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• Purified enzyme isoforms had differential coagulotoxic activity. • This variation was linked to differential ability to activate Factor X vs prothrombin. • This variation was linked to differential antiveom efficacy. Bothrops (lance-head pit vipers) venoms are rich in weaponised metalloprotease enzymes (SVMP). These toxic enzymes are structurally diverse and functionally versatile. Potent coagulotoxicity is particularly important for prey capture (via stroke-induction) and relevant to human clinical cases (due to consumption of clotting factors including the critical depletion of fibrinogen). In this study, three distinct isoforms of P-III class SVMPs (IC, IIB and IIC), isolated from Bothrops neuwiedi venom, were evaluated for their differential capacities to affect hemostasis of prey and human plasma. Furthermore, we tested the relative antivenom neutralisation of effects upon human plasma. The toxic enzymes displayed differential procoagulant potency between plasma types, and clinically relevant antivenom efficacy variations were observed. Of particular importance was the confirmation the antivenom performed better against prothrombin activating toxins than Factor X activating toxins, which is likely due to the greater prevalence of the former in the immunising venoms used for antivenom production. This is clinically relevant as the enzymes displayed differential potency in this regard, with one (IC) in particular being extremely potent in activating Factor X and thus was correspondingly poorly neutralised. This study broadens the current understanding about the adaptive role of the SVMPs, as well as highlights how the functional diversity of SVMP isoforms can influence clinical outcomes. Key Contribution: Our findings shed light upon the hemorrhagic and coagulotoxic effects of three SVMPs of the P-III class, as well as the coagulotoxic effects of SVMPs on human, avian and amphibian plasmas. Antivenom neutralised prothrombin-activating isoforms better than Factor X activating isoforms. [ABSTRACT FROM AUTHOR]

Abstract Envenomation by scorpions of the genus Tityus is an important public health problem in Argentina, involving near 8000 stings and 2 deaths each year. Treatment for envenomation is the use of specific antivenom and intensive hospital care. Antivenom is produced by the Ministry of Health and freely distributed throughout the country. For antivenom production it is necessary to collect scorpion venom, which is a difficult task because although scorpions can be found in Argentina, they are less abundant than in warmer latitudes. For this reason venom collection constitutes a bottleneck for antivenom production. Although in Argentina several species of Tityus can be found, most of the accidents are caused by Tityus trivittatus, and the venom of this scorpion has historically been the venom used for antivenom production. We analyzed retrospectively 26 pools of telson homogenates (6964 telsons) and 37 pools of milked venom obtained by electrical stimulation (equivalent to 6841 milkings). Lethal potencies of samples from different provinces were very similar, although venom from scorpions of Buenos Aires city showed the lowest potency. The venom obtained by milking (median LD 50 12.3 μg), provided batches containing LD 50 s more potent when compared with the venom obtained from telson homogenates (p < 0.0001). Many batches of telson homogenates (30%) showed lower potencies than acceptable for antivenom production and control. In addition to the study of the venom yield, the records of immunization of horses, the potency of the batches and the protein content of each batch of anti-scorpion antivenom produced were analyzed, comparing those produced using milked venom with those using telson homogenates as immunogens. Batches produced using milked venom required a shorter period of immunization (p < 0.0001), rendered higher neutralizing titers (p 0.0350) and possessed lower protein content (p 0.0092). Results clearly showed that the milking of scorpions is a more efficient tool to obtain venom for antivenom production in comparison to the use of telson homogenates. Highlights • We analyzed 26 pools of telson homogenates (6964 telsons) and 37 of milked venom (6841 milkings). • The potency and the number of LD 50 s per milking were higher that those of telson homogenates. • Low toxic variability between regions was found. • 30% of the batches of homogenates did not posses quality enough to be used for antivenom production. • Milked venoms present advantages over telson homogenates for antivenom production. [ABSTRACT FROM AUTHOR]

With the threat of a post-antibiotic era looming, the search for new and effective antibiotics from novel sources is imperative. Not only has crude snake venom been shown to be effective, but specific components within the venoms, such as Phospholipase A 2 s and l -amino acid oxidases have been isolated and demonstrated to be effective as well. Despite numerous studies being completed on snake venoms, there is a heavy bias towards utilizing the venoms from the highly toxic Elapidae and Viperidae species. Very few studies have been conducted on the less toxic, but taxonomically more diverse, Colubridae. Furthermore, an extensive review of the literature examining the efficacy and potential specificity of these venoms has not been completed. Therefore, the aims of this study were to elucidate any similarities in snake venoms as well as investigate the efficacy of snake venom antimicrobial properties towards morphologically and metabolically diverse microbial classes and the prevalence of snake species with antimicrobial properties within each snake family. The results indicate that snake venoms and their isolated components are powerful antimicrobial agents but vary in efficacy towards different microbial classes. Furthermore, due to similarities in venom composition, and limited preliminary studies, the less toxic Colubridae family may be a fruitful area of research to find novel antimicrobial agents that are less harmful to humans. [ABSTRACT FROM AUTHOR]

Background: Honey bee venom is an important toxin which has various useful properties. Bee venom possesses various peptides including melittin, apamin, adolapamin and mast cell degranulating peptide. It appears to stimulate cortisone secretion, enhances antibody production, and affects cytokine production. Aim: The aim of the work is to study changes in levels of complement system proteins, C3 and C4, together with C-reactive proteins and rheumatoid factors (CRP and RF) in response to bee venom in subjects exposed naturally to sting with honey bee workers. Subjects and Methods: Subjects (12) were randomly selected from patients visiting El-Mostafa bee house, Elmarg, cairo, Egypt; to get natural bee venom therapy for various diseases. Blood samples (24) were collected from these volunteers, before and after honey bee sting, at the laboratory of Egypt Air Hospital, Cairo, Egypt following the standard protocol. Serum levels of C3 and C4 were measured by radial immunodiffusion technique. CRP was quantitatively measured by in vitro diagnostic test using auto analyzer (Dimension® EXEL) clinical chemistry system. Serum RF was measured by Rapid latex agglutination test for qualitative screening. Result: After exposure of patients to bee sting: all abnormal levels of C3 returned to normal values while abnormal C4 levels did not change; Half of cases that were showing abnormal high CRP levels have dropped to normal levels and All RF positive cases have become negative. Conclusion: The complement system has been activated, in patients, by both classical and alternative pathways before treatment with bee venom where it became only classically activated after treatment. Improved values of both CRP and RF indicate reduction in the inflammatory immune response after exposure of patients to honey bee venom. [ABSTRACT FROM AUTHOR]

Different types of neuronal nicotinic acetylcholine receptors (nAChRs) are expected to occur in vivo, most structure-activity relationship studies have been carried out for just a few neuronal subtypes. The present review enlightens current aspects of venom modulators of nAChRs. Important electronic databases such as PubMed or Google scholar were explored for the collection of latest studies in the field. Clinical and basic research has shown that cholinergic receptors play a role in several disorders of the nervous system such as chronic pain, Alzheimer's disease and addiction to nicotine, alcohol and drugs. Unfortunately, the lack of selective modulators for each subtype of nAChR makes their pharmacological characterization difficult, which has slowed the development of therapeutic nAChR modulators with high selectivity and absence of off-target side-effects. Animal venoms have proven to be an excellent natural source of bioactive molecules with activity against ion channels. The present review concludes that the presence of small-molecule nAChR modulators in spider venoms support the use of venoms as a potential source of novel modulators. [ABSTRACT FROM AUTHOR]

The encyrtid parasitoid, Diversinervus elegans (Hymenoptera: Encyrtidae), is a natural enemy of the notorious scale pests belonging to the family of Coccidae. Venom containing a rich source of bioactive molecules is a key virulent factor used to regulate host physiology by parasitoids. Although knowledge regarding venom constituents accumulated from limited parasitoids has provided insights into their roles in host-parasitoid interaction, toxins involving in manipulating scale physiology remain sparsely documented. Here, a total number of 48 putative venom proteins were identified from D. elegans using an integrative transcriptomic and proteomic approach. The majority of them such as serine protease, esterase, and major royal jelly protein have been found in venom of other several parasitoid species. Several venom proteins including three novel proteins having unknown function were firstly revealed. Quantitative real time PCR analysis demonstrated that 16 venom genes displayed female-biased expression, which might be important for parasitism success. These data enrich our understanding of parasitoid venom evolution and diversity, and will undoubtedly help deciphering functional venom proteins as potential candidates for pest control. [ABSTRACT FROM AUTHOR]

Small membranous vesicles are small closed fragments of membrane. They are released from multivesicular bodies (exosomes) or shed from the surface membrane (microvesicles). They contains various bioactive molecules and their molecular composition varies depending on their cellular origin. Small membranous vesicles have been identified in snake venoms, but the origin of these small membranous vesicles in the venom is controversial. The aim of this study was to verify the origin of the small membranous vesicles in venom of Crotalus durissus terrificus by morphological analyses using electron microscopy. In addition, the protein composition of the vesicles was analyzed by using a proteome approach. The small membranous vesicles present in the venom were microvesicles, since they originated from microvilli on the apical membrane of secretory cells. They contained cytoplasmic proteins, and proteins from the plasma membrane, endoplasmic reticulum (ER), and Golgi membrane. The release of microvesicles may be a mechanism to control the size of the cell membrane of the secretory cells after intense exocytosis. Microvesicle components that may have a role in envenoming include ecto-5′-nucleotidase, a cell membrane protein that releases adenosine, and aminopeptidase N, a cell membrane protein that may modulate the action of many peptides. [ABSTRACT FROM AUTHOR]

Scorpion envenomation is a common medical problem in many countries; it is an important cause of morbidity and mortality. The venom of Leiurus quinquestriatus (LQ) is responsible for a number of deaths in children and adults. It has been stated that specific pathophysiological conditions such as generation of oxygen free radicals may trigger the onset of multiple organ dysfunction; therefore, the present study aimed to assess the oxidative stress mediated by LQ crude venom and its effect on the biochemical parameters in rats. Adult male Albino rats (250 ± 30 g body weight) were divided into three groups (n = 5). In control group, rats were intraperitoneally (ip) injected with 50 μL saline solution. Groups 2 and 3 were ip injected with 0.1 mg/kg and 0.2 mg/kg body weight of crude venom, respectively. Blood samples and liver tissues were harvested 1, 2 and 4 h post-injection. Serum levels of glucose, cholesterol, creatinine, urea, uric acid and malondialdehyde increased significantly in envenomed animals within 1, 2 and 4 h post-injection, compared to controls. However, the levels of total serum protein, albumin, globulin and triglycerides as well as catalase, glutathione peroxidase and super oxide dismutase in envenomed rats were significantly decreased compared to controls. We can conclude that LQ crude venom induces oxidative stress via reduction of antioxidant systems and alters some biochemical parameters of envenomed rats. [ABSTRACT FROM AUTHOR]

Context: Although rare, coral snake envenomation is a serious health threat in Brazil, because of the highly neurotoxic venom and the scarcely available antivenom. The major bottleneck for antivenom production is the low availability of venom. Furthermore, the available serum is not effective against all coral snake species found in Brazil. An alternative to circumvent the lack of venom for serum production and the restricted protection of the actually available antivenom would be of great value. We compared the Brazilian coral snake and mono and polyvalent Australian antivenoms in terms of reactivity and protection. Methods: The immunoreactivity of venoms from 9 coral snakes species were assayed by ELISA and western blot using the Brazilian Micrurus and the Australian pentavalent as well as monovalent anti-Notechis, Oxyuranus and Pseudechis antivenoms. Neutralization assays were performed in mice, using 3 LD50 of the venoms, incubated for 30 minutes with 100 μL of antivenom/animal. Discussion: All the venoms reacted against the autologous and heterologous antivenoms. Nevertheless, the neutralization assays showed that the coral snake antivenom was only effective against M. corallinus, M. frontalis, M. fulvius, M. nigrocinctus and M. pyrrhocryptus venoms. On the other hand, the Australian pentavalent antivenom neutralized all venoms except the one from M. spixii. A combination of anti-Oxyuranus and Pseudechis monovalent sera, extended the protection to M. altirostris and, partially, to M. ibiboboca. By adding Notechis antivenom to this mixture, we obtained full protection against M. ibiboboca and partial neutralization against M. lemniscatus venoms. Conclusions: Our findings confirm the limited effectiveness of the Brazilian coral snake antivenom and indicate that antivenoms made from Australian snakes venoms are an effective alternative for coral snake bites in South America and also in the United States were coral snake antivenom production has been discontinued. [ABSTRACT FROM AUTHOR]

Venom secretions from snakes, scorpions, spiders and bees, have been widely applied in traditional medicine and current biopharmaceutical research. Possession of anticancer potential is another novel discovery for animal venoms and toxins. An increasing number of studies have shown the anticancer effects of venoms and toxins of snakes, and scorpions in vitro and in vivo, which were achieved mainly through the inhibition of cancer growth, arrest of cell cycle, induction of apoptosis and suppression of cancer metastasis. However, more evidence is needed to support this concept and the mechanisms of anticancer actions are not clearly understood. The present review is focused on the recant updates on anticancer venom research. [ABSTRACT FROM AUTHOR]

Cyanea nozakii Kishinouy e ( C. nozakii), a giant cnidarian of the class Scyphomedusae, order Semaeostomeae and family Cyaneidae, is widely distributed in the East China Sea, the Yellow Sea and the Bohai Sea, and is abundant from late summer to early autumn. Venom produced by C. nozakii during mass agglomerations can contaminate seawater resulting in death of the halobios and seriously damage commercial fisheries. Swimmers and fishermen commonly suff er painful stings from this jellyfish, resulting in local edema, tingling, breathing difficulties, depressed blood pressure and even death. Such effects arise from the complex mixture of biologically active molecules that make up jellyfish venom. In the present study, the hemolytic activity of venom from tentacles of C. nozakii and factors aff ecting its activity were assayed. The HU ( defined as the amount of protein required to lyse 50 % of erythrocytes) of the venom against dove and chicken erythrocytes was 34 and 59 μg/mL, respectively. Carboxylmethyl chitosan and glycerol could increase hemolytic activity at concentrations greater than 0.06% and 0.2 mol/L, respectively. [ABSTRACT FROM AUTHOR]

The primary functions of venoms from solitary and social wasps are different. Whereas most solitary wasps sting their prey to paralyze and preserve it, without killing, as the provisions for their progeny, social wasps usually sting to defend their colonies from vertebrate predators. Such distinctive venom properties of solitary and social wasps suggest that the main venom components are likely to be different depending on the wasps' sociality. The present paper reviews venom components and properties of the Aculeata hunting wasps, with a particular emphasis on the comparative aspects of venom compositions and properties between solitary and social wasps. Common components in both solitary and social wasp venoms include hyaluronidase, phospholipase A2, metalloendopeptidase, etc. Although it has been expected that more diverse bioactive components with the functions of prey inactivation and physiology manipulation are present in solitary wasps, available studies on venom compositions of solitary wasps are simply too scarce to generalize this notion. Nevertheless, some neurotoxic peptides (e.g., pompilidotoxin and dendrotoxin-like peptide) and proteins (e.g., insulin-like peptide binding protein) appear to be specific to solitary wasp venom. In contrast, several proteins, such as venom allergen 5 protein, venom acid phosphatase, and various phospholipases, appear to be relatively more specific to social wasp venom. Finally, putative functions of main venom components and their application are also discussed. [ABSTRACT FROM AUTHOR]

Venomous animals have evolved with sophisticated bio-chemical strategies to arrest prey and defend themselves from natural predators. In recent years, peptide toxins from venomous animals have drawn considerable attention from researchers due to their surprising chemical, biochemical, and pharmacological diversity. Similar to other venomous animals, centipedes are one of the crucial venomous arthropods that have been used in traditional medicine for hundreds of years in China. Despite signifying pharmacological importance, very little is known about the active components of centipede venoms. More than 500 peptide sequences have been reported in centipede venomous glands by transcriptome analysis, but only a small number of peptide toxins from centipede has been functionally described. Like other venomous animals such as snakes, scorpions, and spiders, the venom of centipedes could be an excellent source of peptides for developing drugs for treatments as well as bio-insecticides for agrochemical applications. Although centipede venoms are yet to be adequately studied, the venom of centipedes as well as their components described to date, should be compiled to help further research. Therefore, based on previous reports, this review focusses on findings and possible therapeutic applications of centipede venoms as well as their components. [ABSTRACT FROM AUTHOR]

The present study evaluated the acute bio-physiological effects of Lethal dose 50 (LD 50) of crude venom of Black Pakistani Cobra, Naja naja karachiensis, collected from Southern Punjab, Pakistan. Adult mice (n=20) were injected intramuscularly with normal saline (n/s) or LD50 of crude venom (1.2mg/kg). Blood was collected by cardiac puncture after anesthesia at intervals 0 (n/s), and 1, 1.5, 2 (LD50 venom) hours. Complete blood count, glucose, serum Alanine aminotransferase (ALT), albumin, total proteins and urea were measured to assess the physiology. One way ANOVA was used to analyze the data (P<0.05). A significant increase in erythrocyte count, its indices and thrombocytosis was observed during the study. Leukocyotsis was observed after a transient leucopenia at 1 hour post envenomation. No significant difference was observed for lymphocytes and monocytes. Serum concentrations of urea, total proteins, albumin and ALT were also increased significantly with all time intervals. Hypoglycemia was observed initially followed by hyperglycemia at 1.5 and 2 hours. The changes indicate that venom has severe hemotoxic potential and disrupts physiological processes by affecting vital organs like kidneys and liver in mice. [ABSTRACT FROM AUTHOR]

Within the Apidae, the largest family of bees with over 5600 described species, the honeybee is the sole species with a well studied venom proteome. So far, only little research has focused on bumblebee venom. Recently, the genome sequence of the European large earth bumblebee ( Bombus terrestris ) became available and this allowed the first in-depth proteomic analysis of its venom composition. We identified 57 compounds, with 52 of them never described in bumblebee venom. Remarkably, 72% of the detected compounds were found to have a honeybee venom homolog, which reflects the similar defensive function of both venoms and the high degree of homology between both genomes. However, both venoms contain a selection of species-specific toxins, revealing distinct damaging effects that may have evolved in response to species-specific attackers. Further, this study extends the list of potential venom allergens. The availability of both the honeybee and bumblebee venom proteome may help to develop a strategy that solves the current issue of false double sensitivity in allergy diagnosis, which is caused by cross-reactivity between both venoms. A correct diagnosis is important as it is recommended to perform an immunotherapy with venom of the culprit species. [ABSTRACT FROM AUTHOR]

Scorpions have been shown to control their venom usage in defensive encounters, depending on the perceived threat. Potentially, the venom amount that is injected could be controlled by reducing the flow speed, the flow duration, or both. We here investigated these variables by allowing scorpions to sting into an oil-filled chamber, and recording the accreting venom droplets with high-speed video. The size of the spherical droplets on the video can then be used to calculate their volume. We recorded defensive stings of 20 specimens representing 5 species. Significant differences in the flow rate and total expelled volume were found between species. These differences are likely due to differences in overall size between the species. Large variation in both venom flow speed and duration are described between stinging events of single individuals. Both venom flow rate and flow duration correlate highly with the total expelled volume, indicating that scorpions may control both variables in order to achieve a desired end volume of venom during a sting. [ABSTRACT FROM AUTHOR]

Parasitoid wasps inject insect hosts with a cocktail of venoms to manipulate the physiology, development, and immunity of the hosts and to promote development of the parasitoid offspring. The jewel wasp Nasonia vitripennis is a model parasitoid with at least 79 venom proteins. We conducted a high-throughput analysis of Nasonia venom effects on temporal changes of 249 metabolites in pupae of the flesh fly host ( Sarcophaga bullata), over a 5-day time course. Our results show that venom does not simply arrest the metabolism of the fly host. Rather, it targets specific metabolic processes while keeping hosts alive for at least 5 days post venom injection by the wasp. We found that venom: (a) activates the sorbitol biosynthetic pathway while maintaining stable glucose levels, (b) causes a shift in intermediary metabolism by switching to anaerobic metabolism and blocking the tricarboxylic acid cycle, (c) arrests chitin biosynthesis that likely reflects developmental arrest of adult fly structures, (d) elevates the majority of free amino acids, and (e) may be increasing phospholipid degradation. Despite sharing some metabolic effects with cold treatment, diapause, and hypoxia, the venom response is distinct from these conditions. Because Nasonia venom dramatically increases sorbitol levels without changing glucose levels, it could be a useful model for studying the regulation of the sorbitol pathway, which is relevant to diabetes research. Our findings generally support the view that parasitoid venoms are a rich source of bioactive molecules with potential biomedical applications. [ABSTRACT FROM AUTHOR]

This study reports on a cytotoxic toxin derived from the venom of the crown-of-thorns starfish Acanthaster planci (CAV). The protein toxin was isolated through both ion-exchange and gel-filtration chromatography, and characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrum analyzes. The CAV was identified as plancitoxin I protein. The mechanistic role of the CAV toxin was explored in human malignant melanoma A375.S2 cell death. The results indicated that after incubation with CAV toxin, cells significantly decreased in A375.S2 cell viability and increased in the lactate dehydrogenase (LDH) level in a dose-dependent manner. The assays indicated that CAV toxin promoted reactive oxygen species (ROS) production, induced nitric oxide (NO) formation, lost mitochondrial membrane potential (ΔΨm) and induced inter-nucleosomal DNA fragmentation in A375.S2 cells. The molecular cytotoxicity of the CAV toxin was tested through evaluation of the apoptosis/necrosis ratio by double staining with annexin V-FITC and a propidium iodide (PI) assay. The results suggested that CAV toxin induced a cytotoxic effect in A375.S2 cells via the apoptotic procedure, and may be associated with the regulation of the p38 pathways. Copyright © 2014 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Abstract: Spiders of the Loxosceles genus represent a risk to human health due to the systemic and necrotic effects of their bites. The main symptoms of these bites vary from dermonecrosis, observed in the majority of cases, to occasional systemic hemolysis and coagulopathy. Although the systemic effects are well characterized, the mechanisms of cell death triggered by the venom of these spiders are poorly characterized. In this study, we investigated the cell death mechanisms induced by the whole venom of the spider Loxosceles similis in human skin fibroblasts. Our results show that the venom initiates an apoptotic process and a caspase cascade involving the initiator caspase-9 and the effector caspases-3, -6, and -7. [Copyright &y& Elsevier]

The members of Scorpaenidae family, including stonefish (genus Synanceja) are among the most dangerous and venomous fishes in the word that inhibit in the tropical shallow waters. The toxinology of the venom of stonefish (genus Synanceja) showed that it is a mixture of proteins, containing several enzymes like hyaluronidase. The crude venom can produce cytolytic, hemolytic, neurotoxic and hypotensive effects in vivo. Immediate increasing intensive pain, local ischemia followed by cyanosis, edema and swelling are the local clinical presentations. Systemic manifestations are not uncommon. Hot water immersion of the injured limb, a local anesthetic agent without adrenaline, infiltrated into and around the wound, debridement of the necrotic tissue and the prescription of prophylactic antibiotic are the main therapeutic measures. The administration of antivenom may be considered. [ABSTRACT FROM AUTHOR]

Abstract: Venoms contain active substances with highly specific physiological effects and are increasingly being used as sources of novel diagnostic, research and treatment tools for human disease. Experimental characterisation of individual toxin activities is a severe rate-limiting step in the discovery process, and in-silico tools which allow function to be predicted from sequence information are essential. Toxins are typically members of large multifunctional families of structurally similar proteins that can have different biological activities, and minor sequence divergence can have significant consequences. Thus, existing predictive tools tend to have low accuracy. We investigated a classification model based on physico-chemical attributes that can easily be calculated from amino-acid sequences, using over 250 (mostly novel) viperid phospholipase A2 toxins. We also clustered proteins by sequence profiles, and carried out in-vitro tests for four major activities on a selection of isolated novel toxins, or crude venoms known to contain them. The majority of detected activities were consistent with predictions, in contrast to poor performance of a number of tested existing predictive methods. Our results provide a framework for comparison of active sites among different functional sub-groups of toxins that will allow a more targeted approach for identification of potential drug leads in the future. [Copyright &y& Elsevier]

Context. Limited information exists on the coagulopathy caused by hump-nosed pit viper ( Hypnale hypnale) envenoming. Objectives. This study aimed to characterise the coagulopathy in hump-nosed pit viper bites by measuring laboratory clotting times and factor studies. Materials and methods. Cases of hump-nosed pit viper envenoming were included from a prospective cohort study of Sri Lankan snake-bite patients. Patient age, sex, snake identification, time of bite and clinical effects were recorded. Patients did not receive anti-venom because no specific anti-venom to hump-nosed vipers exists. All patients received supportive care and serial 20-min whole blood clotting tests (WBCT20). The prothrombin time (PT), international normalised ratio (INR), activated partial thromboplastin time (aPTT), coagulation factors I, II, V, VII, VIII, IX and X, von Willebrand factor (vWF) antigen and D-Dimer concentrations were measured. The median of highest or lowest test result for each patient was reported with interquartile range (IQR). Results. There were 80 hump-nosed pit viper bites, median age was 37 years (IQR: 26-51 years) and 48 were male. The WBCT20 was positive in one patient. The median highest INR was 1.9 (1.5-2.2; Range: 1.3 to > 12) and median highest aPTT was 54 s (46-72 s; Range: 35-170 s). There was low fibrinogen [median: 1.3 g/L;1, -1.8 g/L; Range: < 0.2-2.9], low factor VIII levels [median: 23%; 16-37%] and low factor V levels [median: 43%; 23-74%]. D-Dimer concentrations [median: 3.4 mg/L; 2-7.4 mg/L] were slightly elevated. Factors II, VII and X and vWF antigen concentrations were normal. Discussion and Conclusions. Hump-nosed pit viper bites result in a mild coagulopathy which is usually not detected by a WBCT20. It is characterised by mild elevation of INR, low fibrinogen and Factors V and VIII which may be consistent with the venom containing a thrombin-like enzyme. [ABSTRACT FROM AUTHOR]

Abstract: Animal venoms are complex chemical mixtures that typically contain hundreds of proteins and non-proteinaceous compounds, resulting in a potent weapon for prey immobilization and predator deterrence. However, because venoms are protein-rich, they come with a high metabolic price tag. The metabolic cost of venom is sufficiently high to result in secondary loss of venom whenever its use becomes non-essential to survival of the animal. The high metabolic cost of venom leads to the prediction that venomous animals may have evolved strategies for minimizing venom expenditure. Indeed, various behaviors have been identified that appear consistent with frugality of venom use. This has led to formulation of the “venom optimization hypothesis” (Wigger et al. (2002) Toxicon 40, 749–752), also known as “venom metering”, which postulates that venom is metabolically expensive and therefore used frugally through behavioral control. Here, we review the available data concerning economy of venom use by animals with either ancient or more recently evolved venom systems. We conclude that the convergent nature of the evidence in multiple taxa strongly suggests the existence of evolutionary pressures favoring frugal use of venom. However, there remains an unresolved dichotomy between this economy of venom use and the lavish biochemical complexity of venom, which includes a high degree of functional redundancy. We discuss the evidence for biochemical optimization of venom as a means of resolving this conundrum. [Copyright &y& Elsevier]

Abstract: An investigation into the cardiotoxic effects in human cardiomyocytes of different fractions (as produced from an FPLC) of the venom from Chironex fleckeri showed that whole venom caused cardiac cell death in minutes, measured as cell detachment using xCELLigence technology. However, only one fraction of the venom was responsible for this effect. When all extracted venoms were recombined a similar result was seen for the toxic fraction, however these effects were slower than unfractionated venom alone even though the concentrations were similar. The difference in the results between fractioned and unfractionated venom may have been caused by compounds remaining in the FPLC column, which may interact with the toxic fraction to cause rapid cell detachment or death. [Copyright &y& Elsevier]

In this article, the cytotoxicity of the venom from the nematocysts of jellyfish Cyanea nozakii Kishinouye on human hepatoma cells (Bel-7402, SMMC-7721) and human colon cancer cells (H630) was investigated first. Of the three kinds of cells, the venom had the strongest cytotoxicity on H630 cells with the 50% lethal concentration (IC50) of 5.1 μg/ml. However, the IC50 on Bel-7402 and SMMC-7721 was approximate 17.9 and 24.3 μg/ml, respectively. The cytotoxicity of the venom was affected by pH, temperature and storage conditions. At the pH 4.5–8.5, the venom displayed obvious cytotoxicity and the percentage of survival was about 50%. When pre-incubated at temperatures over 60°C for as short as 10 min, the percentage of survival sharply improved from 4.6% up to 80%. The venom was stored in a more stable condition at −80°C and in lyophilized state compared to other storage conditions used in this study. Lactate dehydrogenase release assay performed on H630 cells indicated that exposure to the venom could result in damage to the cell membrane. [ABSTRACT FROM PUBLISHER]

Abstract: Caterpillar envenomation has been an emergent health issue. Lonomia obliqua is a medically important animal that causes a hemorrhagic syndrome that can progress to acute renal failure, intracranial hemorrhage and death. In the past few years the molecular characterization of L. obliqua venom in addition to experimental models has provided fundamental information to the understanding of the envenomation syndrome. Herein studies from several authors which characterized the complex toxic-pharmacological actions of whole venom are reviewed. [ABSTRACT FROM AUTHOR]

Abstract: Loxoscelism is caused by envenomation by spiders from Loxosceles genus. Clinical symptoms only appear a few hours after envenomation and can evolve in local reactions, such as dermonecrosis, and systemic reactions, such as intravascular haemolysis, intravascular coagulation and renal failure. Current therapies are not effective, often not based in scientific research and can be even detrimental. A lack of understanding of the mechanism of action of the venom of the Loxosceles spider had thus far prevented development of effective therapies. In this review we aim to give an overview of our contributions to the understanding of the mechanism of action of the Loxosceles venom and propose targets and therapeutics for medical intervention. [ABSTRACT FROM AUTHOR]

Abstract: Camelid IgG has been reported to be less immunogenic, less able to activate the complement cascade and more thermostable than IgG from other mammals, and has the ability to bind antigens that are unreactive with other mammalian IgGs. We are investigating whether these attributes of camelid IgG translate into antivenom with immunological and venom-neutralising efficacy advantages over conventional equine and ovine antivenoms. The objective of this study was to determine the preclinical venom-neutralising effectiveness of IgG from camels immunised with venoms, individually or in combination, of the saw-scaled viper, Echis ocellatus, the puff adder, Bitis arietans and the spitting cobra, Naja nigricollis – the most medically-important snake species in West Africa. Neutralisation of the pathological effects of venoms from E. ocellatus, B. arietans and N. nigricollis by IgG from the venom-immunised camels, or commercial antivenom, was compared using assays of venom lethality (ED50), haemorrhage (MHD) and coagulopathy (MCD). The E. ocellatus venom ED50, MHD and MCD results of the E. ocellatus monospecific camel IgG antivenom were broadly equivalent to comparable ovine (EchiTAbG®, MicroPharm Ltd, Wales) and equine (SAIMR Echis, South African Vaccine Producer, South Africa) antivenoms, although the equine antivenom required half the amount of IgG. The B. arietans monospecific camel IgG neutralised the lethal effects of B. arietans venom at one fourth the concentration of the SAIMR polyspecific antivenom (a monospecific B. arietans antivenom is not available). The N. nigricollis camel IgG antivenom was ineffective (at the maximum permitted dose, 100 μl) against the lethal effects of N. nigricollis venom. All the equine polyspecific antivenoms required more than 100 μl to be effective against this venom. The polyspecific camel IgG antivenom, prepared from five camels, was effective against the venom-induced effects of E. ocellatus but not against that of B. arietans and N. nigricollis venoms. No direct correlation was evident between either camel IgG relative avidity or titre and the effectiveness of venom neutralisation in preclinical assays. [Copyright &y& Elsevier]

Testing whether venoms may aid in digestion of the prey, eleven snake venoms were compared for the presence of proteases and endopeptidases that function in alkaline pH conditions. In vitro experiments examined the relative protease and endopeptidase activity of the venoms, which involved combining bovine muscle and snake venom in a buffered solution, encased within dialysis tubing. This mixture was then incubated at room temperature (~20°C) for 24hr, with constant shaking. Bicinchoninic acid (BCA) assay and ninhydrin assay were used to determine peptide and amino acid concentrations. Histological and immunohistochemical investigations using N. kaouthia venom confirmed in vitro findings. Results show that B. arietans venom generated the highest amount of protein/peptides and amino acids in the dialysates, while O. scutellatus, N. ater niger and P . textilis venom did not show any significant protein degradation under alkaline conditions. Histological examination revealed varying degrees of muscle cell damage for each of the venom investigated, and the immunohistochemical study on N. kaouthia venom showed that the venom penetrated the muscle tissue to a significant degree. In vitro assays and histological results indicate that particular venoms may possess the ability to enhance digestion of bovine muscle tissue. [ABSTRACT FROM AUTHOR]

Abstract: Three venom peptides (OdVP1, OdVP2 and OdVP3) were isolated from the venom of the solitary wasp Orancistrocerus drewseni (Hymenoptera: Eumenidae). The mature venom peptide sequences were determined via ESI-Q-TOF MS/MS and by searching the O. drewseni venom gland/sac-specific EST library. All of the OdVPs shared the typical characteristics of amidated C-termini proteins and contained a high content of hydrophobic and positively charged amino acids, suggesting that they adopt an amphipathic α-helical secondary structure, as is the case for mastoparan from Vespula lewisii. The cDNA sequence of the OdVP1 precursor was obtained by 5′- and 3′-rapid amplification of cDNA ends (RACE), and the OdVP2 and OdVP3 precursor transcripts were identified from the venom gland/sac-specific EST library. While the mature peptide sequences were distinct from one another, the overall transcript structure of the OdVPs showed a high homology to that of mastoparan-B from Vespa basalis in that they contained a signal sequence, a prosequence, a mature peptide and a C-terminal glycine. The OdVPs, particularly OdVP2 and OdVP2L (an analog of OdVP2), exhibited strong antifungal activities, but poor antibacterial activities. OdVP2L, which possessed additional Glu–Pro residues, did not have antimicrobial activity against bacteria or Gram-positive yeast but retained activity against Botrytis cinerea. [Copyright &y& Elsevier]

Abstract: Envenomation syndromes induced by Atlantic cnidaria have been tabulated and their therapy discussed. The pharmacokinetics of the venom has been emphasized. Pain occurs instantly and reaches a zenith rapidly in the surf. The nematocysts have already penetrated the dermis and disease proceeds before help can arrive. This fact plus the relative impermeability of human epidermis hampers the efficacy of topical agents. Oral analgesics, the agents of choice, are seldom offered and systemic administration of these drugs is usually not necessary. The use of hot or cold applications has not been settled. Measures to inactivate nematocysts within tentacles adherent to skin post-sting have been demonstrated in vitro, but the clinical significance of these actions has not been shown. Using an abrader (sand, crystalline papain) to counter-irritate nearby skin might provide relief. Antivenoms for Atlantic cnidaria are not available. The instances when verapamil might be employed are very rare. The reasons for the increasing case load of these injuries are discussed and the need to discover more effective therapeutic agents is emphasized. [Copyright &y& Elsevier]

Abstract: Class Cubozoa includes several species of box jellyfish that are harmful to humans. The venoms of box jellyfish are stored and discharged by nematocysts and contain a variety of bioactive proteins that are cytolytic, cytotoxic, inflammatory or lethal. Although cubozoan venoms generally share similar biological activities, the diverse range and severity of effects caused by different species indicate that their venoms vary in protein composition, activity and potency. To date, few individual venom proteins have been thoroughly characterised, however, accumulating evidence suggests that cubozoan jellyfish produce at least one group of homologous bioactive proteins that are labile, basic, haemolytic and similar in molecular mass (42–46kDa). The novel box jellyfish toxins are also potentially lethal and the cause of cutaneous pain, inflammation and necrosis, similar to that observed in envenomed humans. Secondary structure analysis and remote protein homology predictions suggest that the box jellyfish toxins may act as α-pore-forming toxins. However, more research is required to elucidate their structures and investigate their mechanism(s) of action. The biological, biochemical and molecular characteristics of cubozoan venoms and their bioactive protein components are reviewed, with particular focus on cubozoan cytolysins and the newly emerging family of box jellyfish toxins. [Copyright &y& Elsevier]

Abstract: Inflammation is a major characteristic of envenomation by snakes from viperine and crotaline species. Bothrops asper snake venom elicits, among other alterations, a pronounced inflammatory response at the site of injection both in humans and experimental animals. This review describes the current status of our understanding of the inflammatory reaction, including pain, triggered by B. asper venom. The experimental studies on the action of this venom as well as the complex network of chemical mediators involved are summarized. Moreover, aspects of the molecular mechanisms orchestrating this important response to envenomation by B. asper are presented. Considering that isolated toxins are relevant tools for understanding the actions of the whole venom, studies dealing with the mechanisms of inflammatory and nociceptive properties of phospholipases A2, a metalloproteinase and serine proteinases isolated from B. asper venom are also described. [Copyright &y& Elsevier]

Abstract: Monovalent antivenoms have a lower volume of specific antibodies that may reduce reactions but require accurate snake identification to be used. Polyvalent antivenoms are larger volume and may have a higher reaction rate. However, they avoid the problem of snake identification and may be more cost-effective to manufacture. We have previously shown cross-neutralisation of two Australian elapid venoms, tiger snake (Notechis scutatus) and brown snake (Pseudonaja textilis) venoms, by their respective monovalent antivenoms. In this study enzyme immunoassays were used to quantify the amount of monovalent antivenom (quantity of monovalent antibodies to a specific snake venom) in vials of commercially produced antivenom in Australia. All antivenoms tested appeared to be polyvalent and contain varying amounts of all five terrestrial snake monovalent antibodies based on their binding to the five representative venoms. Redback spider antivenom did not have any measurable binding affinity for any of the five snake venoms, showing that the observed binding is not due to non-specific interactions with equine protein. The antivenoms had expiry dates over a 15year period, suggesting that the antivenoms have been mixtures for at least this time. This study cannot be used to rationalise hospital stocks of antivenom in Australia because there is no guarantee that the antivenoms will remain as mixtures. However, it would be possible for the manufacturer to reduce the number of types of snake antivenoms available in Australia to two polyvalent antivenoms which would simplify treatment of snakebite. [Copyright &y& Elsevier]

Abstract: The recent bloom of a giant jellyfish Nemopilema nomurai has caused a danger to sea bathers and fishery damages in the waters of China, Korea, and Japan. The present study investigated the cytotoxic and hemolytic activities of crude venom extract of N. nomurai using a number of in vitro assays. The jellyfish venom showed a much higher cytotoxic activity in H9C2 heart myoblast than in C2C12 skeletal myoblast (LC50 =2 µg/mL vs. 12 µg/mL, respectively), suggesting its possible in vivo selective toxicity on cardiac tissue. This result is consistent with our previous finding that cardiovascular function is a target of the venom. In order to determine the stability of N. nomurai venom, its cytotoxicity was examined under the various temperature and pH conditions. The activity was relatively well retained at low environmental temperature (≤20 °C) and dramatically lost at high temperature (≥60 °C). In pH stability test, the venom has abruptly lost its activity at low pH environment (pH≤4). Interestingly enough, however, its activity was not significantly affected even at the highest pH environment tested (pH≤12) in the present study. Additionally, hemolytic activity of the venom was examined using the erythrocytes of cat, dog, human, rabbit and rat. Venom concentration-dependent hemolysis could be observed from 10 µg/mL of protein equivalents or higher with variable potencies in different species, among which dog erythrocyte was the most susceptible to the venom (EC50 =151 µg/mL). SDS-PAGE analysis of N. nomurai venom showed the molecules of 20–40 kDa and 10–15 kDa appeared to be the major protein components of the venom. [Copyright &y& Elsevier]

Abstract: We report the first documented death due to a coral snake (Micrurus species) in the United States (U.S.) in over 40 years. The victim failed to seek medical care following the bite of an eastern coral snake (Micrurus fulvius) and succumbed within hours. Post-mortem proof of envenomation was obtained using an ELISA (enzyme-linked immunosorbent assay) developed specifically for this investigation. U.S. coral snakes are briefly reviewed in terms of their venom compositions, their clinical effects, and proper pre-hospital and hospital management. The clinical significance of the impending absence of commercially available antivenom for coral snake bites in the U.S. is highlighted. [Copyright &y& Elsevier]

Snake venoms are complex mixtures of proteins, which affect the vital biologic systems of prey, as well as humans. Envenomation leads to immobilization by paralysis, cardiac, and circulatory failure. These same venom proteins that cause havoc in the physiologic system could be used as therapeutic agents. Disintegrins and disintegrin-like proteins are molecules found in the venom of four snake families (Atractaspididae, Elapidae, Viperidae, and Colubridae). The disintegrins are non-enzymatic proteins that inhibit cell–cell interactions, cell–matrix interactions, and signal transduction. These proteins may have potential in the treatment of strokes, heart attacks, cancers, osteoporosis, and diabetes. The present study describes the isolation and characterization of a disintegrin (colombistatin) found in the venom of the Venezuelan snake mapanare ( Bothrops colombiensis). Colombistatin was purified by a two-step high-performance liquid chromatography procedure, which included reverse phase C18 and size exclusion protein Pak 60. Colombistatin inhibited ADP-induced platelet aggregation, human urinary (T24) and skin melanoma (SK-Mel-28) cancer cell adhesion to fibronectin, and cell migration. Colombistatin contained 72 amino acids with a mass of 7.778 kDa as determined by mass spectrometry. Colombistatin could be used as a therapeutic tool in the treatment of melanoma cancers and also thrombotic diseases. [ABSTRACT FROM AUTHOR]

Abstract: This study aimed to determine the biochemical, insecticidal and neurotoxic properties of venom from both sexes of the Australian spider Coremiocnemis tropix (Araneae, Theraphosidae). Insecticidal properties were tested in crickets, while in vitro neurotoxicity was determined in an avian skeletal muscle preparation. Some intersexual differences in venom composition were identified by rp-HPLC and by LC–MS, but the majority of components were found in venoms of both sexes. Injecting the venom into crickets revealed that venom from male specimens was slightly more potent, while female venom induced more prominent effects in the chick biventer cervicis nerve–muscle preparation. The results from the chick assay suggest the presence of at least two vertebrate-active neurotoxins. A pre-synaptic neurotoxin may explain the reversible inhibition of muscle twitches and the unaffected response to nicotinic agonists at medium concentrations of female and medium to high concentrations of male venom. In addition, the presence of a neurotoxin that blocks post-synaptic nicotinic receptors might explain the irreversible inhibition of muscle twitches and the reduced response to nicotinic agonists at high concentrations (5–10μg/ml) of venom from female specimens only. [Copyright &y& Elsevier]

Spiders have been incriminated as causes of human suffering for centuries, but few species worldwide cause medically significant envenomation. Widow spiders (Latrodectus spp.) occur worldwide and cause latrodectism, which is characterized by pain (local and generalized) associated with nonspecific systemic effects, diaphoresis, and less commonly other autonomic and neurological effects. Recluse spiders (Loxosceles spp.) are distributed mostly through the tropical and subtropical Western Hemisphere and can cause severe skin lesions and rarely systemic effects; most bites are unremarkable. Highly dangerous spiders in South America (armed spiders) and Australia (funnel-web spiders) cause rare but severe envenomation requiring medical intervention and sometimes antivenom. Most other spiders involved in verified bites cause minor, transient effects. Many spiders blamed for causing medical mischief have been elevated to medical significance via circumstantial evidence, poor reporting, and repetitive citation in the literature; several species have been shown to be harmless with more stringent scientific evidence involving verified bites in humans. [ABSTRACT FROM AUTHOR]

Background: Sol i 1, the venom phospholipase of imported fire ant venom is an important allergen and exhibits some cross-reactivity with IgE antibodies from patients sensitized to other Hymenoptera venoms. Objective: To determine the primary structure of Sol i 1 and evaluate the roles of protein and carbohydrate epitopes in its cross-reactivity. Methods: Sol i 1 was purified from venom, proteolytic peptides prepared and amino acid sequences obtained. The cDNA for Sol i 1 was cloned, sequenced, and compared with sequences of other wasp venom phospholipases. The role of carbohydrate epitopes in the cross-reactivity with other Hymenoptera venoms was studied by RAST inhibition. Results: The sequence identified Sol i 1 as a lipase of the GX class, lipoprotein lipase superfamily, pancreatic lipase homologous family and RP2 subgroup phospholipases as are the vespid venom phospholipases. The 148 residues identified by amino acid sequencing represent about 48% of the translated cDNA sequence. Sol i 1 was 31-32% identical to yellow jacket phospholipases. The identical regions of sequence were clustered in the domain which forms the serine hydrolase active site. Mannosylated N-glycans could completely inhibit binding of IgE from honeybee venom sensitized patients to Sol i 1. Inhibition by glycan of IgE binding from yellow jacket venom sensitized patients was low or absent for three of eight sera and substantial, but not complete for five sera. Conclusions: Sol i 1 is related to wasp venom phospholipases. Cross-reactivity with honeybee venom is caused by carbohydrate, whereas cross-reactivity with yellow jacket venom involves reactivity with both carbohydrate determinants of hyaluronidase and high molecular weight proteins and phospholipase protein determinants. [ABSTRACT FROM AUTHOR]

A woman stung by the box jellyfish Carybdea marsupialis (Cnidaria, Cubozoa) at a Spanish Mediterranean beach showed systemic manifestations over several months [pain far from the inoculation point, arthralgia, paresthesia, hyperesthesia, increase in eosinophils and immunoglobulin E (IgE)] in addition to the skin condition. [ABSTRACT FROM AUTHOR]