Thamnophis Sirtalis Research Articles

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.

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.

Use of a Restored Prairie/Wetland Complex in Ohio byThamnophis sirtalis(Eastern Garter Snake) and Small Mammals during the First Five Years

Use of a Restored Prairie/Wetland Complex in Ohio by<i>Thamnophis sirtalis</i>(Eastern Garter Snake) and Small Mammals during the First Five Years

Effects of invasive jumping worms (Amynthas spp.) on microhabitat and trophic interactions of native herpetofauna

Invasive species can affect native communities through multiple mechanisms, including habitat modification and trophic interactions. In North America, invasive jumping worms (Amynthas spp.) may alter microhabitats used by native herpetofauna or serve as a novel prey resource for herpetofauna predators. We investigated effects of recently introduced Amynthas and previously established Lumbricus spp. earthworms on leaf litter microhabitat and trophic interactions of native herpetofauna using an outdoor mesocosm experiment, laboratory feeding trials, and field surveys of predator stomach contents in southern Wisconsin, USA. In mesocosms, Amynthas and Lumbricus reduced leaf litter biomass but did not have strong effects on soil conditions (pH, moisture, and temperature) or survival of American toad metamorphs (Anaxyrus americanus). In laboratory trials, American toads preyed on Amynthas but were less successful at capturing Amynthas than Lumbricus or crickets (Acheta domesticus). Attack success differences were likely due to unique defensive behaviors of Amynthas. Amynthas spent less time moving than Lumbricus, which was a behavior associated with delayed prey detection times by toads. In our diet surveys, we found novel evidence of Amynthas consumption by common garter snakes (Thamnophis sirtalis). We did not find Amynthas in stomach contents of American toads or red-bellied snakes (Storeria occipitomaculata), though additional surveys would help definitively determine whether these taxa are consuming Amynthas. Our findings highlight the importance of studying multiple mechanisms by which invasive species affect native communities and suggest that unique anti-predator behaviors may influence how Amynthas are incorporated into food webs as a novel prey resource.

Integrating growth and survival models for flexible estimation of size-dependent survival in a cryptic, endangered snake.

Estimates of demographic rates for animal populations and individuals have many applications for ecological and conservation research. In many animals, survival is size‐dependent, but estimating the form of the size–survival relationship presents challenges. For elusive species with low recapture rates, individuals’ size will be unknown at many points in time. Integrating growth and capture–mark–recapture models in a Bayesian framework empowers researchers to impute missing size data, with uncertainty, and include size as a covariate of survival, capture probability, and presence on‐site. If there is no theoretical expectation for the shape of the size–survival relationship, spline functions can allow for fitting flexible, data‐driven estimates. We use long‐term capture–mark–recapture data from the endangered San Francisco gartersnake (Thamnophis sirtalis tetrataenia) to fit an integrated growth–survival model. Growth models showed that females reach longer asymptotic lengths than males and that the magnitude of sexual size dimorphism differed among populations. The capture probability and availability of San Francisco gartersnakes for capture increased with snout–vent length. The survival rate of female snakes exhibits a nonlinear relationship with snout–vent length (SVL), with survival flat between 300 mm and 550 mm SVL before decreasing for females between 550 mm and 700 mm SVL. For male snakes, survival decreased for adult males >550 mm SVL. The survival rates of the smallest and largest San Francisco gartersnakes were highly uncertain because recapture rates were very low for these sizes. By integrating growth and survival models and using penalized splines, we found support for size‐dependent survival in San Francisco gartersnakes. Our results have applications for devising management activities for this endangered subspecies, and our methods could be applied broadly to the study of size‐dependent demography among animals.

Role of Melatonin in Temperature-Induced Activation of the Neuroendocrine Reproductive Axis in Garter Snakes

An animal’s ability to respond optimally to changing environmental conditions is paramount to successfully reproducing and thus maximizing fitness. Studies on photoperiod-induced changes in neural thyroid hormone metabolism have conclusively linked environmental cues to the neuroendocrine reproductive axis of birds and mammals. Whether this conserved mechanism also transduces changes in environmental temperature, however, has not been fully addressed. We investigated whether the hormone melatonin mediates the effects of low-temperature dormancy on thyroid hormone metabolism within the hypothalamus of red-sided garter snakes (Thamnophis sirtalis parietalis). To address this question, we used immunohistochemistry to assess changes in thyroid-stimulating hormone (TSH) in the infundibulum of the pituitary and deiodinase 3 (Dio3) and gonadotropin-releasing hormone (GnRH) in the hypothalamus. We also asked if changes in TSH, Dio3, and/or GnRH immunoreactivity are associated with changes in male courtship behavior. In contrast to our predictions, 6 weeks of dormancy at 4°C significantly decreased the number of TSH-labeled cells in the infundibulum. It is possible that the observed decrease in TSH is related to the release of snakes from temperature refractoriness, but this idea needs further testing. Treatment of snakes with the melatonin precursor 5-hydroxytryptophan during dormancy at 4°C both reversed the temperature-induced change in TSH immunoreactivity and disrupted the temporal pattern of male courtship behavior. These results suggest that TSH cells within the infundibulum are both modulated by temperature and sensitive to changes in melatonin. As predicted, male snakes hibernated at an elevated temperature of 12°C for 6 weeks and treated with vehicle showed no change in TSH-, Dio3-, or GnRH-immunoreactive cell number. Treatment of snakes with the melatonin receptor antagonist luzindole was not sufficient in rescuing the effects of dormancy at 12°C on TSH immunoreactivity or courtship behavior. However, luzindole-treated snakes showed a significant increase in GnRH-immunoreactive cell number, suggesting that melatonin exerts an inhibitory effect on GnRH in garter snakes. In summary, our results provide critical insights into the mechanisms that mediate the effects of temperature on reproductive physiology and behavior.

Temperature‐induced multi‐species cohort effects in sympatric snakes

In reptiles, reproductive maturity is often determined by size rather than age. Consequently, growth early in life may influence population dynamics through effects on generation time and survival to reproduction. Because reproductive phenology and pre‐ and post‐natal growth are temperature dependent, environmental conditions may induce multi‐species cohort effects on body size in sympatric reptiles. I present evidence of this using 10 years of neonatal size data for three sympatric viviparous snakes, Dekay's Brown snakes (Storeria dekayi), Red‐bellied Snakes (S. occipitomaculata), and Common Garter snakes (Thamnophis sirtalis). End‐of‐season neonatal size varied in parallel across species such that snout–vent length was 36%–61% greater and mass was 65%–223% greater in years when gestating females could achieve higher April–May (vs. June–July or August–September) operative temperatures. Thus, temperature had a larger impact during follicular enlargement and ovulation than during gestation or post‐natal growth. Multi‐species cohort effects like these may affect population dynamics and the magnitude of these effects may increase with climate change.

Dehydrated males are less likely to dive into the mating pool

AbstractThe hydration state of animals vying for reproductive success may have implications for the tempo and mode of sexual selection, which may be salient in populations that experience increasing environmental fluctuations in water availability. Using red-sided garter snakes as a model system, we tested the effect of water supplementation on courtship, mating behavior, and copulatory plug (CP) production during a drought year. Over 3 days of mating trials, water-supplemented males (WET males, n = 45) outperformed a control group that was not supplemented with water (DRY males, n = 45). Over 70% of WET males mated but just 33% of DRY males did so. As a group, WET males mated 79 times versus 28 times by DRY males. On the last day of mating trials, over 70% of WET males were still courting, with 19 of them mating, whereas less than 25% of DRY males were courting and only one mated. CP deposition accounted for 4–6% of the mass lost by mating males, but hydration did not affect CP mass or water content. These findings suggest that, in years of low water availability, the number of courting males and the intensity of their courtship declines, thereby affecting sexual selection and conflict, at least within that year.

Comparing the population structure of the specialist Butler’s Gartersnake (Thamnophis butleri) and the generalist Eastern Gartersnake (Thamnophis sirtalis sirtalis) in Ontario (Canada) and Michigan (USA)

Species differing in life-history attributes vary in their responses to features within a shared landscape. We evaluated genetic structure of sympatric gartersnake species in southwestern Ontario (Canada) and southeastern Michigan (USA), where habitat fragmentation is high due to agriculture and urbanization. We surveyed genetic structure of a habitat specialist, Butler’s Gartersnake (Thamnophis butleri (Cope, 1889)), and a habitat generalist, Eastern Gartersnake (Thamnophis sirtalis sirtalis (Linnaeus, 1758)), using DNA microsatellites. Bayesian clustering, discriminant analysis of principal components, and pairwise population comparisons revealed genetic differentiation among three major regional clusters of Butler’s Gartersnake with evidence of further division within one. Genetic clustering of Butler’s Gartersnake suggest that inhospitable habitat limits dispersal. Eastern Gartersnakes showed less structure, with assignment tests implying a single genetic cluster. We found positive significant Mantel’s r for both species in the smallest distance class (&lt;15 km), but significant isolation by distance for Butler’s Gartersnake only. These findings together imply that connectivity for Eastern Gartersnakes is less impacted by habitat loss and fragmentation or that we were less able to detect their effects. Our study shows the value of multispecies comparisons in studies seeking to understand the underlying causes of genetic structure in natural populations.

Unfinished business.

At the time of my retirement there were two topics that I considered unfinished business. The first is the Evolution of Sex Differences and the second, the she-male controversy in the Canadian red-sided snake (Thamnophis sirtalis parietalis). These questions are developed in this perspective.

Chemically mediated self-recognition in sibling juvenile common gartersnakes (Thamnophis sirtalis) reared on same or different diets: evidence for a chemical mirror?

Abstract Although self-recognition or self-awareness has been studied with the visually-based mirror test, passed by several species, primarily apes, the possibility of a chemically-based analogue is controversial. Prior studies suggested that chemical self-recognition may occur in some squamate reptiles. To evaluate this possibility, we studied 24 individually housed gartersnakes, Thamnophis sirtalis, raised from birth on either earthworm or fish diets and tested 12 male and 12 female snakes with cage liners that were either clean, their own, or from same-sex siblings fed their own or the opposite diet. Tongue flicking and activity were recorded in 30-minute video-recorded trials in a balanced design. After initial habituation to the stimuli, male, but not female, snakes discriminated between their own stimuli and those from littermates fed the same diet. Combined with other data and studies, the possibility that a chemical ‘mirror’ form of self-recognition exists in squamate reptiles is supported.

Decreased attractivity in female garter snakes treated with an aromatase inhibitor.

Most experimental studies on sexual signal regulation via hormone manipulation have focused on male signals, yet female signals demonstrate substantial phenotypic variation and hormone-dependent expression. Female red-sided garter snakes (Thamnophis sirtalis parietalis) produce a skin-based sex pheromone used by males in mate selection. The principle female sex steroid, 17 β-estradiol, controls pheromone production in snakes, but studies manipulating female garter snakes have produced conflicting results, relied on behavioral tests with males in the laboratory, and did not quantify pheromone expression. Because aromatase is the terminal enzyme in estradiol biosynthesis, we hypothesized that female garter snakes rely on aromatase to ultimately control pheromone production during the annual cycle of this species. To test this, we used a known pharmacological inhibitor of aromatase, fadrozole (FAD). Wild-caught female garter snakes were chronically treated via subcutaneous injections of either FAD (1.0 mg kg-1 ) or saline (control) for six months in the laboratory during the active period of the annual cycle then hibernated. In two separate field bioassays the next spring at the den site, FAD females received approximately 50% less courtship from wild, sexually active male garter snakes compared to SHAM females. Pheromone analysis revealed that four of the largest, unsaturated methyl ketones were specifically downregulated in FAD females, indicating that aromatase action is a crucial, permissive step in the maintenance of female attractivity.

Staying warm is not always the norm: behavioural differences in thermoregulation of two snake species

Thermal biology research compares field with laboratory data to elucidate the evolution of temperature-sensitive traits in ectotherms. The hidden challenge of many of these studies is discerning whether animals actively thermoregulate, since motivation is not typically assessed. By studying the behaviours involved in thermoregulation, we can better understand the mechanisms underlying body temperature control. Using an integrative approach, we assessed the thermoregulatory and thermotactic behaviours of two sympatric snake species with contrasting life histories: the generalist Eastern Garter Snake (Thamnophis sirtalis sirtalis ( Linnaeus, 1758 )) and the semi-fossorial Northern Red-bellied Snake (Storeria occipitomaculata occipitomaculata ( Storer, 1839 )). We expected that thermoregulatory behaviours would be optimized based on life history, in that T. s. sirtalis would show higher evidence for thermally oriented behaviours than S. o. occipitomaculata due to its active nature. Thamnophis sirtalis sirtalis actively thermoregulated, had higher thermal preferences (29.4 ± 2.5 vs. 25.3 ± 3.6 °C), and was more active than S. o. occipitomaculata, which showed relatively low evidence for thermotaxis. Our results build on the notion that evaluating movement patterns and rostral orientation towards a heat source can help ascertain whether animals make thermally motivated choices. Our data provide insight into the thermoregulatory strategies used by snakes with different life histories and maximize the information provided by behavioural thermoregulation experiments.

Uncovering the genomic and metagenomic research potential in old ethanol-preserved snakes.

Natural history museum collections worldwide represent a tremendous resource of information on past and present biodiversity. Fish, reptiles, amphibians and many invertebrate collections have often been preserved in ethanol for decades or centuries and our knowledge on the genomic and metagenomic research potential of such material is limited. Here, we use ancient DNA protocols, combined with shotgun sequencing to test the molecular preservation in liver, skin and bone tissue from five old (1842 to 1964) museum specimens of the common garter snake (Thamnophis sirtalis). When mapping reads to a T. sirtalis reference genome, we find that the DNA molecules are highly damaged with short average sequence lengths (38-64 bp) and high C-T deamination, ranging from 9% to 21% at the first position. Despite this, the samples displayed relatively high endogenous DNA content, ranging from 26% to 56%, revealing that genome-scale analyses are indeed possible from all specimens and tissues included here. Of the three tested types of tissue, bone shows marginally but significantly higher DNA quality in these metrics. Though at least one of the snakes had been exposed to formalin, neither the concentration nor the quality of the obtained DNA was affected. Lastly, we demonstrate that these specimens display a diverse and tissue-specific microbial genetic profile, thus offering authentic metagenomic data despite being submerged in ethanol for many years. Our results emphasize that historical museum collections continue to offer an invaluable source of information in the era of genomics.

Taxonomy, not locality, influences the cloacal microbiota of two nearctic colubrids: a preliminary analysis.

The gut microbiota is an emerging frontier in wildlife research and its importance to vertebrate health and physiology is becoming ever more apparent. Reptiles, in particular snakes, have not received the same attention given to other vertebrates and the composition of their wild gut microbiome remains understudied. The primary goal of this work was to describe the cloacal microbiota of two Colubrids, the Eastern Gartersnake (Thamnophis sirtalis sirtalis) and the Northern Watersnake (Nerodia sipedon sipedon), and if their cloacal microbiota differed as well as if it did between a wetland and upland population of the former species. We utilized next-generation sequencing of cloacal swabs-a non-destructive proxy for the gut microbiota. The cloacal microbiome of Eastern Gartersnakes (N = 9) was like those of other snakes being comprised of Proteobacteria, Bacteroidetes, and Firmicutes, while that of Northern Watersnakes (N = 6) was dominated by Tenericutes. Seven microbial operational taxonomic units (OTUs), all members of Proteobacteria, were shared among all individuals and were indicative of a core microbiome in Eastern Gartersnakes, but these OTUs were not particularly relevant to Northern Watersnakes. The latter had greater OTU richness than did Eastern Gartersnakes, and habitat did not have any apparent effect on the microbial community composition in Eastern Gartersnakes. Our findings suggest host taxonomy to be a determining factor in the cloacal microbiota of snakes and that Tenericutes are associated with aquatic habitats. This is the first report to examine the cloacal microbiome of these species and provides a useful foundation for future work to build upon.

Evidence of Intraguild Predation in a Snake Assemblage: Bigger Seems Better

Intraguild predation can play a significant role in structuring species composition with intraguild prey under both competitive and predatory pressure from intraguild predators. How intraguild prey are able to persist at sites with their predators is a complex issue with empirical evidence not always supporting theoretical models, indicating a need for more in situ research. A prominent grassland predator in the south central region of Pennsylvania, Coluber constrictor constrictor (Northern Black Racer), co-occurs with potential prey snakes including Lampropeltis triangulum (Eastern Milksnake) and Thamnophis sirtalis sirtalis (Eastern Gartersnake). Starting in 2008, this 7-year study examined how intraguild predation may differentially influence the body size and demographic traits of prey species in south-central Pennsylvania at sites with and without Northern Black Racers. The body size of adult Eastern Milksnakes was unaffected by the presence of racers. Eastern Milksnakes were more abundant, but with fewer juveniles, at racer-present sites. Adult female Eastern Gartersnakes were larger in body size, and adult males were less frequently encountered, in racer-present sites. These results indicate that intraguild predation in a snake community may affect the body size and demographic traits of prey species differentially, likely through predatory exclusion. However, the exact mechanisms behind our findings, and the resulting dynamics of snake diversity, are unclear and require further analyses.

Every breath you take: assessing metabolic costs of toxin resistance in garter snakes (Thamnophis).

Trait specialization often comes at the expense of original trait function, potentially causing evolutionary tradeoffs that may render specialist populations vulnerable to extinction. However, many specialized adaptations evolve repeatedly, suggesting selection favors specialization in specific environments. Some garter snake (Thamnophis) populations possess specialized mutations in voltage-gated sodium channels that allow them to consume Pacific newts (Taricha) defended by a highly potent neurotoxin (tetrodotoxin). These mutations, however, also decrease protein and muscle function, suggesting garter snakes may suffer evolutionary tradeoffs. We measured a key physiological process, standard metabolic rate (SMR), to investigate whether specialized adaptations in toxin-resistant garter snakes affect baseline energy expenditure. In snakes, skeletal muscles influence metabolism and power ventilation, so inefficiencies of sodium channels in these muscles might impact whole-animal energy expenditure. Further, because sodium channels are membrane-bound proteins, inefficiencies of channel kinetics and performance might be exacerbated at suboptimal temperatures. We measured SMR in 2 species, Thamnophis atratus and Thamnophis sirtalis, that independently evolved tetrodotoxin resistance through unique mutations, providing replicate experiments with distinct underlying genetics and potential physiological costs. Despite our expectations, neither resistance phenotype nor sodium channel genotype affected metabolism and resistant snakes did not perform worse under suboptimal body temperature. Instead, T. atratus and T. sirtalis show nearly identical rates of mass-adjusted energy expenditure at both temperatures, despite differing eco-morphologies, life histories, and distant phylogenetic positions. These findings suggest SMR may be a conserved feature of Thamnophis, and that any organismal tradeoffs may be compensated to retain whole-animal function.

Trans-seasonal activation of the neuroendocrine reproductive axis: Low-temperature winter dormancy modulates gonadotropin-releasing hormone neurons in garter snakes.

All animals use external cues from the environment to accurately time life-history events. How the brain decodes environmental stimuli to effect changes in physiology and behavior, however, is poorly understood, particularly with regard to supplementary environmental cues such as temperature. We asked if low-temperature dormancy alters the synthesis and/or release of gonadotropin-releasing hormone (GnRH). We used the well-studied red-sided garter snake (Thamnophis sirtalis) for this study, as low-temperature exposure is both necessary and sufficient to induce reproduction in northern populations of this species. Snakes were collected from the field and hibernated at 4°C or 10°C in complete darkness for up to 16 weeks. In males, increasing duration of low-temperature dormancy significantly increased GnRH-immunoreactive cell number and GnRH soma size (a proxy for relative cell activity) in the forebrain. These changes mirrored those in male reproductive behavior (reported previously) and plasma androgen concentrations. The changes in GnRH cell area observed in males were specific to the neuroendocrine population of cells in the medial preoptic area; soma size in the rostral GnRH cells did not change. Finally, temperature-induced changes in GnRH were sexually dimorphic: neither hibernation temperature nor the duration of winter dormancy significantly modulated GnRH cell number or soma size in females, despite the fact that plasma estradiol and corticosterone increased significantly in response to both. These data demonstrate that the neuroendocrine GnRH system is sensitive to environmental temperature and suggest that GnRH neurons play a conserved but trans-seasonal role in mediating changes in reproductive physiology and behavior in dissociated breeders.

Bithynian snake bombs

Bithynian snake bombs

Postcopulatory sexual selection as a driver of sex- and population-specific kidney mass in garter snakes?

Abstract In lizards and snakes, the kidneys produce seminal fluid in addition to having osmoregulatory functions. Therefore, in response to polyandry, kidney mass should be under selection regimes such as those influencing testes. Male red-sided garter snakes deposit a kidney-derived copulatory plug that functions in sperm competition. We first tested for intersexual differences in kidney mass and allometry in one population and found that males had kidneys twice as heavy as those of females, consistent with stronger selection on male kidney mass. Previous studies have shown that male size enhances mating success in one-on-one competition prevalent in small mating aggregations. We then examined the relationship between body size, kidney mass and testes mass in two populations with low (LD) and high (HD) mating aggregation densities. Males from the HD population had heavier testes and heavier kidneys compared with males from the LD population; HD males were also smaller in body size. Our results suggest that the strength of sexual selection, and possibly the balance between pre- and postcopulatory selection, varies in response to population size or density. However, more replication is needed to confirm the generality of these results within red-sided garter snakes and other squamate reptiles.