Life-History Modeling Reveals the Ecological and Evolutionary Significance of Autotomy.

AbstractAutotomy, the self-amputation of body parts, serves as an antipredator defense in many taxonomic groups of animals. However, its adaptive value has seldom been quantified. Here, we propose a novel modeling approach for measuring the fitness advantage conferred by the capability for autotomy in the wild. Using a predator-prey system where a land snail autotomizes and regenerates its foot specifically in response to snake bites, we conducted a laboratory behavioral experiment and a 3-year multievent capture-mark-recapture study. Combining these empirical data, we developed a hierarchical model and estimated the basic life-history parameters of the snail. Using samples from the posterior distribution, we constructed the snail's life table as well as that of a snail variant incapable of foot autotomy. As a result of our analyses, we estimated the monthly encounter rate with snake predators at 3.3% (95% credible interval: 1.6%-4.9%), the contribution of snake predation to total mortality until maturity at 43.3% (15.0%-95.3%), and the fitness advantage conferred by foot autotomy at 6.5% (2.7%-11.5%). This study demonstrated the utility of the multimethod hierarchical-modeling approach for the quantitative understanding of the ecological and evolutionary processes of antipredator defenses in the wild.

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.

One Bite, Two Patients: Disparate Clinical Courses Following Simultaneous Crotalus oreganus abyssus Envenomation.

A number of crotaline species have been associated with neurotoxic envenomation in North America. One clinical sign that can occur is myokymia: fine, involuntary, wave-like muscle movements occurring at regular intervals. We report an unusual scenario in which a single snakebite resulted in simultaneous envenomation of 2 patients. Both developed myokymia, with 1 having respiratory compromise. One patient also developed a hypersensitivity reaction to antivenom. Envenomation by the Grand Canyon rattlesnake, Crotalus oreganus abyssus, can produce significant neurotoxicity and resultant respiratory compromise. Antivenom may be helpful but can produce hypersensitivity reactions.

Canine ocular and periocular snakebites requiring enucleation: A report of 19 cases.

PURPOSE: To describe the clinical and histopathologic features secondary to ocular and periocular snakebites in dogs requiring enucleation. METHODS: Retrospective review of patients with recorded snakebite envenomation from the archives of the Comparative Ocular Pathology Laboratory of Wisconsin (COPLOW) and Texas A&M Veterinary Medical Teaching Hospital (1997-2017). The cases included in this study required witnessed snakebites to the dog by the owner, clinical signs supportive of periocular or ocular envenomation, and/or histopathologic lesions compatible with snakebites. Two groups were established: ocular bites (OB) and periocular bites (PB). RESULTS: Nineteen cases were included in the study (OB = 16/19; PB = 3/19). Dogs affected were typically older (median 8 years; range 1-18), and both sexes were equally represented. Left eyes (14/19) were more likely to sustain snake-induced trauma compared to right eyes (5/19). Fifteen breeds were identified, with terriers (9/19) commonly represented. Snakes bites occurred in six US states, with the majority of cases from Texas (7/19), California (5/19), and Arizona (4/19). Common clinical signs included facial edema, corneal ulceration, keratomalacia, uveitis, hyphema, and secondary glaucoma. All eyes demonstrated vision loss prior to enucleation. Histologically, the ocular and periocular tissues contained extensive necrosis associated with envenomation. Retinal detachment, lens capsule rupture, and intraocular hemorrhage/inflammation were commonly found. CONCLUSIONS: Snakebite envenomation is a largely necrotizing disease process that can result in profound infiltrative and destructive ocular changes presumed to be related to the proteolytic factors and necrotoxins in venom. Ocular alterations secondary to snakebites may be irreversible regardless of supportive therapy instituted.

Perspective on the Therapeutics of Anti-Snake Venom.

Snakebite envenomation is a life-threatening disease that was recently re-included as a neglected tropical disease (NTD), affecting millions of people in tropical and subtropical areas of the world. Improvement in the therapeutic approaches to envenomation is required to palliate the morbidity and mortality effects of this NTD. The specific therapeutic treatment for this NTD uses snake antivenom immunoglobulins. Unfortunately, access to these vital drugs is limited, principally due to their cost. Different ethnic groups in the affected regions have achieved notable success in treatment for centuries using natural sources, especially plants, to mitigate the effects of snake envenomation. The ethnopharmacological approach is essential to identify the potential metabolites or derivatives needed to treat this important NTD. Here, the authors describe specific therapeutic snakebite envenomation treatments and conduct a review on different strategies to identify the potential agents that can mitigate the effects of the venoms. The study also covers an increased number of literature reports on the ability of natural sources, particularly plants, to treat snakebites, along with their mechanisms, drawbacks and future perspectives.

Retrospective Documentation of a Confirmed White-Lipped Green Pit Viper (Trimeresurus albolabris Gray, 1842) Bite in the South-Central Hills of Nepal.

This case report documents envenomation by an arboreal white-lipped green pit viper (Trimeresurus albolabris), a species found in South and Southeast Asia that causes the majority of venomous snakebites among Southeast Asian pit vipers. Clinical features vary from asymptomatic to serious coagulopathy that may progress into life-threatening or fatal hemorrhage. The proven life-threatening cases described in published literature, however, are sparse. Practically, no specific antivenom targeted to pit viper bites is available in Nepal. We report a case (managed with several non-evidence-based interventions) of noticeable coagulopathic envenomation due to confirmed T albolabris bite in Nepal. This is the first known reported case of such a bite in Nepal. This case highlights the urgent need to improve diagnosis, monitoring, and supportive care for bite victims and to study the effectiveness of Thai pit viper antivenoms for the treatment of T albolabris envenomations.

Severe Systemic Envenomation Following Vipera berus Bite Managed with ViperaTAb Antivenom.

Bites by the European adder (Vipera berus) are a rare medical emergency in the UK with 20 to 50% of an estimated 50 to 200 cases per year necessitating treatment with antivenom. We present a case demonstrating both severe systemic and local effects necessitating out of hospital resuscitation, vasopressor support, and prolonged rehabilitation. Hemodynamic stability was restored promptly after administration of ViperaTAb antivenom, the first published case of its use in the UK.

Proteomic Analysis and Immuno-Profiling of Eastern India Russell's Viper ( Daboia russelii) Venom: Correlation between RVV Composition and Clinical Manifestations Post RV Bite.

The proteomes of Russell's viper venom (RVV) from Burdwan (RVV B) and Nadia (RVV N), the two districts of West Bengal, eastern India (EI), were investigated by gel-filtration chromatography (GFC) followed by tandem mass spectrometry of tryptic fragments of the fractions. A total of 73 and 69 proteins belonging to 15 snake venom protein families were identified in RVV B and RVV N, respectively, by MS/MS search against Viperidae (taxid 8689) protein entries of the nonredundant NCBI database. The minor differences in venom composition of both the EI RV were established unequivocally by their biochemical and pharmacological properties and by SDS-PAGE, gel filtration chromatography, and LC-MS/MS analyses. The composition of EI RVVs was well correlated with published reports on the pathophysiology of RV-envenomed patients from this part of the country. Venom-antivenom cross-reactivity determined by ELISA, Western blotting, and antivenomics approaches demonstrated poor recognition of low molecular mass (<20 kDa) RVV proteins by commercial polyvalent antivenoms, which was substantiated by neutralization of RVV enzymes by antivenom.

Metalloproteinases in disease: identification of biomarkers of tissue damage through proteomics.

INTRODUCTION: Metalloproteinases play key roles in health and disease, by generating novel proteoforms with variable structure and function. Areas covered: This review focuses on the role of endogenous [a Disintegrin and Metalloproteinase (ADAMs), ADAMs with thrombospondin motifs (ADAMTS), and matrix metalloproteinases (MMPs)] and exogenous metalloproteinases in various disease conditions, and describes the application of mass spectrometry-based proteomics to detect qualitative and quantitative changes in protein profiles in tissues and body fluids in disease. Emphasis is placed on the proteomic analysis of exudates collected from affected tissues, including methods that enrich newly generated protein fragments derived from proteolysis in cells, stroma, or extracellular matrix. The use of proteomic analysis of exudates in the study of the local tissue damage induced by metalloproteinases derived from viperid snake venoms is discussed, particularly in relation to extracellular matrix degradation and to the overall pathology of these envenomings. Expert commentary: The information provided by these proteomics approaches is paving the way for the identification of biomarkers based on particular proteolytic signatures associated with different pathologies. Together with other methodological approaches, a comprehensive view of the mechanisms and dynamics of diseases can be achieved. Such basis of knowledge allows for the design of novel diagnostic and therapeutic approaches within the frame of 'precision' or 'personalized' medicine.

Inactivation of Venom PLA2 Alleviates Myonecrosis and Facilitates Muscle Regeneration in Envenomed Mice: A Time Course Observation.

Snake venom is a complex cocktail of toxins which induces a series of clinical and pathophysiological manifestations in victims, including severe local tissue damage and systemic alterations. Deinagkistrodon acutus (D. acutus) ranks among the "big four" life-threatening venomous species in China, whose venom possesses strong myotoxicity and hematotoxicity that often lead to permanent disability or muscle atrophy. Varespladib, an inhibitor of mammalian phospholipase A2 (PLA2), has been recently reproposed as an effective antidote against snakebite envenomation. The present study aimed at evaluating the protective role of varespladib on muscle regeneration in envenomed mice. Mice were grouped and subjected to inoculation with D. acutus venom or a mixture of venom and varespladib or control vehicle in the gastrocnemius muscle. Local injuries including hemorrhage, myonecrosis, ulceration, and systemic damages including general dysfunction, visceral failure, and inflammatory responses were observed at 1, 3, 7, 14, and 21 days. The results indicated that most of the muscle myonecrosis and hemorrhage were alleviated by varespladib. Besides, the pretreated mice recovered rapidly with lesser atrophy and muscle fibrosis. In conclusion, the findings of the present study suggested that varespladib is an effective antidote that could neutralize D. acutus venom and allow for earlier and improved rehabilitation outcome.

Effects of photobiomodulation therapy on Bothrops moojeni snake-envenomed gastrocnemius of mice using enzymatic biomarkers.

Bothropic venom contains a range of biologically active substances capable of causing severe local and systemic envenoming symptomatology within its victims. The snake anti-venom is effective against systemic effects but has no neutralizing effect against the fast developing local effects. Herein, mice gastrocnemius injected with Bothrops moojeni venom (40 µg/kg) or saline solution were irradiated with HeNe (632.8 nm) and GaAs (904 nm) lasers (daily energy density of 4 J/cm2; 0.03/0.21 power density; 0.07/0.16 spot size; 1.2/0.04 total energy, 1 cm off contact, for HeNe and GaAs lasers, respectively) and euthanized in periods ranging from 3 h to 21 days. Blood biochemistry for creatine kinase (CK), alkaline phosphatase (ALP), acid phosphatase (AP), lactate dehydrogenase (LDH), aspartate transaminase (AST), and myoglobin and histopathological analysis, for assessing the degree of myonecrosis and regeneration of gastrocnemius, were done at every time interval. GaAs laser promoted faster photobiomodulation therapy (PBMT) effects, and the GaAs group exhibited a better clinical outcome than the HeNe group. Within the GaAs group, the serum levels of CK, LDH, AP, AST, and myoglobin, which were increased by the physiological effects of the venom, were reduced to initial baseline before snake envenomation in less time than those irradiated by the HeNe laser. However, the group receiving irradiation from the HeNe laser returned the levels of ALP activity to baseline faster than those of the GaAs group. Histopathological analysis revealed enhanced muscle regeneration in mice groups treated with both lasers. PBM promoted by GaAs and HeNe showed well-developed centrally nucleate regenerating cells and an increased number of newly formed blood vessels when compared to unirradiated muscle. We therefore suggest that GaAs had the best outcomes likely derived from a deeper penetrating longer wavelength. We conclude that PMBT is a promising, non-invasive approach to be further tested in pre-clinical studies with a goal to further its clinical use in skeletal muscle recovery in snakebite victims.

PRESUMPTIVE PIT VIPER ENVENOMATION IN PSITTACINES IN A BRAZILIAN ZOO.

Snake bites represent a serious public health risk in many regions of the globe, especially in tropical areas. Clinical signs and postmortem changes are well described in human and other mammalian species. However, detailed case reports about venomous snake attacks in avian species are limited. This report describes presumptive fatal envenomations in three psittacines caused by pit vipers in a Brazilian zoo. In one case, a Brazilian lancehead (Bothrops moojeni) was captured in the aviary. In all three cases the dermis around the suspected snake bite area exhibited hemorrhages and edema. Histologically, degeneration and necrosis of subcutaneous musculature were observed. Lung, heart, and kidneys displayed focal hemorrhages. The local changes are similar to those described for mammalian patients including humans. However, except for the parenchymatous hemorrhages, additional external and internal gross and histopathological lesions were missing. After ruling out other causes, such as aggression and dicoumarinic intoxication, the presumptive diagnosis of snake envenomation was made. The smaller size and variabilities of pathophysiological effects of the venom in parrots might explain the different lesion patterns observed, compared with mammals. Possibly, the birds may have reacted differently to envenomation by pit vipers and died before the venom could cause macroscopic and histological changes often observed in mammals.

Histopatholological Findings in a Fatal Case of Rinkhals Envenomation: A New Forensic Pathology Finding?

Although cobra envenomation continues to pose a threat to life and therefore constitutes a major cause of death in Africa, India, and Sri Lanka, the forensic pathology reports are not always histologically well documented or illustrated. The reports in the literature, although providing graphic evidence of the macroscopic appearances of the bite and its ensuing tissue necrosis, should the victim survive, often do not illustrate the histopathological findings to a sufficient degree. We present an unusual case of fatal envenomation by a rinkhals (Hemachatus haemachatus) in which the victim had been bitten a second time after a previous rinkhals' bite. Two issues therefore arose: (a) whether the histopathological findings in the spleen were an acute reaction to the snake bite or were due to immunostimulation as a consequence of the previously inflicted bite, and (b) the previously undocumented finding of extravasation of erythrocytes into the surrounding Virchow-Robin spaces in the brain, a finding usually associated with blunt head trauma and therefore interpreted as a cortical contusion hemorrhage by forensic pathologists.

Comparative Studies of Structural and Functional Properties of Snake Venom Metalloproteinases.

Snake venom metalloproteinases (SVMPs) induces local and systemic effects on patients suffering from snakebite, degrading extracellular matrix (ECM) proteins such as collagen, gelatin, elastin, laminin, fibronectin, nidogen (entactin), and thrombospondin that cause local hemorrhage and tissue damage. They cleave or activate coagulation factors such as fibrinogen, fibrin, prothrombin, factor V, factor IX, factor X and protein C that bring about systemic coagulopathy. SVMPs and their truncated forms cleave or interfere with platelet adhesive proteins such as vWF, fibrinogen and collagen, and cleave or interfere with platelet receptors such as GPVI, alpha2beta1, GPIb, GPIX, and GPIIbIIIa that result in platelet aggregation defect. SVMPs induce cancer cell line to form morphological changes and apoptosis in vitro concordant with skin necrosis after snakebite in some cases. These local effects caused by SVMPs have no certain treatments, even with commercial anti-venom. SVMPs researches are focusing on their inhibitors, measurement and replacement of blood coagulation factor defects, or anti-cancer drug.

Marine envenomations.

BACKGROUND: Marine stings are common but most are minor and do not require medical intervention. Severe and systemic marine envenoming is uncommon, but includes box jellyfish stings, Irukandji syndrome, major stingray trauma and blue-ringed octopus envenoming. Almost all marine injuries are caused by jellyfish stings, and penetrating injuries from spiny fish, stingrays or sea urchins. OBJECTIVE: This article describes the presentation and management of marine envenomations and injuries that may occur in Australia. DISCUSSION: First aid for jellyfish includes tentacle removal, application of vinegar for box jellyfish, and hot water immersion (45°C for 20 min) for bluebottle jellyfish stings. Basic life support is essential for severe marine envenomings that result in cardiac collapse or paralysis. Irukandji syndrome causes severe generalised pain, autonomic excess and minimal local pain, which may require large amounts of analgesia, and, uncommonly, myocardial depression and pulmonary oedema occur. Penetrating marine injuries can cause significant trauma depending on location of the injury. Large and unclean wounds may have delayed healing and secondary infection if not adequately irrigated, debrided and observed.

Snake Envenomation to the Face of a Child--Rare Case.

Snakebites are seen in summer season in the southern part of Turkey, including Hatay province. In average of 40 patients with snakebites are admitted to our hospital every year. Viper is the most common venomous snakes in our region. Their hemotoxins and necrotoxins lead to local or systemic tissue damage and is responsible for the mortality and morbidity. In this report, we described a rare pediatric case, a six-year-old boy having been bitten on the left side of his face when he was looking around from their home's balcony. The patient was orotracheally intubated and mechanically ventilated because of airway obstruction due to severe edema. 12 flacon of anti-snake venom, mannitol infusion, fresh frozen plasma, erythrocytes suspension and antibiotherapy were administered to the patient. Seven days after the admission, clinical and laboratory findings were improved and the patient was discharged in a good condition. Snakebites inflicted on face and neck areas may cause rapidly progressive edema in respiratory tract and lead to life-threatening conditions. Therefore early orotracheal intubation is very important to prevent mortality.

Effect of externally applied jidesheng anti-venom on skin and soft-tissue necrosis after Chinese cobra bite: a retrospective study.

OBJECTIVE: To evaluate the effects of Jidesheng anti-venom used externally for skin and soft-tissue necrosis from Chinese cobra bite. METHODS: A retrospective review was performed according to the clinical data recorded from January 2002 to December 2012. A total of 126 patients (116 females and 10 males) with skin and soft-tissue necrosis due to Chinese cobra bite were divided into two groups. The control group was treated externally with 40% glyceride magnesium sulfate (n = 52), and the treatment group was given Jidesheng anti-venom externally (n = 74). The data collected included maximum local necrotic area of skin and soft tissues, de-tumescence time, healing time, and skin-grafting rate. RESULTS: There were no significant differences in gender, age, and disease condition between the control and treatment groups (P > 0.05). No statistically significant difference was found in de-tumescence time between the two groups (P > 0.05). The maximum local necrotic area of skin and soft tissues was (19.9 +/- 7.3) cm2 in the treatment group, while it was (23.3 +/- 6.4) cm2 in the control group. The healing time of the treatment group was shorter than that of the control group [(32.1 +/- 3.7) vs (34.4 +/- 4.5) days)]. The skin-grafting rate in the treatment group was lower than that of the control group (10.81% vs 25.00%). There were statistically significant differences in maximum local necrotic area of skin and soft tissues, healing time, and skin-grafting rate between the control and treatment groups (all P < 0.05). CONCLUSION: External application of Jidesheng anti-venom may help to promote wound healing and reduce the skin-grafting rate in cases of skin and soft-tissue necrosis due to Chinese cobra bite.

The establishment and evaluation of a swine model of deinagkistrodon acutus snakebite envenomation.

OBJECTIVE: To establish a preclinical large-animal model of Deinagkistrodon acutus snakebite envenomation and evaluate its feasibility. METHODS: The venom of D. acutus (0 mg/kg, 1 mg/kg, 2 mg/kg, 5 mg/kg, or 10 mg/kg) was injected into the left biceps femoris of 11 male pigs. Then, the circumferences of the limbs were regularly measured, and changes in muscle injury biomarkers, blood parameters, coagulation function, vital organ function and injury biomarkers were regularly detected. At 24 h after venom injection, the animals were euthanized, and the pathological damage to the vital organs mentioned above was evaluated. RESULTS: The two pigs receiving 10 mg/kg and 5 mg/kg snake venom died at 8 h and 12 h after injection, respectively. The remaining pigs were equally divided into 0 mg/kg, 1 mg/kg, and 2 mg/kg snake venom groups, and all of them survived to 24 h after injection. Compared with the pigs receiving 0 mg/kg snake venom, the pigs receiving 1 mg/kg or 2 mg/kg snake venom exhibited significant abnormities, including limb swelling; increased muscle injury biomarker creatine kinase (CK) and coagulation function indicators prothrombin time and D-dimer; and decreased blood routine indicator platelet and coagulation function indicator fibrinogen. Moreover, significant abnormalities in myocardial and cerebral function and injury biomarkers in the heart, brain, liver, kidney and intestine were also observed. In particular, the abnormalities mentioned above were significantly obvious in those pigs receiving 2 mg/kg snake venom. Pathological evaluation revealed that the morphology of muscle, heart, brain, liver, kidney, and intestine in those pigs receiving 0 mg/kg snake venom was normal; however, pathological damage was observed in those pigs receiving 1 mg/kg and 2 mg/kg snake venom. Similarly, the pathological damage was more severe in those pigs receiving 2 mg/kg snake venom. CONCLUSION: The intramuscular injection of 2 mg/kg D. acutus venom seems to be an optimal dose for examining the preclinical efficacy of existing and novel therapeutics for treating D. acutus envenomation in pigs.

Investigating Snake-Venom-Induced Dermonecrosis and Inflammation Using an Ex Vivo Human Skin Model.

Snakebite envenoming is a neglected tropical disease that causes >100,000 deaths and >400,000 cases of morbidity annually. Despite the use of mouse models, severe local envenoming, defined by morbidity-causing local tissue necrosis, remains poorly understood, and human-tissue responses are ill-defined. Here, for the first time, an ex vivo, non-perfused human skin model was used to investigate temporal histopathological and immunological changes following subcutaneous injections of venoms from medically important African vipers (Echis ocellatus and Bitis arietans) and cobras (Naja nigricollis and N. haje). Histological analysis of venom-injected ex vivo human skin biopsies revealed morphological changes in the epidermis (ballooning degeneration, erosion, and ulceration) comparable to clinical signs of local envenoming. Immunostaining of these biopsies confirmed cell apoptosis consistent with the onset of necrosis. RNA sequencing, multiplex bead arrays, and ELISAs demonstrated that venom-injected human skin biopsies exhibited higher rates of transcription and expression of chemokines (CXCL5, MIP1-ALPHA, RANTES, MCP-1, and MIG), cytokines (IL-1ß, IL-1RA, G-CSF/CSF-3, and GM-CSF), and growth factors (VEGF-A, FGF, and HGF) in comparison to non-injected biopsies. To investigate the efficacy of antivenom, SAIMR Echis monovalent or SAIMR polyvalent antivenom was injected one hour following E. ocellatus or N. nigricollis venom treatment, respectively, and although antivenom did not prevent venom-induced dermal tissue damage, it did reduce all pro-inflammatory chemokines, cytokines, and growth factors to normal levels after 48 h. This ex vivo skin model could be useful for studies evaluating the progression of local envenoming and the efficacy of snakebite treatments.