In Vitro Toxic Effects of Puff Adder (Bitis arietans) Venom, and Their Neutralization by Antivenom (original) (raw)
Related papers
2014
This study investigated the in vitro toxic effects of Bitis arietans venom and the ability of antivenom produced by the South African Institute of Medical Research (SAIMR) to neutralize these effects. The venom (50 µg/mL) reduced nerve-mediated twitches of the chick biventer muscle to 19% ± 2% of initial magnitude (n = 4) within 2 h. This inhibitory effect of the venom was significantly attenuated by prior incubation of tissues with SAIMR antivenom (0.864 µg/µL; 67% ± 4%; P < 0.05; n = 3-5, unpaired t-test). Addition of antivenom at t 50 failed to prevent further inhibition or reverse the inhibition of twitches and responses to agonists. The myotoxic action of the venom (50 µg/mL) was evidenced by a decrease in direct twitches (30% ± 6% of the initial twitch magnitude) and increase in baseline tension (by 0.7 ± 0.3 g within 3 h) of the chick biventer. Antivenom failed to block these effects. Antivenom however prevented the venom induced cytotoxic effects on L6 skeletal muscle cells. Venom induced a marginal but significant reduction in plasma clotting times at concentrations above 7.8 µg/100 µL of plasma, indicating poor procoagulant effects. In addition, the results of western immunoblotting indicate strong immunoreactivity with venom proteins, thus warranting further detailed studies on the
Revista do Instituto de …, 2011
BACKGROUND: Serpente das Montanhas da Etiópia (Bitis parviocula) é um viperídeo conhecido somente em poucas localizações do sudoeste da Etiópia. MÉTODOS: Um total de 30 µg de veneno de B. arietans e B. parviocula foram corridos em gel de 10 a 20% de tricina. Para se estabelecer a quinquagésima dose de letalidade (LD50) foram usados cinco grupos de oito camundongos para cada veneno. A atividade hemorrágica para o veneno cru foi testada. A atividade fibrogenolítica do veneno cru foi medida usando 2,5 mg/mL de solução de fibrinogênio e 0,03 mg/mL de veneno cru. A atividade de gelatinase do veneno foi testada em um filme KODAK X-OMATTM. Venenos crus de B. parviocula e B. arietans foram testados no que diz respeito à sua capacidade de afetar o tempo de coagulação, a velocidade de coagulação e a função plaquetogênica em sangue humano total. RESULTADO: o antiveneno SAIMR foi confirmado neste estudo no que diz respeito à neutralização da atividade letal do veneno de Bitis parviocula. ED50s do antiveneno SAIMR sobre a B. parviocula e B. arietans neutralizou metade de 18,2 e 66,7 mg respectivamente do veneno. As atividades hemorrágicas (MHDs) de B. parviocula e B. arietans foram respectivamente 0,88 e 1,7 µg. Os venenos de B. arietans e B. parviocula degradaram cadeias α e β em tempos diferentes. A cadeia Γ permaneceu não afetada. O veneno da B. parviocula não mostrou atividade de gelatinase, enquanto o de B. arietans teve um MGD de 6,9 µg. A nível de 3 mg/mL os venenos crus de B. parviocula e B. arietans não afetaram significantemente o tempo e a velocidade de coagulação. CONCLUSÕES: O antiveneno SAIMR é bastante efetivo para neutralizar o veneno da B. parviocula e deveria ser considerado para o tratamento de envenenamentos por estas serpentes.
Toxicon, 1989
F . CI-IAVFS, J. M. GuTdRREZ, B. LOMONTE and L. CsRDi~s. Histopathological and biochemical alterations induced by intramuscular injection of Bothrops riper (terciopelo) venom in mice . Toxicon 27, 1081093, 1989 .-The local and systemic pathological changes induced by an i.m . injection of 100 ug of Bothrops riper venom in mice were studied histologically and by following th ; changes in serum levels of enzymes, proteins, ATP and lactate, as well as alterations in hematocrit and clotting time. B. aspen venom induced a rapid and marked increase in serum levels of creatine kinase, asparta,te aminotransferase and lactate dehydrogenase, but not alanine aminotransferase or alkaline phosphatase. A local myonecrosis and hemorrhage was observed, with the lungs collapsing by 24 hr and the kidneys showing glomerular congestion and vacuolar degeneration of tubular cells. Only minor histopathological changes were observed in cardiac muscle and liver. Both ATP and lactate blood levels decreased after venom injection, whereas there were no changes in serum protein concentration . Blood incoagulability was observed 1 and 3 hr after envenomation . Antivenom neutralized venom-induced increases in serum enzyme levels following preincubation with venom, indicating that antivenom contains antibodies against tissue-damaging toxins. However, when antivenom was administered i.v . at different time intervals after venom injection, neutralization was only partial, with the exception of defibrinating activity, which was totally neutralized even after a delay of 1 hr in administering antivenom.
Toxicon : official journal of the International Society on Toxinology
A murine model of venom-induced myotoxicity was used to assess the antimyotoxic capacity of a polyvalent antivenom (PAV), rich in F(ab')2 fragments, obtained from horses immunized with Bitis venoms. Intramuscular (i.m.) injection of Bitis rhinoceros, Bitis arietans or Bitis nasicornis into mice induced a time- and dose-dependent increase in plasma CK activity. The area under the plasma CK activity vs. time curve (AUC) between 0 and 48 h was used to quantify the data. Pre-incubation with PAV neutralized the venoms' myotoxicity, in a concentration-dependent manner: 80-100% neutralization occurred when the ratio of the PAV volume to the venom mass was 3-fold that recommended for use in human envenomation. Intravenous administration of PAV 1 h before the i.m. venom injection, afforded significant protection against myotoxicity, especially in the case of B. arietans. An antimyotoxic effect was also observed, albeit reduced, when the PAV treatment was applied 1 h after the venom i...
Toxicon, 2001
Five-month-old white leghorn chickens were immunized with 50 mg of Common Cobra (Naja naja) and 30 mg of Krait venoms (Bungarus caeruleus) to generate antivenom antibodies against the venom antigen. Chickens received booster doses of increasing concentrations of venom at 14 days time intervals to raise the antivenom level in egg yolk. The antivenom from immunized chicken egg yolk was extracted by polyethylene glycol (PEG) and ammonium sulphate precipitation method which was further purified by DEAE cellulose ion exchange column chromatography. A high molecular weight protein of 180 kDa was detected by electrophoretic analysis which shows the purity of antivenom generated in chicken. Antibodies generated were specific and sensitive to the venom antigen. Various pharmacological activities of Cobra and Krait venoms were carried out by both in-vivo and in-vitro methods. The neutralization of lethality, hemorrhagic, edema, PLA 2 and procoagulant activity was evaluated in assays involving pre-incubation of venom and antivenom prior to testing. The antivenom was effective in neutralizing the toxic and enzymatic activities of venom. The LD 50 of venom for 18 g of mice was found to be 10 mg for Cobra and 3 mg for Krait venoms. The median effective dose (ED 50 ) of anti-Cobra venom was 4.48 mg/ 5LD 50 and 1.0 ml neutralized 0.127 mg of Cobra venom and the median effective dose (ED 50 ) of anti-Krait venom was 3.18 mg/5LD 50 and 1.0 ml neutralized 0.051 mg of Krait venom. The results indicate that antivenom generated in chicken could be used for therapeutic purposes in case of snakebite envenomation.
Toxins, 2014
Bungarus candidus and Bungarus fasciatus are two species of krait found in Southeast Asia. Envenoming by these snakes is often characterized by neurotoxicity and, without treatment, causes considerable morbidity and mortality. In this study, the in vitro neurotoxicity of each species, and the effectiveness of two monovalent antivenoms and a polyvalent antivenom, against the neurotoxic effects of the venoms, were examined in a skeletal muscle preparation. Both venoms caused concentration-dependent inhibition of indirect twitches, and attenuated responses to exogenous nicotinic receptor agonists, in the chick biventer preparation, with B. candidus venom being more potent than B. fasciatus venom. SDS-PAGE and western blot analysis indicated different profiles between the venoms. Despite these differences, most proteins bands were recognized by all three antivenoms. Antivenom, added prior to the venoms, attenuated the neurotoxic effect of the venoms. Interestingly, the respective monovalent antivenoms did not neutralize the effects of the venom from the other Bungarus species indicating a relative absence of cross-neutralization. Addition of a high concentration of polyvalent antivenom, at the t90 time point after addition of venom, partially reversed the neurotoxicity of B. fasciatus venom but not B. candidus venom. The monovalent antivenoms had no significant effect when added at the t90 time point. This study showed that B. candidus and B. fasciatus venoms display marked in vitro neurotoxicity in the chick biventer preparation and administration of antivenoms at high dose is necessary to prevent or reverse neurotoxicity
PLOS ONE
Naja sumatrana and Naja kaouthia are medically important elapids species found in Southeast Asia. Snake bite envenoming caused by these species may lead to morbidity or mortality if not treated with the appropriate antivenom. In this study, the in vitro neurotoxic and myotoxic effects N. sumatrana and N. kaouthia venoms from Malaysian specimens were assessed and compared. In addition, the neutralizing capability of Cobra Antivenom (CAV), King Cobra Antivenom (KCAV) and Neuro Polyvalent Antivenom (NPAV) from Thailand were compared. Both venoms produced concentration-dependent neurotoxic and myotoxic effects in the chick biventer cervicis nerve-muscle preparation. Based on the time to cause 90% inhibition of twitches (i.e. t90) N. kaouthia venom displayed more potent neurotoxic and myotoxic effects than N. sumatrana venom. All three of the antivenoms significantly attenuated venom-induced twitch reduction of indirectly stimulated tissues when added prior to venom. When added after N. ...
Species-dependent variations in the in vitro myotoxicity of death adder (Acanthophis) venoms
Toxicological Sciences, 2003
Based on early studies on Acanthophis antarcticus (common death adder) venom, it has long been thought that death adder snake venoms are devoid of myotoxicity. However, a recent clinical study reported rhabdomyolysis in patients following death adder envenomations, in Papua New Guinea, by a species thought to be different to A. antarcticus. Subsequently, a myotoxic phospholipase A 2 component was isolated from A. rugosus (Irian Jayan death adder) venom. The present study examined the venoms of
Toxicology and applied …, 2001
Although viperlike in appearance and habit, death adders belong to the Elapidae family of snakes. Systemic envenomation represents a serious medical problem with antivenom, which is raised against Acanthophis antarcticus venom, representing the primary treatment. This study focused on the major Acanthophis variants from Australia and islands in the Indo-Pacific region. Venoms were profiled using liquid chromatography-mass spectrometry, and analyzed for in vitro neurotoxicity (0.3-10 g/ml), as well as the effectiveness of antivenom (1-5 units/ml; 10 min prior to the addition of 10 g/ml venom). The following death adder venoms were examined: A. antarcticus (from separate populations in New South Wales, Queensland, South Australia, and Western Australia), hawkei venoms, although it was markedly less effective against venoms from A. antarcticus (NSW, SA, WA), A. rugosus, A. wellsi, and A. sp. Seram. However, at 5 units/ml, antivenom was effective against all venoms tested. Death adder venoms, including those from A. antarcticus geographic variants, differed not only in their venom composition but also in their neurotoxic activity and susceptibility to antivenom. For the first time toxicological aspects of A. hawkei, A. wellsi, A. rugosus, and A. sp. Seram venoms were studied.