Activity Of Snake Venoms Research Articles

A functional and thromboelastometric-based micromethod for assessing crotoxin anticoagulant activity and antiserum relative potency against Crotalus durissus terrificus venom

The assessment of the capacity of antivenoms to neutralize the lethal activity of snake venoms still relies on traditional rodent in vivo lethality assay. ED50 and LD50 assays require large quantities of venoms and antivenoms, and besides leading to animal suffering. Therefore, in vitro tests should be introduced for assessing antivenom neutralizing capacity in intermediary steps of antivenom production. This task is facilitated when one key lethal toxin is identified. A good example is crotoxin, a β-neurotoxin phospholipase A2-like toxin that presents anticoagulant activity in vitro and is responsible for the lethality of venoms of Crotalus durissus snakes. By using rotational thromboelastometry, we reported recently one sensitive coagulation assay for assessing relative potency of the anti-bothropic serum in neutralizing procoagulant activity of Bothrops jararaca venom upon recalcified factor-XII-deficient chicken plasma samples (CPS). In this study, we stablished conditions for determining relative potency of four batches of the anti-crotalic serum (ACS) (antagonist) in inactivating crotoxin anticoagulant activity in CPS (target) simultaneously treated with one classical activator of coagulation (agonists). The correlation coefficient (r) between values related the ACS potency in inactivating both in vitro crotoxin anticoagulant activity and the in vivo lethality of whole venom (ED50) was 0.94 (p value < 0.05). In conclusion, slowness in spontaneous thrombin/fibrin generation even after recalcification elicit time lapse sufficient for elaboration of one dose-response curve to pro- or anti-coagulant agonists in CPS. We propose this methodology as an alternative and sensitive assay for assessing antivenom neutralizing ability in plasma of immunized horses as well as for in-process quality control.

Characterization of the Rabbit as an In Vitro and In Vivo Model to Assess the Effects of Fibrinogenolytic Activity of Snake Venom on Coagulation.

Several invitro investigations have demonstrated that anticoagulant effects of fibrinogenolytic snake venom metalloproteinases have been abrogated in human plasma by modifying fibrinogen with iron (Fe) and carbon monoxide (CO) to prevent catalysis or by directly inhibiting these enzymes with CO. To translate these findings, we chose to assess the rabbit as a model of envenomation with Crotalus atrox venom. It was determined with thrombelastography that 15 times the concentration of venom noted to compromise coagulation in plasma invitro was required to cause coagulopathy invivo, likely secondary to venom binding to blood cells and being cleared from the circulation rapidly. Unlike human plasma, rabbit plasma pre-treated with Fe/CO was not protected from fibrinogenolysis by venom. Consequently, the administration of purified human fibrinogen (with or without Fe/CO) would be required before venom administration to rabbits. Of greater interest, venom exposed to CO had complete loss of fibrinogenolytic effect in rabbit plasma and partial loss of activity in whole blood, indicative of unbinding of CO from venom and binding to haemoglobin. Thus, venom exposed to CO could remain partially or completely inhibited in whole blood long enough for clearance from the circulation, allowing rabbits to be a useful model to test the efficacy of regional CO administration to the bite site. Future investigations are planned to test these novel approaches to attenuate venom-mediated coagulopathy in the rabbit.

Both non-covalent and covalent interactions were involved in the mechanism of detoxifying effects of persimmon tannin on Chinese cobra PLA2

Persimmon tannin (PT) has been shown to inhibit snake venom activities and toxicities both in vitro and in vivo. To clarify the detoxifying mechanism of PT on snake venom, the interaction of characteristic structural elements of PT (EGCG, ECG, EGCG dimer and ECG dimer) and Chinese cobra phospholipase A2 (PLA2) was studied. The results revealed that except non-covalent bonds like hydrogen bonds, hydrophobic bonds and iron bonds were formed between PT and PLA2, covalent interaction was also occurred. PT could bind with the key active residues of PLA2, such as lysine, histidine, tryptophan and tyrosine, restraining their activity and disturbing the structure of PLA2, thus showing detoxifying effects on snake venom.

Cross-Neutralisation of In Vitro Neurotoxicity of Asian and Australian Snake Neurotoxins and Venoms by Different Antivenoms.

There is limited information on the cross-neutralisation of neurotoxic venoms with antivenoms. Cross-neutralisation of the in vitro neurotoxicity of four Asian and four Australian snake venoms, four post-synaptic neurotoxins (α-bungarotoxin, α-elapitoxin-Nk2a, α-elapitoxin-Ppr1 and α-scutoxin; 100 nM) and one pre-synaptic neurotoxin (taipoxin; 100 nM) was studied with five antivenoms: Thai cobra antivenom (TCAV), death adder antivenom (DAAV), Thai neuro polyvalent antivenom (TNPAV), Indian Polyvalent antivenom (IPAV) and Australian polyvalent antivenom (APAV). The chick biventer cervicis nerve-muscle preparation was used for this study. Antivenom was added to the organ bath 20 min prior to venom. Pre- and post-synaptic neurotoxicity of Bungarus caeruleus and Bungarus fasciatus venoms was neutralised by all antivenoms except TCAV, which did not neutralise pre-synaptic activity. Post-synaptic neurotoxicity of Ophiophagus hannah was neutralised by all antivenoms, and Naja kaouthia by all antivenoms except IPAV. Pre- and post-synaptic neurotoxicity of Notechis scutatus was neutralised by all antivenoms, except TCAV, which only partially neutralised pre-synaptic activity. Pre- and post-synaptic neurotoxicity of Oxyuranus scutellatus was neutralised by TNPAV and APAV, but TCAV and IPAV only neutralised post-synaptic neurotoxicity. Post-synaptic neurotoxicity of Acanthophis antarcticus was neutralised by all antivenoms except IPAV. Pseudonaja textillis post-synaptic neurotoxicity was only neutralised by APAV. The α-neurotoxins were neutralised by TNPAV and APAV, and taipoxin by all antivenoms except IPAV. Antivenoms raised against venoms with post-synaptic neurotoxic activity (TCAV) cross-neutralised the post-synaptic activity of multiple snake venoms. Antivenoms raised against pre- and post-synaptic neurotoxic venoms (TNPAV, IPAV, APAV) cross-neutralised both activities of Asian and Australian venoms. While acknowledging the limitations of adding antivenom prior to venom in an in vitro preparation, cross-neutralization of neurotoxicity means that antivenoms from one region may be effective in other regions which do not have effective antivenoms. TCAV only neutralized post-synaptic neurotoxicity and is potentially useful in distinguishing pre-synaptic and post-synaptic effects in the chick biventer cervicis preparation.

Application of Gold Nanoparticle Conjugation with 2-Hydroxy-4-Methoxy Benzoic Acid (HMBA) from Hemidesmus indicus Root Enhancing Neutralization of Snake (Viper) Venom Activity

Application of Gold Nanoparticle Conjugation with 2-Hydroxy-4-Methoxy Benzoic Acid (HMBA) from <i>Hemidesmus indicus</i> Root Enhancing Neutralization of Snake (Viper) Venom Activity

­­Ethnobotany, phytochemistry, pharmacognostic and pharmacological aspects of Cordia macleodii Hook.f. &amp; Thomson - A review

Cordia macleodii Hook.f. &amp; Thomson is an important ethnomedicinal plant of India. It is found in the moist and dry deciduous forests of peninsular India such as Odisha, Madhya Pradesh, Chhattisgarh, Tamil Nadu etc. The plant is used ethnomedicinally for various purposes like healing wounds, mouth sores, treating jaundice and also as an aphrodisiac by the tribal people where it is found. Its pharmacological activities include anti-microbial, wound healing, anti-oxidant and hepatoprotective, anti snake venom, analgesic and anti-inflammatory activity. Many phytocompounds like quercetin, p-hydroxyphenylacetic acid and phytosterols such as β-sitosterol, stigmasterol, camphesterol and Cholest-5-EN-3OL (3β)-Carbonyl chlorinated are isolated from this plant. The present work aims at presenting a comprehensive of the plants botany, ethnomedicinal uses, chemical constituents, pharmacognostic and pharmacological uses. The information reported in this work contributes scientifically to recognizing the importance of Cordia macleodii as a target in the search for new biotechnological investments.

Integrative characterization of the venom of the coral snake Micrurus dumerilii (Elapidae) from Colombia: Proteome, toxicity, and cross-neutralization by antivenom

In Colombia, nearly 2.8% of the 4200 snakebite accidents recorded annually are inflicted by coral snakes (genus Micrurus). Micrurus dumerilii has a broad distribution in this country, especially in densely populated areas. The proteomic profile of its venom was here studied by a bottom-up approach combining RP-HPLC, SDS-PAGE and MALDI-TOF/TOF. Venom proteins were assigned to eleven families, the most abundant being phospholipases A2 (PLA2; 52.0%) and three-finger toxins (3FTx; 28.1%). This compositional profile shows that M. dumerilii venom belongs to the ‘PLA2-rich’ phenotype, in the recently proposed dichotomy for Micrurus venoms. Enzymatic and toxic venom activities correlated with protein family abundances. Whole venom induced a conspicuous myotoxic, cytotoxic and anticoagulant effect, and was mildly edematogenic and proteolytic, whereas it lacked hemorrhagic activity. Some 3FTxs and PLA2s reproduced the lethal effect of venom. A coral snake antivenom to Micrurus nigrocinctus demonstrated significant cross-recognition of M. dumerilii venom proteins, and accordingly, ability to neutralize its lethal effect. The combined compositional, functional, and immunological data here reported for M. dumerilii venom may contribute to a better understanding of these envenomings, and support the possible use of anti-M. nigrocinctus coral snake antivenom in their treatment. Biological significanceCoral snakes represent a highly diversified group of elapids in the New World, with nearly 70 species within the genus Micrurus. Owing to their scarce yields, the biochemical composition and toxic activities of coral snake venoms have been less well characterized than those of viperid species. In this work, an integrative view of the venom of M. dumerilii, a medically relevant coral snake from Colombia, was obtained by a combined proteomic, functional, and immunological approach. The venom contains proteins from at least eleven families, with a predominance of phospholipases A2 (PLA2), followed by three-finger toxins (3FTx). According to its compositional profile, M. dumerilii venom can be grouped with those of several Micrurus species from North and Central America that present a PLA2-predominant phenotype, to date it is the most southerly coral snake species to do so. Other coral snake species that a ‘PLA2-rich’ venom, M. dumerilii venom contains both components that form MitTx, a pain-inducing heterodimeric complex recently characterized from the venom of Micrurus tener, also present in Micrurus mosquitensis and M. nigrocinctus venoms. In addition to a lethal three-finger toxin, PLA2s participate in the toxicity of M. dumerilii venom, some of them displaying ability to induce cytolysis, muscle necrosis, and lethality to mice. An antivenom to M. nigrocinctus demonstrated significant cross-recognition of M. dumerilii venom proteins, and accordingly, ability to neutralize its lethal effect, being of potential therapeutic usefulness in these envenomings.

In Vitro Activity Screening of Snake Venom against Multi Drug Resistant Tuberculosis

The re-emergence of multidrug-resistant tuberculosis (MDR-TB) events has brought to light the importance of screening effective novel drugs. In the present study, in vitro activities of different snake (Naja naja, Bungarus fasciatus, Daboia russelli russelli, Naja kaouthia) venoms have been investigated against clinical isolate of MDR-TB strains. All the venoms inhibited the mycobacterial growth for at least a week in common and two of them (Naja naja, Naja kaouthia) showed significantly longer inhibition up to two weeks against the MDR-TB strain with single dose and one repetition of those two venoms exhibited inhibition up to more than 4 weeks.

A Study of Snake Oral Flora-An Aid in Management of Snake Bite

Since ancient times snake bite is a rural and occupational public health problem in many countries. In the present study variety of microorganisms, including both aerobic and anaerobic organisms has been isolated from snake oral cavity. Snake bite may result in venom toxicity and secondary infections due to microorganisms from snake oral cavity. Since few snake oral cavity did not show any growth and the antibacterial activity of snake venom makes to reconsider the use of prophylactic antibiotics. Use of antibiotics is better supported with culture sensitivity examination.

In vitro screening of snake venom against multidrug-resistant tuberculosis

The re-emergence of multidrug-resistant tuberculosis (MDR-TB) has brought to light the importance of screening effective novel drugs. In the present study, in vitro activities of different snake (Naja naja, Bungarus fasciatus, Daboia russelli russelli, Naja kaouthia) venoms have been investigated against clinical isolate of MDR-TB strains. The treatment with all the venoms inhibited the mycobacterial growth for at least a week in common and two of them (Naja naja and Naja kaouthia) showed significantly longer inhibition up to two weeks against the MDR-TB strain with single dose and a repetition of those two venoms exhibited inhibition up to more than four weeks.

Assessment of the Antimicrobial Activity of Few Saudi Arabian Snake Venoms.

BackgroundVenoms of two cobras, four vipers, a standard antibiotic and an antimycotic, were evaluated comparatively, as antimicrobials.Methods:Six venom concentrations and three of the standard antibiotic and the antimycotic were run in micro-dilution and diffusion plates against the microorganisms.RESULTS: Echis pyramidum, Echis coloratus and Cerastes cerastes gasperettii highest venom concentrations gave significant growth inhibition zones (GIZ) with respect to a negative control, except Bitis arietans, whose concentrations were significant. The cobra Walterinnesia aegyptia had significant venom concentrations more than Naja haje arabica. The Staphylococcus aureus Methicillin Resistant (MRSA) bacterium was the most susceptible, with a highly (P < 0.001) significant GIZ mean difference followed by the Gram positive Staphylococcus aureus, (P < 0.001), Escherichia coli (P < 0.001), Enterococcus faecalis (P < 0.001) and Pseudomonas aeruginosa which, had the least significance (P < 0.05). The fungus Candida albicans was resistant to both viper and cobra venoms (P > 0.05). The antibiotic Vancomycin was more effective than snake venoms though, they were more efficient in inhibiting growth of the resistant Pseudomonas aeruginosa. This antibiotic was also inactive against the fungus, whilst its specific antifungal Fungizone was highly efficient with no antibacterial activity.Conclusions:These findings showed that snake venoms had antibacterial activity comparable to antibiotics, with a directly proportional relationship of venom concentration and GIZ, though, they were more efficient in combatting resistant types of bacteria. Both venoms and the standard antibiotic, showed no antifungal benefits.

In vitro Antibacterial Activity of Snake Venom, Naja naja from Bangladesh

Aim: Snake venom is a source of antimicrobial peptides. Composition as well as activities of venom may vary based on geographic region and notably the antimicrobial activity of the venom of Naja naja from Bangladesh has not yet been tested. Thus, in this study, investigated the antibacterial activity of the venom of snake Naja naja from Bangladesh against two bacterial strains. Place and Duration of Study: The study was carried out in protein science laboratory, Department of Genetic Engineering and Biotechnology, University of Rajshahi, Bangladesh, between July 2012 to December 2012. Methods: Luria agar medium was used to culture the E. coli and B. thuringiensis strains of bacteria. Disc diffusion method was used to carry out the test of antibacterial sensitivity. Discs prepared by Whatman No. 1 filter paper were soaked with different doses (25, 50, 75 and 100 μg) of crude venom and placed on the cultured bacterial plates along with two standard antimicrobial antibiotic discs. After overnight incubation at 37oC, antibacterial activity of venom was measured by inhibition zone observed around the discs in the plates. Short Research Article Hakim and Reza; BBJ, 8(2): 1-5, 2015; Article no.BBJ.18374 2 Results: Application of 75 and 100 μg of crude venom showed antibacterial activity against E. coli, whereas only 100 μg crude venom showed little antibacterial activity against B. thuringiensis. Conclusion: The results of this study revealed that the crude venom of Naja naja may contain some proteins responsible for antibacterial activity and the quantity of antibacterial peptides might be lower in venom than other components.

Evaluation of cytotoxic activities of snake venoms toward breast (MCF-7) and skin cancer (A-375) cell lines.

Snake venoms are mixtures of bioactive proteins and peptides that exhibit diverse biochemical activities. This wide array of pharmacologies associated with snake venoms has made them attractive sources for research into potentially novel therapeutics, and several venom-derived drugs are now in use. In the current study we performed a broad screen of a variety of venoms (61 taxa) from the major venomous snake families (Viperidae, Elapidae and "Colubridae") in order to examine cytotoxic effects toward MCF-7 breast cancer cells and A-375 melanoma cells. MTT cell viability assays of cancer cells incubated with crude venoms revealed that most venoms showed significant cytotoxicity. We further investigated venom from the Red-bellied Blacksnake (Pseudechis porphyriacus); venom was fractionated by ion exchange fast protein liquid chromatography and several cytotoxic components were isolated. SDS-PAGE and MALDI-TOF mass spectrometry were used to identify the compounds in this venom responsible for the cytotoxic effects. In general, viper venoms were potently cytotoxic, with MCF-7 cells showing greater sensitivity, while elapid and colubrid venoms were much less toxic; notable exceptions included the elapid genera Micrurus, Naja and Pseudechis, which were quite cytotoxic to both cell lines. However, venoms with the most potent cytotoxicity were often not those with low mouse LD50s, including some dangerously venomous viperids and Australian elapids. This study confirmed that many venoms contain cytotoxic compounds, including catalytic PLA2s, and several venoms also showed significant differential toxicity toward the two cancer cell lines. Our results indicate that several previously uncharacterized venoms could contain promising lead compounds for drug development.

An alternative micromethod to access the procoagulant activity of Bothrops jararaca venom and the efficacy of antivenom

The assessment of the capacity of antivenoms to neutralize the lethal activity of snake venoms still relies largely on traditional rodent lethality assay (LD50). However, adequately validated in vitro tests should be introduced for assessing antivenom neutralizing capacity in plasma of immunized horses as well as for in-process quality control. The dynamic of fibrin formation in recalcified avian plasma samples is extremely slow, when compared to that presented by mammalian plasmas. In this study, we present one new coagulant assay, by performing dose–response curve after plotting the clotting time (CT) parameter of the ROTEM profile of recalcified chicken plasma samples (target) against semi-logarithmic doses of Bothrops jararaca venom (agonist), either in absence or in presence of the semi-logarithmic doses of anti-bothropic serum (ABS) (antagonist).The mean coagulant dose 50% (CD50) was defined as the quantity of venom (in μg) which reduces CT to 900 s, between minimum and maximum responses. The CT induced by 5CD50 of the venom was used as the control for calculating the effective dose (ED) of each batch of ABS. ED was defined as the ABS dose (nanoliters, nL) at which CT induced by one amount of venom corresponding to 5CD50 is displaced to the maximum threshold (1800 s). Five batches of the ABS, previously assayed for their lethality neutralizing activity (ED50) were assayed. The correlation coefficient (r) between both in vitro (ED) and in vivo (ED50) values was 0.87 (p value < 0.05). We propose this micro method as highly sensitive for characterization and quantification of possible procoagulant activity of small doses of snake venoms (nanograms) and for detecting small doses (nanoliters) of specific antibodies against this effect in little volume samples of biological fluids.

Inhibitory effect of the red seaweed Plocamium brasiliense against the toxic effects of Lachesis muta snake venom

The ability of extracts and fractions of the red seaweed Plocamium brasiliense to inhibit hemorrhagic, edematogenic, hemolytic, clotting and proteolytic activities of Lachesis muta snake venom was analyzed. In Brazil, snakebites by L. muta are low (2 %) when compared to Bothrops genus (90 %); however, their lethality indexes are three times higher than Bothrops .E nvenomation byL. muta venom results in hemorrhage, pain, necrosis, hemolysis, myotoxicity, and death. Since antivenom does not efficiently neutralize local effects, a large number of researchers have attempted to identify molecule(s) from natural sources to inhibit such activities to use them as an alternative treatment for snakebite. We tested four extracts of seaweed P. brasiliense obtained with solvents of increasing polarities: n-hexane (HEX), dichloromethane (DCM), ethyl acetate (ACE), and hydroalcohol (HYD). Extracts of alga or fractions were incubated with L. muta venom, and then, biological assays were performed. The extracts, except the HYD, inhibited all the assays but with different potencies. The DCM extract fully inhibited all activities. Moreover, DCM and HEX extracts inhibited hemolysis induced by a phospho- lipase A2 isolated from L. muta venom (LM-PLA2-I). A fraction from HEX enriched in cholesterol isolated from HEX extract inhibited proteolysis by L. muta venom and hemolysis by LM-PLA2-I; in contrast, monoterpenes isolated from DCM extract did not inhibit both activities. Seaweeds may be a promising source of natural inhibitors of the toxic effects caused by snakebite by L. muta venom, and they may be used to develop new strategies for antivenom treatment.

Inhibitory Effect of the Plant Clusia fluminensis against Biological Activities of Bothrops jararaca Snake Venom

The ability of extracts of the plant Clusia fluminensis Planch & Triana (Clusiaceae Lindl.) to neutralize proteolysis, clotting, hemolysis, hemorrhagic and lethality activities of Bothrops jararaca snake venom was studied. Clusianone and lanosterol from the flower and fruit extracts, respectively, were also tested. The extracts of different organs of C. fluminensis inhibited proteolysis and hemolysis induced by B. jararaca venom, but with different potencies. Only the stems prevented blood clotting. Only the acetone extract of the fruit protected mice from hemorrhage while the acetone or methanol extracts prevented mice from death. Clusianone and lanosterol did not inhibit clotting or hemorrhage, but the former inhibited proteolysis and the latter hemolysis.

Inhibitory effect of plant Manilkara subsericea against biological activities of Lachesis muta snake venom.

Snake venom is composed of a mixture of substances that caused in victims a variety of pathophysiological effects. Besides antivenom, literature has described plants able to inhibit injuries and lethal activities induced by snake venoms. This work describes the inhibitory potential of ethanol, hexane, ethyl acetate, or dichloromethane extracts and fractions from stem and leaves of Manilkara subsericea against in vivo (hemorrhagic and edema) and in vitro (clotting, hemolysis, and proteolysis) activities caused by Lachesis muta venom. All the tested activities were totally or at least partially reduced by M. subsericea. However, when L. muta venom was injected into mice 15 min first or after the materials, hemorrhage and edema were not inhibited. Thus, M. subsericea could be used as antivenom in snakebites of L. muta. And, this work also highlights Brazilian flora as a rich source of molecules with antivenom properties.

Anti-hemorrhagic effect of hydro-alcoholic extract of the leaves of Mikania glomerata in lesions induced by Bothrops jararaca venom in rats

To investigate the effects of hydroalcoholic leaf extract of Mikania glomerata Spreng (Asteraceae) on the activity of Bothrops jararaca snake venom in Wistar rats. Fifty four rats Wistar were divided into six groups of nine animals in each: control treated with saline; control treated with B. jararaca venom; control treated with M. glomerata extract; B. jararaca venom incubated with M. glomerata extract at proportions of 1:1, 1:2, and 1:4. Histopathological and morphometric analysis showed that intradermal administration of snake venom incubated with the hydroalcoholic extract at proportions of 1:1, 1:2 and 1:4 promoted a significant reduction in the number of inflammatory cells and a marked decrease in edema after the third hour. There was also a significant reduction in the intensity of the hemorrhagic halo in animals receiving the snake venom incubated with the extract, with the observation of a progressive and parallel inhibition with increasing proportion of M. glomerata. The Mikania glomerata hydroalcoholic extract exerted effective anti-inflammatory and antihemorrhagic activity against the effects induced by Bothrops jararaca snake venom.

Anti Snake Venom Potential of Securidaca longepedunculata Leaf and Root Bark on Spitting Cobra (Naja nigricollis Hallowel) in Envenomed Wister Rats

This study was undertaken to investigate the anti snake venom potential of aqueous extracts of Securidaca longepedunculata leaves and root bark extracts against Snake venom of Spitting cobra (Naja nigricollis). The venom of Naja nigricollis snake was used to test the anti snake venom efficacy of the extracts, using experimental rats. The anti snake venom activity revealed that root bark extract are able to neutralize snake venom at 300mg/kg body weight with 100% survival. And the leaf extract recorded lower anti venom activity of 0% at 200mg/ml survival and 33.33% of mortality at 300mg/ml. But in the combined root bark and leaves extract it shows higher activity of 66.67% survival at 300mg/ml and 33.33% mortality at 200mg/ml. The result indicate that phytochemical constituents of S. longepedunculata leaves and root bark extracts could also be used to enhance the treatment of the snake venom or bites as it shows a significant anti venom effect (p<0.05) against Naja nigricollis venom. Sub-acute toxicity and determination of LD50 of the plant extracts should be carried out and to enlighten pharmacists on the use of such plant extract for the treatment of various snake bites.

In vivo and in vitro antileishmanial activity of Bungarus caeruleus snake venom through alteration of immunomodulatory activity

Leishmaniasis threatens more than 350 million people worldwide specially in tropical and subtropical region. Antileishmanial drugs that are currently available have various limitations. The search of new drugs from natural products (plants, animals) possessing antileishmanial activity is ventured throughout the world. The present study deals with the antileishmanial activity of Bungarus caeruleus snake venom (BCV) on in vitro promastigotes and amastigotes of Leishmania donovani parasite and leishmania infected BALB/c mice. The effect of BCV on peritoneal macrophage, release of cytokines from the activated macrophages, production of nitric oxide, reactive oxygen species and cytokines were studied in vivo and in vitro. IC50 value of BCV on L. donovani promastigote was 14.5μg/ml and intracellular amastigote was 11.2μg/ml. It activated peritoneal macrophages, significantly increased cytokines and interleukin production. BCV (20μg/kg and 40μg/kg body weight of mice) decreased parasite count by 54.9% and 74.2% in spleen and 41.4% and 60.4% in liver of infected BALB/c mice. BCV treatment significantly increased production of TNF-α, IFN-γ, ROS, NO in infected mice. Histological studies showed decreased granuloma formation in treated liver as compared with control. Liver and spleen structure was partially restored due to BCV treatment in infected mice. The present study revealed that BCV possessed antileishmanial activity against L. donovani parasite in vivo and in vitro and this activity was partly mediated through immunomodulatory activity involving macrophages.