Sistrurus Catenatus Research Articles

Initial impacts of wildfire on overwintering conditions for a Species-at-Risk snake

Peatlands are important ecosystems that are becoming increasingly vulnerable to climate-mediated disturbances such as wildfire, which can threaten peatland hydrological, biogeochemical, and ecological function. However, the magnitude of these changes and their impacts to peatland-dependent species worldwide is a key knowledge gap. Peatlands in the eastern Georgian Bay, Ontario, region provide overwintering habitat for the eastern massasauga rattlesnake (Sistrurus c. catenatus), a species considered at-risk across its North American range. Overwintering habitat is considered suitable when peat temperature is above 0°C and the water table position provides moisture without risk of prolonged flooding. This combination of suitable ecohydrological conditions, also known as the life zone or resilience zone, commonly occurs in hummocks which are raised microforms on the peatland surface. Due to a changing climate, peatlands are at risk of increased wildfire frequency and burn severity which may reduce overwintering habitat availability and suitability through changes in peat thermal and hydrological properties. In 2018, a wildfire burned over 11,000 ha of the eastern Georgian Bay landscape which supports critical habitat for the massasauga at the northern limit of the species range. To assess the potential impact of wildfire on massasauga overwintering habitat, we monitored water table position, snow depth, and peat thermal dynamics in hummocks across a burn severity gradient (unburned to severely burned) in three burned and three unburned peatlands across three winters (2019–2022). We found that hummocks were able to provide unflooded habitat above 0°C regardless of peat burn severity; however, there was moderate evidence that hummock burn severity influenced mean daily resilience zone size. Overall, hummock overwintering suitability appears to be dominantly controlled by peatland surface topography and interannual winter weather. With the frequency and intensity of wildfires predicted to increase globally under a changing climate, it is critical to understand how interannual variability of winter weather conditions influences overwintering habitat suitability after wildfire, to identify peatland ecosystems that provide resilient species at risk habitat.

Occupancy and Detection of Eastern Massasauga Rattlesnakes (Sistrurus Catenatus): Implications for Evaluating Population Recovery Efforts

Occupancy and Detection of Eastern Massasauga Rattlesnakes (Sistrurus Catenatus): Implications for Evaluating Population Recovery Efforts

Selection and validation of release sites for conservation translocations of temperate-zone snakes

Poor habitat quality is one of the most important reasons for reintroduction failure of reptiles; therefore, release site suitability ought to be evaluated prior to conducting conservation translocations. In temperate zone snakes, translocations have failed due to high overwinter mortality, so practitioners have recommended that release sites be located near suitable hibernacula. The presence of a Life Zone (LZ), the underground space above the groundwater table and below the frost line, may indicate the presence of suitable hibernation habitat. Identification and validation of sites with LZ, however, is challenged by the dynamic nature of groundwater and frost levels, coupled with the secretive nature of hibernating snakes. In this study, our goals were to 1) identify potential reintroduction sites for the globally imperiled Eastern Massasauga (Sistrurus catenatus) in southern Canada based on the presence of a LZ, 2) to validate the suitability of those reintroduction sites by hibernating a surrogate species (Eastern Gartersnake, Thamnophis sirtalis) in constructed hibernacula, and, 3) to determine if two separate measures of LZ were associated with the survival of individual gartersnakes to the end of hibernation. Four 1-ha study grids, each consisting of multiple groundwater and frost monitoring stations, were surveyed from 2015 to 19. Release sites were identified in each grid where a LZ of ≥ 10 cm was observed consistently for at least 2 full winters, and gartersnakes were successfully hibernated therein and at 2 reference sites over 3 winters. Overall, survival of subadult/adult snakes was very high (100 %; n = 20), regardless of site, whereas juvenile survival was lower (78 %; n = 93). A Kaplan-Meier test indicated that juvenile survival differed significantly among sites, and ranged from 60 % to 100 %. GLMM analysis indicated that mass at ingress had a significant positive effect on snake survival during hibernation. The temporary loss of a LZ did not impact snake survival. Contrary to our expectations, snake survival in hibernacula was negatively associated with both minimum LZ size and LZ frequency (i.e., the % of sampling occasions when LZ size ≥ 10 cm). Hibernating snakes can tolerate periods with a very small or non-existent LZ, but a large LZ may become unsuitable to juveniles, possibly due to a lack of moisture. Our results will guide the selection of release sites for Massasauga reintroductions, and for the development of a rigorous release site selection process for temperate zone snakes.

A plethora of rodents: Rattlesnake predators generate unanticipated patterns of venom resistance in a grassland ecosystem

Predation has the potential to impart strong selective pressures on organisms within their environments, resulting in adaptive changes in prey that minimize risk of predation. Pressures from venomous snakes present an exceptional challenge to prey, as venom represents a unique chemical arsenal evolutionarily tailored to incapacitate prey. In response, venom resistance has been detected in various snake prey species, and to varying degrees. This study analyzes venom resistance in an eastern Colorado grassland habitat, where the Prairie Rattlesnake (Crotalus viridis) and Desert Massasauga Rattlesnake (Sistrurus tergeminus edwardsii) co-occur with a suite of grassland rodents. We test for venom resistance across rodent and snake pairings using two geographically distant field sites to determine the role of 1) predation pressure and trophic ecology, and 2) sympatric and allopatric patterns of venom resistance. Resistance was measured using serum-based metalloproteinase inhibition assays to determine potential inhibition of proteolytic activity, augmented by median lethal dose (LD50) assays on rodent species to assess toxicity of crude venoms. Resistance is present in several rodent species, with strong resistance present in populations of Eastern Woodrat (Neotoma floridana), Ord's Kangaroo Rat (Dipodomys ordii), and Northern Grasshopper Mouse (Onychomys leucogaster). Resistance is less developed in other species, including the House Mouse (Mus musculus) and Plains Pocket Mouse (Perognathus flavescens). An unexpected differential is present, where Lincoln County Kangaroo Rats are highly resistant to venom of co-occurring Prairie Rattlesnakes yet are sensitive to an allopatric population of Prairie Rattlesnakes in Weld County. Lincoln Co. Northern Grasshopper Mice also demonstrate extremely elevated resistance to Weld Co. Prairie Rattlesnake venoms, and they may possess resistance mechanisms for myotoxin a, an abundant component of Weld Co. C. v viridis venoms. This study illustrates the complexity of venom resistance in biological communities that can exist when incorporating multiple species interactions. Future studies aimed at characterizing resistance mechanisms at the molecular level will provide a more detailed physiological context for understanding mechanisms by which resistance to venoms occurs.

Functional genomic diversity is correlated with neutral genomic diversity in populations of an endangered rattlesnake

Theory predicts that genetic erosion in small, isolated populations of endangered species can be assessed using estimates of neutral genetic variation, yet this widely used approach has recently been questioned in the genomics era. Here, we leverage a chromosome-level genome assembly of an endangered rattlesnake (Sistrurus catenatus) combined with whole genome resequencing data (N = 110 individuals) to evaluate the relationship between levels of genome-wide neutral and functional diversity over historical and future timescales. As predicted, we found positive correlations between genome-wide estimates of neutral genetic diversity (π) and inferred levels of adaptive variation and an estimate of inbreeding mutation load, and a negative relationship between neutral diversity and an estimate of drift mutation load. However, these correlations were half as strong for projected future levels of neutral diversity based on contemporary effective population sizes. Broadly, our results confirm that estimates of neutral genetic diversity provide an accurate measure of genetic erosion in populations of a threatened vertebrate. They also provide nuance to the neutral-functional diversity controversy by suggesting that while these correlations exist, anthropogenetic impacts may have weakened these associations in the recent past and into the future.

Modeling occupancy and detection probabilities to update the status of threatened eastern massasauga rattlesnake populations

Modeling occupancy and detection probabilities to update the status of threatened eastern massasauga rattlesnake populations

Diet of a threatened rattlesnake (eastern massasauga) revealed by DNA metabarcoding.

Characterizing the diet of imperiled species using minimally invasive methods is crucial to understanding their ecology and conservation requirements. Here, we apply a DNA metabarcoding approach to study the diet of the eastern massasauga rattlesnake (Sistrurus catenatus), a Federally Threatened snake found throughout the Great Lakes region. Eighty-three fecal samples collected across 10 different massasauga populations located in Michigan, USA, were sequenced, with 70 samples containing prey DNA. We used universal metazoan primers and developed a host-specific oligonucleotide blocker to characterize their diet. We identified at least 12 different prey species, with eastern massasaugas exhibiting opportunistic feeding and a strong preference towards small mammals. Meadow voles (Microtus pennsylvanicus) were the most common prey item (70% of diet) followed by the northern short-tailed shrew (Blarina brevicauda) and masked shrew (Sorex cinereus; 15.7% of diet each), along with occasional bird and snake prey. Adult individuals exhibited a more generalized diet, consuming a larger number of prey taxa on average. Younger snakes consumed a smaller variety of prey items and tended to consume smaller-sized mammals such as masked shrews (Sorex cinereus) and northern short-tailed shrews (Blarina brevicauda). We conclude that small mammals are a crucial part of eastern massasauga rattlesnake diet and recommend this be taken into consideration when conservation strategies are developed. The methods developed in this study can be applied to other reptile species, providing an accurate, minimally invasive, and thorough diet assessment for at-risk reptile species.

Inferring population connectivity in eastern massasauga rattlesnakes (Sistrurus catenatus) using landscape genetics.

Assessing the environmental factors that influence the ability of a threatened species to move through a landscape can be used to identify conservation actions that connect isolated populations. However, direct observations of species' movement are often limited, making the development of alternate approaches necessary. Here we use landscape genetic analyses to assess the impact of landscape features on the movement of individuals between local populations of a threatened snake, the eastern massasauga rattlesnake (Sistrurus catenatus). We linked connectivity data with habitat information from two landscapes of similar size: a large region of unfragmented habitat and a previously studied fragmented landscape consisting of isolated patches of habitat. We used this analysis to identify features of the landscape where modification or acquisition would enhance population connectivity in the fragmented region. We found evidence that current connectivity was impacted by both contemporary land-cover features, especially roads, and inherent landscape features such as elevation. Next, we derived estimates of expected movement ability using a recently developed pedigree-based approach and least-cost paths through the unfragmented landscape. We then used our pedigree and resistance map to estimate resistance polygons of the potential extent for S. catenatus movement in the fragmented landscape. These polygons identify possible sites for future corridors connecting currently isolated populations in this landscape by linking the impact of future habitat modification or land acquisition to dispersal ability in this species. Overall, our study shows how modeling landscape resistance across differently fragmented landscapes can identify habitat features that affect contemporary movement in threatened species in fragmented landscapes and how this information can be used to guide mitigation actions whose goal is to connect isolated populations.

Community Structure of Terrestrial Vertebrates in a Restored Meadow Habitat in Pennsylvania: Assessing the Potential Prey Base for Eastern Massasauga Rattlesnakes

We sampled the vertebrate community of a 20-ha conservation easement in Venango County, PA, from April through August 2021. The restoration and management efforts on this easement were designed to improve the meadow habitat for one of the few remaining populations of Sistrurus catenatus (Eastern Massasauga Rattlesnake) in Pennsylvania, and our sampling efforts were focused on evaluating the potential prey base available to the rattlesnakes at this site. We identified 9 amphibian species, 10 reptile species, and 11 small mammal species. For amphibians, Anaxyrus americanus (American Toad) was the most abundant species, accounting for 32.6% of the total, followed by Notophthalmus viridescens (Red-Spotted Newt) with 30.6% of total abundance. Thamnophis brachystoma (Short-Headed Gartersnake) comprised more than half (53.7%) of all reptiles sampled, followed by Thamnophis sirtalis (Eastern Gartersnake; 22.7%). Peromyscus leucopus (White-Footed Mouse) was the most abundant mammal species, accounting for 37.4% of all mammals surveyed, followed by Microtus pennsylvanicus (Eastern Meadow Vole; 24.8%). Mammals had the greatest species richness (S = 11), while reptiles had the greatest total abundance (TA = 361), and amphibians had the greatest species evenness (E = 0.799). Species accounts for Venango County indicate that we sampled 40.9% (9/22), 37.0% (10/27), and 21.2% (11/52) of the known amphibian, reptilian, and mammalian species, respectively, within this restored meadow habitat. Our results suggest that the restoration efforts on this easement study site have been extremely effective in creating meadow habitat that supports a diversity of prey species for the Eastern Massasauga Rattlesnake.

Correction: Identifcation of the Eastern Massasauga Rattlesnake (Sistrurus catenatus) through genetic analysis of shed skin

Correction: Identifcation of the Eastern Massasauga Rattlesnake (Sistrurus catenatus) through genetic analysis of shed skin

Identification of the Eastern Massasauga Rattlesnake (Sistrurus catenatus) through genetic analysis of shed skin

Identification of the Eastern Massasauga Rattlesnake (Sistrurus catenatus) through genetic analysis of shed skin

Limited gene flow and pronounced population genetic structure of Eastern Massasauga (Sistrurus catenatus) in a Midwestern prairie remnant.

As anthropogenic changes continue to ecologically stress wildlife, obtaining measures of gene flow and genetic diversity are crucial for evaluating population trends and considering management and conservation strategies for small, imperiled populations. In our study, we conducted a molecular assessment to expand on previous work to elucidate patterns of diversity and connectivity in the remaining disjunct Eastern Massasauga Rattlesnake (Sistrurus catenatus) hibernacula in Illinois. We assayed genetic data for 327 samples collected during 1999-2015 from the Carlyle Lake study area across 21 microsatellite loci. We found hibernacula formed distinct genetic clusters corresponding to the three main study areas (Dam Recreation Areas, Eldon Hazlet State Park, and South Shore State Park). Genetic structuring and low estimates of dispersal indicated that connectivity among these study areas is limited and each is demographically independent. Hibernacula exhibited moderate levels of heterozygosity (0.60-0.73), but estimates of effective population size (5.2-41.0) were low and track census sizes generated via long-term mark-recapture data. Hibernacula at Carlyle Lake, which represent the only Eastern Massasauga remaining in Illinois, are vulnerable to future loss of genetic diversity through lack of gene flow as well as demographic and environmental stochastic processes. Our work highlights the need to include population-level genetic data in recovery planning and suggests that recovery efforts should focus on managing the three major study areas as separate conservation units in order to preserve and maintain long-term adaptive potential of these populations. Specific management goals should include improving connectivity among hibernacula, maintaining existing wet grassland habitat, and minimizing anthropogenic sources of mortality caused by habitat management (e.g., mowing, prescribed fire) and recreational activities. Our molecular study provides additional details about demographic parameters and connectivity at Carlyle Lake that can be used to guide recovery of Eastern Massasauga in Illinois and throughout its range.

In cold blood: Observational descriptive review of Eastern Massasauga rattlesnake bites reported to a single poison center over time

The Eastern Massasauga rattlesnake (Sistrurus catenatus catenatus) is a pit viper indigenous to the Great Lakes region and the only venomous snake native to Michigan. It is small-to-medium, thick-bodied with dark brown, bow-tie shaped blotches. Its behavior is described as reclusive and docile and it prefers damp habitats. The venom of the Eastern Massasauga is primarily cytotoxic and hemotoxic. Previous literature describes severe coagulopathies following Eastern Massasauga envenomings, with some resulting in death. The objective of this study was to characterize Eastern Massasauga envenomings in humans reported to the Michigan Poison & Drug Information Center from 2003 to 2020, including a description of clinical manifestations, incidence and characterization of coagulopathies, and medical outcome severities. This was a retrospective review of Eastern Massasauga snakebites reported to our state poison center over time. Coagulopathies were classified according to previous toxicological snakebite literature. The degree of envenoming was scored using an institutional guideline, representing a modified version of validated snakebite severity score system. Our longitudinal review demonstrated Eastern Massasauga bites led to clinically significant toxicity, including persistent, recurrent, and late coagulopathies, though with low incidence of bleeding events. Cases typically resolved with use of antivenom. This, to our knowledge, is the largest descriptive case series characterizing Eastern Massasauga snakebites.

Genomic signatures of inbreeding and mutation load in a threatened rattlesnake.

Theory predicts that threatened species living in small populations will experience high levels of inbreeding that will increase their genetic load, but recent work suggests that the impact of load may be minimized by purging resulting from long-term population bottlenecks. Empirical studies that examine this idea using genome-wide estimates of inbreeding and genetic load in threatened species are limited. Here we use individual genome resequencing data to compare levels of inbreeding, levels of genetic load (estimated as mutation load) and population history in threatened Eastern massasauga rattlesnakes (Sistrurus catenatus), which exist in small isolated populations, and closely related yet outbred Western massasauga rattlesnakes (Sistrurus tergeminus). In terms of inbreeding, S.catenatus genomes had a greater number of runs of homozygosity of varying sizes, indicating sustained inbreeding through repeated bottlenecks when compared to S.tergeminus. At the species level, outbred S.tergeminus had higher genome-wide levels of mutation load in the form of greater numbers of derived deleterious mutations compared to S.catenatus, presumably due to long-term purging of deleterious mutations in S.catenatus. In contrast, mutations that escaped species-level drift effects within S.catenatus populations were in general more frequent and more often found in homozygous genotypes than in S.tergeminus, suggesting a reduced efficiency of purifying selection in smaller S.catenatus populations for most mutations. Our results support an emerging idea that the historical demography of a threatened species has a significant impact on the type of genetic load present, which impacts implementation of conservation actions such as genetic rescue.

Epidemiology of Eastern Massasauga (Sistrurus catenatus) Snakebites in the Great Lakes States, 1800-2015.

The eastern massasauga rattlesnake (Sistrurus catenatus) has the northernmost distribution of any venomous snake in the United States and presents a potential, but little-studied, risk to humans. We quantified the reported incidence of massasauga bites to humans in the Great Lakes states by searching historic and contemporary media reports between 1800 and 2015. We identified 848 reported massasauga bites across the region, most of which were concentrated along the border of Indiana and Michigan. The number of bites per decade increased into the late 1800s as human population increased; it then declined sharply in the 1900s, likely owing to declining massasauga population and habitat. The majority of bites were to adult males. There was no relationship between victim sex and age or activity when bitten. Most bites resulted from snakes the victims were unaware of, usually when individuals accidentally put their hands or feet near an unseen snake. Many bites, however, resulted from people intentionally interacting with a massasauga, such as attempting to capture or kill it. Fatality rates were lower among men than among women and were lower in adolescents than older or younger victims. No fatalities from massasaugas were reported after 1935. Fatality rates from massasauga bites reported herein should be interpreted with caution owing to the nature of the data we collected. In the modern era, massasauga bites are generally considered neither common nor life-threatening, although our findings suggest that historically they may have been both more frequent and potentially more dangerous.

Intact landscape promotes gene flow and low genetic structuring in the threatened Eastern Massasauga Rattlesnake.

Genetic structuring of wild populations is dependent on environmental, ecological, and life‐history factors. The specific role environmental context plays in genetic structuring is important to conservation practitioners working with rare species across areas with varying degrees of fragmentation. We investigated fine‐scale genetic patterns of the federally threatened Eastern Massasauga Rattlesnake (Sistrurus catenatus) on a relatively undisturbed island in northern Michigan, USA. This species often persists in habitat islands throughout much of its distribution due to extensive habitat loss and distance‐limited dispersal. We found that the entire island population exhibited weak genetic structuring with spatially segregated variation in effective migration and genetic diversity. The low level of genetic structuring contrasts with previous studies in the southern part of the species’ range at comparable fine scales (~7 km), in which much higher levels of structuring were documented. The island population's genetic structuring more closely resembles that of populations from Ontario, Canada, that occupy similarly intact habitats. Intrapopulation variation in effective migration and genetic diversity likely corresponds to the presence of large inland lakes acting as barriers and more human activity in the southern portion of the island. The observed genetic structuring in this intact landscape suggests that the Eastern Massasauga is capable of sufficient interpatch movements to reduce overall genetic structuring and colonize new habitats. Landscape mosaics with multiple habitat patches and localized barriers (e.g., large water bodies or roads) will promote gene flow and natural colonization for this declining species.

Low Bottleneck Detection in Long-Lived Species Despite Lost Genetic Diversity: A Case Study of Tuatara and Eastern Massasauga Rattlesnakes.

Population bottlenecks can reduce genetic diversity and may lead to inbreeding depression. However, some studies have provided evidence that long lifespans buffer negative genetic effects of bottlenecks. Others have cautioned that longevity might merely mask the effects of genetic drift, which will still affect long-term population viability. We used microsatellite data from actual populations of tuatara (Sphenodon punctatus) and eastern massasaugas (Sistrurus catenatus) as a starting point for simulated population declines to evaluate the performance of bottleneck tests under a range of scenarios. We quantified losses in genetic diversity for each scenario and assessed the power of commonly used tests (i.e., M-ratio, heterozygosity excess, and mode-shift) to detect known bottlenecks in these moderate- to long-lived species. Declines in genetic diversity were greater in bottlenecks simulated for eastern massasaugas, the shorter-lived species, and mode-shift and heterozygosity excess tests were more sensitive to population declines in this species. Conversely, M-ratio tests were more sensitive to bottlenecks simulated in tuatara. Despite dramatic simulated population declines, heterozygosity excess and mode-shift tests often failed to detect bottlenecks in both species, even when large losses in genetic diversity had occurred (both allelic diversity and heterozygosity). While not eliminating type II error, M-ratio tests generally performed best and were most reliable when a critical value (Mc) of 0.68 was used. However, in tuatara simulations, M-ratio tests had high rates of type I error when Mc was calculated assuming θ = 10. Our results suggest that reliance on these tests could lead to misguided species management decisions.

Pedigree-based assessment of recent population connectivity in a threatened rattlesnake.

Managing endangered species in fragmented landscapes requires estimating dispersal rates between populations over contemporary timescales. Here, we developed a new method for quantifying recent dispersal using genetic pedigree data for close and distant kin. Specifically, we describe an approach that infers missing shared ancestors between pairs of kin in habitat patches across a fragmented landscape. We then applied a stepping-stone model to assign unsampled individuals in the pedigree to probable locations based on minimizing the number of movements required to produce the observed locations in sampled kin pairs. Finally, we used all pairs of reconstructed parent-offspring sets to estimate dispersal rates between habitat patches under a Bayesian model. Our approach measures connectivity over the timescale represented by the small number of generations contained within the pedigree and so is appropriate for estimating the impacts of recent habitat changes due to human activity. We used our method to estimate recent movement between newly discovered populations of threatened Eastern Massasauga rattlesnakes (Sistrurus catenatus) using data from 2996 RAD-based genetic loci. Our pedigree analyses found no evidence for contemporary connectivity between five genetic groups, but, as validation of our approach, showed high dispersal rates between sample sites within a single genetic cluster. We conclude that these five genetic clusters of Eastern Massasauga rattlesnakes have small numbers of resident snakes and are demographically isolated conservation units. More broadly, our methodology can be widely applied to determine contemporary connectivity rates, independent of bias from shared genetic similarity due to ancestry that impacts other approaches.

Spatial Heterogeneity of Surface Topography in Peatlands: Assessing Overwintering Habitat Availability for the Eastern Massasauga Rattlesnake

The eastern massasauga rattlesnake (Sistrurus catenatus) relies on small-scale differences in peatland surface elevation to survive harsh overwintering conditions at the northern limit of its range. We characterized the spatial heterogeneity of surface topography in peatlands within the eastern Georgian Bay rock barrens landscape of Ontario, Canada, to assess available snake overwintering habitat. At six peatlands, we used a differential global positioning system to collect surface-elevation data. We created spatially-explicit surface models to map peatland surface topography, quantify habitat that was likely to remain unflooded during the overwintering period, and identify key characteristics associated with greater habitat availability. While surface elevations were spatially heterogeneous within and among sites, larger peatlands were associated with greater surface spatial variability relative to the lowest elevation measured within each site. However, even peatlands with very little spatial heterogeneity (average of 0.24 m above lowest elevation), provided unflooded overwintering habitat. Inter-annual weather conditions and peatland and watershed characteristics likely control the availability and distribution of unflooded overwintering habitat. We found that trees, specifically white pine (Pinus strobus) and maple (Acer spp.), were spatially associated with higher surface elevations and could be used to identify areas of unflooded winter habitat. Our findings are useful for landscape-scale assessments of available overwintering habitat to prioritize conservation and management efforts.

Identifying Potential Connectivity for an Urban Population of Rattlesnakes (Sistrurus catenatus) in a Canadian Park System

In the face of ongoing habitat loss and fragmentation, maintaining an adequate level of landscape connectivity is needed to both encourage dispersal between habitat patches and to reduce the extinction risk of fragmented wildlife populations. In a developing region of southwestern Ontario, Canada, a declining population of Eastern Massasauga rattlesnakes (Sistrurus catenatus) persists in fragmented remnants of tallgrass prairie in an urban park system. The goal of this study was to identify potential connectivity pathways between habitat patches for this species by using a GIS least-cost permeability swath model, and to evaluate the outputs with snake road mortality data. Results identified seven pathways between five core habitat blocks, a subset of which were validated with aerial imagery and mortality data. Four high-ranking pathways intersected roads through or near road mortality hotspots. This research will guide conservation interventions aimed at recovering endangered reptiles in a globally rare ecosystem, and will inform the use of permeability swaths for the identification of locations most suitable for connectivity interventions in dynamic, urbanizing landscapes.