Anti-snake Venom Activity Research Articles

Exploring the inhibitory potential of phytochemicals from Vernonia glaberrima leaves against snake venom toxins through computational simulation and experimental validation

Phospholipase A2 (PLA2) is an enzyme present in appreciable quantity in snake venoms which catalyze the hydrolysis of glycerophospholipids at sn-2 position and promote the release of lysophospholipids and fatty acids. 5-methylcoumarin-4-β-glucoside (5MC4BG) and lupeol were previously isolated from the leaves of V. glaberrima. The aim of this research was to evaluate effect of these compounds as potential inhibitors of snake venom toxins of Naja nigricollis using an in vitro and in silico studies. Antisnake venom studies was conducted using acidimetry while the molecular docking analysis against PLA2 enzyme from N. nigricollis was performed using Auto Dock Vina and ADME-Tox analysis was evaluated using swissADME and ProTox-II online servers. The two compounds (5MC4BG and Lupeol) were able to inhibit the hydrolytic actions of PLA2 enzyme with percentage inhibition ranging from 23.99 to 72.36 % and 21.97–24.82 % at 0.0625–1.00 mg/mL respectively while the standard ASV had 82.63 % at 1.00 mg/mL after 10 min incubation at 37 °C. Similar effects were observed after 30 min incubation, although there was significant increase in percentage inhibition of 5MC4BG and lupeol ranging from 66.51 to 83.73 % and 54.87–59.60 % at similar concentrations. Furthermore, the compounds were able to bind to the active site of PLA2 enzyme with high affinity (−7.7 to −6.3 kcal/mol); the standard ligand, Varespladib had a docking score of −6.9 kcal/mol and they exhibited favorable drug-likeness and pharmacokinetic properties and according to toxicity predictions, the two compounds are toxic. In conclusion, the leaf of V. glaberrima contains phytoconstituents with antisnake activity and thus, validates the hypothesis that, the phytoconstituents of V. glaberrima leaves has antisnake venom activity against N. nigricollis venom and thus, should be studied further for the development as antisnake venom agents.

Evaluation of anti-snake venom activity of ethanolic leaf extract of Euphorbia hirta, Ageratum conyzoide and Anogeisus leiocarpus on West African Carpet Viper (Echis ocellatus) in envenomed Sprague Dawley rats.

Evaluation of anti-snake venom activity of ethanolic leaf extract of Euphorbia hirta, Ageratum conyzoide and Anogeisus leiocarpus on West African Carpet Viper (Echis ocellatus) in envenomed Sprague Dawley rats.

Azadirachta indica A. Juss.: Ethnobotanical knowledge, phytochemical studies, pharmacological aspects future prospects

Azadirachta indica, commonly known as Neem, it is a member of the Meliaceae family, a fast-growing evergreen popular tree found commonly in India, Africa and America. All parts of the neem tree, including the seeds, fruits, twigs, leaves, flowers, roots, and bark, have traditionally been used to treat inflammation, infections, fever, skin ailments, and dental diseases. Azadirachtin, nimbolinin, nimbin, nimbidin, nimbidol, salannin, and quercetin are all useful active chemicals extracted from various plant parts. The aim of this review article provides information mainly on various pharmacological activities like Antioxidant Activity, Anti-cancer, Anti-inflammatory, Anti-bacterial, Antiviral Activity, Antifungal Activity, Antihelmethic effects, Antidiabetic action, Wound healing effect, Activity against ulcer, Orodental protection, Contraceptive, Hepatoprotective activity, Immunomodulatory and Growth Promoting, Antinephrotoxicity Effect and Antisnake venom activity of neem plant and medicinal uses. Plant extracts function as both a reducing and capping agent. Neem is a well-known medicinal plant and has been studied for the biosynthesis of nanoparticles. A. indica possesses many phytochemicals that can reduce metal ions. The nanoparticles are reported to exhibit good antimicrobial, and antioxidant and cytotoxicity effects against MCF-7 and HeLa cells in vitro.

In vitro Detoxifying Effects of Catunaregam nilotica(STAPF) Root Extracts against Naja nigricollis Phospholipase A2 (PLA2) Enzyme

Snakebite is a severe public health problem across the globe, especially in sub-Saharan Africa where most victims live in rural areas where their livelihood depends on farming and rearing animals. Catuneragamniloticahas been used in ethnomedicine to treat snakebites, and diarrhea among others. The study aims to evaluate the phospholipase A2 inhibition potential of C. niloitca using in vitro model. The plant was collected, authenticated, and sequentially extracted using solvents of increasing polarity starting from n-hexane, ethyl acetate, and methanol. The extracts were screened in vitro for their inhibitory activity against N. nigricollisvenom using PLA2 assay, and polyvalent serum-base antivenin was used as a standard. The phytochemical screening conducted using standard procedures revealed the presence of flavonoids, alkaloids, saponins, polyphenols, tannins, terpenoids, cardiac glycosides, anthraquinones, and steroids which varies in the extracts. The extracts of the plant have demonstrated significant (p<0.05) and concentration-dependent antisnake venom activity by combating the hydrolytic actions of N. nigricollisPLA2 enzyme; methanol extract was the most active with PLA2 inhibition of 83.01 to 63.01% at1.0mg/mL to 0.125mg/mL respectively while hexane extract was the least active with percentage inhibition of 45.04 to 30.00% at 1.0mg/mL to 0.125mg/mL. In conclusion, the plant C. niloticahas demonstrated good antisnake venom activity and could be studied further for development as safe and affordable antivenom by subjecting it to bioassay-guided isolation and purification of bioactive compound(s) responsible for the observed activity.

Ethnobotany, Phytochemistry, Ethnopharmacology, and Toxicity of Euclea divinorum Hern (Ebenaceae): A Review

Euclea divinorum Hiern (Ebenaceae) is a medicinal plant widely used in Yemen, Eastern and Southern Africa in traditional phytotherapy. This review was an attempt to compile a comprehensive report on its ethnomedicinal uses, phytochemicals, pharmacological activities, and toxicity, lending credence to the use of its various parts in herbal medicine. The literature encountered indicated that herbal formularies from different parts of Euclea divinorum (E. divinorum) are majorly used for traditional treatment of odontological, dermatological, respiratory, reproductive, and gastrointestinal ailments. At least 18 compounds have been isolated and characterized in extracts of E. divinorum, while 31 others have been identified in the crude extracts analyzed by GC-MS. Extracts, as well as isolated compounds from E. divinorum stem, stem barks, leaves, and roots, showed renoprotective, antiproliferative, antinociceptive, contractile, proteolytic, diuretic, antiprotozoal, anti-giardial, antioxidant, anti-inflammatory, antimicrobial, molluscicidal, and insecticidal activities. Further research is warranted to explore other pharmacological properties such as antisnake venom, aphrodisiac, antidiabetic, analgesic, and antimycobacterial activities and the responsible bioactive compounds in the different parts of E. divinorum claimed in herbal medicine. The composition and bioactivities of essential oils from this species also warrant further studies.

Anti-Snake Venom Property of Medicinal Plants: A Comprehensive Review of Literature

Snakebite is one of the major health issues posing considerable morbidity and mortality. According to an estimate of World Health Organization (WHO) (World health organization, 2021) approximately 5 million people are bitten by several species of snakes resulting in up to 2.5 million envenomation cases annually. The mainstay of treatment for envenomation is intravenous administration of anti-snake venom. Although antivenom neutralizes the systemic effects but it does not relieve the symptoms such as venom-induced hemorrhage, necrosis and nephrotoxicity. Moreover, the use of antivenoms is associated with hypersensitivity reactions including urticaria, anaphylaxis, or serum sickness due to their heterologous property. Furthermore, stringent storage conditions and narrow specificity of antivenoms limit their use in both developed as well as developing countries. In this context, researchers have been searching for natural products and plant extracts to explore their antivenom activity along with anti-myotoxic, anti-hemorrhagic and anti-inflammatory properties. Plant remedies may prove to be an effective alternate for antivenom sera with less adverse events and better tolerability. To the best of our knowledge, this is the first comprehensive review of medicinal plants possessing anti-snake venom activities against certain species of snakes. The current review highlights the investigated plants with their phytochemical analysis to integrate the available information for future research and development of antivenom sera.

A Review of Aristolochia indica: Ethnomedicinal Uses, Phytochemistry, Pharmacological and Toxicological Effects

Background: Aristolochia indica L. of the family Aristolochiaceae is a twining perennial herb with an ancient history of medicinal use. In the Indian Ayurvedic system, it is used for the treatment of snake venom, scorpion venom, pimple, fever and worm infections. Objective: This studay aimed to make accessible the current information on the traditional uses, phytochemistry, pharmacology and toxicology of Aristolochia indica. Additionally, the purpose was to emphasize the potential uses of this plant to treat various diseases and to bring a foundation for further research. Methods: The present review is carried out by compiling literature from 1935 to 2020, concerning the morphology, traditional uses, phytochemistry, pharmacological activities, and toxicological aspects of Aristolochia indica. Results: Diverse chemical compounds including aristolochic acids, aristolactam, Phenanthrenes, alkaloids, lignans, steroids and terpenes have been isolated from this plant. Mostly in-vivo models indicated several pieces of evidence on the use of this plant particularly to regulate fertility. Few in-- vivo studies also proved the usefulness of this plant in inflammation and diabetes. In some in-vitro studies, the anti-snake venom, larvicidal, and anti-oxidant properties have been proved. Conclusion: Preclinical studies have demonstrated remarkable activity that supports the conventional use of the plant as an antivenum, antifertility, antimicrobial and anti-inflammatory agent. Although few phytochemicals isolated (aristolic acid, (12S)-7,12-secoishwaran-12-ol, aristololactam- I N-β-D-glucoside, aristolochic acids, β-sitosterol, (-)hinokinin and aristolactam-I) from the plant exhibited remarkable biological activity, it was only confined to preclinical studies. Even though the isolated aristolochic acids showed significant anti-snake venom activity, but it was found to be nephrotoxic and mutagenic. More detailed safety data pertaining to the dose of crude extracts or pure compounds needs to be generated.

Phytochemical composition, antisnake venom and antibacterial activities of ethanolic extract of Aegiphila integrifolia (Jacq) Moldenke leaves

Snakebites are considered a major neglected tropical disease, resulting in around 100,000 deaths per year. The recommended treatment by the WHO is serotherapy, which has limited effectiveness against the toxins involved in local tissue damage. In some countries, patients use plants from folk medicines as antivenoms. Aegiphila species are common plants from the Brazilian Amazon and are used to treat snakebites. In this study, leaves from Aegiphila integrifolia (Jacq) Moldenke were collected from Roraima state, Brazil and its ethanolic extract was evaluated through in vitro and in vivo experiments to verify their antiophidic activity against Bothrops atrox crude venom. The isolated compounds from A. integrifolia were analyzed and the chemical structures were elucidated on the basis of infrared, ultraviolet, mass, 1H and 1³C NMR spectrometry data. Among the described compounds, lupeol (7), betulinic acid (1), β-sitosterol (6), stigmasterol (5), mannitol (4), and the flavonoids, pectolinarigenin (2) and hispidulin (3), were identified. The ethanolic extract and flavonoids (2 and 3) partially inhibited the proteolytic, phospholipase A2 and hyaluronidase activities of B. atrox venom, and the skin hemorrhage induced by this venom in mice. Antimicrobial activity against different bacteria was evaluated and the extract partially inhibited bacterial growth. Thus, taken together, A. integrifolia ethanolic extract has promising use as an antiophidic and antimicrobial.

In vitro anti-venom potential of various solvent based leaf extracts of Andrographis serpyllifolia (Rottler ex Vahl) Wight against Naja naja and Daboia russelli

Ethnopharmacological relevanceSnake bite is a major occupational hazard in tropical and subtropical countries including India as per the World Health Organization. Naja naja (Indian cobra) and Daboia russelli (Russell's viper) are the two poisonous snakes commonly associated with human mortality in India. Andrographis serpyllifolia (Rottler ex Vahl) Wight has been documented in ethnobotanical records as a plant possessing potent anti-snake venom activity. Aim of the studyThe present study is aimed for systematic evaluation of in vitro anti-venom potential of various solvent based leaf extracts of A. serpyllifolia against toxic venom enzymes of Naja naja and Daboia russelli. Materials and methodsDifferent solvent based leaf extracts of A. serpyllifolia were tested against the snake venoms of Naja naja and Daboia russelli obtained from Irula Snake Catchers Industrial Co-operative Society Limited, Kancheepuram, Tamil nadu, India. Three different in vitro neutralization assays such as indirect hemolysis, procoagulent and lytic activities and seven in vitro enzyme inhibition assays such as protease, acetylcholinesterase, phosphomonoesterase, phosphodiesterase, 5′nucleotidase, phospholipase A2, hyaluronidase and post synaptic acetylcholine receptor binding activity were carried out according to standard protocols. The results were analyzed using the standard ANOVA procedures. ResultsAmong various solvent based leaf extracts of A. serpyllifolia tested, aqueous extract showed maximum neutralizing and inhibitory activities against Naja naja and Daboia russelli venoms. ConclusionsThe various in vitro enzymatic studies reveal that the aqueous leaf extract of A. serpyllifolia plant could inhibit most of the toxic enzymes of the Naja naja and Daboia russelli venoms which could be further confirmed by in vivo studies.

Anti-Snake Venom, Anti-Arthritic and Cytotoxic Activities of Tectona grandis L. f. Stem Bark (Lamiaceae)

Tectona grandis L. f. (Lamiaceae) is famous for timber production and has been used in traditional medicine for treating bronchitis, liver-related roubles, urinary discharge amongst other diseases. Methanol extract of the stem bark was investigated for in vivo antiophidian assay against Bitis arietans and Naja nigricollis. Also, the extract and chromatographic fractions were subjected to cytotoxicity tests using tadpole model and antiarthritic assay by proteinase inhibition. Chromatography of crude methanol extract afforded three highly polar vacuum liquid chromatography (VLC) fractions (BVLC-1, BVLC-2 and BVLC-3). BVLC-2 further gave four semi-pure uncharacterized isolates (I, II, III, IV). Only BVLC-2 and BVLC-3 were cytotoxic at 10 - 80 mg/ml, with BVLC-3 being most potent (100% mortality, LC50 40 mg/ml). Concentration-dependent proteinase enzyme inhibition (24 - 71%) at 200 - 1000 μg/ml of BVLC-2 was observed, and this was less (IC50 659.24 μg/ml) than the activity of the standard drug, acetyl salicylic acid. However, neutralization of B. arietans and N. nigricollis snake venoms using methanol extract was not dose-dependent, but the extract atthe least dose, 50 mg/kg offered better protection (75%) on Naja nigricollis envenomed-mice in 48 h. Its activity was comparable to that of the positive antivenin tested at 0.2 mg/kg. These findings justify the folkloric use of T. grandis in the treatment of snake bites, arthritic conditions, and oxidative stress-induced diseases.
 Keywords: Tectona grandis, methanol extract, anti-snake venom activity, tadpole cytotoxicity, anti-arthritic activity

Evaluation of Snake Venom’s PhospholipaseA2 Enzyme Inhibition Activity of Cyphostemma adenocoule

Background: A snake bite is fundamentally an injury often resulting in puncture wounds meted out by the animal's fangs and occasionally resulting in envenomation. Rate of snake bites around 5,400,000 bites per year leads to over 2,500,000 envenomings and around 125,000 fatal cases annually. Snake venom enzymes are rich in metalloproteinases, phospholipaseA2, proteinases, acetylcholinesterases and hyaluronidases. Objective: Cyphostemma adenocoule is traditionally being used for the treatment of snake bites in Eritrea. The present research was aimed at evaluating the snake venom enzyme inhibition activity of C. adenocoule against puff adder venom and developing a base for the traditional use of the plant against snakebites in Eritrea. Methods: The anti-venom activity of C. adenocoule was assessed in-vitro through phospholipaseA2 enzyme inhibition assay using egg yolk as a cell. The ethanol and chloroform extracts of C. adenocoule showed in vitro anti phospholipase A2 activity, whereas the water extracts of the plant showed no activity. Results: Among the extracts of C. adenocoule, the highest percentage of inhibition was obtained from chloroform extract (95.55% at 100mg/ml). The extract showed prominent activity at different concentrations (34.7% at10mg/ml, 48.8% at 20mg/ml, 54.8% at 40mg/ml, 60.9% at 60mg/ml, 80.5% at 80mg /ml). The ethanol extract also showed certain activity at various concentrations (25.22% at10mg/ml, 14.78% at 20mg/ml, 2.6% at40mg/ml). The activity of the chloroform extracts increases as concentration increases, whereas the activity of the ethanol extracts decreases as concentration increases. The aqueous extract of C. adenocoule did not show any activity at all concentrations. Conclusion: In this study, the chloroform and ethanol extracts of the plant inhibited the enzyme of interest and thus proved the efficacy of anti-snake venom activity of the plant.

In vitro Phytochemical Screening and Anti-snake Venom Activity of the Methanol Leaf and Stem Bark Extracts of Leptadenia hastata (Asclepiadaceae) against Naja nigricollis

Snake envenomation is a major cause of death and morbidity in many developing countries. Leptadenia hastata (Pers.) Decne (Asclepiadaceae) has been reportedly used in traditional medicine as an antivenom, antiulcer, antidiabetic, analgesic, cardiovascular disorders, bacterial and viral infections. This research design is to investigate the phytochemical analysis and phospholipase A2 enzyme inhibition potential of L. hastata leaf and stem bark extracts using standard procedures. Preliminary phytochemical screening revealed the presence of key constituents such as carbohydrates, tannins, flavonoids, alkaloids, triterpenes, steroids, saponins, and diterpenes. The methanol leaf and stem extracts were able to inhibit the hydrolytic action of phospholipase A2 enzyme in a concentration-dependent manner. The research findings lay credence to the folkloric claim of the leaf and stem of L. hastata as an anti-snake venom.

Anti-snake venom and analgesic activities of extracts and betulinic and oleanolic acids isolated from Parinari curatellifolia

Background: Parinari curatellifolia is used in Nigeria by traditional medicine practitioners (TMPs) for the treatment of snakebite and other conditions. Aim: This study is aimed at evaluating the anti-snake venom (ASV) and analgesic activities of n-hexane, ethyl acetate and methanol extracts as well as that of betulinic and oleanolic acids. Setting: The extracts and the isolates were subjected to acute toxicity studies, anti-snake venom and analgesic activities. Methods: Acute toxicity studies for the extracts were performed according to Lorke’s method whilst that of betulinic and oleanolic acids were performed according to Organization for Economic Co-operation Development 425 at the limit dose of 2000 mg/kg in mice. The ASV activity of the extracts and the betulinic and oleanolic acids were screened against Naja nigricollis venom phospholipases using the phospholipase-A2 acidimetric assay. The analgesic activity of ethyl acetate extract (35 mg/kg, 70 mg/kg, 140 mg/kg), betulinic acid (1 mg/kg, 5 mg/kg, 10 mg/kg) and oleanolic acid (1 mg/kg, 5 mg/kg, 10 mg/kg) were evaluated using acetic acid-induced abdominal constrictions in mice. Results: The LD 50 of the methanol and ethyl acetate extracts were 113 mg/kg and 471.17 mg/kg, respectively, whilst that of the betulinic and oleanolic acids were greater than 2000 mg/kg. In the ASV study, the extracts inhibited the hydrolytic action of phospholipases with a percentage inhibition of 24.1% (n-hexane), 29.3% (ethyl acetate) and 20.7% (methanol), whilst the ASV (standard) caused 72.9% inhibition. On the other hand, the betulinic and oleanolic acids inhibited the hydrolytic action of phospholipases with a percentage inhibition of 35.6% and 31.4% respectively. For the analgesic evaluation, the ethyl acetate extract exhibited analgesic activity by producing a significant ( p < 0.05) reduction in the number of writhes (7.75 ± 1.20, 11.82 ± 3.10 and 14.98 ± 2.72) at 140 mg/kg, 70 mg/kg and 35 mg/kg, respectively. Betulinic acid at 10 mg/kg, 5 mg/kg and 1 mg/kg showed a mean number of writhes of 3.40 ± 0.8, 4.20 ± 1.4 and 5.60 ± 2.1, respectively. Oleanolic acid at 10 mg/kg and 5 mg/kg also showed 7.50 ± 2.6 and 6.40 ± 1.8 mean number of writhes. Conclusion: This study has validated scientifically why P. curatellifolia is used traditionally in the treatment of snakebite, as the ethyl acetate extract and the isolated compounds moderately inhibited the hydrolytic action of phospholipase-A2. Also, betulinic and oleanolic acids exhibited analgesic activity in acetic acid-induced writhing in mice.

Morphology, Phytochemistry and Pharmacological Activity of Vitex negundo: An Overview

Now days a herbal and traditional medicines mostly used in all over the world according to WHO, about 60 to 65% world population practicing herbal and traditional medicines. Vitex negundo Linn. is an aromatic herb belong to family Verbenaceae. It is also known as Nirgundi, Five leaved chaste tree. Plants have many chemical constituents as flavonoids, volatile oil, triterpenes, diterpenes, sesquiterpenes, lignan, flavones, glycosides, iridoid glycosides and stilbene derivative. These chemical constituents are present in each part of the plant. Vitex negundo extract have various pharmacological activities such as anti-inflammatory, antipyretic, anti-arthritic, antioxidant, analgesic activity, antibacterial activity, antitumor activity, anti-amnesic activity, anxiolytic activity, Nephroprotective activity, anti-HIV activity, Antitubercular activity, antieosinophilic activity, anti-snake venom activity. This review article provides depth information related to plant morphology, cultivation, chemical constitutes, pharmacological activities and their medicinal uses.
 Keywords: Vitex negundo, Nirgundi, Casticin, Anti-Gout Activity

Neutralization Effect of Tithonia diversifolia (Hemsl.) A. Gray against Bitis arietans and Naja nigricollis venoms-induced toxicity

Leaf methanol extract and fractions of Tithonia diversifolia (Hemsl.) A. Gray (Asteraceae) used in traditional medicine was evaluated for neutralization of toxicity induced by Bitis arietans and Naja nigricollis snake venoms in mice and rats. Inhibition of adenosine diphosphate-induced platelet aggregation, antioxidant activity, total phenol content and neutralization of venom-induced toxicity were evaluated by standard methods. The methanol extract was the most active inhibitor of platelet aggregation (66.7 - 84.8%) in the B. arietans model at 100 - 500 μg/ml. The methanol extract and fractions appear to be equipotent in inhibiting platelet aggregation (68 - 74%) induced by N. nigricollis. The aqueous fraction was more active against B. arietans venom at 500 μg/ml as an inhibitor of platelet aggregation (59.4% inhibition), while the dichloromethane (DCM) fraction was more potent against N. nigricollis (70.97% inhibition). The methanol extract showed highest DPPH radical scavenging activity followed by the aqueous and DCM fractions. Total phenolic content was in the order: methanol extract > DCM fraction > aqueous fraction. The aqueous fraction was a better antivenin agent in both B. arietans (complete protection at 24 h) and N. nigricollis-envenomed mice and rat models. Anti-snake venom activity was more prominent at 24 h. The results herein, being the very first report, indicate the potential antiophidic activity of T. diversifolia leaf particularly against B. arietans-induced toxicity.

A bioactive flavan-3-ol from the stem bark of Neocarya macrophylla

Antisnake venom activity of the ethylacetate and n-butanol soluble fractions of the methanol stem bark extract of Neocarya macrophylla were evaluated against Naja nigricollis venom. There was maximum protection to mice against venom-induced action after administration of the pre-incubated mixture of the venom and the fractions compared with the venom only. The fractions were also able to significantly inhibit phospholipase A2 enzyme activity. Based on the above findings, the ethylacetate fraction was subjected to purification using column chromatography and gel filtration; the procedures led to the isolation of a flavan-3-ol, catechin. The structure of the compound was identified based on chemical test, NMR analysis (1D- & 2D-NMR) and comparison with existing data in the literature. The catechin have demonstrated significant (p<0.05) antisnake venom activity in vitro with 100% survival rate; it was also found to be 42.0% effective in combating the hydrolytic action of N. nigricollis PLA2 enzyme. Catechin significantly (p<0.05) reduced the nociceptive response induced by formalin in both phases and there was remarkable inhibition of formalin-induced inflammation at the 1st, 2nd and 3rd hours, though the effect was not statistically significant compared to the control. The compound, catechin could serve as a lead in the development of novel antisnake venoms, analgesics and anti-inflammatory agents.

Inhibition of snake venom induced sterile inflammation and PLA2 activity by Titanium dioxide Nanoparticles in experimental animals

Sterile inflammation (SI) is an essential process in response to snakebite and injury. The venom induced pathophysiological response to sterile inflammation results into many harmful and deleterious effects that ultimately leads to death. The available treatment for snakebite is antiserum which does not provide enough protection against venom-induced pathophysiological changes like haemorrhage, necrosis, nephrotoxicity and often develop hypersensitive reactions. In order to overcome these hindrances, scientists around the globe are searching for an alternative therapy to provide better treatment to the snake envenomation patients. In the present study TiO2 (Titanium dioxide)-NPs (Nanoparticles) has been assessed for antisnake venom activity and its potential to be used as an antidote. In this study, the synthesis of TiO2-NPs arrays has been demonstrated on p-type Silicon Si < 100 > substrate (∼30 ohm-cm) and the surface topography has been detected by Field-emission scanning electron microscopy (FESEM). The TiO2-NPs successfully neutralized the Daboia russelii venom (DRV) and Naja kaouthia venom (NKV)-induced lethal activity. Viper venom induced haemorrhagic, coagulant and anticoagulant activities were effectively neutralized both in in-vitro and in vivo studies. The cobra and viper venoms-induced sterile inflammatory molecules (IL-6, HMGB1, HSP70, HSP90, S100B and vWF) were effectively neutralised by the TiO2-NPs in experimental animals.

Strategy for early callus induction and identification of anti-snake venom triterpenoids from plant extracts and suspension culture of Euphorbia hirta L.

Euphorbia hirta L. from the family of Euphorbiaceae is an annual herb, which grows as a roadside weed in most tropical countries. It is prominently used by the traditional healers in rural India for the treatment of snakebites. However, the mechanisms and the major bioactive compounds behind its inhibition activity are relatively unknown. From our preliminary in silico studies, it was found that a group of pentacyclic triterpenoids from this plant are playing a major role in inhibiting the snake venom proteins. The present study was aimed at standardizing methods for obtaining callus from this medicinal plant at a much faster rate by hormone pretreatment of explants and, thus, by developing suspension cultures to obtain bioactive secondary metabolites in vitro. The results were promising that longer incubation of explants with hormone treatment showed early induction of callus. The major bioactive compounds responsible for the anti-snake venom activity were characterized from natural plant material as well as from suspension cultures, and the efficiency was found to be relatively high. The secondary metabolite analysis from suspension culture and natural plant extracts revealed that a major compound 'Taraxerol' and its derivatives was found abundant along with few other triterpenoids. This compound showed high inhibitory activity against pit viper snake venoms from our in silico studies with molecular docking tools. Hence, this study with identification of potential bioactive compounds against snake venom with standardization of In vitro culture methods would help in developing natural alternative medicine for snakebites in near future.

Antisnake venom activity and isolation of quercetin from the leaf of Neocarya macrophylla (Sabine) Prance ex F. White (Malpighiales: Chrysobalanaceae)

Snake envenomation is a major cause of death and disability in many developing countries. Neocarya macrophylla (Sabine) Prance ex F. White (Malpighiales: Chrysobalanaceae) have been reportedly used in traditional medicine to treat snake envenomation. Bioassay-guided isolation of antivenom principles was carried out on the leaf of N. macrophylla against Naja nigricollis venom. The methanol extract of N. macrophylla leaf and its ethylacetate and n-butanol fraction significantly (P &lt; 0.05) protected mice against venom-induced lethality with 100% survival rate and there was remarkable inhibition of the poisonous effects of PLA2 enzyme by the extracts and the fractions. Encouraged by this result, the ethylacetate soluble fraction was subjected to purification using vacuum liquid chromatography and gel filtration which led to the isolation of quercetin as the bioactive principle. The identity of the compound was determined on the basis of chemical tests, and by comparison of its 1H-NMR data with literature, this is the first report of isolation of this compound from the leaf of the plant. However, the results of the study suggests that the leaf of N. macrophylla possess significant antisnake venom activity which provide the scientific basis for its use in traditional treatment of snakebites.

Anti-snake venom potential of Clerodendrum serratum extracts on Bungarus caeruleus and Daboia russelii venom

Anti-snake venom therapy is the only treatment for snake bite but leads to acute and chronic conditions which may be severe. The medicinal plants have gained importance over years to find an effective alternative to anti-snake venom. The present study focused on evaluating the potential of Clerodendrum serratum for the anti-snake venom activity. Phytochemicals were extracted from the C. serratum with different solvents. The ethyl acetate and methanolic extracts were found to neutralize the major enzyme toxins (phospholipase A2, protease and hyaluronidase) of Bungarus caeruleus and Daboia russelii venom at a concentration of 100 µg/mL. The fibrinogenolytic activity of both the venoms were neutralized. The study proves that the plant C. serratum possesses certain compounds which inhibit the toxins present in the venom of B. caeruleus and D. russelii.&#x0D; Video Clip of Methodology:&#x0D; Hyaluronidase assay: 3 min 30 sec Click to watch