Recombinant snake antivenoms get closer to the clinic.

Snakebite envenomings kill ~100 000 victims each year and leave many more with permanent sequelae. Antivenoms have been available for more than 125 years but are in need of innovation. A new study by Khalek et al. highlights broadly neutralizing human monoclonal antibodies (mAbs) that might be used to develop recombinant antivenoms with superior therapeutic benefits.

Dermonecrosis caused by a spitting cobra snakebite results from toxin potentiation and is prevented by the repurposed drug varespladib.

Snakebite envenoming is a neglected tropical disease that causes substantial mortality and morbidity globally. The venom of African spitting cobras often causes permanent injury via tissue-destructive dermonecrosis at the bite site, which is ineffectively treated by current antivenoms. To address this therapeutic gap, we identified the etiological venom toxins in Naja nigricollis venom responsible for causing local dermonecrosis. While cytotoxic three-finger toxins were primarily responsible for causing spitting cobra cytotoxicity in cultured keratinocytes, their potentiation by phospholipases A2 toxins was essential to cause dermonecrosis in vivo. This evidence of probable toxin synergism suggests that a single toxin-family inhibiting drug could prevent local envenoming. We show that local injection with the repurposed phospholipase A2-inhibiting drug varespladib significantly prevents local tissue damage caused by several spitting cobra venoms in murine models of envenoming. Our findings therefore provide a therapeutic strategy that may effectively prevent life-changing morbidity caused by snakebite in rural Africa.

A Bite by Shaw's Short Sea Snake (Hydrophis curtus): A Case of Mild Myotoxicity or a Dry Bite?

Although sea snakes (Elapidae) are commonly encountered by fishermen, accurately authenticated envenomings by them are uncommon in clinical literature. We report an authenticated case of Shaw's short, or spine-bellied, sea snake (Hydrophis curtus) bite in a young fisherman from northern Sri Lanka. The patient had clinical and biochemical evidence of mild transient myotoxicity but no evidence of neuromuscular paralysis or significant renal injury. Consideration of the clinical manifestations suggests either a mild envenoming or a dry bite. The patient completely recovered without any antivenom therapy and was discharged on the fourth day. Prolonged observation may be beneficial to exclude complications of sea snake envenoming.

Experience of using expired lyophilized snake antivenom during a medical emergency situation in Lao People's Democratic Republic--A possible untapped resource to tackle antivenom shortage in Southeast Asia.

OBJECTIVES: To evaluate the safety and efficacy of expired lyophilized snake antivenom of Thai origin during a medical emergency in 2020/2021 in Lao People's Democratic Republic. METHODS: Observational case series of patients with potentially life-threatening envenoming who consented to the administration of expired antivenom between August 2020 and May 2022. RESULTS: A total of 31 patients received the expired antivenom. Malayan pit vipers (Calloselasma rhodostoma) were responsible for 26 (84%) cases and green pit vipers (Trimeresurus species) for two cases (6%). In three patients (10%) the responsible snake could not be identified. Of these, two presented with signs of neurotoxicity and one with coagulopathy. A total of 124 vials of expired antivenom were administered. Fifty-nine vials had expired 2-18 months earlier, 56 vials 19-36 months and nine vials 37-60 months before. Adverse effects of variable severity were observed in seven (23%) patients. All 31 patients fully recovered from systemic envenoming. CONCLUSIONS: Under closely controlled conditions and monitoring the use of expired snake antivenom proved to be effective and safe. Discarding this precious medication is an unnecessary waste, and it could be a valuable resource in ameliorating the current shortage of antivenom. Emergency use authorization granted by health authorities and preclinical testing of expired antivenoms could provide the support and legal basis for such an approach.

Development of an Inhibition Enzyme-Linked Immunosorbent Assay (ELISA) Prototype for Detecting Cytotoxic Three-Finger Toxins (3FTxs) in African Spitting Cobra Venoms.

The administration of toxin-specific therapy in snake envenoming is predicated on improved diagnostic techniques capable of detecting specific venom toxins. Various serological tests have been used in detecting snakebite envenoming. Comparatively, enzyme-linked immunosorbent assay (ELISA) has been shown to offer a wider practical application. We report an inhibition ELISA for detecting three-finger toxin (3FTx) proteins in venoms of African spitting cobras. The optimized assay detected 3FTxs in N. ashei (including other Naja sp.) venoms, spiked samples, and venom-challenged mice samples. In venoms of Naja sp., the assay showed inhibition, implying the detection of 3FTxs, but showed little or no inhibition in non-Naja sp. In mice-spiked samples, one-way ANOVA results showed that the observed inhibition was not statistically significant between spiked samples and negative control (p-value = 0.164). Similarly, the observed differences in inhibition between venom-challenged and negative control samples were not statistically significant (p-value = 0.9109). At an LOD of 0.01 µg/mL, the assay was able to confirm the presence of 3FTxs in the samples. Our results show a proof of concept for the use of an inhibition ELISA model as a tool for detecting 3FTxs in the venoms of African spitting cobra snakes.

Proximity between humans and a highly medically significant snake, Russell's viper, in a tropical rural community.

Snakebite envenoming is a major neglected tropical health issue. The high incidence of snakebites in tropical rural communities suggests that venomous snakes and people are often in proximity but quantitative evidence is lacking. I used radio-telemetry on a population of Russell's vipers (Daboia russelii), one of the most medically important snakes in the world, to quantify proximity between this venomous snake and people and estimate susceptibility to snakebite envenoming in India. I observed people ≤50 m of a radio-equipped viper in ~17% of 2,066 snake relocations. People were more frequently observed in proximity to Russell's vipers in January and July compared to March, but all other contrasts were statistically similar. This pattern indicates that snakebite incidence, which peaks in summer in the study area, is not particularly linked to the encounter frequency between people and vipers. However, consistent with epidemiological data plantation workers were the most at-risk part of the population. By integrating information about the locations of humans and snakes in space and time, this pioneering research highlights the need to include snake ecology into the study of the human-venomous-snake conflict, and provides a model approach to help mitigate the burden caused by venomous snakes in the rural Tropics.

A Genus-Wide Bioactivity Analysis of Daboia (Viperinae: Viperidae) Viper Venoms Reveals Widespread Variation in Haemotoxic Properties.

The snake genus Daboia (Viperidae: Viperinae; Oppel, 1811) contains five species: D. deserti, D. mauritanica, and D. palaestinae, found in Afro-Arabia, and the Russell's vipers D. russelii and D. siamensis, found in Asia. Russell's vipers are responsible for a major proportion of the medically important snakebites that occur in the regions they inhabit, and their venoms are notorious for their coagulopathic effects. While widely documented, the extent of venom variation within the Russell's vipers is poorly characterised, as is the venom activity of other species within the genus. In this study we investigated variation in the haemotoxic activity of Daboia using twelve venoms from all five species, including multiple variants of D. russelii, D. siamensis, and D. palaestinae. We tested the venoms on human plasma using thromboelastography, dose-response coagulometry analyses, and calibrated automated thrombography, and on human fibrinogen by thromboelastography and fibrinogen gels. We assessed activation of blood factors X and prothrombin by the venoms using fluorometry. Variation in venom activity was evident in all experiments. The Asian species D. russelii and D. siamensis and the African species D. mauritanica possessed procoagulant venom, while D. deserti and D. palaestinae were net-anticoagulant. Of the Russell's vipers, the venom of D. siamensis from Myanmar was most toxic and D. russelli of Sri Lanka the least. Activation of both factor X and prothrombin was evident by all venoms, though at differential levels. Fibrinogenolytic activity varied extensively throughout the genus and followed no phylogenetic trends. This venom variability underpins one of the many challenges facing treatment of Daboia snakebite envenoming. Comprehensive analyses of available antivenoms in neutralising these variable venom activities are therefore of utmost importance.

First Authenticated Case of a Bite by Rare and Elusive Blood-Bellied Coral Snake (Calliophis haematoetron).

Blood-bellied coral snake (Calliophis haematoetron) is a recently discovered forest-dwelling species of elapid hitherto known from 3 specimens found from central Sri Lanka. Herein we describe the first authenticated case of blood-bellied coral snakebite. The victim, an 11-mo-old infant who received the bite while handling the snake at her home, had mild transient swelling at the bite site. The patient had no clinical or laboratory evidence of systemic envenoming. We highlight the importance of clinicians being aware of the occurrence of this potentially medically important elapid snake in anthropogenic habitats.

The 'Snake song': a pilot study of musical intervention in Eswatini.

INTRODUCTION: In Eswatini in Southern Africa, rural populations experience unnecessary snakebite-inflicted injuries and deaths. Children are at the highest risk because of their small size and curious nature. This qualitative study explores the current knowledge and attitudes about snakebite, and the perceptions of a musical intervention, titled Iculo ngenyoka ('Snake song' in Zulu), as an educational tool aimed to raise awareness about snakes in the Lubombo region, Eswatini. METHODS: Semi-structured interviews with community members (n=56), parents/guardians/key informant (n=11) and children aged 7-17 years (n=45) were conducted between May and June 2018. Participants were selected from four communities within the Lubombo region. Data were analyzed using a framework analysis approach. RESULTS: The current sources of snake education evolved from information learned in the homesteads, schools, and through personal experiences. The majority of interviewees perceived music as a culturally appropriate, engaging and memorable method to learn about snakes. Iculo ngenyoka was perceived as an effective tool to raise awareness about snakes in the community. CONCLUSION: This study is the first to explore the importance of musical interventions in educating vulnerable communities about snakes. The Iculo ngenyoka song offers a portable medium for communicating messages about snakebite prevention, affirming the value of snakebite awareness and promoting cooperative efforts to address the burden of snakebite envenoming in the region. The results emphasize the demand for education and the potential use of Iculo ngenyoka and similar musical tools to raise awareness about snakebite in Eswatini. Re-translation and other customizations of structured musical education tools for children could be applied broadly if shown to be effective.

Low-dose ketamine provides poor analgesia for pain in redback spider envenoming.

Redback spider envenoming causes severe pain lasting several days. A recent clinical trial found that antivenom is not effective. We investigated ketamine for pain in redback spider envenoming. Ten adult patients with severe pain from redback spider envenoming were administered 15 mg intravenous ketamine after standard analgesia, then up to 4 oral doses of ketamine 25- 50 mg. Three patients had a clinically significant improvement in pain compared to baseline after intravenous ketamine. Five patients had a minimal decrease in pain and 2 had no improvement. Eight patients received oral ketamine: 4 doses in 5 and 2 doses in 3. At 24 h, 3/6 patients assessed had clinically significant improvement in pain and 4/5 patients assessed at 48 h, had clinically significant improvement in pain. Six patients reported side effects, including dissociation (4) and hallucinations (2). Five patients required rescue opioids and 2 were readmitted to hospital. We found that ketamine provided no additional pain relief in redback spider envenoming, compared to standard analgesia, and resulted in unacceptable adverse effects.

Analysis of hemocoagulation tests for prediction of venom-induced consumption coagulopathy development after Viperidae bite.

OBJECTIVE: Viperidae snakes are responsible for 95 % of the bites caused by exotic-bred snakes in our country. Their envenoming may be associated with a severe acute coagulation disorder - venom-induced consumption coagulopathy (VICC). Thus, its early prediction is vital for an adequate therapy including antivenom delivery. MATERIAL AND METHODS: Laboratory coagulation tests of 14 patients suffering from VICC were processed and statistically analyzed to evaluate the importance of individual parameters in the time after the bite. RESULTS: The pathological values of D-dimer (D-dim) and fibrinogen (FBG) were found to be the first indicators of VICC development, with a median time of 4.55 hours since the bite, while median values for prothrombin time and international normalized ratio (PT/INR), activated partial thromboplastin time (APTT), and thrombin time (TT) were 5.9 h, 8.15 h, and 5.5 h, respectively. In the first samples, the values of D-dim were found to be pathologically increased in all 14 patients, while pathological levels of FBG were seen only in 11 cases. PT/INR and APTT were prolonged in 8 and 6 cases, respectively. CONCLUSION: An increase in D-dim values was found to be the first parameter signaling developing VICC in all analyzed cases (Tab. 2, Ref. 12).

Haemotoxic snake venoms: their functional activity, impact on snakebite victims and pharmaceutical promise.

Snake venoms are mixtures of numerous proteinacious components that exert diverse functional activities on a variety of physiological targets. Because the toxic constituents found in venom vary from species to species, snakebite victims can present with a variety of life-threatening pathologies related to the neurotoxic, cytotoxic and haemotoxic effects of venom. Of the 1·8 million people envenomed by snakes every year, up to 125 000 die, while hundreds of thousands survive only to suffer with life-changing long-term morbidity. Consequently, snakebite is one of the world's most severe neglected tropical diseases. Many snake venoms exhibit strong haemotoxic properties by interfering with blood pressure, clotting factors and platelets, and by directly causing haemorrhage. In this review we provide an overview of the functional activities of haemotoxic venom proteins, the pathologies they cause in snakebite victims and how their exquisite selectivity and potency make them amenable for use as therapeutic and diagnostic tools relevant for human medicine.

Injury trends from envenoming in Australia, 2000-2013.

BACKGROUND: Accidental injury is a major public health problem in developed countries with 20 years elapsed since a national overview of venomous bites undertaken in Australia. AIM: Provide the first contemporary epidemiological insight into venomous injuries based on demographics and geography nationally in Australia in the period 2000-2013. METHODS: An analysis of national hospitalisation and mortality data was undertaken to examine the incidence of injury and death due to envenoming in Australia. Rates were calculated using the intercensal population for all Australian age groups. RESULTS: Over the study period, deaths occurred due to an anaphylactic event (0.16 per 100 000), snake envenoming (0.13 per 100 000) or box jellyfish envenoming (0.01 per 100 000). Only 44% of cases involving anaphylaxis reached medical care prior to death, compared to 74% of those envenomed by snakes. Over half of all deaths (52%) occurred at home, and 64% of these occurred within a major city or inner regional area, with 48% of work-related anaphylaxis deaths. Hospital admission rates of 199 per 100 000 persons over the 11 years were caused by contact with wasps or bees (31%), spiders (30%) and snakes (15%), with a predominant age range of 30-44 years. CONCLUSIONS: The greatest burden of injury due to envenoming was caused by arthropods and snakes. Causes of death were led by anaphylaxis subsequent to an arthropod bite or sting, followed by death from snake envenoming. Over half the incidents resulting in death occurred at home, in areas where healthcare is accessible. Operational data routinely collected are informative, with variations of injury incidence between the States and Territories, indicating a need for a more localised approach to the management of this injury.

Adverse reactions to snake antivenom, and their prevention and treatment.

Antivenom is the mainstay of treatment of snakebite envenoming. However, adverse reactions to snake antivenom that is available are common in many parts of the world where snakebite is prevalent. Both acute (anaphylactic or pyrogenic) and delayed (serum sickness type) reactions occur. Acute reactions are usually mild but severe systemic anaphylaxis may develop, often within an hour or so of exposure to antivenom. Serum sickness after antivenom has a delayed onset between 5 and 14 days after its administration. Ultimately, the prevention reactions will depend mainly on improving the quality of antivenom. Until these overdue improvements take place, doctors will have to depend on pharmacological prophylaxis, where the search for the best prophylactic agent is still on-going, as well as careful observation of patients receiving antivenom in preparation for prompt management of acute as well as delayed reactions when they occur.

Severe Snakebite Envenoming in Intensive Care.

Snakebites by exotic venomous snakes can cause serious or even life-threatening envenoming. In Europe and North America most victims are breeders, with a few snakebites from wild native American rattlesnakes. The envenomed victims may present in organ and/or system failure with muscle paralysis, respiratory failure, circulatory instability, acute kidney injury, severe coagulation disorder, and local disability - compartment syndrome and necrosis. Best managed by close collaboration between clinical toxicology and intensive care, most severe envenomings are managed primarily by intensive care physicians. Due to the low incidence of severe envenoming, the clinical course and correct management of these cases are not intrinsically familiar to most physicians. This review article summarizes the clinical syndromes caused by severe envenoming and the therapeutic options available in the intensive care setting.

Management of a Pediatric Snake Envenomation After Presentation With a Tight Tourniquet.

We describe an illustrative case of pediatric snake envenomation presenting with a tightly wound tourniquet. A 10-year-old boy presented after a snake bite to the right calf. A tourniquet was in place just below the right knee. The species of snake was unknown. The patient was hemodynamically stable, but the entirety of the right leg distal to the tourniquet was discolored. Over concern for a potential venom bolus effect upon tourniquet removal, the decision was made to start a crotaline Fab antivenom infusion and gradually loosen the tourniquet. The patient tolerated the infusion and removal of the tourniquet without signs of anaphylaxis or decompensation. Dynamic improvements were observed in the right leg and wound site that appeared to be the result of vascular congestion. Tourniquets are generally not recommended for snakebites; however, if a tourniquet is already placed, we avoid removal until prepared to manage acute toxicity or immediate hypersensitivity.

Next-generation snake venomics: protein-locus resolution through venom proteome decomplexation.

Venom research has been continuously enhanced by technological advances. High-throughput technologies are changing the classical paradigm of hypothesis-driven research to technology-driven approaches. However, the thesis advocated in this paper is that full proteome coverage at locus-specific resolution requires integrating the best of both worlds into a protocol that includes decomplexation of the venom proteome prior to liquid chromatography-tandem mass spectrometry matching against a species-specific transcriptome. This approach offers the possibility of proof-checking the species-specific contig database using proteomics data. Immunoaffinity chromatography constitutes the basis of an antivenomics workflow designed to quantify the extent of cross-reactivity of antivenoms against homologous and heterologous venom toxins. In the author's view, snake venomics and antivenomics form part of a biology-driven conceptual framework to unveil the genesis and natural history of venoms, and their within- and between-species toxicological and immunological divergences and similarities. Understanding evolutionary trends across venoms represents the Rosetta Stone for generating broad-ranging polyspecific antivenoms.

Use of Antibiotics following Snakebite in the Era of Antimicrobial Stewardship.

Even though there are guidelines for the management of snakebite envenoming (SBE), the use of antibiotics in this pathology remains controversial. The aim of this study is to provide a narrative review of the literature and recommendations based on the best available evidence regarding antibiotic use in SBE. We performed a narrative review of relevant literature regarding SBE and antibiotic use as prophylaxis or treatment. A total of 26 articles were included. There is wide use of antibiotics in SBE; nevertheless, infection was not necessarily documented. The antibiotics used varied according to the study, from beta lactams to lincosamide and nitroimidazoles, and from monotherapy to combined antimicrobials. The most common recommendations were to manage skin and soft tissue infections and avoid infectious complications, but these suggestions are not necessarily based on bacteriological findings. Prophylactic use of antibiotics in SBE is discouraged in most studies. Antibiotic prescription in SBE should be based on the susceptibility of microorganisms isolated from the affected tissue or identified in snakes' oral cavities. Antibiotics should be reserved only for patients with a demonstrated infection, or those at a high risk of developing an infection, i.e., presenting severe local envenoming, local signs of infection, or those with incorrect manipulation of wounds. Prospective studies are needed to correlate microbiological findings at the wound site and the response to antibiotic use.

Lack of controlled studies on snakebite prevention: a rapid review.

Globally, snakebites cause an estimated 80 000-140 000 deaths annually. While there are evidence-based recommendations for managing snakebite victims, recommendations on the prevention of snakebites are limited to expert opinions. We conducted a rapid review to summarise evidence from human studies with a control group on preventing snakebites. Searching PubMed, Web of Science, Scopus, CINAHL and EMBASE with inclusive search terms without language or time limits only yielded three eligible studies (one case control study and two prospective controlled clinical studies), highlighting a knowledge gap. Two studies in Nepal by the same group showed that health education of stakeholders and sleeping under a bednet can significantly reduce snakebite incidence (p<0.05), but these observations are not confirmed elsewhere, and because of the high risk of bias the certainty of evidence was low. The third study from Sri Lanka, which assessed if sleeping above ground would prevent snakebites, had inconclusive results. This demonstrates an urgent need for studies with a control group to guide evidence-based recommendations for snakebite prevention. Potential interventions tested can range from low-cost measures such as wearing appropriate footwear in resource-limited settings to testing the efficacy of chemical, biological (e.g. rodent control) or device-based methods and community-supported platforms tracking snakebite sightings with real-time geolocation data in highly resourced settings.

African polyvalent antivenom can maintain pharmacological stability and ability to neutralise murine venom lethality for decades post-expiry: evidence for increasing antivenom shelf life to aid in alleviating chronic shortages.

INTRODUCTION: Antivenom is a lifesaving medicine for treating snakebite envenoming, yet there has been a crisis in antivenom supply for many decades. Despite this, substantial quantities of antivenom stocks expire before use. This study has investigated whether expired antivenoms retain preclinical quality and efficacy, with the rationale that they could be used in emergency situations when in-date antivenom is unavailable. METHODS: Using WHO guidelines and industry test requirements, we examined the in vitro stability and murine in vivo efficacy of eight batches of the sub-Saharan African antivenom, South African Institute for Medical Research polyvalent, that had expired at various times over a period of 30 years. RESULTS: We demonstrate modest declines in immunochemical stability, with antivenoms older than 25 years having high levels of turbidity. In vitro preclinical analysis demonstrated all expired antivenoms retained immunological recognition of venom antigens and the ability to inhibit key toxin families. All expired antivenoms retained comparable in vivo preclinical efficacy in preventing the lethal effects of envenoming in mice versus three regionally and medically important venoms. CONCLUSIONS: This study provides strong rationale for stakeholders, including manufacturers, regulators and health authorities, to explore the use of expired antivenom more broadly, to aid in alleviating critical shortages in antivenom supply in the short term and the extension of antivenom shelf life in the longer term.