Thamnophis Sirtalis Research Articles

The Age of Snakes: a comparison of three Canadian species at their northern peripheries

ABSTRACT Aging animals enables an understanding of life-history strategies, population dynamics, and conservation concerns. Such information is particularly wanting for reptiles at their range peripheries where life-history trade-offs make body size an unreliable surrogate for age. Using skeletochronology we quantified the ages of three species sampled near their northern range limits (Common gartersnake – Thamnophis sirtalis, Western Rattlesnake – Crotalus oreganus, Great Basin Gophersnake – Pituophis catenifer). Considerable range in length was seen for snakes of the same age, and the relationship between age and size was stronger in males. Male gartersnakes reached a size associated with sexual maturity in about half the time of females, but for the other two species there was no significant difference between sexes. The oldest snakes sampled were 10–12 years, with one outlier gartersnake estimated at 15 years. Given that females in northern populations cannot reproduce annually, our estimated ages of these snakes suggest reproductive bouts occur only a small number of times during their lifespan, reflecting the precarious existence of northern snake populations. Age data improves our understanding of how life-history varies with locations (including latitude), and further studies that collect contemporary and widespread data on this basic population parameter are required.

Occurrence and molecular characterization of Giardia spp. in snakes from China

Giardia intestinalis is a major diarrhea-causing parasite that colonizes the proximal small intestine of humans and various other mammalian species, including pets and livestock. Despite its global occurrence, there is limited information about the epidemiology of Giardia in reptiles, particularly snakes. The aim of this study was to amplify the beta-giardin (bg) gene of Giardia by polymerase chain reaction (PCR), followed by a comparative evolutionary tree analysis to determine the occurrence and molecular characterization of Giardia in snakes. We collected 603 asymptomatic samples from 26 provinces in China, representing species such as Pantherophis guttatus, Pantherophis obsoletus, Pituophis melanoleucus, Thamnophis sirtalis, Lampropeltis getula, and Heterodon nasicus. Ultimately, a occurrence of Giardia infection of 4.15 % was detected in these snakes, with corn snakes (Pantherophis guttatus) having a significantly higher occurrence than other species. Molecular analysis identified assemblage F as the predominant genotype, and also identified an assemblage B that can infect humans and a variety of mammals. The results of this study illustrate the potential risk of transmission of Giardia from snakes to humans, especially in environments where close contact occurs. The present epidemiological study examines epidemiological investigations of Giardia in reptiles, provides data to understand the zoonotic risk of Giardia, and emphasizes the need for targeted surveillance, stringent hygiene measures, and public awareness campaigns to reduce these risks.

Increased offspring size and reduced gestation length in an ectothermic vertebrate under a worst-case climate change scenario

As global temperatures continue to rise, understanding the impacts of warming environments has become increasingly important. Temperature is especially relevant for ectothermic organisms which depend upon consistent and predictable annual temperature cycles for reproduction and development. However, additional research is required in this area to elucidate the potential impacts of climate change on future generations. To understand how projected increases in environmental temperatures may impact reproductive outcomes within natural populations of ectothermic vertebrates, we manipulated minimum ambient temperatures during gestation in Red-sided garter snakes (Thamnophis sirtalis parietalis). Wild snakes were collected in the Interlake region of Manitoba, Canada during their spring mating season and allowed to mate in controlled conditions. For the duration of gestation, mated females were placed into one of two ambient thermal conditions: temperatures emulating those found in the species’ natural habitat or temperatures with a consistent 5 °C increase to match end-of-century climate change projections. We recorded observations for each litter and all neonates resulting from controlled mating trials. We observed no difference in litter sizes or birth rates between thermal conditions. However, we observed a significant reduction in gestation length and significant increase to neonate body mass and body condition associated with increased ambient temperatures. These results suggest that increased minimum temperatures during gestation may confer reproductive benefits for the northern populations of this species even under the most extreme current modeled warming predictions. We discuss the broader implications of this effect, including possible negative ecological outcomes.

Princes and Paupers: Effects of Annual and Early-Life Resource Variability on Demography in Populations of Common Garter Snakes, Thamnophis sirtalis

Princes and Paupers: Effects of Annual and Early-Life Resource Variability on Demography in Populations of Common Garter Snakes, Thamnophis sirtalis

Earthworms as a prey source for the insular snake Thamnophis sirtalis (Linnaeus, 1758)

Insular organisms typically evolve in closed, isolated systems; thus, changes caused by introduced species can drastically affect their ecology. Isle Royale National Park (Michigan, USA) is a remote island in Lake Superior that, until relatively recently, was void of earthworms. Its two native natricine snakes are known to feed on earthworms on the mainland; here, we confirm that eastern gartersnakes (Thamnophis sirtalis) are also feeding on earthworms on Isle Royale. It is unknown how this new important dietary source will impact the snake ecology on Isle Royale.

A novel cathelicidin TS-CATH derived from Thamnophis sirtalis combats drug-resistant gram-negative bacteria in vitro and in vivo

Antimicrobial peptides are promising therapeutic agents for treating drug-resistant bacterial disease due to their broad-spectrum antimicrobial activity and decreased susceptibility to evolutionary resistance. In this study, three novel cathelicidin antimicrobial peptides were identified from Thamnophis sirtalis, Balaenoptera musculus, and Lipotes vexillifer by protein database mining and sequence alignment and were subsequently named TS-CATH, BM-CATH, and LV-CATH, respectively. All three peptides exhibited satisfactory antibacterial activity and broad antibacterial spectra against clinically isolated E. coli, P. aeruginosa, K. pneumoniae, and A. baumannii in vitro. Among them, TS-CATH displayed the best antimicrobial/bactericidal activity, with a rapid elimination efficiency against the tested drug-resistant gram-negative bacteria within 20 min, and exhibited the lowest cytotoxicity toward mammalian cells. Furthermore, TS-CATH effectively enhanced the survival rate of mice with ceftazidime-resistant E. coli bacteremia and promoted wound healing in meropenem-resistant P. aeruginosa infection. These results were achieved through the eradication of bacterial growth in target organs and wounds, further inhibiting the systemic dissemination of bacteria and the inflammatory response. TS-CATH exhibited direct antimicrobial activity by damaging the inner and outer membranes, resulting in leakage of the bacterial contents at super-MICs. Moreover, TS-CATH disrupted the bacterial respiratory chain, which inhibited ATP synthesis and induced ROS formation, significantly contributing to its antibacterial efficacy at sub-MICs. Overall, TS-CATH has potential for use as an antibacterial agent.

The relative contributions of habitat area, configuration, and vegetative diversity on snake and lizard presence in agricultural landscapes

AbstractNearly one in five reptile species is at risk of extinction. Changes in habitat area, its configuration, and vegetation diversity could affect habitat use, but their relative importance is understudied. We assessed how these factors affected reptile presence in agricultural landscapes figure in Iowa, United States, using 695 cover boards visited 16,441 times in 2015–2020. Species‐wise encounter rates ranged 0.0001–0.012. Eight of 11 species and 54.2% of individuals were species of greatest conservation need. Habitat area, configuration, and vegetation diversity influenced reptile presence similarly. Mean patch occupancy was 0.18 for common garter snake (CG, Thamnophis sirtalis) and 0.45 for all snakes (AS). Naïve presence was explained by effort (odds ratio [OR]AS = 1.83, ORCG = 1.79), vegetation diversity (ORAS = 1.28, ORCG = 1.28), woody cover (ORAS = 1.24, ORCG = 1.41), and patch size (ORAS = 1.30). Large patch prairies were more likely to contain snakes than other conservation practices (encounter = 0.291), and more likely to contain CG (0.098) than prairie contour strips (0.031), waterways (0.018), grass contour strips (0.016), or terraces (0.015). While we documented low overall reptile presence, their higher presence in large prairie patches underscores the importance of core nature reserves for reptile conservation.

Bolder together: conformity drives behavioral plasticity in eastern gartersnakes

Personality traits drive individual differences in behaviour that are consistent across time and context. Personality limits behavioural plasticity, which could lead to maladaptive choices if animals cannot adapt their behavior to changing conditions. Here, we assessed consistency and flexibility in one personality trait, boldness, across non-social and social contexts in eastern gartersnakes (Thamnophis sirtalis sirtalis). Snakes explored a novel open arena either alone or in a pair. Pairs were assigned based on the data from the solo trials, such that each snake was paired once with a bolder and once with a less bold partner. We predicted that snakes would conform when in a social context, displaying plasticity in their personality, and causing boldness scores to converge. We found that snakes were consistent within contexts (solo or paired), but changed their behavior across contexts (from solo to paired). Plasticity in boldness resulted from an interaction between conformity and repeatable individual differences in plasticity. In line with some data on other species, snakes conformed more when they were the less bold partner. Personality reflects a consistent bias in decision-making, but our results highlight that the cognitive processes that drive the expression of personality traits in behavior are flexible and sensitive to social context. We show that both consistency and plasticity combine to shape snake social behavior in ways that are responsive to competition. This pattern of behavior may be particularly beneficial for species in which group-living is seasonal.

Cryptosporidium spp. in captive snakes from 26 provinces in China: Prevalence, molecular characterization, and symptoms.

Snakes are sometimes regarded as pets and are used in traditional Chinese medicine. Cryptosporidium spp. are frequently identified in snakes, representing an important pathogen and causing gastrointestinal diseases. Current data indicate that risk factors for infection and patterns of clinical symptom presentation may differ among Cryptosporidium spp. To better understand the infection status by Cryptosporidium spp., fecal samples were collected from 603 asymptomatic and 147 symptomatic snakes in 26 provinces of China. These samples came from Elaphe guttata, Elaphe obsoleta, Pituophis melanoleucus, Thamnophis sirtalis, Lampropeltis getulus, and Heterodon nasicus. The partial small subunit (SSU) rRNA gene was amplified using nested polymerase chain reaction (PCR) to investigate the infection rate of Cryptosporidium spp., and to assess evolutionary relationships and genetic characterization. A prevalence of 20% was recorded in asymptomatic snakes, with age identified as a significant risk factor. In contrast, 70% of symptomatic snakes were positive for Cryptosporidium spp., with Cryptosporidium serpentis and Cryptosporidium varanii (syn. C. saurophilum). Further analysis revealed a potential association between C. serpentis and regurgitation, and C. varanii and diarrhea, while neither species was linked to flatulence. To our knowledge, this is the first study to report Cryptosporidium spp. and associated clinical signs in symptomatic snakes in China. This study aims to enhance the understanding of Cryptosporidium infections, risk factors, and clinical manifestations in snakes, providing data crucial for the control and prevention of cryptosporidiosis.

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.

Comparing reintroduction strategies for the endangered San Francisco gartersnake (Thamnophis sirtalis tetrataenia) using demographic models.

For endangered species persisting in a few populations, reintroductions to unoccupied habitat are a popular conservation action to increase viability in the long term. Identifying the reintroduction strategy that is most likely to result in viable founder and donor populations is essential to optimally use resources available for conservation. The San Francisco gartersnake (Thamnophis sirtalis tetrataenia) is an endangered sub-species that persists in a small number of populations in a highly urbanized region of California. Most of the extant populations of San Francisco gartersnakes have low adult abundance and effective population size, heightening the need for establishment of more populations for insurance against the risk of extinction. We used simulations from demographic models to project the probability of quasi-extinction for reintroduced populations of San Francisco gartersnakes based on the release of neonate, juvenile, adult, or mixed-age propagules. Our simulation results indicated that the release of head-started juveniles resulted in the greatest viability of reintroduced populations, and that releases would need to continue for at least 15 years to ensure a low probability of quasi-extinction. Releasing captive-bred juvenile snakes would also have less effect on the viability of the donor population, compared to strategies that require more adult snakes to be removed from the donor population for translocation. Our models focus on snake demography, but the genetic makeup of donor, captive, and reintroduced populations will also be a major concern for any proposed reintroduction plan. This study demonstrates how modeling can be used to inform reintroduction strategies for highly imperiled species.

Biogeographic barriers and historic climate shape the phylogeography and demography of the common gartersnake

AbstractAimCurrent distributions of widespread North American (NA) species have been shaped by Pleistocene glacial cycles, latitudinal temperature gradients, sharp longitudinal habitat transitions and the vicariant effects of major mountain and river systems that subdivide the continent. Within these transcontinental species, genetic diversity patterns might not conform to established biogeographic breaks compared to more spatially restricted taxa due to intrinsic differences or spatiotemporal differences. In this study, we highlight the effects of these extrinsic variables on genetic structuring by investigating the phylogeographic history of a widespread generalist squamate found throughout NA.LocationNorth America.TaxonCommon gartersnake, Thamnophis sirtalis.MethodsWe evaluate the effects of major river basins and the forest‐grassland transition into the Interior Plains on genetic structure patterns using phylogenetic, spatially informed population structure and demographic analyses of single nucleotide polymorphism data and address range expansion history with ecological niche modelling using locality and historic climate data.ResultsWe identify four phylogeographic lineages with varying degrees of connectivity between them. We find discordant population structure patterns between sex‐linked and autosomal loci with respect to the relationship between the central NA lineage relative to coastal lineages. We find support for southeast Pleistocene refugia where recent secondary contact occurred during the Last Glacial Maximum and evidence for both northern and southern refugia in western NA.Main ConclusionOur results provide strong evidence for a Pliocene origin for T. sirtalis in central‐southeastern NA preceding its rapid expansion across the continent prior to middle Pleistocene climate‐mediated lineage formation. We implicate major riverine networks within the Mississippi watershed in likely repeated westward expansion events across the Interior Plains. Finally, we corroborate prior conclusions that phenotypic differences between subspecies do not reflect shared evolutionary history and note that the degree of separation between inferred lineages warrants further investigation before any taxonomic revisions are proposed.

The ripple effects of clines from coevolutionary hotspots to coldspots.

Coevolution has the potential to alter not only the ecological interactions of coevolving partners, but also their interactions with yet other species. The effects of coevolution may ripple throughout networks of interacting species, cascading across trophic levels, swamping competitors, or facilitating survival or reproduction of yet other species linked only indirectly to the coevolving partners. These ripple effects of coevolution may differ among communities, amplifying how the coevolutionary process produces geographic mosaics of traits and outcomes in interactions among species. In a From the Cover article in this issue of Molecular Ecology, Hague et al.(2022) provide a clear example, using the well-studied interactions between Pacific newts (Taricha spp.) and their common garter snake (Thamnophis sirtalis) predators in western North America. Pacific newts harbour tetrodotoxin (TTX), which is highly toxic to vertebrate predators. In coevolutionary hotspots, extreme escalation of toxicity in the newts and resistance to toxicity in the snakes have resulted in snake populations that retain high levels of TTX. In two geographic regions, snakes in these hotspot populations have evolved bright, aposematic colours that may act as warning signals to their own vertebrate predators. The warning signals and toxin-resistance alleles in the snake populations decrease clinally away from the coevolutionary hotpots, shaped by a geographically variable mix of selection imposed by the snakes' prey and by their own predators.

Phenotypic outcomes of predator–prey coevolution are predicted by landscape variation in climate and community composition

Abstract Landscape patterns of phenotypic coevolution are determined by variation in the outcome of predator–prey interactions. These outcomes may depend not only on the functional phenotypes that mediate species interactions, but also on aspects of the environment that enable encounters between coevolutionary partners. Exploring the relationship between coevolutionary traits and the environment requires extensive sampling across the range of the interaction to determine the relationship between local ecological variation and coevolution. In this study, we synthesized >30 years of data on predator–prey interactions between toxic newts (Taricha granulosa) and their snake predators (Thamnophis sirtalis) to explore the environmental predictors of arms race escalation. We found that geographic variation in phenotypes at the interface of coevolution was best predicted by a combination of community and climatic variation. Coevolutionary phenotypes were greatest in environments with climate favourable for newt–snake overlap. We found prey toxicity was elevated in regions with more predator species, and predator resistance was higher in regions with more prey species. Our results suggest specific environmental conditions reinforce the process of coevolution, signifying the phenotypic outcomes of coevolutionary arms races are sensitive to local ecological contexts that vary across the landscape. Read the free Plain Language Summary for this article on the Journal blog.

Reconsidering reproductive patterns in a model dissociated species, the red-sided garter snake: Sex-specific and seasonal changes in gonadal steroidogenic gene expression.

Sex steroid hormones are powerful regulators of reproductive behavior and physiology in vertebrates, and steroidogenesis has distinct sex- and season-specific patterns ultimately dictated by the expression of key enzymes. Most comparative endocrinology studies, however, focus only on circulating levels of sex steroids to determine their temporal association with life-history events in what are termed associated reproductive patterns. The red-sided garter snake (Thamnophis sirtalis parietalis) is a notable exception; this species exhibits maximal sex behavior decoupled from maximal sex steroid production and gametogenesis in what is termed a dissociated reproductive pattern. And while this is true for male red-sided garter snakes and their production of testosterone, females have maximal estradiol production during peak breeding (spring) but only immediately after mating. Here, we demonstrate that expression of ovarian aromatase (conversion of androgens to estrogens) matches the established seasonal hormone pattern in females. Additionally, steroidogenic gene expression in the ovary is broadly reduced if not suppressed compared to the testis throughout the active year. Bizarrely, male red-sided garter snakes demonstrate an unexplained pattern of steroidogenic gene expression in the testis. StAR (import of cholesterol to steroidogenesis) is maximally expressed in spring, yet Hsd17b3 expression (conversion of androstenedione to testosterone) is highest in summer, with the latter matching the established summer peak in male testosterone. The function of elevated StAR in spring is unknown, but our results suggest a decoupling between maximal StAR expression and testosterone biosynthesis (Hsd17b3 expression). We also purport that the reproductive pattern binary should be reassessed given its lack of fit for many vertebrate species that demonstrate seasonal, mixed patterns of (a)synchrony between circulating sex hormones and reproductive behavior.

Eastern red-backed salamanders (Plethodon cinereus) avoid chemical cues from snake predators regardless of colour morph or snake individual and size

Abstract Plethodontid salamanders often possess the ability to detect and avoid hazards in the environment which may be important for survival. We examined whether the two main colour morphs of Plethodon cinereus differ in their avoidance of cues from a potential predator (Thamnophis sirtalis) and whether P. cinereus avoided cues from T. sirtalis of different sizes (i.e., small vs. large). Plethodon cinereus avoided chemical cues deposited by T. sirtalis on a substrate; but avoidance did not differ between colour morphs of P. cinereus, snake sizes, or individual donor snakes.

Do roads affect the abundance of garter (Thamnophis sirtalis) and redbelly snakes (Storeria occipitomaculata)?

The greatest driver of the current biodiversity crisis is habitat loss. Roads are a major contributor to habitat loss because they destroy and fragment habitat, in addition to causing direct mortality. Animals may respond to roads either by avoiding them, thus leading to population isolation, or by attempting to cross them, thus potentially leading to increased mortality and, if so, also to population isolation. We studied the impact of road density on abundance of two snake species: redbelly snakes ( Storeria occipitomaculata Storer, 1839) and garter snakes ( Thamnophis sirtalis Linnaeus, 1758) around Ottawa, Canada. We hypothesized that roads are detrimental to snake populations due to road avoidance and mortality. Therefore, we predicted that snakes should be less abundant at sites with higher road density in their surroundings. We deployed cover boards at 28 sites along a gradient of road density in 2020 and 2021. We visited sites weekly, counted the number of individuals of both species, and measured snout–vent length (SVL) of all individuals captured. We captured fewer garter snakes at sites surrounded by more roads and fewer redbelly snakes at sites surrounded by more urban habitat. Snakes at sites surrounded by more roads were not smaller. The effects of roads and urbanization on the number of snakes were modest, but indicate decreasing population sizes that could lead to loss of ecological function.

Material properties of skin in the flying snake Chrysopelea ornata.

In snakes, the skin serves for protection, camouflage, visual signaling, locomotion, and its ability to stretch facilitates large prey ingestion. The flying snakes of the genus Chrysopelea are capable of jumping and gliding through the air, requiring additional functional demands: its skin must accommodate stretch in multiple directions during gliding and, perhaps more importantly, during high-speed, direct-impact landing. Is the skin of flying snakes specialized for gliding? Here, we characterized the material properties of the skin of Chrysopelea ornata and compared them with two nongliding species of colubrid snakes, Thamnophis sirtalis and Pantherophis guttatus, as well as with previously published values. The skin was examined using uniaxial tensile testing to measure stresses, and digital image correlation methods to determine strains, yielding metrics of strength, elastic modulus, strain energy, and extensibility. To test for loading orientation effects, specimens were tested from three orientations relative to the snake's long axis: lateral, circumferential, and ventral. Specimens were taken from two regions of the body, pre- and pos-tpyloric, to test for regional effects related to the ingestion of large prey. In comparison with T. sirtalis and P. guttatus, C. ornata exhibited higher post-pyloric and lower pre-pyloric extensibility in circumferential specimens. However, overall there were few differences in skin material properties of C. ornata compared to other species, both within and across studies, suggesting that the skin of flying snakes is not specialized for gliding locomotion. Surprisingly, circumferential specimens demonstrated lower strength and extensibility in pre-pyloric skin, suggesting less regional specialization related to large prey.

SEASONAL AND INTERSPECIFIC VARIATION IN THE PREVALENCE OF OPHIDIOMYCES OPHIDIICOLA AND OPHIDIOMYCOSIS IN A COMMUNITY OF FREE-RANGING SNAKES.

Ophidiomycosis in snakes is caused by the fungus Ophidiomyces ophidiicola. Clinical signs associated with the disease range from minor skin lesions to severe swelling of the face. In some cases, the fungus invades the snake's underlying muscle and bone and internal organs; disease severity appears to peak during brumation. We quantified the prevalence of O. ophidiicola and ophidiomycosis in free-ranging snakes to explore seasonal variation in detection of the pathogen and disease. We collected skin swabs (n=464 samples) from seven species of free-ranging snakes (n=336) from Rondeau Provincial Park (Ontario, Canada) and tested the swabs for O. ophidiicola using quantitative PCR. We also assessed individuals for lesions consistent with ophidiomycosis and monitored changes in gross lesions over time in recaptured individuals. Eastern foxsnakes (Pantherophis vulpinus) had the highest prevalence of O. ophidiicola (24/84) and of lesions consistent with ophidiomycosis (34/84). On other species (Nerodia sipedon, Storeria dekayi, Thamnophis sirtalis, and Thamnophis sauritus), we detected the pathogen on only 4/229 snakes and observed gross lesions consistent with ophidiomycosis on 24/229 snakes. Body length of eastern foxsnakes was associated with detection of O. ophidiicola, suggesting that eastern foxsnakes' large size increases the risk of pathogen exposure relative to the other, smaller, species at our study site. Ophidiomyces ophidiicola and lesions consistent with ophidiomycosis were detected most frequently in eastern foxsnakes soon after emergence from brumation and less frequently later in the active season (O. ophidiicola: April=29.8%, October=3.9%; lesions: April=36.1%, October=5.5%). This decrease corresponded with resolution of lesions in 6/13 resampled eastern foxsnakes. Considering the seasonal cycle of O. ophidiicola and ophidiomycosis when planning disease surveillance research may improve detection probabilities for ophidiomycosis in Nearctic snakes.

Conspicuous coloration of toxin-resistant predators implicates additional trophic interactions in a predator-prey arms race.

Antagonistic coevolution between natural enemies can produce highly exaggerated traits, such as prey toxins and predator resistance. This reciprocal process of adaptation and counter-adaptation may also open doors to other evolutionary novelties not directly involved in the phenotypic interface of coevolution. We tested the hypothesis that predator-prey coevolution coincided with the evolution of conspicuous coloration on resistant predators that retain prey toxins. In western North America, common garter snakes (Thamnophis sirtalis) have evolved extreme resistance to tetrodotoxin (TTX) in the coevolutionary arms race with their deadly prey, Pacific newts (Taricha spp.). TTX-resistant snakes can retain large amounts of ingested TTX, which could serve as a deterrent against the snakes' own predators if TTX toxicity and resistance are coupled with a conspicuous warning signal. We evaluated whether arms race escalation covaries with bright red coloration in snake populations across the geographic mosaic of coevolution. Snake colour variation departs from the neutral expectations of population genetic structure and covaries with escalating clines of newt TTX and snake resistance at two coevolutionary hotspots. In the Pacific Northwest, bright red coloration fits an expected pattern of an aposematic warning to avian predators: TTX-resistant snakes that consume highly toxic newts also have relatively large, reddish-orange dorsal blotches. Snake coloration also seems to have evolved with the arms race in California, but overall patterns are less intuitively consistent with aposematism. These results suggest that interactions with additional trophic levels can generate novel traits as a cascading consequence of arms race coevolution across the geographic mosaic.