools, a landscape classification (LC) model and flow accumulation (FLOW) model, both based on 100-m2 pixels. The LC model assigned each pixel to one of eight categories defined by slope, aspect and landform. The FLOW model assigned each pixel to one of five categories based on the number of 100-m2 cells that drained into a given cell. Salamander data had been collected in the Allegheny Mountains of West Virginia during 2001–2003 on 543 100-m2 plots located along random transects. Using area-constrained searches we collected 1879 individuals of eleven species. Plethodon cinereus, Desmognathus ochrophaeus, D. monticola and D. fuscus represented 92.8% of all captures. All...