North American Pit Viper Research Articles

Efficacy and safety of two Antivenoms in the treatment of eastern copperhead (Agkistrodon contortrix) envenomations in Southeast Texas.

There are two Food and Drug Administration-approved antivenoms for the treatment of North American pitviper envenomations: Crotalidae polyvalent immune fab (ovine) (CroFab, henceforth FabAV) and Crotalidae immune F(ab')2 (equine) (Anavip, henceforth Fab2AV). This study compared the effectiveness and safety of the two products in eastern copperhead envenomations. We retrospectively reviewed eastern copperhead bites on which our toxicology service consulted between January 1, 2021 and November 1, 2023. Collected data included patient demographics, bite location, clinical features, antivenom use, response to treatment, and adverse reactions to antivenom. There were 134 patients with confirmed copperhead envenomations. We administered antivenom to 89 patients, of which 36 (40%) were female. The median age was 42 years old (range: 2-89 years). Fifty-nine patients received Fab2AV and 30 were treated with FabAV. Initial control was achieved in 30 (100%) patients treated with FabAV. In the Fab2AV group, 53 (89.8%) achieved control; six patients with acute adverse reactions declined further treatment despite persistent symptoms. The median FabAV dose required for initial control was six vials (range: 4-6). A median dose of 10 vials (range: 10-30) was used in the Fab2AV group. Repeat doses of Fab2AV were required in 15 (25.4%) cases. There were no acute adverse reactions to FabAV. Acute adverse reactions were seen in seven (11.9%) of patients treated with Fab2AV. Fewer patients treated with FabAV required repeat antivenom dosing to attain initial control following eastern copperhead envenomation. A higher percentage of patients treated with Fab2AV developed acute adverse reactions.

Platelet and red cell responses to three North American pit vipers

We investigated the hemotoxic effects of three North American pit vipers in healthy human donor blood. Using experiments focusing on platelet and red blood cell activity, we found differential effects of these venoms on these cellular components. Platelet aggregation was most induced by C. adamanteus. Platelet activation was highest with C. atrox. Red blood cells had calcium expression and erythrocyte formation most induced by C. adamanteus and A. piscivorus. These results demonstrate the complex interplay of individual cellular effects with clinical presentations seen in envenomings from these species.

Fasciotomy following North American pit viper envenomation in Texas 2004–2021

Introduction North American pit viper envenomation occurs over 4,000 times annually in the United States, with polyvalent Fab antivenom being the primary treatment. Fasciotomy is occasionally performed due to concerns about compartment syndrome. We utilized our direct access to Texas Poison Center Network data to create a new snakebite abstraction form and database on relevant available information between 2004 and 2021 and to identify, describe, and estimate the incidence of fasciotomy following pit viper envenomation in Texas. Methods We searched the Texas Poison Center Network database for cases during 2004–2021 using keywords such as fasciotomy, surgery, compartment pressure, and compartment syndrome. Descriptive statistics summarized the data. Results Of 16,911 reported envenomations, 0.69 percent involved fasciotomies (n = 117). Most common bite sites were digits/hands and lower extremities. Patients who underwent fasciotomy were typically male, aged 20–59, and 10 years younger than the total snakebite population. Only 6 percent of reported compartment syndrome cases had a compartment pressure measurement. Antivenom was administered in 101 (86.3 percent) cases, 92 (91.1 percent) of which received only Fab antivenom product. Patients with bites from rattlesnakes (47.9 percent) were associated with most fasciotomies Discussion Our findings suggest a potential increase in snakebite exposures, accompanied by a decrease in fasciotomies. Overall, copperheads constituted the majority of snakebites, but most fasciotomies were from rattlesnake envenomations (47.9 percent). In this cohort, compartment syndrome diagnosis and decisions regarding fasciotomy were primarily based on clinical evaluation/surgeon expertise without compartment pressure measurements. Despite the efficacy of antivenom, only 86.3 percent of patients in our study received antivenom. Conclusions Fasciotomy after North American pit viper envenomation in Texas is uncommon (0.69 percent) and has decreased over time, possibly due to increased antivenom use or surgeon comfort with nonsurgical management.

Snake venom cysteine-rich secretory protein from Mojave rattlesnake venom (Css-CRiSP) induces acute inflammatory responses on different experimental models

Snake venoms contain various molecules known for activating innate immunity and causing local effects associated with increased vascular permeability, such as vascular leakage and edema, common symptoms seen in snakebite envenomings. We have demonstrated that snake venom cysteine-rich secretory proteins (svCRiSPs) from North American pit vipers increase vascular permeability. This study aimed to explore the functional role of CRiSP isolated from Mojave rattlesnake (Crotalus scutulatus scutulatus) venom (Css-CRiSP) on the activation of inflammatory responses in different models. We measured the release of inflammatory mediators in cultured human dermal blood endothelial cells (HDBEC), lymphatic endothelial cells (HDLEC) and monocyte-derived macrophages (MDM) at 0.5, 1, 3, 6, and 24 h after treatment with Css-CRiSP (1 μM). We also determined the acute inflammatory response in BALB/c mice 30 min after intraperitoneal injection of the toxin (2 μg/mouse). Css-CRiSP induced the production of IL-8 and IL-6, but not TNF-α, in HDBEC and HDLEC in a time-dependent manner. In addition, Css-CRiSP significantly enhanced the production of IL-6, TNF-α, IL-8, and IL-1β in MDM. Moreover, it caused a remarkable increase of chemotactic mediators in the exudates of experimental mice. Our results reveal that Css-CRiSPs can promote a sustained release of inflammatory mediators on cell lines and an acute activation of innate immunity in a murine model. These findings contribute to the growing body of evidence supporting the involvement of svCRiSPs in the augmentation of envenomation effects, specifically, the role of svCRiSPs in inducing vascular dysfunction, initiating early inflammatory responses, and facilitating the activation of leukocytes and releasing mediators. These findings will lead to a better understanding of the pathophysiology of envenoming by Mojave rattlesnakes, allowing the development of more efficient therapeutic strategies.

Fission-fusion dynamics in the social networks of a North American pitviper.

Many animal species exist in fission-fusion societies, where the size and composition of conspecific groups change spatially and temporally. To help investigate such phenomena, social network analysis (SNA) has emerged as a powerful conceptual and analytical framework for assessing patterns of interconnectedness and quantifying group-level interactions. We leveraged behavioral observations via radiotelemetry and genotypic data from a long-term (>10 years) study on the pitviper Crotalus atrox (western diamondback rattlesnake) and used SNA to quantify the first robust demonstration of social network structures for any free-living snake. Group-level interactions among adults in this population resulted in structurally modular networks (i.e., distinct clusters of interacting individuals) for fidelis use of communal winter dens (denning network), mating behaviors (pairing network), and offspring production (parentage network). Although the structure of each network was similar, the size and composition of groups varied among them. Specifically, adults associated with moderately sized social groups at winter dens but often engaged in reproductive behaviors-both at and away from dens-with different and fewer partners. Additionally, modules formed by individuals in the pairing network were frequently different from those in the parentage network, likely due to multiple mating, long-term sperm storage by females, and resultant multiple paternity. Further evidence for fission-fusion dynamics exhibited by this population-interactions were rare when snakes were dispersing to and traversing their spring-summer home ranges (to which individuals show high fidelity), despite ample opportunities to associate with numerous conspecifics that had highly overlapping ranges. Taken together, we show that long-term datasets incorporating SNA with spatial and genetic information provide robust and unique insights to understanding the social structure of cryptic taxa that are understudied.

Proteomic Profiling of Extracellular Vesicles Isolated from Plasma and Peritoneal Exudate in Mice Induced by Crotalus scutulatus scutulatus Crude Venom and Its Purified Cysteine-Rich Secretory Protein (Css-CRiSP).

Increased vascular permeability is a frequent outcome of viperid snakebite envenomation, leading to local and systemic complications. We reported that snake venom cysteine-rich secretory proteins (svCRiSPs) from North American pit vipers increase vascular permeability both in vitro and in vivo. They also induce acute activation of several adhesion and signaling molecules that may play a critical role in the pathophysiology of snakebites. Extracellular vesicles (EVs) have gained interest for their diverse functions in intercellular communication, regulating cellular processes, blood-endothelium interactions, vascular permeability, and immune modulation. They also hold potential as valuable biomarkers for diagnosing, predicting, and monitoring therapeutic responses in different diseases. This study aimed to identify proteins in peritoneal exudate and plasma EVs isolated from BALB/c mice following a 30 min post-injection of Crotalus scutulatus scutulatus venom and its purified CRiSP (Css-CRiSP). EVs were isolated from these biofluids using the EVtrap method. Proteomic analysis of exudate- and plasma-derived EVs was performed using LC-MS/MS. We observed significant upregulation or downregulation of proteins involved in cell adhesion, cytoskeleton rearrangement, signal transduction, immune responses, and vesicle-mediated transports. These findings suggest that svCRiSPs play a crucial role in the acute effects of venom and contribute to the local and systemic toxicity of snakebites.

Epidemiology and Characteristics of North American Crotalid Bites Reported to the National Poison Data System 2006-2020.

North American pit viper, or crotalid bites, remain a low-incidence and potentially high-consequence medical event. Although the venom of these snakes is known to cause tissue, hematologic, and neurologic toxicity, the published literature on North American crotalid bites remains limited. The National Poison Data System, the data repository for the 55 poison control centers in the United States, offers a unique opportunity to examine nationwide trends involving venomous snake bites. National Poison Data System cases involving North American crotalids from 2006 to 2020 were analyzed. Data collected included age and type of snake, date, geographic location, pertinent clinical characteristics, treatments administered, and medical outcomes including incidence of "dry" bites and death. A total of 55,914 cases were identified during the 15-year study period. Cases, especially those involving copperheads, increased during the study period. Most of the cases were reported in July. Cases were reported in all 50 states and Washington, DC, with Texas having the most cases (n = 9115). North Carolina had the largest increase in bites during the study period. Moderate or major medical outcomes were documented in 58% (n = 32,584) of cases, with 25% (n = 14,195) being admitted to a critical care unit. Puncture wound, edema, and pain were the most commonly documented symptoms. Antivenom was documented as being administered in 25% (n = 14,151) of cases. Dry bites were reported in <1.5% of cases. Thirty-two deaths were reported, 23 involving rattlesnakes. This study demonstrates that reported North American crotalid bites appear to be increasing over time and are associated with potentially significant morbidity. Mortality, however, remains low.

Mountain Emergency Medicine Review, 1st edition

Mountain Emergency Medicine Review, 1st edition

Structural and bioinformatic analyses of Azemiops venom serine proteases reveal close phylogeographic relationships to pitvipers from eastern China and the New World

The semi-fossil and pit-less Azemiops feae is possibly the most primitive crotalid species. Here, we have cloned and sequenced cDNAs encoding four serine proteases (vSPs) from the venom glands of Chinese A. feae. Full amino-acid sequences of the major vSP (designated as AzKNa) and three minor vSPs (designated as AzKNb, AzKNc and Az-PA) were deduced. Using Protein-BLAST search, the ten most-similar vSPs for each Azemiops vSP have been selected for multiple sequence alignment, and all the homologs are crotalid vSPs. The results suggest that the A. feae vSPs are structurally most like those of eastern-Chinese Gloydius, Viridovipera, Protobothrops and North American pitvipers, and quite different from more-specialized vSPs such as Agkistrodon venom Protein-C activators. The vSPs from Chinese A. feae and those from Vietnamese A. feae show significant sequence variations. AzKNa is acidic and contains six potential N-glycosylation sites and its surface-charge distribution differs greatly from that of AzKNb, as revealed by 3D-modeling. AzKNb and AzKNc do not contain N-glycosylation sites although most of their close homologs contain one or two. Az-PA belongs to the plasminogen-activator subtype with a conserved N20-glycosylation site. The evolution of this subtype of vSPs in Azemiops and related pitvipers has been traced by phylogenetic analysis.

Caring for patients with venomous Crotalinae snakebites

Venomous snakebites are surprisingly common in the US. This article provides an overview of what to do when a patient has been bitten by a North American pit viper, a venomous subset of indigenous snakes.

Review of North American pit viper antivenoms.

For the first time in nearly 20 years, 2 antigen-binding fragment (Fab) antivenoms are available to treat patients who incur North American pit viper snakebites: Crotalidae polyvalent immune Fab (ovine), or simply FabAV; and Crotalidae immune F(ab')2 (equine), or simply F(ab')2. Pharmacists are in a key position for the selection, dosing, reconstitution, administration, and monitoring of antivenom therapy; however, they encounter inconsistent exposure and experience with these drugs. Thus, an updated review of the literature is necessary. The search strategy and selection incorporated both controlled vocabulary terms and keywords to describe concepts relevant to the search. Retrieval was limited to literature published from 1997 to the present in English, Portuguese, or Spanish. Given the paucity of available prospective literature, the authors elected to include all prospective evidence to best describe the role of antivenom. For the primary literature review, manuscripts were excluded if they were observational studies, conference abstracts, narrative or opinion articles, letters to the editor, or in-progress studies. While there is limited evidence-based guidance on the superiority of F(ab')2 to FabAV, or vice versa, individual and regional considerations should contribute to formulary decisions. Pharmacists must play a role in the development of clinical pathways to ensure appropriate evaluation, supportive care, and antivenom procurement, administration, and monitoring.

Prevalence of hematologic toxicity from copperhead envenomation: an observational study

Background: Copperhead snakes (Agkistrodon contortrix) are considered as the least toxic of the North American pit vipers. The reported incidence of coagulopathy from copperhead envenomation is variable, possibly secondary to regional variation in subspecies and venom potency. Coagulation studies are often obtained when evaluating for the coagulopathic effects of copperhead venom, but the clinical utility of these indices is unclear. The aim of this study was to determine the prevalence of hematologic toxicity due to copperhead envenomation in hospitalized patients.Methods: This was a multi-center retrospective chart review study using electronic hospital data between January 1, 2006 and December 31, 2016 evaluating prevalence of coagulopathy following copperhead envenomation. Patients presenting to one of three major academic tertiary care centers in Virginia with suspected copperhead envenomation were identified using medical billing codes. The primary outcome was to summarize the prevalence of hematologic toxicity including thrombocytopenia, elevated prothrombin or partial thromboplastin times, or hypofibrinogenemia.Results: There were 244 cases used for final analysis. Hematologic toxicity occurred in 14% (95% CI 10–18%) of patients. Specific indices included thrombocytopenia in 1.2% (95% CI 0.4–3.6%), hypofibrinogenemia in 0.7% (95% CI 0.0–3.8%), elevated PT in 10.0% (95% CI 6.8–14.5%), and aPTT in 3.9% (95% CI 2.1–7.2%) of patients. There was no clinically significant bleeding reported in any case.Conclusions: Subtle hematologic abnormalities due to copperhead envenomation in patients treated in the Commonwealth of Virginia were relatively common, but do not appear to be clinically significant in this study population.

Early administration of Fab antivenom resulted in faster limb recovery in copperhead snake envenomation patients

Background: No previous research has studied whether early snake antivenom administration leads to better clinical outcomes than late antivenom administration in North American pit viper envenomation.Methods: A secondary analysis of data from a clinical trial of Fab antivenom (FabAV) versus placebo for copperhead snake envenomation was conducted. Patients treated before the median time to FabAV administration were classified as receiving early treatment and those treated after the median time were defined as the late treatment group. A Cox proportional hazards model was used to compare time to full recovery on the Patient-Specific Functional Scale (PSFS) instrument between groups. Secondary analyses compared estimated mean PSFS scores using a generalized linear model and the estimated proportion of patients with full recovery at each time point using logistic regression. To evaluate for confounding, the main analysis was repeated using data from placebo-treated subjects.Results: Forty-five subjects were treated with FabAV at a median of 5.47 h after envenomation. Patients in the early treatment group had a significantly shorter time to full recovery than those treated late (median time: 17 versus 28 days, p = .025). Model-estimated PSFS scores were numerically higher at each time point in the early group. No difference was found between patients treated early versus late with placebo.Conclusions: In this secondary analysis of trial data, recovery of limb function was faster when Fab antivenom was administered soon after envenomation, as opposed to late administration.

Population Genetics of the Copperhead at Its Most Northeastern Distribution

Population genetic data are an important standard with which to gauge demography and gene flow within and among biodiversity units, and are often gathered on species of conservation concern. Yet an exclusive focus on ‘conservation immediacy' can also have negative consequences. For example, it can shift monitoring efforts away from more abundant and/or widely distributed clades and, by so doing, promote a more myopic approach to conservation and management. A current example concerns North American pitvipers (Viperidae: Crotalinae), within which listed species of Crotalus and Sistrurus receive considerable population genetic attention whereas broadly distributed Agkistrodon is largely overlooked. To address this disparity, we used 22 polymorphic tetra-nucleotide microsatellite loci to explore genetic structure, diversity, and relatedness in a Connecticut population of Copperhead (A. contortrix). Three admixed genetic clusters were identified across five winter dens, with overall and sex-specific relatedne...

North American Pit Viper Envenomation

Abstract Acute snake envenomation is an uncommon but potentially serious envenomation causing local tissue injury, hematologic derangements, and systemic effects. The purpose of this simulation is ...

Post-birth separation affects the affiliative behaviour of kin in a pitviper with maternal attendance

The relatedness of individuals can have pronounced effects on behavioural interactions, as engaging in mutually beneficial behaviours with kin can increase inclusive fitness. Parental care can be particularly important for kin discrimination in birds and mammals, but similar studies have not been conducted on species exhibiting more rudimentary forms of care. Maternal attendance of young is ubiquitous among viviparous temperate pitvipers, but the adaptive value of this behaviour has received little attention. We sought to determine if being deprived of a maternal attendance period as neonates altered how cottonmouths (Agkistrodon piscivorus Lacépède), a common North American pitviper, responded to kin vs. non-kin. We measured the affiliative behaviour of related and unrelated juvenile–juvenile and mother–juvenile pairs that had been allowed a maternal attendance period or had been separated since birth. We found that maternal attendance was not required for sibling or mother–offspring recognition, but did enhance female affiliative behaviour overall, and particularly that of sisters. In contrast, related juveniles that were separated at birth showed a reversal of the sex-specific affiliative behaviour observed in maternally attended juveniles. Post-birth separation had only a modest effect on mother–juvenile affiliative behaviour, and no effect on the strong affiliation between mothers and daughters. The patterns of affiliative behaviour observed in maternally attended snakes corresponded to patterns that have emerged from previous captive and field studies of pitvipers; however, the behaviour of juveniles separated at birth was atypical. Thus, it is possible that maternal attendance plays some role in the development of adaptive sex-specific and kin-directed affiliative behaviour in pitvipers.

Putative Pair-Bonding inAgkistrodon contortrix(Copperhead)

Pair-bonding between sexes is common in vertebrate taxa, yet it has been noted far less frequently in some groups such as reptiles, and snakes in particular. Evidence to date indicates that many snake mating-systems are polyandrous, with both males and females having multiple partners in a single breeding season, and thus unlikely to exhibit lengthy pair-bonds. Wittenberger and Tilson (1980) suggested that pair-bonding exists when pairs remain intact for a consecutive period equaling at least 25% of the breeding season. Using this criterion, we present evidence of pair-bond formation in a North American pitviper, Agkistrodon contortrix (Copperhead), a species with a polyandrous mating system.

Male mating success in a North American pitviper: influence of body size, testosterone, and spatial metrics

Males with enhanced traits relative to conspecifics often show increased mating and reproductive success and thus have a fitness advantage. The opportunity or potential for sexual selection is predicted to occur under these conditions. Here, we investigated proximate determinants of mating success in male copperhead snakes (Agkistrodon contortrix), a medium-sized pitviper of North America. Specifically, we investigated the relationships of body size (snout-vent length, body mass), body condition index, spatial metrics (total distance moved, home range size), and plasma testosterone concentration on mating success in males. The single mating season lasts from August through September. We compared a set of candidate linear mixed models and selected the best-fitting one using the adjusted Akaike Information Criterion (AICc). The AICc-selected model (model 2), with testosterone, body condition index, and home range size as predictor variables, showed that male mating success was positively correlated with testosterone. To our knowledge, this is the first report to show the relationship of testosterone and individual mating success in any snake species. A parallel study conducted on male fitness in A. contortrix of the same population used microsatellite markers to assign parentage of fathers (known mothers). Unlike our study, they found that snout-vent length was positively correlated with reproductive success and that males were experiencing greater sexual selection. This relationship has been detected under natural conditions in other species of snakes. Although behavioural data are important in any mating system analysis, they should not stand alone to infer parentage, relationships or selection metrics (e.g. Bateman gradients). Long-term sperm storage by females, female cryptic choice, and other factors contribute to the complexity of mating success of males. Accordingly, we thus conclude that estimates of reproductive success and fitness in cryptic species, such as copperheads and other snakes, require robust molecular methods to draw accurate conclusions regarding proximate and evolutionary responses. © 2015 The Linnean Society of London, Biological Journal of the Linnean Society, 2015, 115, 185‐194.

Incipient speciation with biased gene flow between two lineages of the Western Diamondback Rattlesnake (Crotalus atrox)

We used mitochondrial DNA sequence data from 151 individuals to estimate population genetic structure across the range of the Western Diamondback Rattlesnake (Crotalus atrox), a widely distributed North American pitviper. We also tested hypotheses of population structure using double-digest restriction site associated DNA (ddRADseq) data, incorporating thousands of nuclear genome-wide SNPs from 42 individuals. We found strong mitochondrial support for a deep divergence between eastern and western C. atrox populations, and subsequent intermixing of these populations in the Inter-Pecos region of the United States and Mexico. Our nuclear RADseq data also identify these two distinct lineages of C. atrox, and provide evidence for nuclear admixture of eastern and western alleles across a broad geographic region. We identified contrasting patterns of mitochondrial and nuclear genetic variation across this genetic fusion zone that indicate partially restricted patterns of gene flow, which may be due to either pre- or post-zygotic isolating mechanisms. The failure of these two lineages to maintain complete genetic isolation, and evidence for partially-restricted gene flow, imply that these lineages were in the early stages of speciation prior to secondary contact.

Considering gene flow when using coalescent methods to delimit lineages of North American pitvipers of the genusAgkistrodon

Examining species diversity and mechanisms of speciation using coalescent models provides a framework for how regional diversity is accrued, even in well-studied areas such as the Nearctic. It is likely, that gene flow among closely-related species with adjacent distributions may be common. However, the absence of gene flow is a primary assumption of many phylogeographical methods that produce species trees and delimit species using Bayesian or likelihood functions in a coalescent framework. In the present study, we examine delimitation when gene flow between species is present using empirical datasets from two species of North American pitvipers of the genus Agkistrodon. We also use niche modelling to determine whether these young lineages occur in distinct environmental niches. To manage the problem of gene flow between species, we first identify admixed individuals, demonstrate that gene flow has occurred, and then identify the impact of alternative population assignments of admixed individuals on delimitation posterior probabilities. In addition, we examine the influence of mitochondrial genes relative to other loci combined in coalescent analyses that delimit species. Here, we find that the copperheads (Agkistrodon contortrix) and the cottonmouths (Agkistrodon piscivorus) are each composed of two distinct species, with each occupying different niches. Importantly, we find that species can be delimited when the amount of gene flow between lineages is low, although the methods are acutely sensitive to population assignment of individuals. © 2014 The Linnean Society of London