Injection Of Snake Venom Research Articles

Oxidative-Stress and Hematological Alterations Induced by the Egyptian Naja nubiae Venom in Rats

Background: Snake venoms have been the subject of intense studies to understand the mechanisms involved in toxicity. Limited information is available regarding the Egyptian Spitting Cobra’s oxidative stress and hematological profile (Naja nubiae). Objectives: The present study aimed to evaluate the oxidative stress produced by the venom of N. nubiae and determine its hematotoxic effects in rats. Methods: The adult male Albino rats (N=30) were subcutaneously (SC) injected with a physiological saline solution in the control group. The SC injection of snake venom in groups 2 and 3 was 1/4 and 1/2 of the LD50 (0.32 mg/kg and 0.65 mg/kg body weight, respectively). Blood samples were collected at 30, 120, and 360 minutes post-injection for biochemical and hematological assays in both control and treated groups. Levels of oxidative stress biomarkers, including lipid peroxidation (LPO), protein carbonyl content (PCC), and nitric oxide (NO) were estimated. Antioxidants agents comprising glutathione (GSH) level, activities of superoxide dismutase (SOD), and catalase (CAT) were also evaluated. Results: The results showed that the effects of snake venom on blood cells are dose-dependent. Furthermore, significant alterations (P≤0.05) were observed in the hemoglobin (Hb) concentration, packed cell volume (PCV), and the number of red blood cells (RBCs) in response to a low dose of venom at the 30-minute time. In contrast to the control group, the venom induced a substantial elevation in LPO, NO, and PCC levels, indicating a disturbance in redox equilibrium. Additionally, significant reductions were detected in the GSH levels as well as SOD and CAT activities in all treated groups. Conclusion: Overall, the cytotoxicity’s potential to induce oxidative stress may reduce its antioxidant systems, leading to redox disturbance and hematological alteration.

A Review of the Proteomic Profiling of African Viperidae and Elapidae Snake Venoms and Their Antivenom Neutralisation.

Snakebite envenoming is a neglected tropical disease (NTD) that results from the injection of snake venom of a venomous snake into animals and humans. In Africa (mainly in sub-Saharan Africa), over 100,000 envenomings and over 10,000 deaths per annum from snakebite have been reported. Difficulties in snakebite prevention and antivenom treatment are believed to result from a lack of epidemiological data and underestimated figures on snakebite envenoming-related morbidity and mortality. There are species- and genus-specific variations associated with snake venoms in Africa and across the globe. These variations contribute massively to diverse differences in venom toxicity and pathogenicity that can undermine the efficacy of adopted antivenom therapies used in the treatment of snakebite envenoming. There is a need to profile all snake venom proteins of medically important venomous snakes endemic to Africa. This is anticipated to help in the development of safer and more effective antivenoms for the treatment of snakebite envenoming within the continent. In this review, the proteomes of 34 snake venoms from the most medically important snakes in Africa, namely the Viperidae and Elipdae, were extracted from the literature. The toxin families were grouped into dominant, secondary, minor, and others based on the abundance of the protein families in the venom proteomes. The Viperidae venom proteome was dominated by snake venom metalloproteinases (SVMPs-41%), snake venom serine proteases (SVSPs-16%), and phospholipase A2 (PLA2-17%) protein families, while three-finger toxins (3FTxs-66%) and PLA2s (16%) dominated those of the Elapidae. We further review the neutralisation of these snake venoms by selected antivenoms widely used within the African continent. The profiling of African snake venom proteomes will aid in the development of effective antivenom against snakebite envenoming and, additionally, could possibly reveal therapeutic applications of snake venom proteins.

MO1030THE FIRST CASE OF ATYPICAL HEMOLYTIC UREMIC SYNDROME (AHUS) FEATURES IN AN EXPERIMENTAL MODEL

Abstract Background and Aims Thrombotic Microangiopathies (TM) are three distinct clinical syndromes presenting the same histological renal pattern: typical and atypical Hemolytic Uremic Syndrome (SEU- aSEU) and Thrombotic Thrombocytopenic Purpura (TTP). So far, the first report of a TM has been generally attributed to Eli Moschowitz. In 1922 he described the case of a 16 years old girl who died after acute onset of fever with petechial lesions and autoptic findings of hyaline thrombosis of terminal arterioles and capillaries, thus profiling the first case of TTP. Only in the 1955, Conrad Gasser described the first medical record of HUS, describing the case of a patient with a manifestation of bilateral necrosis of the renal cortex. We describe the first reported case of a Thrombotic Microangiopathy, in particular an experimentally induced aHUS by Richard M. Pearce in 1909. In this case, the trigger leading to aHUS was represented by snake venom injection in an experimental rabbit model. Method Pearce described acute glomerular lesions produced in the rabbit using dried venom of rattlesnake Crotalus Adamanteus. It was dissolved in salt solution in the proportion of 0.25 of a milligram to one cubic centimeter, rising gradually to two milligrams, and then followed by doses of 0.5 of a milligram of fresh venom at various intervals. The intervals between injections depended on the general condition of the animal and the amount of albuminuria. Results Rabbit kidneys showed well marked hemorrhagic and exudative lesions in the glomeruli; hyaline, granular, blood, and hemoglobin casts in both convoluted and collecting tubules; and granular degeneration of the epithelium of the convoluted tubules and loops of Henle. Pearce also described a “penetration of the cells of the compressed glomerular tuft into the mass of hemorrhage lying either in the tuft itself or in the capsular space” (Figure 1). In animals surviving 20-30 days after the first injection of the venom, the acute lesions were demonstrated to subside at the microscopic examination and their earlier presence was marked by “occasional casts and compressed masses of red cells in the glomerular spaces and tufts”. At the same time other models also showed “extensive granular degeneration of the convoluted tubules and many casts”. The renal autoptic findings presented the features of a vascular nephritis with severe endothelial changes. Conclusion Glomerular and tubular lesions of rabbits’ kidneys induced by crotalus’s venom showed typical features of what is today defined as aHUS. In this first experiment, the author described a glomerular tuft as “more analogous to the organization of a red thrombus than it is to any form of glomerular lesion known in man”, so we can affirm that Pearce described, ante litteram TMA histological features many years before other scientists.

Venom Ophthalmia and Ocular Complications Caused by Snake Venom.

Little is known about the detailed clinical description, pathophysiology, and efficacy of treatments for ocular envenoming (venom ophthalmia) caused by venom of the spitting elapid and other snakes, as well as ocular complications caused by snake venom injection. In this paper, we review clinical information of case reports regarding venom ophthalmia and snake venom injection with associated ocular injuries in Asia, Africa, and the United States. We also review the literature of snake venom such as their compositions, properties, and toxic effects. Based on the available clinical information and animal studies, we further discuss possible mechanisms of venom ophthalmia derived from two different routes (Duvernoy’s gland in the mouth and nuchal gland in the dorsal neck) and the pathophysiology of snake venom injection induced ocular complications, including corneal edema, corneal erosion, cataract, ocular inflammation, retinal hemorrhage, acute angle closure glaucoma, as well as ptosis, diplopia, and photophobia. Finally, we discuss the appropriate first aid and novel strategies for treating venom ophthalmia and snake envenoming.

Systemic and Local Phenotypes of Barium Chloride Induced Skeletal Muscle Injury in Mice.

Skeletal muscle regeneration in mice has traditionally been studied using local freeze burn or snake venom injection models. More recently, a barium chloride (BaCl2)-induced muscle injury model has been established and is gaining popularity due to the relatively simple procedure and accessibility to required reagents. Here we sought to characterize the local and systemic effects of BaCl2-induced muscle injury. For this study, a 1.2% BaCl2 solution was locally administered to the tibialis anterior (TA) muscle and local and systemic phenotypes were analyzed at different timepoints. When 50 μL of the solution was injected unilaterally in the TA muscle, no mortality was observed. However, when 100 μL of the solution was injected, 50% of the mice died within 24 h. Serum analysis of the mice injected with 50 μL of BaCl2 solution at days 1 and 7 revealed changes resembling rhabdomyolysis. At day 1 post-injection of 50 μL of the BaCl2 solution, acute suppurative inflammation was observed in gross examination of the TA muscle, while extensive hemorrhagic necrosis was revealed on histological examination. At day 7, regenerated myofibers with centralized nuclei appeared with the resolution of acute inflammatory infiltration and the muscle tissue displayed molecular signatures consistent with myofiber differentiation. The overall muscle injury and regeneration phenotypes in the BaCl2-induced muscle injury model were similar to those of the well-established freeze burn or snake venom injection models. Taken together, the BaCl2-induced muscle injury model is comparable to conventional muscle injury and regeneration models, with considerations for possible systemic effects.

Attempted suicide by snake bite: A case study.

Snake bite is an important public health issue in India and is almost always accidental in manner. Suicide by snake bite or injection of snake venom is extremely rare. Suicidal ideation and behavior is known to be influenced by various socio-economic and psychological factors. The method employed for suicide is also influenced by the occupation of the victim. We report a case where a snake charmer had attempted suicide by inflicting a bite by a monocled cobra.

Immediate active movement following Bitis arietans or Naja mossambica venom injection diminishes or prevents necrosis in mice

Tourniquet use, which localises necrotising snake venom to the bite site, is suspected to exacerbate local necrosis. In view of this, the effect of immediate active movement on the outcome of necrotising snake venom injection was investigated in mice. A minimal macroscopic necrotic dose of Bitis arietans venom (2.5 μg) was administered subcutaneously into the dorsum of the right hind feet of 40 mice of which 20 were previously anaesthetised. The 20 non-anaesthetised mice swam for 10 min. All mice were euthanased on day 4. A further 40 mice underwent an identical experiment using 20 μg Naja mossambica venom and were euthanased on day 6. The areas of resulting macroscopic necrosis were measured in the B. arietans venom injected mice, while the largest diameter of macroscopic necrosis was determined in the N. mossambica venom injected mice. Immediate active movement (swimming) significantly prevented or reduced the area of necrosis in comparison to non-ambulant mice.

Effects of anabolic/androgenic steroids on regenerating skeletal muscles in the rat.

We have examined the effect of male sexual hormones on the regeneration of skeletal muscles. Degeneration/regeneration of the left soleus and extensor digitorum longus muscles (EDL) of Wistar male rats was induced by an injection of snake venom (2 microg, Notechis scutatus scutatus). During the muscle regeneration (25 days), rats were treated with either oil (CON), nandrolone (NAN), NAN combined with exercise (NAN + EXE) or were castrated (CAS). Muscle growth and myosin heavy chain (MyHC) isoform content of regenerating muscles were studied. Castration altered the concentrations of MyHC in venom-treated EDL (P < 0.01) and soleus (P < 0.05). NAN increased the mass (P < 0.01) of regenerating soleus and decreased the relative amount of fast MyHC protein (% of total, P < 0.05). The effect of NAN + EXE on the fast MyHC proteins of venom-treated soleus was opposite (P < 0.05). NAN and NAN + EXE were without effect on the regenerating EDL (P > 0.05). In conclusion, it is possible that male sexual hormones play a role in the growth (synthesis of contractile proteins) of regenerating muscles in rat. In addition, contrary to NAN + EXE, NAN could be beneficial to soleus regeneration.

Effects of glucocorticoid treatment on regenerating skeletal muscles in the rat

Summary Introduction. Regeneration of the soleus and extensor digitorum longus muscles of Wistar male rats was induced by an injection of snake venom (2 μ g, Notechis scutatus scutatus). Methods and results. Rats were treated for 14 to 16 days with dexamethasone (sub-cutaneous, 1 mg/kg on a daily basis) or not (control). Twenty five days after venom injection, the mass of the venom-treated extensor digitorum longus in dexamethasone rats was decreased when compared to control rats ( P P Conclusion. The results of this preliminary study suggest that dexamethasone treatment inhibits both the growth of extensor digitorum longus and the maturation of the soleus during regeneration.

Myosin heavy chain composition of regenerated soleus muscles during hindlimb suspension.

The aim of this study was to determine the MHC profile of regenerated soleus muscles in control (C, n = 8) and hindlimb suspended rats (HS, n = 8). After muscle degeneration was induced by injection of snake venom containing notexin, male rats were either tail suspended for 21 days or submitted to normal weight-bearing activity. Separation and detection of MHCs by sodium dodecyl sulphate-polyarcylamide gel electrophoresis (SDS-PAGE) showed that regenerated soleus muscles from C rats contained only type I and type IIa MHCs. The relative amount of type I MHC was higher in regenerated (93.9 +/- 1.7%) than in untreated muscles (86.5 +/- 2.3%)(P < 0.01). In the HS group, the immunohistochemical analysis showed that the majority of regenerated myofibres reacted positively with the antibody against fast MHCs. SDS-PAGE analysis demonstrated that HS resulted in a shift toward faster MHCs in both intact and regenerated myofibres. Regenerated soleus muscle from HS rats contained approximately 34% type IIa MHC, approximately 37% type IIx/d MHC and approximately 18% type IIb MHC, when type I MHC contributed to only approximately 12% of total myosin. The proportions of fast MHC isoforms in regenerated muscles were higher than those recorded in untreated muscles. Collectively, these results suggest that the shift in the MHC profile associated with hindlimb unweighting in adult undamaged soleus muscles is also related to the heterogeneity of early myoblasts.

Effect of electric shock therapy on local tissue reaction to poisonous snake venom injection in rabbits

Effect of electric shock therapy on local tissue reaction to poisonous snake venom injection in rabbits

Suicide by Snake Venom Injection

A case of apparent suicide by injection of snake venom was investigated. Evidence found at the scene and presumably used for self-injection was positive for nonspecific venom but had the characteristics of a viper venom.

Suicide by injection of snake venom

An unusual suicide is described where the deceased, a schizophrenic young man in perfect physical health, deliberately injected himself subcutaneously with snake venom, by means of a syringe. The circumstances, autopsy findings and laboratory investigations are recorded.

Enzymic hydrolysis of various lecithin monolayers employing surface pressure and potential technique

Enzymic hydrolysis of dipalmitoyl, egg, soybean, and dioleoyl lecithin monolayers was studied by measuring the changes in surface pressure (π) and surface potential (Δ V) upon injection of snake venom ( Naja Naja) into the subsolution. The presence of Ca ++, which could not be substituted by Mg ++, Sr ++, or Ba ++, was essential for hydrolysis. Thin layer chromatography of monolayers removed after injection of venom into the subsolution showed that the snake venom indeed hydrolyzed lecithin monolayers into lysolecithin and free fatty acid. The rates of hydrolysis of different lecithins were in the order: dioleoyl lecithin > soybean lecithin > egg lecithin > dipalmitoyl lecithin, which is also the order of their molecular areas and hence their intermolecular spacings in monolayers. The solubility of lysolecithin molecules in the subsolution depends upon the chain length and unsaturation of the fatty acyl chains, which affects the changes in Δ V and π upon hydrolysis. The presence of citrate buffer or chelating agents with excess of Ca ++ in the subsolution increased the rate of hydrolysis by decreasing the binding of Ca ++ to lecithin monolayers. The hydrolysis of egg lecithin monolayers on different buffers was in the order, tris > veronal > collidine; which is the reverse of that found in the bulk reaction. Curves of π -area and Δ V - area of egg lecithin on these buffers indicated penetration of veronal and collidine ions into monolayers; this prevents the formation of the enzyme-substrate complex at the surface. In bulk reactions, these buffer ions act as lipid dispersants increasing the lipid/water interfacial area and consequently increase the rate of hydrolysis. Anionic, cationic, and neutral spacer molecules (15 mole %) were introduced in egg lecithin monolayers. The rate of hydrolysis increased in the presence of eicosanyl trimethylammonium, decreased in the presence of dicetyl phosphate or dipalmitin, and remained unchanged in the presence of cholesterol, indicating the importance of surface charge and state of the monolayer.

Hydroxyproline excretion in rat urine following injection of snake venom

In rats, a statistically significant increase in hydroxyproline excretion occurs following injection of crotalus toxin (Agkistrodon piscivorus Lacepede). The significance of this finding is briefly discussed.

Effects of the injection of snake venom containing lecithinase A into Physarum polycephalum.

Journal of Cellular and Comparative PhysiologyVolume 44, Issue 2 p. 203-210 Article Effects of the injection of snake venom containing lecithinase a into Physarum polycephalum Howard R. Engel, Howard R. Engel Department of Anatomy, University of ChicagoSearch for more papers by this author Howard R. Engel, Howard R. Engel Department of Anatomy, University of ChicagoSearch for more papers by this author First published: October 1954 https://doi.org/10.1002/jcp.1030440205Citations: 3AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Citing Literature Volume44, Issue2October 1954Pages 203-210 RelatedInformation

Lyso-lecithin fragility in blackwater fever and haemolytic jaundice

Red cells transfused into a case of haemolysing blackwater fever are destroyed just as readily as the patient's own cells, making it appear that it is not the red cells that are at fault in this condition, but that there is some circulating haemolysin. By an improved spectrophotometric technique it has now been shown that, like those of haemolytic jaundice, the red cells from blackwater fever have an increased fragility to lyso-lecithin, although their fragility to saline is normal, unlike that of the cells of haemolytic jaundice. These two facts taken together make it seem that in blackwater fever the cells may possess some abnormal feature, but that this is probably secondary to more fundamental changes that take place in the cells' environment. By using van Boros' formula it is possible to demonstrate that spherocytosis occurs in blackwater fever and is accompanied by decreases in diameter-thickness ratio, volumes and areas; that these changes in blackwater fever are intermediate between those taking place in haemolytic jaundice and normal controls, and are not related in blackwater fever to changes in hypotonic saline fragility. It is probable that the initial stage in the destruction of the cells is a change in the permeability of the cell membrane, which allows haemoglobin to escape. The cells have later been seen to swell, become “transparent,” and then to disappear. That they have not been broken up can be shown by resuspending them in saline when the “ghosts” reappear. This phenomenon takes place in both lyso-lecithin and snake venom haemolysis. If the process is allowed to continue the cells are finally disrupted, and will not reappear on saline resuspension. In haemolytic jaundice it seems that there may be some defect in the red cell as well as some “splenic factor,” shown by the fact that splenectomy fails to alter the abnormal fragility of the cells to saline, but does stop the periodic haemolysis. A combination of both abnormal cells and splenic factors would seem to be necessary for the destruction of the red cells in this condition. The same cannot be determined for blackwater fever. The enlargement of the spleen in so many of these haemolytic conditions is regarded by many as of considerable significance. The production of lyso-lecithin as a result of the separation of the cells and plasma in this organ has led to the suggestion that this powerfully haemolytic substance may play a part in the haemolyses of certain of these conditions. It is possible that lyso-lecithin fragility is in some way related to the action of this substance on the lipoids or lipo-protein complex in the red cell membrane. It has been shown that the amount of lipoid present in the cell envelope is closely related to the dimensions of the cell, and that the ability of the cell to increase its volume without being disrupted is related to the lipo-protein ratio in its membrane. The kinetics of lyso-lecithin haemolysis have not been worked out, but there is some ground for regarding the process as one of enzyme action where the enzyme lecithinase acting on a substrate of red cells or plasma lecithin results in the production of lyso-lecithin or some allied phosphatide. Lecithinase has been shown to be present in many conditions where haemolytic reactions take place, such as in favism, snake poisoning, Cl. welchii filtrates as well as in the spleen and peripheral blood. The degradation products of the action of lecithinase on lecithin are not always the same, nor are the various lecithinases alike. Although the end product of the action of snake venom lecithinase on lecithin is lyso-lecithin, yet this substance when injected into baboons in relatively huge amounts produces no haemolysis, and only a slight shift of the Price-Jones curve to the right. This is in sharp contrast to the haemolysis that follows the injection of snake venom freed of its neurotoxic factor. It is suggested that this difference may be due to the fact that in the former case the lecithin in the cells and/or plasma is not disturbed, whilst in the case of snake venom injection of the lecithinase present in the venom will split the lecithin of the cells and/or plasma and so disturb the intracellular lipo-protein complex. The part played by the reticulo-endothelial system in the haemolytic process is briefly discussed—the resulting decision being that no precise data are available to enable any conclusions to be reached either upon the part that this system plays in actually destroying red cells, or elaborating haemolysins that later lyse the cells. Proteolytic enzymes may be produced in the macrophages of the reticulo-endothelial system and these have been shown to be capable of destroying red and white cells.

A Toxic Ocular Reaction

Summary and Conclusions The work embodied in this paper demonstrates that Shwartzman toxins are capable of producing a primary ocular reaction in rabbits, named the toxic ocular reaction. The reaction consists of intense iridoconjunctival hyperemia and coagulation of the aqueous humor after its aspiration. It appears several minutes after the intravenous injection of toxin, reaches its maximal intensity in a few hours, and is notably reduced or has completely disappeared in about twelve hours. The smallest amount of toxin which, by intravenous injection, induces aqueous humor coagulation is termed the minimal coagulating dose. The reaction was also obtained using the intra-arterial and intraabdominal routes and a highly vascularized area of the skin. However, the intestinal and conjunctival routes proved non-effective. Protection against the ocular reaction was afforded by previous intracutaneous or intravenous injection of rabbits with Shwartzman filtrates or by moccasin snake venom injection. Toxins kept their potency for several months when undiluted preparations were stored in the refrigerator. The toxic ocular principles did not pass through dialyzing membranes, showed resistance to heat at high pressure, were definitely weakened by the addition of formaldehyde, and were specifically neutralized by Shwartzman antitoxins. This reaction was reproduced in cats and dogs but failed to appear in other usual laboratory animals, even guinea-pigs in which, in a certain percentage, the Shwartzman phenomenon may be elicited. The toxic ocular reaction was obtained in approximately 95 per cent of a large number of rabbits injected intravenously with 10 or more minimal coagulating doses per kilo of body weight. It thus represents a new property of the Shwartzman toxins, and indicates their marked primary toxicity for the rabbit's uveal tract.