WorldWideScience

Sample records for bothrops jararaca jararaca

  1. Evaluation of antivenoms in the neutralization of hyperalgesia and edema induced by Bothrops jararaca and Bothrops asper snake venoms

    OpenAIRE

    Picolo, G.; Chacur, M.; Gutiérrez, J.M.; Teixeira, C.F.P.; Cury, Y.

    2002-01-01

    Neutralization of hyperalgesia induced by Bothrops jararaca and B. asper venoms was studied in rats using bothropic antivenom produced at Instituto Butantan (AVIB, 1 ml neutralizes 5 mg B. jararaca venom) and polyvalent antivenom produced at Instituto Clodomiro Picado (AVCP, 1 ml neutralizes 2.5 mg B. aspar venom). The intraplantar injection of B. jararaca and B. asper venoms caused hyperalgesia, which peaked 1 and 2 h after injection, respectively. Both venoms also induced edema with a simil...

  2. Proteomic identification of gender molecular markers in Bothrops jararaca venom.

    Science.gov (United States)

    Zelanis, André; Menezes, Milene C; Kitano, Eduardo S; Liberato, Tarcísio; Tashima, Alexandre K; Pinto, Antonio F M; Sherman, Nicholas E; Ho, Paulo L; Fox, Jay W; Serrano, Solange M T

    2016-04-29

    Variation in the snake venom proteome is a well-documented phenomenon; however, sex-based variation in the venom proteome/peptidome is poorly understood. Bothrops jararaca shows significant sexual size dimorphism and here we report a comparative proteomic/peptidomic analysis of venoms from male and female specimens and correlate it with the evaluation of important venom features. We demonstrate that adult male and female venoms have distinct profiles of proteolytic activity upon fibrinogen and gelatin. These differences were clearly reflected in their different profiles of SDS-PAGE, two-dimensional electrophoresis and glycosylated proteins. Identification of differential protein bands and spots between male or female venoms revealed gender-specific molecular markers. However, the proteome comparison by in-solution trypsin digestion and label-free quantification analysis showed that the overall profiles of male and female venoms are similar at the polypeptide chain level but show striking variation regarding their attached carbohydrate moieties. The analysis of the peptidomes of male and female venoms revealed different contents of peptides, while the bradykinin potentiating peptides (BPPs) showed rather similar profiles. Furthermore we confirmed the ubiquitous presence of four BPPs that lack the C-terminal Q-I-P-P sequence only in the female venom as gender molecular markers. As a result of these studies we demonstrate that the sexual size dimorphism is associated with differences in the venom proteome/peptidome in B. jararaca species. Moreover, gender-based variations contributed by different glycosylation levels in toxins impact venom complexity. Bothrops jararaca is primarily a nocturnal and generalist snake species, however, it exhibits a notable ontogenetic shift in diet and in venom proteome upon neonate to adult transition. As is common in the Bothrops genus, B. jararaca shows significant sexual dimorphism in snout-vent length and weight, with females being

  3. Evaluation of antivenoms in the neutralization of hyperalgesia and edema induced by Bothrops jararaca and Bothrops asper snake venoms.

    Science.gov (United States)

    Picolo, G; Chacur, M; Gutiérrez, J M; Teixeira, C F P; Cury, Y

    2002-10-01

    Neutralization of hyperalgesia induced by Bothrops jararaca and B. asper venoms was studied in rats using bothropic antivenom produced at Instituto Butantan (AVIB, 1 ml neutralizes 5 mg B. jararaca venom) and polyvalent antivenom produced at Instituto Clodomiro Picado (AVCP, 1 ml neutralizes 2.5 mg B. aspar venom). The intraplantar injection of B. jararaca and B. asper venoms caused hyperalgesia, which peaked 1 and 2 h after injection, respectively. Both venoms also induced edema with a similar time course. When neutralization assays involving the independent injection of venom and antivenom were performed, the hyperalgesia induced by B. jararaca venom was neutralized only when bothropic antivenom was administered iv 15 min before venom injection, whereas edema was neutralized when antivenom was injected 15 min or immediately before venom injection. On the other hand, polyvalent antivenom did not interfere with hyperalgesia or edema induced by B. asper venom, even when administered prior to envenomation. The lack of neutralization of hyperalgesia and edema induced by B. asper venom is not attributable to the absence of neutralizing antibodies in the antivenom, since neutralization was achieved in assays involving preincubation of venom and antivenom. Cross-neutralization of AVCP or AVIB against B. jararaca and B. asper venoms, respectively, was also evaluated. Only bothropic antivenom partially neutralized hyperalgesia induced by B. asper venom in preincubation experiments. The present data suggest that hyperalgesia and edema induced by Bothrops venoms are poorly neutralized by commercial antivenoms even when antibodies are administered immediately after envenomation.

  4. Evaluation of antivenoms in the neutralization of hyperalgesia and edema induced by Bothrops jararaca and Bothrops asper snake venoms

    Directory of Open Access Journals (Sweden)

    Picolo G.

    2002-01-01

    Full Text Available Neutralization of hyperalgesia induced by Bothrops jararaca and B. asper venoms was studied in rats using bothropic antivenom produced at Instituto Butantan (AVIB, 1 ml neutralizes 5 mg B. jararaca venom and polyvalent antivenom produced at Instituto Clodomiro Picado (AVCP, 1 ml neutralizes 2.5 mg B. aspar venom. The intraplantar injection of B. jararaca and B. asper venoms caused hyperalgesia, which peaked 1 and 2 h after injection, respectively. Both venoms also induced edema with a similar time course. When neutralization assays involving the independent injection of venom and antivenom were performed, the hyperalgesia induced by B. jararaca venom was neutralized only when bothropic antivenom was administered iv 15 min before venom injection, whereas edema was neutralized when antivenom was injected 15 min or immediately before venom injection. On the other hand, polyvalent antivenom did not interfere with hyperalgesia or edema induced by B. asper venom, even when administered prior to envenomation. The lack of neutralization of hyperalgesia and edema induced by B. asper venom is not attributable to the absence of neutralizing antibodies in the antivenom, since neutralization was achieved in assays involving preincubation of venom and antivenom. Cross-neutralization of AVCP or AVIB against B. jararaca and B. asper venoms, respectively, was also evaluated. Only bothropic antivenom partially neutralized hyperalgesia induced by B. asper venom in preincubation experiments. The present data suggest that hyperalgesia and edema induced by Bothrops venoms are poorly neutralized by commercial antivenoms even when antibodies are administered immediately after envenomation.

  5. Functional and proteomic comparison of Bothrops jararaca venom from captive specimens and the Brazilian Bothropic Reference Venom.

    Science.gov (United States)

    Farias, Iasmim Baptista de; Morais-Zani, Karen de; Serino-Silva, Caroline; Sant'Anna, Sávio S; Rocha, Marisa M T da; Grego, Kathleen F; Andrade-Silva, Débora; Serrano, Solange M T; Tanaka-Azevedo, Anita M

    2018-03-01

    Snake venom is a variable phenotypic trait, whose plasticity and evolution are critical for effective antivenom production. A significant reduction of the number of snake donations to Butantan Institute (São Paulo, Brazil) occurred in recent years, and this fact may impair the production of the Brazilian Bothropic Reference Venom (BBRV). Nevertheless, in the last decades a high number of Bothrops jararaca specimens have been raised in captivity in the Laboratory of Herpetology of Butantan Institute. Considering these facts, we compared the biochemical and biological profiles of B. jararaca venom from captive specimens and BBRV in order to understand the potential effects of snake captivity upon the venom composition. Electrophoretic analysis and proteomic profiling revealed few differences in venom protein bands and some differentially abundant toxins. Comparison of enzymatic activities showed minor differences between the two venoms. Similar cross-reactivity recognition pattern of both venoms by the antibothropic antivenom produced by Butantan Institute was observed. Lethality and neutralization of lethality for B. jararaca venom from captive specimens and BBRV showed similar values. Considering these results we suggest that the inclusion of B. jararaca venom from captive specimens in the composition of BBRV would not interfere with the quality of this reference venom. Snakebite envenomation is a neglected tropical pathology whose treatment is based on the use of specific antivenoms. Bothrops jararaca is responsible for the majority of snakebites in South and Southeastern Brazil. Its venom shows individual, sexual, and ontogenetic variability, however, the effect of animal captivity upon venom composition is unknown. Considering the reduced number of wild-caught snakes donated to Butantan Institute in the last decades, and the increased life expectancy of the snakes raised in captivity in the Laboratory of Herpetology, this work focused on the comparative

  6. Bothrops asper and Bothrops jararaca snake venoms trigger microbicidal functions of peritoneal leukocytes in vivo.

    Science.gov (United States)

    Zamuner, S R; Gutiérrez, J M; Muscará, M N; Teixeira, S A; Teixeira, C F

    2001-10-01

    Venoms from snakes of the genus Bothrops cause pronounced local effects in the victims. These alterations result not only from the direct toxic action of venom components, but also from the prominent inflammatory reaction associated with these envenomations. In this study we investigated the ability of Bothrops asper (BaV) and Bothrops jararaca (BjV) venoms to induce cellular influx and microbicidal functions in leukocytes. BaV and BjV (5 microg/animal) caused a long lasting infiltration of leukocytes (3-48 h) when injected into mouse peritoneal cavity. Both venoms increased phagocytosis and production of hydrogen peroxide (H2O2) by polymorphonuclear (PMN) and mononuclear (MN) peritoneal leukocytes. In addition, nitric oxide (NO) production by macrophages was also enhanced after the venom injections. This effect was inhibited by treating animals with L-NAME and aminoguanidine, thus suggesting the induction of iNOS synthesis by the venoms. Western blot analysis confirmed the expression of iNOS in macrophages. BaV and BjV injection led to increased levels of IFN-gamma at the site of inflammation. Since IFN-gamma is an effective inducer of iNOS expression, an indirect action of the venoms on iNOS expression can be proposed. A marked formation of nitrotyrosine-containing proteins was also observed in macrophage homogenates. Based on these results, we suggest that reactive oxygen and nitrogen-derived species are involved in the pathogenesis of the local tissue damage characteristic of Bothrops sp envenomations.

  7. Individual variability in the venom proteome of juvenile Bothrops jararaca specimens.

    Science.gov (United States)

    Dias, Gabriela S; Kitano, Eduardo S; Pagotto, Ana H; Sant'anna, Sávio S; Rocha, Marisa M T; Zelanis, André; Serrano, Solange M T

    2013-10-04

    Snake venom proteomes/peptidomes are highly complex and subject to ontogenetic changes. Individual variation in the venom proteome of juvenile snakes is poorly known. We report the proteomic analysis of venoms from 21 juvenile specimens of Bothrops jararaca of different geographical origins and correlate it with the evaluation of important venom features. Individual venoms showed similar caseinolytic activities; however, their amidolytic activities were significantly different. Rather intriguingly, plasma coagulant activity showed remarkable variability among the venoms but not the prothrombin-activating activity. LC-MS analysis showed significant differences between venoms; however, an interesting finding was the ubiquitous presence of the tripeptide ZKW, an endogenous inhibitor of metalloproteinases. Electrophoretic profiles of proteins submitted to reduction showed significant variability in total proteins, glycoproteins, and in the subproteomes of proteinases. Moreover, identification of differential bands revealed variation in most B. jararaca toxin classes. Profiles of venoms analyzed under nonreducing conditions showed less individual variability and identification of proteins in a conserved band revealed the presence of metalloproteinases and l-amino acid oxidase as common components of these venoms. Taken together, our findings suggest that individual venom proteome variability in B. jararaca exists from a very early animal age and is not a result of ontogenetic and diet changes.

  8. Estrutura tridimensional da bothropasina, uma metaloprotease/desintegrina do veneno de bothrops jararaca

    OpenAIRE

    João Renato Carvalho Muniz

    2007-01-01

    A bothropasina é uma proteína hemorrágica de 48 kDa, pertencente à classe P-III das metaloproteases, isolada a partir do veneno bruto da serpente brasileira Bothrops jararaca, e que possui os domínios adesivos desintegrina (D) e rico em cisteína (C). Neste trabalho, nós apresentamos a estrutura cristalográfica da bothropasina complexada ao inibidor POL647. O domínio catalítico , metaloprotease (M), pode ser dividido em dois subdomínios, dispostos de maneira muito similar aos descritos para es...

  9. Paper electrophoretic and enzimatic studies on blood serum, venom and liver of "Bothrops jararaca"

    Directory of Open Access Journals (Sweden)

    Luiz P. Ribeiro

    1955-12-01

    Full Text Available Fígado, veneno e sôro sanguíneo de "Bothrops jararaca" foram estudados por meio da eletroforese em papel e determinação de atividades enzimáticas. Xantina oxidase e deshidrogenase foram encontradas sòmente no fígado das cobras. A análise espectrográfica do veneno e do sôro confirmam os resultados negativos obtidos para xantina oxidase uma vez que não foi encontrado molibdêneo. L-amino ácido oxidase foi determinada no fígado, sÔro e veneno. A eletroforese em papel do sôro sanguíneo mostrou que existem 7 frações proteicas, sendo que duas apresentam fluorescência característica de flavinas, quando expostas à luz ultra-violeta. Em vista dos resultados obtidos é concluido que as flavinas do sôro e do veneno de Bothrops jararaca estão na maior parte ligadas às proteínas. Estas flavinas combinadas parecem estar sob a forma de FAD (flavina adenina dinucleotídeo fazendo parte do grupo prostético da L-amino ácido oxidase, uma vez que não foi encontrada nenhuma atividade de xantina oxidase.

  10. Anti-hemorrhagic Activity of Four Brazilian Vegetable Species Against Bothrops jararaca Venom

    Directory of Open Access Journals (Sweden)

    2009-03-01

    Full Text Available Around 20,000 snakebites are reported annually in Brazil and 90% of them are inflicted by species of the genus Bothrops. Intravenous administration of antibothropic antivenom neutralizes the systemic actions, but it is of little effect on the reversal of local symptoms and often induces adverse reactions, a context that drives the search for complementary treatments for snakebite accidents. Vegetable extracts with a range of antiophidian activities constitute an excellent alternative. In this study, we investigated the anti-hemorrhagic effects of Mouriri pusa Gardn. (Melastomataceae, Byrsonima crassa Niedenzu (Malpighiaceae, Davilla elliptica St. Hill. (Dilleniaceae and Strychnos pseudoquina St. Hil. (Loganiaceae against Bothrops jararaca venom. The methanolic extracts from M. pusa (leaves, B. crassa (leaves and D. elliptica (leaves showed total neutralization capacity against local hemorrhages. The amenthoflavone and quercetin fractions from B. crassa and the flavonoids fractions (quercetin and myricetin from M. pusa and D. elliptica also showed total neutralization capacity. We conclude that flavonoids derived from myricetin, quercetin and amenthoflavone play an important role in the anti-hemorrhagic potential of these Brazilian vegetables species against B. jararaca venom.

  11. Anti-hemorrhagic activity of four Brazilian vegetable species against Bothrops jararaca venom.

    Science.gov (United States)

    Nishijima, Catarine Massucato Nishijima; Rodrigues, Clenilson Martins; Silva, Marcelo Aparecido; Lopes-Ferreira, Mônica; Vilegas, Wagner; Hiruma-Lima, Clélia Akiko

    2009-03-09

    Around 20,000 snakebites are reported annually in Brazil and 90% of them are inflicted by species of the genus Bothrops. Intravenous administration of antibothropic antivenom neutralizes the systemic actions, but it is of little effect on the reversal of local symptoms and often induces adverse reactions, a context that drives the search for complementary treatments for snakebite accidents. Vegetable extracts with a range of antiophidian activities constitute an excellent alternative. In this study, we investigated the anti-hemorrhagic effects of Mouriri pusa Gardn. (Melastomataceae), Byrsonima crassa Niedenzu (Malpighiaceae), Davilla elliptica St. Hill. (Dilleniaceae) and Strychnos pseudoquina St. Hil. (Loganiaceae) against Bothrops jararaca venom. The methanolic extracts from M. pusa (leaves), B. crassa (leaves) and D. elliptica (leaves) showed total neutralization capacity against local hemorrhages. The amenthoflavone and quercetin fractions from B. crassa and the flavonoids fractions (quercetin and myricetin) from M. pusa and D. elliptica also showed total neutralization capacity. We conclude that flavonoids derived from myricetin, quercetin and amenthoflavone play an important role in the anti-hemorrhagic potential of these Brazilian vegetables species against B. jararaca venom.

  12. Antivenom Effects of 1,2,3-Triazoles against Bothrops jararaca and Lachesis muta Snakes

    Science.gov (United States)

    Domingos, Thaisa F. S.; Moura, Laura de A.; Carvalho, Carla; Campos, Vinícius R.; Jordão, Alessandro K.; Cunha, Anna C.; Ferreira, Vitor F.; de Souza, Maria Cecília B. V.; Sanchez, Eladio F.; Fuly, André L.

    2013-01-01

    Snake venoms are complex mixtures of proteins of both enzymes and nonenzymes, which are responsible for producing several biological effects. Human envenomation by snake bites particularly those of the viperid family induces a complex pathophysiological picture characterized by spectacular changes in hemostasis and frequently hemorrhage is also seen. The present work reports the ability of six of a series of 1,2,3-triazole derivatives to inhibit some pharmacological effects caused by the venoms of Bothrops jararaca and Lachesis muta. In vitro assays showed that these compounds were impaired in a concentration-dependent manner, the fibrinogen or plasma clotting, hemolysis, and proteolysis produced by both venoms. Moreover, these compounds inhibited biological effects in vivo as well. Mice treated with these compounds were fully protected from hemorrhagic lesions caused by such venoms. But, only the B. jararaca edema-inducing activity was neutralized by the triazoles. So the inhibitory effect of triazoles derivatives against some in vitro and in vivo biological assays of snake venoms points to promising aspects that may indicate them as molecular models to improve the production of effective antivenom or to complement antivenom neutralization, especially the local pathological effects, which are partially neutralized by antivenoms. PMID:23710441

  13. An in-depth snake venom proteopeptidome characterization: Benchmarking Bothrops jararaca.

    Science.gov (United States)

    Nicolau, Carolina A; Carvalho, Paulo C; Junqueira-de-Azevedo, Inácio L M; Teixeira-Ferreira, André; Junqueira, Magno; Perales, Jonas; Neves-Ferreira, Ana Gisele C; Valente, Richard H

    2017-01-16

    A large-scale proteomic approach was devised to advance the understanding of venom composition. Bothrops jararaca venom was fractionated by OFFGEL followed by chromatography, generating peptidic and proteic fractions. The latter was submitted to trypsin digestion. Both fractions were separately analyzed by reversed-phase nanochromatography coupled to high resolution mass spectrometry. This strategy allowed deeper and joint characterizations of the peptidome and proteome (proteopeptidome) of this venom. Our results lead to the identification of 46 protein classes (with several uniquely assigned proteins per class) comprising eight high-abundance bona fide venom components, and 38 additional classes in smaller quantities. This last category included previously described B. jararaca venom proteins, common Elapidae venom constituents (cobra venom factor and three-finger toxin), and proteins typically encountered in lysosomes, cellular membranes and blood plasma. Furthermore, this report is the most complete snake venom peptidome described so far, both in number of peptides and in variety of unique proteins that could have originated them. It is hypothesized that such diversity could enclose cryptides, whose bioactivities would contribute to envenomation in yet undetermined ways. Finally, we propose that the broad range screening of B. jararaca peptidome will facilitate the discovery of bioactive molecules, eventually leading to valuable therapeutical agents. Our proteopeptidomic strategy yielded unprecedented insights into the remarkable diversity of B. jararaca venom composition, both at the peptide and protein levels. These results bring a substantial contribution to the actual pursuit of large-scale protein-level assignment in snake venomics. The detection of typical elapidic venom components, in a Viperidae venom, reinforces our view that the use of this approach (hand-in-hand with transcriptomic and genomic data) for venom proteomic analysis, at the specimen

  14. Ocorrência de híbridos não naturais entre Bothrops jararaca e B. neuwiedi (Serpentes, Viperidae Occurrence of non natural hybridism between Bothrops jararaca and B. neuwiedi (Serpentes, Viperidae

    Directory of Open Access Journals (Sweden)

    Rafael Lucchesi Balestrin

    Full Text Available In May 1987, a female of Bothrops jararaca (Wied, 1824, from Carazinho, Rio Grande do Sul (RS, Brazil, was placed in the same vivarium with a male of Bothrops neuwiedi Wagler, 1824 coming from Guaíba, RS. There, they stayed for aproximately ten months. In March 1988, it was observed a delivery of five live and two still born, among them six presented morphologic characteristics of B. neuwiedi and one of B. jararaca. After the female died, in April 1988, through necropsy, two fetusus were found, one near the cloaca and, both identified as B. neuwiedi. The morphologic analysis and the origin of the progenitors suggest the hypothesis that the litter was resulted of cross-breeding.

  15. Evaluation of the interaction of surfactants with stratum corneum model membrane from Bothrops jararaca by DSC.

    Science.gov (United States)

    Baby, André Rolim; Lacerda, Aurea Cristina Lemos; Velasco, Maria Valéria Robles; Lopes, Patrícia Santos; Kawano, Yoshio; Kaneko, Telma Mary

    2006-07-06

    The interaction of surfactants sodium dodecyl sulfate (SDS), cetyl trimethyl ammonium chloride (CTAC) and lauryl alcohol ethoxylated (12 mol ethylene oxide) (LAE-12OE) was evaluated on the stratum corneum (SC) of shed snake skins from Bothrops jararaca, used as model membrane, and thermal characterized by differential scanning calorimetry (DSC). Surfactant solutions were employed above of the critical micellar concentration (CMC) with treatment time of 8h. The SDS interaction with the SC model membrane has increased the characteristic transition temperature of 130 degrees C in approximately 10 degrees C for the water loss and keratin denaturation, indicating an augmentation of the water content. Samples treated with CTAC have a decrease of the water loss temperature, while, for the LAE-12OE treated samples, changes on the transition temperature have not been observed.

  16. Cross-reactivity and phospholipase A2 neutralization of anti-irradiated Bothrops jararaca venom antibodies

    International Nuclear Information System (INIS)

    Spencer, P.J.; Nascimento, N. do; Paula, R.A. de; Cardi, B.A.; Rogero, J.R.

    1995-01-01

    The detoxified Bothrops jararaca venom, immunized rabbits with the toxoid obtained and investigated cross-reactivity of the antibodies obtained against autologous and heterelogous venoms was presented. It was also investigated the ability of the IgGs, purified by affinity chromatography, from those sera to neutralize phospholipase. A 2 , an ubiquous enzyme in animal venoms. Results indicate that venom irradiation leads to an attenuation of toxicity of 84%. Cross-reactivity was investigated by ELISA and Western blot and all venoms were reactive to the antibodies. On what refers to phospholipase A 2 activity neutralization, the antibodies neutralized autologous venoms efficiently and, curiously, other venoms from the same genus were not neutralized, while Lachesis muta venom, a remote related specier, was neutralized by this serum. These data suggest that irradiation preserve important epitopes for induction of neutralizing antibodies and that these epitopes are not shared by all venoms assayed. (author). 8 refs, 2 figs, 3 tabs

  17. Oral fibrosarcoma in jararaca (Bothrops pubescens: anatomopathological and immunohistochemical aspects

    Directory of Open Access Journals (Sweden)

    Ezequiel D. Santos

    2015-07-01

    Full Text Available Abstract A 4-year-old female captive-bred snake of the genus Bothrops showed swelling on the left side of the oral cavity, suggesting the development of neoplasia. The mass was removed surgically and sent for pathological examination. Two months later a new increase in volume in the same site was observed, suggesting recurrence. The lesion was completely removed and sent for pathological analysis. Histologically, the two-samples consisted of a mass with highly-cell density composed of spindle-shaped anaplastic cells arranged in interwoven bundles, distributed throughout the tissue extension and, occasionally, polygonal cells arranged in irregular fascicles. The Masson trichrome staining showed modest amount of collagen supporting the neoplastic cells. PAS-positive content was not observed in the cytoplasm of neoplastic cells. Histological and histochemical findings indicated that it was a spindle cell neoplasm, but the classification was not possible. Immunohistochemistry was requested and performed using the streptavidin-biotin-peroxidase method. The markers used were anti-vimentin, anti-PCNA, anti-EMA, anti-melan A and anti-melanosome, anti-desmin, anti-actin, anti-CD68 and anti- S100protein. The neoplastic cells were immunoreactive for vimentin and PCNA and negative for the other antibodies. The morphology characterization, histochemical and immunohistochemical analysis of neoplastic cells allowed the definitive diagnosis of oral fibrosarcoma.

  18. The pharmacological effect of Bothrops neuwiedii pauloensis (jararaca-pintada snake venom on avian neuromuscular transmission

    Directory of Open Access Journals (Sweden)

    C.R. Borja-Oliveira

    2003-05-01

    Full Text Available The neuromuscular effects of Bothrops neuwiedii pauloensis (jararaca-pintada venom were studied on isolated chick biventer cervicis nerve-muscle preparations. Venom concentrations of 5-50 µg/ml produced an initial inhibition and a secondary increase of indirectly evoked twitches followed by a progressive concentration-dependent and irreversible neuromuscular blockade. At venom concentrations of 1-20 µg/ml, the responses to 13.4 mM KCl were inhibited whereas those to 110 µM acetylcholine alone and cumulative concentrations of 1 µM to 10 mM were unaffected. At venom concentrations higher than 50 µg/ml, there was pronounced muscle contracture with inhibition of the responses to acetylcholine, KCl and direct stimulation. At 20-24ºC, the venom (50 µg/ml produced only partial neuromuscular blockade (30.7 ± 8.0%, N = 3 after 120 min and the initial inhibition and the secondary increase of the twitch responses caused by the venom were prolonged and pronounced and the response to KCl was unchanged. These results indicate that B.n. pauloensis venom is neurotoxic, acting primarily at presynaptic sites, and that enzyme activity may be involved in this pharmacological action.

  19. Variação anual do sistema reprodutor de fêmeas de Bothrops jararaca (Serpentes, Viperidae Annual variation of the reproductive system in females of Bothrops jararaca (Serpentes, Viperidae

    Directory of Open Access Journals (Sweden)

    Thélia R. F. Janeiro-Cinquini

    2004-09-01

    Full Text Available The morphological changes of the ovary and oviduct of 238 tropical snakes Bothrops jararaca (Wied, 1824 were determined. The ovarian mass presented a remarkable decrease in October, after ovulation, staying in low levels from November to March, during the gestational period. From April to September, it increased because of the ovarian follicles maturation. A gradual increase in oviduct weight was observed from October to March due to stages of embryonic development. A significant difference was observed between right and left ovary weight, and oviduct length, independently of the months considered.

  20. The influence of circadian rhythms on the metabolism of the snake Bothrops jararaca (Serpentes, Viperidae = A influência dos ritmos circadianos no metabolismo da serpente Bothrops jararaca (Serpentes, Viperidae

    Directory of Open Access Journals (Sweden)

    José Geraldo Pereira da Cruz

    2008-07-01

    Full Text Available The thermoregulatory activity has led to an extensive search for correlations between physiological variables, including metabolic functions, and the ideal level of body temperature. Snakes were also often seen basking, when their body temperatures were relatively independent of ambient temperature, indicating successful thermoregulation. Bothrops jararaca were exposed to two different ambient temperatures (20 and 30oC over a time course of three weeks and oxygen consumption and body temperature were measured. The snakes exhibited a freerunning rhythm of body temperature. Metabolic rate wasincreased at the same circadian phase as the increase in body temperature in the 30oC. The increase of body temperature and oxygen consumption of B. jararaca occurs in the scotophase of the photoperiod, consistent with that of nocturnal species. However, prior to a scotophase period the snakes under 20oC maintain body temperature and oxygen consumption at higher levels during the day. These results demonstrate for the first time that ectothermic animals may display physiologically generated circadian rhythms of bodytemperature similar to those recorded in endotherms. Circadian rhythms allow animals to anticipate environmental changes: physiological parameters such as body temperature andmobilization of energy reserves have to be adjusted before the expected environmental changes actually take place.A atividade termorreguladora conduziu a uma busca extensiva para o entendimento das correlações entre as variáveis fisiológicas, incluindo as funções metabólicas e a temperatura corporal. Frequentes observações mostram que algumas serpentes podem se aquecer, sendo este aumento de temperatura independente da temperatura ambiente, indicando a termorregulação bem sucedida. Bothrops jararaca foramexpostas a dois ambientes com diferentes temperaturas (20 e 30oC durante três semanas, sendo mensuradas a temperatura corporal e o consumo de oxigênio. O aumento

  1. Effect of Diterpenes Isolated of the Marine Alga Canistrocarpus cervicornis against Some Toxic Effects of the Venom of the Bothrops jararaca Snake

    Directory of Open Access Journals (Sweden)

    Thaisa Francielle Souza Domingos

    2015-02-01

    Full Text Available Snake venoms are composed of a complex mixture of active proteins and peptides which induce a wide range of toxic effects. Envenomation by Bothrops jararaca venom results in hemorrhage, edema, pain, tissue necrosis and hemolysis. In this work, the effect of a mixture of two secodolastane diterpenes (linearol/isolinearol, previously isolated from the Brazilian marine brown alga, Canistrocarpus cervicornis, was evaluated against some of the toxic effects induced by B. jararaca venom. The mixture of diterpenes was dissolved in dimethylsulfoxide and incubated with venom for 30 min at room temperature, and then several in vivo (hemorrhage, edema and lethality and in vitro (hemolysis, plasma clotting and proteolysis assays were performed. The diterpenes inhibited hemolysis, proteolysis and hemorrhage, but failed to inhibit clotting and edema induced by B. jararaca venom. Moreover, diterpenes partially protected mice from lethality caused by B. jararaca venom. The search for natural inhibitors of B. jararaca venom in C. cervicornis algae is a relevant subject, since seaweeds are a rich and powerful source of active molecules which are as yet but poorly explored. Our results suggest that these diterpenes have the potential to be used against Bothropic envenomation accidents or to improve traditional treatments for snake bites.

  2. Bothrops jararaca venom metalloproteinases are essential for coagulopathy and increase plasma tissue factor levels during envenomation.

    Directory of Open Access Journals (Sweden)

    Karine M Yamashita

    2014-05-01

    Full Text Available BACKGROUND/AIMS: Bleeding tendency, coagulopathy and platelet disorders are recurrent manifestations in snakebites occurring worldwide. We reasoned that by damaging tissues and/or activating cells at the site of the bite and systemically, snake venom toxins might release or decrypt tissue factor (TF, resulting in activation of blood coagulation and aggravation of the bleeding tendency. Thus, we addressed (a whether TF and protein disulfide isomerase (PDI, an oxireductase involved in TF encryption/decryption, were altered in experimental snake envenomation; (b the involvement and significance of snake venom metalloproteinases (SVMP and serine proteinases (SVSP to hemostatic disturbances. METHODS/PRINCIPAL FINDINGS: Crude Bothrops jararaca venom (BjV was preincubated with Na2-EDTA or AEBSF, which are inhibitors of SVMP and SVSP, respectively, and injected subcutaneously or intravenously into rats to analyze the contribution of local lesion to the development of hemostatic disturbances. Samples of blood, lung and skin were collected and analyzed at 3 and 6 h. Platelet counts were markedly diminished in rats, and neither Na2-EDTA nor AEBSF could effectively abrogate this fall. However, Na2-EDTA markedly reduced plasma fibrinogen consumption and hemorrhage at the site of BjV inoculation. Na2-EDTA also abolished the marked elevation in TF levels in plasma at 3 and 6 h, by both administration routes. Moreover, increased TF activity was also noticed in lung and skin tissue samples at 6 h. However, factor VII levels did not decrease over time. PDI expression in skin was normal at 3 h, and downregulated at 6 h in all groups treated with BjV. CONCLUSIONS: SVMP induce coagulopathy, hemorrhage and increased TF levels in plasma, but neither SVMP nor SVSP are directly involved in thrombocytopenia. High levels of TF in plasma and TF decryption occur during snake envenomation, like true disseminated intravascular coagulation syndrome, and might be implicated in

  3. Study of 60 Co gamma radiation effects on the biochemical, biological and immunological properties of the Bothrops jararaca venom

    International Nuclear Information System (INIS)

    Guarnieri, M.C.

    1992-01-01

    Gamma radiation, by including different modifications on the toxic, enzymatic and immunological activities of proteins, could be an useful implement for detoxification of snake venoms. The present work was done to study the mechanism of action and effects of gamma rays on the Bothrops jararaca venom, determining the radiation dose that attenuates the toxic and enzymatic activities maintaining the immunological properties of venom, and also the most important free radicals on this process. The results of immuno diffusion, immunoblotting, immunoprecipitation, immunization of mice and rabbits, and neutralization tests, showed the maintenance of antigenic and immunogenic properties and decrease of neutralizing capacity of antibodies induced by 3,000 and 4,000 Gy irradiated venom. Since the immunological properties were the most radioresistant, it was possible to determine the dose of 2,000 Gy, as the ideal radiation dose in the treatment of venoms aiming the improvement of the immunization schedule to obtain bothropic antisera. (author). 164 refs, 19 tabs, 54 figs

  4. Envenenamento experimental por Bothropoides jararaca e Bothrops jararacussu em ovinos: aspectos clínico-patológicos e laboratoriais Experimental poisoning by Bothropoides jararaca and Bothrops jararacussu in sheep: clinic-pathological and laboratory aspects

    Directory of Open Access Journals (Sweden)

    Ana Paula Aragão

    2010-09-01

    Full Text Available Esse estudo teve como objetivo determinar as alterações clínico-patológicas e laboratoriais em ovinos inoculados com a peçonha de Bothropoides jararaca e Bothrops jararacussu, no intuito de fornecer subsídios que possam facilitar o estabelecimento do diagnóstico e do diagnóstico diferencial dessa condição. Os venenos liofilizados foram diluídos em 1 ml de solução fisiológica e administrados a quatro ovinos por via subcutânea. Três ovinos foram a óbito e um que recebeu a dose de 0,5mg/kg (B. jararaca, recuperou-se. Os sinais clínicos tiveram início entre 7 minutos e 1 hora. O período de evolução variou de 7 horas 9 minutos a 21 horas 59 minutos. O quadro clínico, independentemente das doses, caracterizou-se por aumento de volume no local da inoculação, tempo de sangramento e de preenchimento capilar aumentados, taquicardia, dispnéia, mucosas hipocoradas e apatia. Os exames laboratoriais revelaram acentuada anemia normocítica normocrômica, trombocitopenia, acentuada redução de fibrinogênio e proteínas plasmáticas totais, hematócrito diminuído em dois animais, além de acentuado aumento de creatinaquinase e desidrogenase lática em todos os animais. À necropsia, os principais achados no local da inoculação e tecidos adjacentes eram extensas hemorragias no animal que recebeu o veneno de B. jararaca e edema e acentuado edema pulmonar agudo para os dois animais envenenados por B. jararacussu. Além de hemorragia e edema a principal alteração histopatológica verificada foi necrose das fibras musculares e de vasos, no local de inoculação e adjacências. A necrose tubular renal foi atribuída ao quadro de choque. Nos ovinos deste estudo, o aumento de volume observado no local de inoculação e adjacências era constituído predominantemente por sangue (B. jararaca e por edema (B. jararacussu.The purpose of this study was to establish the clinic-pathological and laboratory changes in sheep inoculated with

  5. Inhibitory Effects of Hydroethanolic Leaf Extracts of Kalanchoe brasiliensis and Kalanchoe pinnata (Crassulaceae) against Local Effects Induced by Bothrops jararaca Snake Venom.

    Science.gov (United States)

    Fernandes, Júlia Morais; Félix-Silva, Juliana; da Cunha, Lorena Medeiros; Gomes, Jacyra Antunes Dos Santos; Siqueira, Emerson Michell da Silva; Gimenes, Luisa Possamai; Lopes, Norberto Peporine; Soares, Luiz Alberto Lira; Fernandes-Pedrosa, Matheus de Freitas; Zucolotto, Silvana Maria

    2016-01-01

    The species Kalanchoe brasiliensis and Kalanchoe pinnata, both known popularly as "Saião," are used interchangeably in traditional medicine for their antiophidic properties. Studies evaluating the anti-venom activity of these species are scarce. This study aims to characterize the chemical constituents and evaluate the inhibitory effects of hydroethanolic leaf extracts of K. brasiliensis and K. pinnata against local effects induced by Bothrops jararaca snake venom. Thin Layer Chromatography (TLC) and High Performance Liquid Chromatography coupled with Diode Array Detection and Electrospray Mass Spectrometry (HPLC-DAD-MS/MS) were performed for characterization of chemical markers of the extracts from these species. For antiophidic activity evaluation, B. jararaca venom-induced paw edema and skin hemorrhage in mice were evaluated. In both models, hydroethanolic extracts (125-500 mg/kg) were administered intraperitoneally in different protocols. Inhibition of phospholipase enzymatic activity of B. jararaca was evaluated. The HPLC-DAD-MS/MS chromatographic profile of extracts showed some particularities in the chemical profile of the two species. K. brasileinsis exhibited major peaks that have UV spectra similar to flavonoid glycosides derived from patuletin and eupafolin, while K. pinnata showed UV spectra similar to flavonoids glycosides derived from quercetin and kaempferol. Both extracts significantly reduced the hemorrhagic activity of B. jararaca venom in pre-treatment protocol, reaching about 40% of inhibition, while only K. pinnata was active in post-treatment protocol (about 30% of inhibition). In the antiedematogenic activity, only K. pinnata was active, inhibiting about 66% and 30% in pre and post-treatment protocols, respectively. Both extracts inhibited phospholipase activity; however, K. pinnata was more active. In conclusion, the results indicate the potential antiophidic activity of Kalanchoe species against local effects induced by B. jararaca snake

  6. Inhibitory Effects of Hydroethanolic Leaf Extracts of Kalanchoe brasiliensis and Kalanchoe pinnata (Crassulaceae against Local Effects Induced by Bothrops jararaca Snake Venom.

    Directory of Open Access Journals (Sweden)

    Júlia Morais Fernandes

    Full Text Available The species Kalanchoe brasiliensis and Kalanchoe pinnata, both known popularly as "Saião," are used interchangeably in traditional medicine for their antiophidic properties. Studies evaluating the anti-venom activity of these species are scarce. This study aims to characterize the chemical constituents and evaluate the inhibitory effects of hydroethanolic leaf extracts of K. brasiliensis and K. pinnata against local effects induced by Bothrops jararaca snake venom. Thin Layer Chromatography (TLC and High Performance Liquid Chromatography coupled with Diode Array Detection and Electrospray Mass Spectrometry (HPLC-DAD-MS/MS were performed for characterization of chemical markers of the extracts from these species. For antiophidic activity evaluation, B. jararaca venom-induced paw edema and skin hemorrhage in mice were evaluated. In both models, hydroethanolic extracts (125-500 mg/kg were administered intraperitoneally in different protocols. Inhibition of phospholipase enzymatic activity of B. jararaca was evaluated. The HPLC-DAD-MS/MS chromatographic profile of extracts showed some particularities in the chemical profile of the two species. K. brasileinsis exhibited major peaks that have UV spectra similar to flavonoid glycosides derived from patuletin and eupafolin, while K. pinnata showed UV spectra similar to flavonoids glycosides derived from quercetin and kaempferol. Both extracts significantly reduced the hemorrhagic activity of B. jararaca venom in pre-treatment protocol, reaching about 40% of inhibition, while only K. pinnata was active in post-treatment protocol (about 30% of inhibition. In the antiedematogenic activity, only K. pinnata was active, inhibiting about 66% and 30% in pre and post-treatment protocols, respectively. Both extracts inhibited phospholipase activity; however, K. pinnata was more active. In conclusion, the results indicate the potential antiophidic activity of Kalanchoe species against local effects induced by B

  7. Flow cytometric characterization of peripheral blood leukocyte populations of 3 neotropical snake species: Boa constrictor, Bothrops jararaca, and Crotalus durissus.

    Science.gov (United States)

    Carvalho, Marcelo P N; Queiroz-Hazarbassanov, Nicolle G T; Massoco, Cristina O; Rossi, Silmara; Sant'Anna, Sávio S; Catão-Dias, José L; Grego, Kathleen F

    2016-06-01

    The reptilian immune system is represented by innate, humoral, and cell-mediated mechanisms, involving different types of blood leukocytes. The development of optimized methods for the advanced study of origin and function of reptilian blood leukocytes is needed. The purpose of the study was to optimize leukocyte density gradient isolation protocols from snake peripheral blood samples, and characterize recovered cells by flow cytometry based on size and internal complexity for a qualitative and semi-quantitative assessment of leukocyte populations in one boa (Boa constrictor), and 2 viper species (Bothrops jararaca, Crotalus durissus). Blood samples from 30 snakes (10 from each species, 5 males and 5 females) were collected in tubes with sodium heparin. Fresh blood was centrifuged with either ficoll-paque PLUS or percoll density gradients for leukocyte isolation. Flow cytometric leukocyte gates were defined based on size (forward scatter [FSC]) and internal complexity (side scatter [SSC]). Relative leukocyte differential counts after sorting the cells in these gates in one snake for each species were compared to conventional light microscopic differential counts on unsorted isolated leukocytes. There was no statistical difference in the relative leukocyte populations, including heterophils, azurophils, and small and large lymphocytes between samples isolated by ficoll or percoll. Four leukocyte gates were identified based on their location in FSC/SSC cytograms. The relative leukocyte differential counts after sorting in single animals showed some agreement with the light microscopy differential count on unsorted cells. Based on FSC and SSC, 4 distinct leukocyte populations were found in ficoll or percoll density gradient isolated leukocytes from peripheral blood from boa and viper species. Further optimization of the technique should allow the performance of functional assays. © 2016 American Society for Veterinary Clinical Pathology.

  8. Venom-related transcripts from Bothrops jararaca tissues provide novel molecular insights into the production and evolution of snake venom.

    Science.gov (United States)

    Junqueira-de-Azevedo, Inácio L M; Bastos, Carolina Mancini Val; Ho, Paulo Lee; Luna, Milene Schmidt; Yamanouye, Norma; Casewell, Nicholas R

    2015-03-01

    Attempts to reconstruct the evolutionary history of snake toxins in the context of their co-option to the venom gland rarely account for nonvenom snake genes that are paralogous to toxins, and which therefore represent important connectors to ancestral genes. In order to reevaluate this process, we conducted a comparative transcriptomic survey on body tissues from a venomous snake. A nonredundant set of 33,000 unigenes (assembled transcripts of reference genes) was independently assembled from six organs of the medically important viperid snake Bothrops jararaca, providing a reference list of 82 full-length toxins from the venom gland and specific products from other tissues, such as pancreatic digestive enzymes. Unigenes were then screened for nontoxin transcripts paralogous to toxins revealing 1) low level coexpression of approximately 20% of toxin genes (e.g., bradykinin-potentiating peptide, C-type lectin, snake venom metalloproteinase, snake venom nerve growth factor) in body tissues, 2) the identity of the closest paralogs to toxin genes in eight classes of toxins, 3) the location and level of paralog expression, indicating that, in general, co-expression occurs in a higher number of tissues and at lower levels than observed for toxin genes, and 4) strong evidence of a toxin gene reverting back to selective expression in a body tissue. In addition, our differential gene expression analyses identify specific cellular processes that make the venom gland a highly specialized secretory tissue. Our results demonstrate that the evolution and production of venom in snakes is a complex process that can only be understood in the context of comparative data from other snake tissues, including the identification of genes paralogous to venom toxins. © The Author 2014. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  9. Differential transcript profile of inhibitors with potential anti-venom role in the liver of juvenile and adult Bothrops jararaca snake

    Directory of Open Access Journals (Sweden)

    Cícera Maria Gomes

    2017-04-01

    Full Text Available Background Snakes belonging to the Bothrops genus are vastly distributed in Central and South America and are responsible for most cases of reported snake bites in Latin America. The clinical manifestations of the envenomation caused by this genus are due to three major activities—proteolytic, hemorrhagic and coagulant—mediated by metalloproteinases, serine proteinases, phospholipases A2 and other toxic compounds present in snake venom. Interestingly, it was observed that snakes are resistant to the toxic effects of its own and other snake’s venoms. This natural immunity may occur due the absence of toxin target or the presence of molecules in the snake plasma able to neutralize such toxins. Methods In order to identify anti-venom molecules, we construct a cDNA library from the liver of B. jararaca snakes. Moreover, we analyzed the expression profile of four molecules—the already known anti-hemorrhagic factor Bj46a, one gamma-phospholipase A2 inhibitor, one inter-alpha inhibitor and one C1 plasma protease inhibitor—in the liver of juvenile and adult snakes by qPCR. Results The results revealed a 30-fold increase of gamma-phospholipase A2 inhibitor and a minor increase of the inter-alpha inhibitor (5-fold and of the C1 inhibitor (3-fold in adults. However, the Bj46a factor seems to be equally transcribed in adults and juveniles. Discussion The results suggest the up-regulation of different inhibitors observed in the adult snakes might be a physiological adaptation to the recurrent contact with their own and even other snake’s venoms throughout its lifespan. This is the first comparative analysis of ontogenetic variation of expression profiles of plasmatic proteins with potential anti-venom activities of the venomous snake B. jararaca. Furthermore, the present data contributes to the understanding of the natural resistance described in these snakes.

  10. A influência dos ritmos circadianos no metabolismo da serpente Bothrops jararaca (Serpentes, Viperidae - DOI: 10.4025/actascibiolsci.v30i3.5030 The influence of circadian rhythms on the metabolism of the snake Bothrops jararaca (Serpentes, Viperidae - DOI: 10.4025/actascibiolsci.v30i3.5030

    Directory of Open Access Journals (Sweden)

    José Geraldo Pereira da Cruz

    2008-10-01

    Full Text Available A atividade termorreguladora conduziu a uma busca extensiva para o entendimento das correlações entre as variáveis fisiológicas, incluindo as funções metabólicas e a temperatura corporal. Frequentes observações mostram que algumas serpentes podem se aquecer, sendo este aumento de temperatura independente da temperatura ambiente, indicando a termorregulação bem sucedida. Bothrops jararaca foram expostas a dois ambientes com diferentes temperaturas (20 e 30oC durante três semanas, sendo mensuradas a temperatura corporal e o consumo de oxigênio. O aumento da temperatura corporal e consumo de oxigênio de Bothrops jararaca ocorreram na fase de escuro do fotoperíodo, consistente para espécies noturnas. Entretanto, antecedendo a fase de escuro, as serpentes em 20oC apresentaram os níveis mais elevados durante o dia para temperatura corporal e consumo de oxigênio. Estes resultados indicam pela primeira vez que animais termodependentes podem controlar a temperatura corporal por meio de ritmos fisiológicos circadianos, semelhante aos observados em termoindependentes. Os ritmos circadianos permitem que os animais antecipem as mudanças no ambiente: parâmetros fisiológicos como a temperatura corporal e as reservas de energia ou sua mobilização podem ser ajustadas antes que as mudanças ambientais previstas ocorram realmente.The thermoregulatory activity has led to an extensive search for correlations between physiological variables, including metabolic functions, and the ideal level of body temperature. Snakes were also often seen basking, when their body temperatures were relatively independent of ambient temperature, indicating successful thermoregulation. Bothrops jararaca were exposed to two different ambient temperatures (20 and 30ºC over a time course of three weeks and oxygen consumption and body temperature were measured. The snakes exhibited a freerunning rhythm of body temperature. Metabolic rate was increased at the same

  11. Investigating possible biological targets of Bj-CRP, the first cysteine-rich secretory protein (CRISP) isolated from Bothrops jararaca snake venom.

    Science.gov (United States)

    Lodovicho, Marina E; Costa, Tássia R; Bernardes, Carolina P; Menaldo, Danilo L; Zoccal, Karina F; Carone, Sante E; Rosa, José C; Pucca, Manuela B; Cerni, Felipe A; Arantes, Eliane C; Tytgat, Jan; Faccioli, Lúcia H; Pereira-Crott, Luciana S; Sampaio, Suely V

    2017-01-04

    Cysteine-rich secretory proteins (CRISPs) are commonly described as part of the protein content of snake venoms, nevertheless, so far, little is known about their biological targets and functions. Our study describes the isolation and characterization of Bj-CRP, the first CRISP isolated from Bothrops jararaca snake venom, also aiming at the identification of possible targets for its actions. Bj-CRP was purified using three chromatographic steps (Sephacryl S-200, Source 15Q and C18) and showed to be an acidic protein of 24.6kDa with high sequence identity to other snake venom CRISPs. This CRISP was devoid of proteolytic, hemorrhagic or coagulant activities, and it did not affect the currents from 13 voltage-gated potassium channel isoforms. Conversely, Bj-CRP induced inflammatory responses characterized by increase of leukocytes, mainly neutrophils, after 1 and 4h of its injection in the peritoneal cavity of mice, also stimulating the production of IL-6. Bj-CRP also acted on the human complement system, modulating some of the activation pathways and acting directly on important components (C3 and C4), thus inducing the generation of anaphylatoxins (C3a, C4a and C5a). Therefore, our results for Bj-CRP open up prospects for better understanding this class of toxins and its biological actions. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  12. Avaliação da influência de tensoativos na pele de muda de cobra (Bothrops jararaca e Spilotis pullatus) por espectroscopia fatoacústica no infravermelho, calorimetria exploratória diferencial e espectroscopia Raman

    OpenAIRE

    Aurea Cristina Lemos Lacerda

    2004-01-01

    A influência dos tensoativos lauril sulfato de sódio, cloreto de cetil trimetil amônio e álcool láurico etoxilado com 12 moles de óxido de etileno sobre o stratum corneum da pele de muda das cobras Bothrops jararaca e Spilotis pullatus foi avaliada através das técnicas biofísicas de PAS-FTIR, FT-Raman e DSC. Foram utilizadas soluções dos tensoativos em concentrações acima e abaixo da cmc e tratamentos por 4 e 8 horas (stratum corneum íntegro) e por 12 horas (stratum corneum após a remoção mec...

  13. Referências hematológicas para a jararaca de rabo branco (Bothrops leucurus recém capturadas da natureza Hematological references for wild recently-caught white-tailed lancehead (Bothrops leucurus

    Directory of Open Access Journals (Sweden)

    K.F. Grego

    2006-12-01

    Full Text Available The hematological values for the specie Bothrops leucurus, recently captured from nature, were determined from blood samples of 29 snakes (11 males and 18 females. The performed hematological tests were: total red blood cell count (TRBCC, total white blood cell count (TWBCC, total trombocyte blood cell count (TTBCC; packed cell volume (PCV; hemoglobin content; and mean corpuscular volume (MCV, mean corpuscular hemoglobin (MCH and mean corpuscular hemoglobin concentration (MCHC. The mean values for TRBCC was 4.23cellsx10(5/mm³, for TWRCC was 8.92cellsx10³/mm³ and for TTBCC was 8.60cellsx10³/mm³. The mean for packed cell volume was 22.3%. The intraerythrocytic hemoparasite Hepatozoon sp was found on 58.6% of the studied animals.

  14. Depletion of Plasma Albumin for Proteomic Analysis of Bothrops jararaca Snake Plasma

    OpenAIRE

    de Morais-Zani, Karen; Grego, Kathleen Fernandes; Tanaka, Aparecida Sadae; Tanaka-Azevedo, Anita Mitico

    2011-01-01

    The proteomic analysis of plasma samples represents a challenge as a result of the presence of highly abundant proteins such as albumin. To enable the detection of biomarkers, which are commonly low-abundance proteins, in complex blood fluids, it is necessary to remove high-abundance proteins efficiently. Moreover, there is a range of about 10 orders of magnitude for the abundance of different protein species in serum. Here, we describe for the first time a study of reptilian albumin depletio...

  15. Human antibody fragments specific for Bothrops jararacussu venom reduce the toxicity of other Bothrops sp. venoms.

    Science.gov (United States)

    Roncolato, Eduardo Crosara; Pucca, Manuela Berto; Funayama, Jaqueline Carlos; Bertolini, Thaís Barboza; Campos, Lucas Benício; Barbosa, José Elpidio

    2013-01-01

    Approximately 20,000 snakebites are registered each year in Brazil. The classical treatment for venomous snakebite involves the administration of sera obtained from immunized horses. Moreover, the production and care of horses is costly, and the use of heterologous sera can cause hypersensitivity reactions. The production of human antibody fragments by phage display technology is seen as a means of overcoming some of these disadvantages. The studies here attempted to test human monoclonal antibodies specific to Bothrops jararacussu against other Bothrops sp. venoms, using the Griffin.1 library of human single-chain fragment-variable (scFv) phage antibodies. Using the Griffin.1 phage antibody library, this laboratory previously produced scFvs capable of inhibiting the phospholipase and myotoxic activities of Bothrops jararacussu venom. The structural and functional similarities of the various forms of phospholipase A2 (PLA₂) in Bothrops venom served as the basis for the present study wherein the effectiveness of those same scFvs were evaluated against B. jararaca, B. neuwiedi, and B. moojeni venoms. Each clone was found to recognize all three Bothrops venoms, and purified scFvs partially inhibited their in vitro phospholipase activity. In vivo assays demonstrated that the scFv clone P2B7 reduced myotoxicity and increased the survival of animals that received the test venoms. The results here indicate that the scFv P2B7 is a candidate for inclusion in a mixture of specific antibodies to produce a human anti-bothropic sera. This data demonstrates that the human scFv P2B7 represents an alternative therapeutic approach to heterologous anti-bothropic sera available today.

  16. Quantificação de citocinas no soro e homogenato da pata na intoxicação experimental com veneno de Bothropoides jararaca em ratos Wistar tratados com soroterapia e Mikania glomerata

    Directory of Open Access Journals (Sweden)

    Y.P. Motta

    2014-10-01

    Full Text Available O presente estudo teve como objetivo quantificar os níveis de citocinas pró-inflamatórias, entre as quais TNF-α, interleucina-1β (IL-1β, IL-6, e anti-inflamatórias, como IL-10, interferon-γ (INF-γ, bem como comparar o efeito do tratamento convencional com o efeito do tratamento complementado pelo extrato da planta Mikania glomerata, na intoxicação experimental por Bothropoides jararaca. Foram usados ratos Wistar,divididos em três grupos: C - controle, VB - veneno botrópico + soro antiofídico e VBM - veneno botrópico + soro antiofídico + Mikania glomerata. As citocinas foram quantificadas, no soro e no homogenato desses animais, pelo teste ELISA, em três momentos (M1 - 30 minutos, M2 - seis horas e M3 - 24 horas após a inoculação do veneno. Os resultados obtidos evidenciaram que a intoxicação por veneno botrópico estimula principalmente a produção de IL-6 no soro e TNF-α, IL-1β, IL-6 no homogenato da pata de animais experimentalmente intoxicados. O tratamento complementar, com o extrato da planta Mikania glomerata, teve influência principalmente na produção de IL-6, IL-10 e IFN-γ no soro e IL-6, IL-1β e IFN-γ no homogenato. Porém, são necessários novos estudos com o extrato de Mikania glomerata para que se possa entender a ação dessa planta sobre a intoxicação botrópica, bem como verificar qual a melhor via para administrá-lo.

  17. Aqueous Leaf Extract of Jatropha mollissima (Pohl Bail Decreases Local Effects Induced by Bothropic Venom

    Directory of Open Access Journals (Sweden)

    Jacyra Antunes dos Santos Gomes

    2016-01-01

    Full Text Available Snakebites are a serious worldwide public health problem. In Brazil, about 90% of accidents are attributed to snakes from the Bothrops genus. The specific treatment consists of antivenom serum therapy, which has some limitations such as inability to neutralize local effects, difficult access in some regions, risk of immunological reactions, and high cost. Thus, the search for alternative therapies to treat snakebites is relevant. Jatropha mollissima (Euphorbiaceae is a medicinal plant popularly used in folk medicine as an antiophidic remedy. Therefore, this study aims to evaluate the effect of the aqueous leaf extract from J. mollissima on local effects induced by Bothrops venoms. High Performance Liquid Chromatography with Diode Array Detection analysis and Mass Spectrometry analysis of aqueous leaf extract confirmed the presence of the flavonoids isoschaftoside, schaftoside, isoorientin, orientin, vitexin, and isovitexin. This extract, at 50–200 mg/kg doses administered by intraperitoneal route, showed significant inhibitory potential against local effects induced by Bothrops erythromelas and Bothrops jararaca snake venoms. Local skin hemorrhage, local edema, leukocyte migration, and myotoxicity were significantly inhibited by the extract. These results demonstrate that J. mollissima extract possesses inhibitory potential, especially against bothropic venoms, suggesting its potential as an adjuvant in treatment of snakebites.

  18. Preclinical assessment of the neutralizing capacity of antivenoms produced in six Latin American countries against medically-relevant Bothrops snake venoms.

    Science.gov (United States)

    Segura, A; Castillo, M C; Núñez, V; Yarlequé, A; Gonçalves, L R C; Villalta, M; Bonilla, C; Herrera, M; Vargas, M; Fernández, M; Yano, M Y; Araújo, H P; Boller, M A A; León, P; Tintaya, B; Sano-Martins, I S; Gómez, A; Fernández, G P; Geoghegan, P; Higashi, H G; León, G; Gutiérrez, J M

    2010-11-01

    Species of the genus Bothrops induce the vast majority of snakebite envenomings in Latin America. A preclinical study was performed in the context of a regional network of public laboratories involved in the production, quality control and development of antivenoms in Latin America. The ability of seven polyspecific antivenoms, produced in Argentina, Brazil, Peru, Bolivia, Colombia and Costa Rica, to neutralize lethal, hemorrhagic, coagulant, defibrinogenating and myotoxic activities of the venoms of Bothrops neuwiedi (diporus) (Argentina), Bothrops jararaca (Brazil), B. neuwiedi (mattogrossensis) (Bolivia), Bothrops atrox (Peru and Colombia) and Bothrops asper (Costa Rica) was assessed using standard laboratory tests. Despite differences in the venom mixtures used in the immunization of animals for the production of these antivenoms, a pattern of extensive cross-neutralization was observed between these antivenoms and all the venoms tested, with quantitative differences in the values of effective doses. This study reveals the capacity of these antivenoms to neutralize, in preclinical tests, homologous and heterologous Bothrops venoms in Central and South America, and also highlight quantitative differences in the values of Median Effective Doses (ED50s) between the various antivenoms. (c) 2010 Elsevier Ltd. All rights reserved.

  19. Effects of Schizolobium parahyba extract on experimental Bothrops venom-induced acute kidney injury.

    Directory of Open Access Journals (Sweden)

    Monique Silva Martines

    Full Text Available BACKGROUND: Venom-induced acute kidney injury (AKI is a frequent complication of Bothrops snakebite with relevant morbidity and mortality. The aim of this study was to assess the effects of Schizolobium parahyba (SP extract, a natural medicine with presumed anti-Bothrops venom effects, in an experimental model of Bothrops jararaca venom (BV-induced AKI. METHODOLOGY: Groups of 8 to 10 rats received infusions of 0.9% saline (control, C, SP 2 mg/kg, BV 0.25 mg/kg and BV immediately followed by SP (treatment, T in the doses already described. After the respective infusions, animals were assessed for their glomerular filtration rate (GFR, inulin clearance, renal blood flow (RBF, Doppler, blood pressure (BP, intra-arterial transducer, renal vascular resistance (RVR, urinary osmolality (UO, freezing point, urinary neutrophil gelatinase-associated lipocalin (NGAL, enzyme-linked immunosorbent assay [ELISA], lactate dehydrogenase (LDH, kinetic method, hematocrit (Hct, microhematocrit, fibrinogen (Fi, Klauss modified and blinded renal histology (acute tubular necrosis score. PRINCIPAL FINDINGS: BV caused significant decreases in GFR, RBF, UO, HcT and Fi; significant increases in RVR, NGAL and LDH; and acute tubular necrosis. SP did not prevent these changes; instead, it caused a significant decrease in GFR when used alone. CONCLUSION: SP administered simultaneously with BV, in an approximate 10∶1 concentration, did not prevent BV-induced AKI, hemolysis and fibrinogen consumption. SP used alone caused a decrease in GFR.

  20. Biochemical and hematological study of goats envenomed with natural and 60Co-irradiated bothropic venom

    International Nuclear Information System (INIS)

    Lucas de Oliveira, P.C.; Madruga, R.A.; Barbosa, N.P.U.; Sakate, M.

    2007-01-01

    Venoms from snakes of the Bothrops genus are proteolytic, coagulant, hemorrhagic and nephrotoxic, causing edema, necrosis, hemorrhage and intense pain at the bite site, besides systemic alterations. Many adjuvants have been added to the venom used in the sensitization of antiserum-producer animals to increase antigenic induction and reduce the envenomation pathological effects. Gamma radiation from 60 Co has been used as an attenuating agent of the venoms toxic properties. The main objective was to study, comparatively, clinical and laboratory aspects of goats inoculated with bothropic (Bothrops jararaca) venom, natural and irradiated from a 60 Co source. Twelve goats were divided into two groups of six animals: GINV, inoculated with 0.5 mg/kg of natural venom; and GIIV, inoculated with 0.5 mg/kg of irradiated venom. Blood samples were collected immediately before and one, two, seven, and thirty days after venom injection. Local lesions were daily evaluated. The following exams were carried out: blood tests; biochemical tests of urea, creatinine, creatine kinase, aspartate amino-transferase and alanine amino-transferase; clotting time; platelets count; and total serum immunoglobulin measurement. In the conditions of the present experiment, irradiated venom was less aggressive and more immunogenic than natural venom. (author)

  1. Immunological assessment of mice hyperimmunized with native and Cobalt-60-irradiated Bothrops venoms

    International Nuclear Information System (INIS)

    Ferreira Junior, R.S.; Meira, D.A.; Martinez, J.C.

    2005-01-01

    ELISA was used to evaluate, accompany, and compare the humoral immune response of Swiss mice during hyperimmunization with native and Cobalt-60-irradiated ( 60 Co) venoms of Bothrops jararaca, Bothrops jararacussu and Bothrops moojeni. Potency and neutralization were evaluated by in vitro challenges. After hyperimmunization, immunity was observed by in vivo challenge, and the side effects were assessed. The animals immunization with one LD50 of each venom occurred on days 1, 15, 21, 30, and 45, when blood samples were collected; challenges happened on the 60th day. Results showed that ELISA was efficient in evaluating, accompanying and comparing mouse immune response during hyperimmunization. Serum titers produced with natural venom were similar to those produced with irradiated venom. Immunogenic capacity was maintained after 60 Co-irradiation. The sera produced with native venom showed neutralizing potency and capacity similar to those of the sera produced with irradiated venom. All antibodies were able to neutralize five LD50 from these venoms. Clinical alterations were minimum during hyperimmunization with irradiated venom, however, necrosis and death occurred in animals inoculated with native venom. (author)

  2. Biochemical and hematological study of goats envenomed with natural and 60Co-irradiated bothropic venom

    Energy Technology Data Exchange (ETDEWEB)

    Lucas de Oliveira, P.C.; Madruga, R.A.; Barbosa, N.P.U. [Uberaba School of Veterinary Medicine (UNIUBE), MG (Brazil)]. E-mail: pedrolucaso@uol.com.br; Sakate, M. [UNESP, Botucatu, SP (Brazil). School of Veterinary Medicine and Animal Husbandry

    2007-07-01

    Venoms from snakes of the Bothrops genus are proteolytic, coagulant, hemorrhagic and nephrotoxic, causing edema, necrosis, hemorrhage and intense pain at the bite site, besides systemic alterations. Many adjuvants have been added to the venom used in the sensitization of antiserum-producer animals to increase antigenic induction and reduce the envenomation pathological effects. Gamma radiation from {sup 60}Co has been used as an attenuating agent of the venoms toxic properties. The main objective was to study, comparatively, clinical and laboratory aspects of goats inoculated with bothropic (Bothrops jararaca) venom, natural and irradiated from a {sup 60}Co source. Twelve goats were divided into two groups of six animals: GINV, inoculated with 0.5 mg/kg of natural venom; and GIIV, inoculated with 0.5 mg/kg of irradiated venom. Blood samples were collected immediately before and one, two, seven, and thirty days after venom injection. Local lesions were daily evaluated. The following exams were carried out: blood tests; biochemical tests of urea, creatinine, creatine kinase, aspartate amino-transferase and alanine amino-transferase; clotting time; platelets count; and total serum immunoglobulin measurement. In the conditions of the present experiment, irradiated venom was less aggressive and more immunogenic than natural venom. (author)

  3. Preclinical efficacy against toxic activities of medically relevant Bothrops sp. (Serpentes: Viperidae) snake venoms by a polyspecific antivenom produced in Mexico.

    Science.gov (United States)

    Segura, Álvaro; Herrera, María; Vargas, Mariángela; Villalta, Mauren; Uscanga-Reynell, Alfredo; León, Guillermo; Gutiérrez, José María

    2017-03-01

    The assessment of the preclinical neutralizing ability of antivenoms in Latin America is necessary to determine their scope of efficacy. This study was aimed at analyzing the neutralizing efficacy of a polyspecific bothropic-crotalic antivenom manufactured by BIRMEX in Mexico against lethal, hemorrhagic, defibrinogenating and in vitro coagulant activities of the venoms of Bothrops jararaca (Brazil), B. atrox (Perú and Colombia), B. diporus (Argentina), B. mattogrossensis (Bolivia), and B. asper (Costa Rica). Standard laboratory tests to determine these activities were used. In agreement with previous studies with bothropic antivenoms in Latin America, a pattern of cross-neutralization of heterologous venoms was observed. However, the antivenom had low neutralizing potency against defibrinogenating effect of the venoms of B. atrox (Colombia) and B. asper (Costa Rica), and failed to neutralize the in vitro coagulant activity of the venom of B. asper (Costa Rica) at the highest antivenom/venom ratio tested. It is concluded that, with the exception of coagulant and defibrinogenating activities of B. asper (Costa Rica) venom, this antivenom neutralizes toxic effects of various Bothrops sp venoms. Future studies are necessary to assess the efficacy of this antivenom against other viperid venoms.

  4. Duvernoy's gland secretion of Philodryas olfersii and Philodryas patagoniensis (Colubridae): neutralization of local and systemic effects by commercial bothropic antivenom (Bothrops genus).

    Science.gov (United States)

    Rocha, Marisa M Teixeira; Paixão-Cavalcante, Danielle; Tambourgi, Denise V; Furtado, Maria de Fátima D

    2006-01-01

    Colubrids involved in human envenomation in Brazil are mainly from the genera Helicops, Oxyrhopus, Thamnodynastes and Philodryas. There is a relatively large number of clinical descriptions involving the Xenodontinae snakes, Philodryas olfersii and Philodryas patagoniensis, in human accidents. The most common manifestations of envenomation are local pain, swelling, erythema and ecchymosis and regional lymphadenopathy with normal coagulation. The aims of this study were to characterize the biochemical and biological properties of P. olfersii and P. patagoniensis venoms, and to investigate their immunological cross-reactivities by using both specific antisera and anti-Bothrops sp serum used for human serum therapy in Brazil, in neutralizing the lethal and hemorrhagic effects of these venoms. We show here that P. olfersii e P. patagoniensis venoms present proteolytic and haemorrhagic activities but are devoid of phospholipase A2 activity. Haemorrhage and lethality induced by P. olfersii and P. patagoniensis are associated with metal-dependent proteinases, since EDTA could block these toxic activities. P. olfersii and P. patagoniensis venoms were immunogenic and the antisera produced were able to recognize several bands in P. olfersii, P. patagoniensis venoms in Bothrops jararaca venom.

  5. Inhibition of local effects induced by Bothrops erythromelas snake venom: Assessment of the effectiveness of Brazilian polyvalent bothropic antivenom and aqueous leaf extract of Jatropha gossypiifolia.

    Science.gov (United States)

    Félix-Silva, Juliana; Gomes, Jacyra A S; Xavier-Santos, Jacinthia B; Passos, Júlia G R; Silva-Junior, Arnóbio A; Tambourgi, Denise V; Fernandes-Pedrosa, Matheus F

    2017-01-01

    Bothrops erythromelas is a snake of medical importance responsible for most of the venomous incidents in Northeastern Brazil. However, this species is not included in the pool of venoms that are used in the Brazilian polyvalent bothropic antivenom (BAv) production. Furthermore, it is well known that antivenom therapy has limited efficacy against venom-induced local effects, making the search for complementary alternatives to treat snakebites an important task. Jatropha gossypiifolia is a medicinal plant widely indicated in folk medicine as an antidote for snakebites, whose effectiveness against Bothrops jararaca venom (BjV) has been previously demonstrated in mice. In this context, this study assessed the effectiveness of the aqueous extract (AE) of this plant and of the BAv against local effects induced by B. erythromelas venom (BeV). Inhibition of BeV-induced edematogenic and hemorrhagic local effects was assayed in mice in pre-treatment (treatment prior to BeV injection) and post-treatment (treatment post-envenomation) protocols. Inhibition of proteolytic, phospholipase A 2 (PLA 2 ) and hyaluronidase enzymatic activities of BeV were evaluated in vitro. BAv cross-reactivity and estimation of antibody titers against BeV and BjV were assessed by Ouchterlony double diffusion test. The results show that in pre-treatment protocol AE and BAv presented very similar effects (about 70% of inhibition for edematogenic and 40% for hemorrhagic activities). However, BAv poorly inhibited edema and hemorrhage in post-envenomation protocol, whilst, in contrast, AE was significantly active even when used after BeV injection. AE was able to inhibit all the tested enzymatic activities of BeV, while BAv was active only against hyaluronidase activity, which could justify the low effectiveness of BAv against BeV-induced local effects in vivo. Ouchterlony's test showed positive cross-reactivity against BeV, but the antibody titers were slightly higher against BjV. Together, these

  6. Acidente por serpentes do gênero Bothrops: série de 3.139 casos

    Directory of Open Access Journals (Sweden)

    Lindioneza Adriano Ribeiro

    1997-12-01

    Full Text Available Em avaliação dos prontuários médicos de 3.139 pacientes picados por serpentes do gênero Bothrops atendidos no Hospital Vital Brazil (HVB, de 1981 a 1990, observou-se maior acometimento do sexo masculino (75,7%. Em 1.412 casos (45,0% a serpente foi identificada, sendo 1.376 B. jararaca, 20 B. jararacussu, 11 B. neuwiedi, 2 B. moojeni, 2 B. alternatus e 1 B. pradoi. As regiões anatômicas mais comumente picadas foram: pé (47,5% e mão (21,3%. O torniquete foi realizado em 38,2% dos casos e sua freqüência diminuiu durante esse período (p Medical records of 3,139 patients bitten by Bothrops snakes and attended at Vital Brazil Hospital (HVB from 1981 to 1990 were reviwed. They were more frequent in males (75.7%. In 1,412 cases (45.0% the snake was classified by species, and 1,376 were B. jararaca, 20 B. jararacussu, 11 B. neuwiedi, 2 B. moojeni, 2 B. alternatus e 1 B. pradoi. The most frequent bitten anatomic regions were: foot (47.5% and hand (21.3%. Tourniquet was used in 38.2% of the cases and its frequency fell down during the study period (p < 0.05. The clinical features at the bite site were: pain (95.6%, swelling (95.4%, echimosis (56.1%, blisters (13.8%, necrosis (16.5%, and abscess (11.0%. Systemic manifestations were: bleeding (12.3%, acute renal failure (1.6%, and shock (0.7%. There were blood coagulation disorders in 1,730 (57.9% of the 2,990 cases. There were 21 amputations (0.7% and 9 deaths (0.3%. The average serum dose that was used in treatment fell down during the study period (p < 0.001.

  7. The interaction of the antitoxin DM43 with a snake venom metalloproteinase analyzed by mass spectrometry and surface plasmon resonance

    DEFF Research Database (Denmark)

    Brand, Guilherme D; Salbo, Rune; Jørgensen, Thomas J D

    2012-01-01

    DM43 is a circulating dimeric antitoxin isolated from Didelphis aurita, a South American marsupial naturally immune to snake envenomation. This endogenous inhibitor binds non-covalently to jararhagin, the main hemorrhagic metalloproteinase from Bothrops jararaca snake venom, and efficiently...

  8. Factors underlying the natural resistance of animals against snake venoms

    Directory of Open Access Journals (Sweden)

    H. Moussatché

    1989-01-01

    Full Text Available The existence of mammals and reptilia with a natural resistance to snake venoms is known since a long time. This fact has been subjected to the study by several research workers. Our experiments showed us that in the marsupial Didelphis marsupialis, a mammal highly resistant to the venom of Bothrops jararaca, and other Bothrops venoms, has a genetically origin protein, a alpha-1, acid glycoprotein, now highly purified, with protective action in mice against the jararaca snake venom.

  9. Hemiparesia esquerda consecutiva a empeçonhamento: Por Bothrops Jararacussu

    Directory of Open Access Journals (Sweden)

    Napoleão L. Teixeira

    1944-09-01

    Full Text Available O A. apresenta uma observação de hemiparesia esquerda, consecutiva a envenenamento por Bothrops Jararacussu. É o segundo caso que publica, com idêntica etiologia; além das suas, há duas outras observações semelhantes, de Octávio de Magalhães. Depois de discorrer sobre as hemiplegias cerebrais infantis - mais comuns - lembra que estas nem sempre se acompanham de espasticidade, podendo apresentar-se, raramente, com hipotonia muscular, ou mesmo de coréia, atetose, ou movimentos córeo-atetóticos que falam em favor de comprometimento extrapiramidal. Assinala, também, que crises epileptiformes - generalizadas ou parciais - podem ocorrer, o mesmo se podendo dizer de distúrbios psíquicos. Traduzir-se-iam, estes, por déficit intelectual marcado, podendo ir desde a debilidade mental até a idiotia; ou então, mesmo quando não há aparente comprometimento da esfera intelectual, por perturbações da afetividade, da vontade e do pragmatismo. Passando em revista as causas mais frequentes das hemiplegias cerebrais infantis, abre lugar, dentre elas, para os empeçonhamentos ofídico e escorpiônico (Magalhães e Guimarães. Estuda, a seguir, a patogenia das hemiplegias no decurso dos referidos empeçonhamentos, alinhando as hipóteses correntes, da preferência de conceituadas autoridades no assunto. Transcreve, finalmente, sua observação clínica, da qual se pode concluir, sem dúvida possível, haver, no caso "que apresenta, estreita e imediata ligação entre o envenenamento ofídico e a hemiparesia em estudo. O observado seria portador, além do mais, de crises epileptiformes generalizadas, aparecidas logo a seguir ao empeçonhamento. Além da fotografia do caso que apresenta, mostra-nos o A. outra, de caso anteriormente publicado, este, porém, de hemiplegia cerebral infantil típica, instalada aos 11 anos, após picada de Bothrops jararaca.

  10. Characterization of aspercetin, a platelet aggregating component from the venom of the snake Bothrops asper which induces thrombocytopenia and potentiates metalloproteinase-induced hemorrhage.

    Science.gov (United States)

    Rucavado, A; Soto, M; Kamiguti, A S; Theakston, R D; Fox, J W; Escalante, T; Gutiérrez, J M

    2001-04-01

    Thrombocytopenia occurs in a number of patients bitten by Bothrops asper, a species responsible for the majority of snakebites in Central America and southern Mexico. In this work we describe the isolation of a new platelet-aggregating protein, named aspercetin, from the venom of B. asper, which induces thrombocytopenia in mice. Isolation was carried out by a combination of ion-exchange chromatography on DEAE-Sepharose and affinity chromatography on Affi-Gel Blue. Aspercetin is a disulfide-linked heterodimer, with a pI of 4.5 and a molecular mass of 29,759 Da, detemined by MALDI-ESI mass spectrometry. N-terminal sequence shows homology with a number of venom proteins which belong to the C-type lectin family. Aspercetin has functional similarities with botrocetin, from B. jararaca venom, since it induces platelet aggregation only in the presence of plasma or purified von Willebrand factor. Aspercetin-mediated platelet aggregation results from the interaction of von Willebrand factor with platelet receptor GPIb. Aspercetin lacks anticoagulant effect and does not agglutinate erythrocytes, in contrast with other representatives of the C-type lectin family isolated from snake venoms. Moreover, aspercetin is not lethal, nor does it induce myonecrosis, hemorrhage and edema. When injected intravenously or intramuscularly in mice it induces a rapid, dose-dependent drop in platelet counts and prolongs the bleeding time, suggesting that it may play a role in the thrombocytopenia that develops in a number of B. asper envenomations. Moreover, mice injected intravenously with aspercetin and then receiving an intradermal injection of B. asper hemorrhagic metalloproteinase BaP1 develop a larger hemorrhagic lesion than mice receiving only BaP1. This suggests that aspercetin, by reducing platelet numbers, may

  11. Variability of Venom-Neutralizing Properties of Serum from Snakes of the Colubrid Genus Lampropeltis

    Science.gov (United States)

    1992-01-01

    chick in the terciopelo Bothrops asper venom by blood se- zootoxicologic studies on venoms. Copeia 1959: rum of the colubnd snake Cletie dIeia. Toxicon 20...effect of Bothrops jararaca and faded rattlesnake (Crotalus viridis concolor) venom: Waglerophis merremii snake plasma on the coagu- lethal toxicity and

  12. Inflammation induced by Bothrops asper venom.

    Science.gov (United States)

    Teixeira, Catarina; Cury, Yara; Moreira, Vanessa; Picolo, Gisele; Chaves, Fernando

    2009-07-01

    Inflammation is a major characteristic of envenomation by snakes from viperine and crotaline species. Bothrops asper snake venom elicits, among other alterations, a pronounced inflammatory response at the site of injection both in humans and experimental animals. This review describes the current status of our understanding of the inflammatory reaction, including pain, triggered by Bothrops asper venom. The experimental studies on the action of this venom as well as the complex network of chemical mediators involved are summarized. Moreover, aspects of the molecular mechanisms orchestrating this important response to envenomation by Bothrops asper are presented. Considering that isolated toxins are relevant tools for understanding the actions of the whole venom, studies dealing with the mechanisms of inflammatory and nociceptive properties of phospholipases A(2), a metalloproteinase and serine-proteases isolated from Bothrops asper venom are also described.

  13. Preclinical testing of Peruvian anti-bothropic anti-venom against Bothrops andianus snake venom.

    Science.gov (United States)

    Schneider, Francisco S; Starling, Maria C; Duarte, Clara G; Machado de Avila, Ricardo; Kalapothakis, Evanguedes; Silva Suarez, Walter; Tintaya, Benigno; Flores Garrido, Karin; Seraylan Ormachea, Silvia; Yarleque, Armando; Bonilla, César; Chávez-Olórtegui, Carlos

    2012-11-01

    Bothrops andianus is a venomous snake found in the area of Machu Picchu (Peru). Its venom is not included in the antigenic pool used for production of the Peruvian anti-bothropic anti-venom. B. andianus venom can elicit many biological effects such as hemorrhage, hemolysis, proteolytic activity and lethality. The Peruvian anti-bothropic anti-venom displays consistent cross-reactivity with B. andianus venom, by ELISA and Western Blotting and is also effective in neutralizing the venom's toxic activities. Copyright © 2012 Elsevier Ltd. All rights reserved.

  14. Determination of sphingomyelinase-D activity of Loxosceles venoms in sphingomyelin/cholesterol liposomes containing horseradish peroxidase.

    Science.gov (United States)

    Gomes, Marco Túlio R; Guimarães, Gabriela; Frézard, Frédéric; Kalapothakis, Evanguedes; Minozzo, João Carlos; Chaim, Olga Meiri; Veiga, Silvio Sanches; Oliveira, Sergio Costa; Chávez-Olórtegui, Carlos

    2011-03-15

    Based on degradation of sphingomyelin/cholesterol liposomes containing entrapped horseradish peroxidase, we evaluated the Sphingomyelinase-D (SMase-D) activity of scorpion, spider and snake venoms by monitoring spectrophotometrically the product of oxidation of HRP released. The results indicate that Loxosceles crude venoms (Loxosceles intermedia, Loxosceles laeta, Loxosceles gaucho and Loxosceles similis) displayed SMase-D activity in a concentration-dependent manner. Furthermore, this activity was blocked by the anti-loxoscelic antivenom. However, Tityus serrulatus scorpion venom, Phoneutria nigriventer spider venom and Bothrops jararaca, Crotalus durissus, Lachesis muta and Micrurus frontalis snake venoms did not show measurable SMase-D activity. Copyright © 2011 Elsevier Ltd. All rights reserved.

  15. Ação de venenos de serpentes brasileiras sôbre a alexina ou complemento

    Directory of Open Access Journals (Sweden)

    F. da Rocha Lagôa

    1947-06-01

    Full Text Available O autor estudou a ação inativante, sôbre o complemento de cobaia, alguns venenos de serpentes brasileiras pertencentes às famílias dos Elapideos e Crotalideos. Da primeira, foi utilizado veneno de Micrurus frontalis, da segunda, foram usados venenos de espécies pertencentes aos gêneros Crotalus (C. terrificus e Bothrops (B. atrox, B. neuwiedii, B. jararaca, B. ja-raracussú, B. cotiara e B. alternata. O venenos de M. frontalis e C. terri¬ficus se revelaram incapazes de inativar o complemento, ao passo que os diversos de Bothrops empregados se mostraram altamente inativantes, destruindo sempre o 4.° componente do complemento (C4, fração idêntica à afetada pela ação da amônea.The author shows in this paper the results of the inactivation of com¬plement or alexin by some Brazilian snakes venoms of the Elapideae and Cro¬talideae families. The venom of Micrurus frontalis (Elapideae family and of Crotalus terrificus (Crotalideae family did not destroy the complement; but the venoms of Bothrops generus (B. atrox, B. neuwiedii, B. jararaca, B. jararacussú, B. cotiara e B. alternata, (Crotalideae family are highly active in the inactivation complement, always destroing the fourth component, whichis exactly the same component that is detroyed by ammonia.

  16. Analysis of Brazilian snake venoms by neutron activation analysis

    International Nuclear Information System (INIS)

    Saiki, M.; Vasconcellos, M.B.A.; Rogero, J.R.; Cruz, M.C.G.

    1991-01-01

    Instrumental neutron activation analysis (INAA) has been applied to multielemental determinations of Brazilian snake venoms from the species: Bothrops jararacussu, Crotalus durissus terrificus and Bothrops jararaca. Concentrations of Br, Ca, Cl, Cs, K, Mg, Na, Rb, Sb, Se and Zn have been determined in lyophilized venoms by using short and long irradiations in the IEA-RI nuclear reactor under a thermal neutron flux of 10 11 to 10 13 n · cm -2 · s -1 . The reference materials NIST Bovine Liver 1577 and IUPAC Bowen's Kale were also analyzed simultaneously with the venoms to evaluate the accuracy and the reproducibility of the method. The concentrations of the elements found in snake venoms from different species were compared. The Crotalus durissus terrificus venoms presented high concentration of Se but low concentrations of Zn when these results are compared with those obtained from genera Bothrops venoms. (author) 9 refs.; 2 tabs

  17. Somatic pairing, endomitosis and chromosome aberrations in snakes (Viperidae and Colubridae

    Directory of Open Access Journals (Sweden)

    Beçak Maria Luiza

    2003-01-01

    Full Text Available The positioning of macrochromosomes of Bothrops jararaca and Bothrops insularis (Viperidae was studied in undistorted radial metaphases of uncultured cells (spermatogonia and oogonia not subjected to spindle inhibitors. Colchicinized metaphases from uncultured (spleen and intestine and cultured tissues (blood were also analyzed. We report two antagonic non-random chromosome arrangements in untreated premeiotic cells: the parallel configuration with homologue chromosomes associated side by side in the metaphase plate and the antiparallel configuration having homologue chromosomes with antipolar distribution in the metaphase ring. The antiparallel aspect also appeared in colchicinized cells. The spatial chromosome arrangement in both configurations is groupal size-dependent and maintained through meiosis. We also describe, in untreated gonia cells, endomitosis followed by reductional mitosis which restores the diploid number. In B. jararaca males we observed that some gonad regions present changes in the meiotic mechanism. In this case, endoreduplicated cells segregate the diplochromosomes to opposite poles forming directly endoreduplicated second metaphases of meiosis with the suppression of first meiosis. By a successive division, these cells form nuclei with one set of chromosomes. Chromosome doubling in oogonia is known in hybrid species and in parthenogenetic salamanders and lizards. This species also presented chromosome rearrangements leading to aneuploidies in mitosis and meiosis. It is suggested that somatic pairing, endomitosis, meiotic alterations, and chromosomal aberrations can be correlated processes. Similar aspects of nuclei configurations, endomitosis and reductional mitosis were found in other Viperidae and Colubridae species.

  18. Reactividad inmunoquímica de sueros anti- Caiman yacare y Caiman latirostris frente a sueros de diferentes especies

    Directory of Open Access Journals (Sweden)

    de Roodt, Adolfo Rafael

    2010-06-01

    Full Text Available Se estudió la reactividad inmunoquímica entre los sueros de distintas especies de reptiles frente a sueros hiperinmunes experimentales anti-suero de Caiman yacare y anti-suero de Caiman latirostris. Los sueros que se probaron fueron los homólogos de Caiman yacare, Caiman latirostris y los heterólogos de Alligator missisipiensis, Tupinambis merinae, Tupinambis rufescens, Chelonoidis chilensis, Clelia rustica, Waglerophis merremii, Lystrophys dorbignyi, Phyton molurus, Boa constrictor occidentalis, Eunectes notaeus, Crotalus durissus terrificus, Bothrops alternatus, Bothrops diporus, Bothrops jararaca, Bothrops jararacussu, Bothrops moojeni, Pitangus sulphuratus y Gallus gallus. La reactividad inmunoquímica se determinó mediante las técnicas de doble inmunodifusión y ELISA, mostrándose importante entre los sueros de los crocodrílidos y baja entre estos y los de las otras especies de reptiles estudiadas. Se observó mayor reactividad entre los antisueros anti-Caiman respecto a los sueros de Caiman latirostris y Caiman yacare que frente al suero de Alligator missisipiensis. Además, se encontró una fuerte reactividad entre ambos sueros anti-Caiman y el de Gallus gallus poniendo en evidencia la fuerte reactividad entre los sueros de arcosaurios. In order to study the immunochemical reactivity among sera from different species of reptiles regarding sera from Caiman, the immunoreactivity of sera from reptiles against antisera to Caiman yacare or anti-Caiman latirostris sera was studied. These hiperimmune sera were tested against sera from Alligator missisipiensis, Tupinambis merinae, Tupinambis rufescens, Chelonoidis chilensis, Clelia rustica, Waglerophis merremii, Lystrophys dorbignyi, Phyton molurus, Boa constrictor occidentalis, Eunectes notaeus, Crotalus durissus terrificus, Bothrops alternatus, Bothrops neuwiedii, Bothrops jararaca, Bothrops jararacussu, Bothrops moojeni, Pitangus sulphuratus and Gallus gallus. The immunochemical

  19. Hemostatic properties of Venezuelan Bothrops snake venoms with special reference to Bothrops isabelae venom.

    Science.gov (United States)

    Rodríguez-Acosta, Alexis; Sánchez, Elda E; Márquez, Adriana; Carvajal, Zoila; Salazar, Ana M; Girón, María E; Estrella, Amalid; Gil, Amparo; Guerrero, Belsy

    2010-11-01

    In Venezuela, Bothrops snakes are responsible for more than 80% of all recorded snakebites. This study focuses on the biological and hemostatic characteristics of Bothrops isabelae venom along with its comparative characteristics with two other closely related Bothrops venoms, Bothrops atrox and Bothrops colombiensis. Electrophoretic profiles of crude B. isabelae venom showed protein bands between 14 and 100 kDa with the majority in the range of 14-31 kDa. The molecular exclusion chromatographic profile of this venom contains five fractions (F1-F5). Amidolytic activity evaluation evidenced strong thrombin-like followed by kallikrein-like activities in crude venom and in fractions F1 and F2. The fibrinogenolytic activity of B. isabelae venom at a ratio of 100:1 (fibrinogen/venom) induced a degradation of A alpha and B beta chains at 15 min and 2 h, respectively. At a ratio of 100:10, a total degradation of A alpha and B beta chains at 5 min and of gamma chains at 24 h was apparent. This current study evidences one of rarely reported for Bothrops venoms, which resembles the physiologic effect of plasmin. B. isabelae venom as well as F2 and F3 fractions, contain fibrinolytic activity on fibrin plate of 36, 23.5 and 9.45 mm(2)/microg, respectively using 25 microg of protein. Crude venom and F1 fraction showed gelatinolytic activity. Comparative analysis amongst Venezuelan bothropoid venoms, evidenced that the LD(50) of B. isabelae (5.9 mg/kg) was similar to B. atrox-Puerto Ayacucho 1 (6.1 mg/kg) and B. colombiensis-Caucagua (5.8 mg/kg). B. isabelae venom showed minor hemorrhagic activity, whereas B. atrox-Parguasa (Bolivar state) was the most hemorrhagic. In this study, a relative high thrombin-like activity was observed in B. colombiensis venoms (502-568 mUA/min/mg), and a relative high factor Xa-like activity was found in B. atrox venoms (126-294 mUA/min/mg). Fibrinolytic activity evaluated with 10 microg protein, showed that B. isabelae venom contained higher

  20. Neutralization of proteases from Bothrops snake venoms by the aqueous extract from Casearia sylvestris (Flacourtiaceae).

    Science.gov (United States)

    Borges, M H; Soares, A M; Rodrigues, V M; Oliveira, F; Fransheschi, A M; Rucavado, A; Giglio, J R; Homsi-Brandeburgo, M I

    2001-12-01

    Aqueous extract from Casearia sylvestris leaves, a typical plant from Brazilian open pastures, was able to neutralize the hemorrhagic activity caused by Bothrops asper, Bothrops jararacussu, Bothrops moojeni, Bothrops neuwiedi and Bothrops pirajai venoms. It also neutralized two hemorrhagic metalloproteinases from Bothrops asper venom. Proteolytic activity on casein induced by bothropic venoms and by isolated proteases, including Bn2 metalloproteinase from B. neuwiedi venom, was also inhibited by the C. sylvestris extract in different levels. The alpha-fibrinogen chain was partially protected against degradation caused by B. jararacussu venom, when this venom was incubated with C. sylvestris extract. We also observed that this extract partially increased the time of plasma coagulation caused by B. jararacussu, B. moojeni and B. neuwiedi venoms. C. sylvestris extract did not induce proteolysis in any substrate assayed.

  1. Bothrops fonsecai snake venom activities and cross-reactivity with commercial bothropic venom.

    Science.gov (United States)

    Collaço, Rita de Cássia O; Randazzo-Moura, Priscila; Tamascia, Mariana L; da Silva, Igor Rapp F; Rocha, Thalita; Cogo, José C; Hyslop, Stephen; Sanny, Charles G; Rodrigues-Simioni, Léa

    2017-01-01

    In this work, we examined some biochemical and biological activities of Bothrops fonsecai venom, a pitviper endemic to southeastern Brazil, and assessed their neutralization by commercial bothropic antivenom (CAv). Cross-reactivity of venom with CAv was also assessed by immunoblotting and size-exclusion high performance chromatography (SE-HPLC). Bothrops fonsecai venom had PLA 2 , proteolytic and esterase activities that were neutralized to varying extents by venom:antivenom ratios of 5:1 and 5:2 (PLA 2 and esterase activities) or not significantly by either venom:antivenom ratio (proteolytic activity). The minimum hemorrhagic dose (69.2μg) was totally neutralized by both ratios. Clotting time in rat citrated plasma was 33±10.5s (mean±SD; n=5) and was completely neutralized by a 5:2 ratio. Edema formation was dose-dependent (1-30μg/site) and significantly inhibited by both ratios. Venom (10-300μg/mL) caused neuromuscular blockade in extensor digitorum longus preparations; this blockade was inhibited best by a 5:2 ratio. Venom caused myonecrosis and creatine kinase release in vivo (gastrocnemius muscle) and in vitro (extensor digitorum longus) that was effectively neutralized by both venom:antivenom ratios. Immunoblotting showed that venom components of ~25-100kDa interacted with CAv. SE-HPLC profiles for venom incubated with CAv or specific anti-B. fonsecai antivenom raised in rabbits (SAv) indicated that CAv had a higher binding capacity than SAv, whereas SAv had higher affinity than CAv. These findings indicate that B. fonsecai venom contains various activities that are neutralized to different extents by CAv and suggest that CAv could be used to treat envenoming by B. fonsecai. Copyright © 2016. Published by Elsevier Inc.

  2. Hemorrhagic stroke secondary to Bothrops spp. venom: A case report.

    Science.gov (United States)

    Delgado, Anna Beatriz Temoteo; Gondim, Caio Cesar Vaz Lacet; Reichert, Lucas Pereira; da Silva, Pedro Hugo Vieira; Souza, Rodrigo Marmo da Costa E; Fernandes, Thiago Monteiro de Paiva; Calvo, Bernardino Fernandez

    2017-06-15

    The Bothrops spp. venom contain metalloproteinases that contributes to vascular and hemorrhagic effects. This case report describes a 58 years-old patient from the city of Dona Inês, Paraiba, Brazil victim of an ophidian accident by Bothrops spp. The vascular and hemorrhagic effects of venom components have triggered a hemorrhagic stroke. Brazil has about 600 deaths annually due to ophidian accidents. However, as reports have been precarious, the obtaining of epidemiological-clinical data has been affected. This case highlights the importance of prior knowledge of possible neurological and vascular complications in Bothrops spp. venom to increase the effectiveness of an adequate treatment. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. First report of hepatic hematoma after presumed Bothrops envenomation

    Directory of Open Access Journals (Sweden)

    Fernanda Cristina Cunha

    2015-10-01

    Full Text Available ABSTRACTIn Latin America, Bothrops envenomation is responsible for the majority of accidents caused by venomous snakes. Patients usually present local edema, bleeding and coagulopathy. Visceral hemorrhage is extremely rare and considered a challenge for diagnosis and management. We report the first case of hepatic hematoma owing to the bothropic envenomation in a 66-year-old man who was bitten in the left leg. He presented local edema, coagulopathy, and acute kidney injury. Radiological findings suggested hepatic hematoma, with a volume of almost 3 liters. The hepatic hematoma was gradually absorbed without the need for surgical intervention with complete resolution in 8 months.

  4. Ontogenetic variability of Bothrops atrox and Bothrops asper snake venoms from Colombia.

    Science.gov (United States)

    Saldarriaga, Mónica María; Otero, Rafael; Núñez, Vitelbina; Toro, Maria Fabiola; Díaz, Abel; Gutiérrez, José María

    2003-09-15

    The lancehead snakes Bothrops asper and Bothrops atrox inflict 70-90% of the 3000 bites reported every year in Colombia. In this work, the venoms of B. atrox from Meta (Villavicencio, 33 specimens) and B. asper from Antioquia (San Carlos, 45 specimens), all of them born in captivity, were obtained at different ages (0-6 months; 1, 2 and 3-years old) and compared in terms of their pharmacological and immunochemical characteristics. A conspicuous ontogenetic variability was observed in venom samples from both species. Venoms from newborn and juvenile specimens showed higher lethal, hemorrhagic, edema-forming and coagulant activities, whereas venoms from 3-year old specimens showed higher indirect hemolytic, i.e. phospholipase A2 activity, being more significant in the case of B. asper. SDS-polyacrylamide gel electrophoresis of whole venom for both species evidenced a predominance of high mol. mass bands in the venoms from specimens of asper of <6 months and in those from 3-year old specimens. Venom of adult specimens showed a higher number of peaks with indirect hemolytic activity than venom of newborn specimens. Polyvalent antivenom produced in Costa Rica recognized all the bands of both venoms from specimens at all ages tested, when assayed by Western blotting.

  5. Postprandial thermogenesis in Bothrops moojeni (Serpentes: Viperidae

    Directory of Open Access Journals (Sweden)

    DR Stuginski

    2011-01-01

    Full Text Available Snakes that can ingest prey that are proportionally large have high metabolic rates during digestion. This great increase in metabolic rate (specific dynamic action - SDA may create a significant augment in the animal's body temperature. The present study investigated postprandial thermogenesis in Bothrops moojeni. Briefly, two groups of snakes were fed meals equivalent to 17 ± 3% and 32 ± 5% of their body weight and were observed for 72 hours, in which thermal images of each snake were taken with an infrared camera in a thermostable environment with a constant air temperature of 30°C. The results showed a significant increase in snake surface temperature, with a thermal peak between 33 and 36 hours after feeding. The meal size had a great impact on the intensity and duration of the thermogenic response. Such increase in temperature appears to be connected with the huge increase in metabolic rates during digestion of relatively large prey by snakes that feed infrequently. The ecologic implication of the thermogenic response is still not well understood; however, it is possible that its presence could affect behaviors associated with the snake digestion, such as postprandial thermophily.

  6. Body size, reproductive biology and abundance of the rare pseudoboini snakes genera Clelia and Boiruna (Serpentes, Colubridae in Brazil

    Directory of Open Access Journals (Sweden)

    Lígia Pizzatto

    2005-12-01

    Full Text Available Pseudoboini snakes of the genera Clelia and Boiruna are apparently rare in nature and certainly rare in collections. This work presents data on body size, reproduction and abundance of five Brazilian species of these genera, in the largest collection of snakes in Latin America, the Instituto Butantan. Despite scarcity of data, follicular cycle seems to be continuous in most species, except Clelia rustica, which occurs in highlands. Females are largerthan males in all species, and fecundity is low when compared to other pseudoboini. Abundance is very low for all species even considering 100 years of collecting, and it is decreasing in recent decades when compared to other snakes (Bothrops jararaca, Oxyrhopus guibei, O. clathratus, Philodryas patagoniensis, Sibynomorphus mikanii, and Spilotes pullatus. The studied species present at least five traits of commonly threatened species and require more attention in researches and conservation policies.

  7. Snakes killed on the roads in the state of Santa Catarina, southern Brazil

    Directory of Open Access Journals (Sweden)

    Ivo Rohling Ghizoni-Jr

    2009-06-01

    Full Text Available Roads cause diverse impacts to ecosystems, including habitat loss and fragmentation, alteration of the natural landscape and death of animals. However, little is known about the impact of this mortality on the animal populations. The snake fauna of the state of Santa Catarina is the least studied of southern Brazil. In this work we present data on 165 snakes of 38 species found dead on the roads of the state between 2003 and 2008, which corresponds to 50% of the snake species in relation to the expected occurrence for the state of Santa Catarina. The four most frequent species were Philodryas patagoniensis (n = 22; 13.33%, Liophis miliaris (n = 21; 12.72%, P. aestiva (n = 13; 7.87% and Bothrops jararaca (n = 12; 7.27%, which represent together about 41% of the snakes found dead on the roads. We extend the known distribution of Imantodes cenchoa by about 60km southward.

  8. ELISA for the detection of venom antigens in experimental and clinical envenoming by Loxosceles intermedia spiders.

    Science.gov (United States)

    Chávez-Olórtegui, C; Zanetti, V C; Ferreira, A P; Minozzo, J C; Mangili, O C; Gubert, I C

    1998-04-01

    Enzyme linked immunosorbent assays (ELISA) were developed to detect antigens from Loxosceles intermedia spider venom. Hyperimmune horse anti-Loxosceles intermedia IgGs were prepared by immunoaffinity chromatography and used to set up a sandwich-type ELISA. The specificity of the assay was demonstrated by its capacity to correctly discriminate the circulating antigens in mice that were experimentally inoculated with L. intermedia venom from those inoculated with L. gaucho, L. laeta, and Phoneutria nigriventer spider venoms, Tityus serrulatus scorpion venom and Bothrops jararaca, Crotalus durissus terrificus, Lachesis muta muta and Micrurus frontalis snake venoms. Measurable absorbance signals were obtained with 0.8 ng of venom per assay. The ELISA also detected antigens in the sera of patients envenomed by L. intermedia. Therefore, after standardization for clinical use this ELISA may be a valuable tool for clinicians and epidemiologists.

  9. Snake venomics and antivenomics of Bothrops colombiensis, a medically important pitviper of the Bothrops atrox-asper complex endemic to Venezuela: Contributing to its taxonomy and snakebite management

    OpenAIRE

    Calvete, Juan J.; Borges, Adolfo; Segura Ruiz, Álvaro; Flores Díaz, Marietta; Alape Girón, Alberto; Gutiérrez, José María; Diez, Nardy; De Sousa, Leonardo; Kiriakos, Demetrio; Sánchez, Eladio; Faks, José G.; Escolano, José; Sanz, Libia

    2009-01-01

    The taxonomic status of the medically important pitviper of the Bothrops atrox-asper complex endemic to Venezuela, which has been classified as Bothrops colombiensis, remains incertae cedis. To help resolving this question, the venom proteome of B. colombiensis was characterized by reverse-phase HPLC fractionation followed by analysis of each chromatographic fraction by SDS-PAGE, N-terminal sequencing, MALDI-TOF mass fingerprinting, and collision-induced dissociation tandem mass spectrometry ...

  10. Insuficiencia renal aguda inducida por mordedura de serpiente Bothrops

    Directory of Open Access Journals (Sweden)

    Gustavo A. Aroca Martínez

    2014-01-01

    Full Text Available Mujer de 58 años de edad, remitida a urgencias por presentar cuadro clínico de insuficiencia renal aguda (IRA secundaria a mordedura de serpiente (Bothrops Atrox. Ingresa hipotensa con elevación de azoados e hiperkalemia, ecografía renal dentro de parámetros normales. Se maneja terapia dialítica con lo cual presenta mejoría clínica. En este reporte se detallan aspectos del diagnóstico, manejo clínico y posibles mecanismos fisiopatológicos que explican el daño renal.

  11. Neuromuscular action of venom from the South American colubrid snake Philodryas patagoniensis.

    Science.gov (United States)

    Carreiro da Costa, Roberta S; Prudêncio, Luiz; Ferrari, Erika Fonseca; Souza, Gustavo H M F; de Mello, Sueli Moreira; Prianti Júnior, Antonio Carlos Guimarães; Ribeiro, Wellington; Zamunér, Stella Regina; Hyslop, Stephen; Cogo, José Carlos

    2008-07-01

    Snakes of the opisthoglyphous genus Philodryas are widespread in South America and cause most bites by colubrids in this region. In this study, we examined the neurotoxic and myotoxic effects of venom from Philodryas patagoniensis in biventer cervicis and phrenic nerve-diaphragm preparations and we compared the biochemical activities of venoms from P. patagoniensis and Philodryas olfersii. Philodryas patagoniensis venom (40 microg/mL) had no effect on mouse phrenic nerve-diaphragm preparations but caused time-dependent neuromuscular blockade of chick biventer cervicis preparations. This blockade was not reversed by washing. The highest concentration of venom tested (40 microg/mL) significantly reduced (pPhilodryas venoms had low esterase and phospholipase A(2) but high proteolytic activities compared to the pitviper Bothrops jararaca. SDS-PAGE showed that the Philodryas venoms had similar electrophoretic profiles, with most proteins having a molecular mass of 25-80 kDa. Both of the Philodryas venoms cross-reacted with bothropic antivenom in ELISA, indicating the presence of proteins immunologically related to Bothrops venoms. RP-HPLC of P. patagoniensis venom yielded four major peaks, each of which contained several proteins, as shown by SDS-PAGE. These results indicate that P. patagoniensis venom has neurotoxic and myotoxic components that may contribute to the effects of envenoming by this species.

  12. Quantitative evaluation of blood elements by neutron activation analysis in mice immunized with Bothrops snake venoms

    International Nuclear Information System (INIS)

    Zamboni, C.B.; Metairon, S.; Suzuki, M.F.; Furtado, M.F.; Sant'Anna, O.A.; Tambourgi, D.V.

    2009-01-01

    Mice genetically selected for high antibody responsiveness (HIII) were immunized against different Bothrops species snake venoms from distinct region of Brazil. The Neutron Activation Analysis technique was used to evaluate the whole blood concentrations of elements of clinical relevance [Ca, Cl, K, Mg and Na] in order to establish a potential correlation between antibody response and blood constituents after Bothrops venom administration for clinical screening of envenomed patients. (author)

  13. Comparison of Phylogeny, Venom Composition and Neutralization by Antivenom in Diverse Species of Bothrops Complex

    Science.gov (United States)

    Peixoto, Pedro S.; Bernardoni, Juliana L.; Oliveira, Sâmella S.; Portes-Junior, José Antonio; Mourão, Rosa Helena V.; Lima-dos-Santos, Isa; Sano-Martins, Ida S.; Chalkidis, Hipócrates M.; Valente, Richard H.; Moura-da-Silva, Ana M.

    2013-01-01

    In Latin America, Bothrops snakes account for most snake bites in humans, and the recommended treatment is administration of multispecific Bothrops antivenom (SAB – soro antibotrópico). However, Bothrops snakes are very diverse with regard to their venom composition, which raises the issue of which venoms should be used as immunizing antigens for the production of pan-specific Bothrops antivenoms. In this study, we simultaneously compared the composition and reactivity with SAB of venoms collected from six species of snakes, distributed in pairs from three distinct phylogenetic clades: Bothrops, Bothropoides and Rhinocerophis. We also evaluated the neutralization of Bothrops atrox venom, which is the species responsible for most snake bites in the Amazon region, but not included in the immunization antigen mixture used to produce SAB. Using mass spectrometric and chromatographic approaches, we observed a lack of similarity in protein composition between the venoms from closely related snakes and a high similarity between the venoms of phylogenetically more distant snakes, suggesting little connection between taxonomic position and venom composition. P-III snake venom metalloproteinases (SVMPs) are the most antigenic toxins in the venoms of snakes from the Bothrops complex, whereas class P-I SVMPs, snake venom serine proteinases and phospholipases A2 reacted with antibodies in lower levels. Low molecular size toxins, such as disintegrins and bradykinin-potentiating peptides, were poorly antigenic. Toxins from the same protein family showed antigenic cross-reactivity among venoms from different species; SAB was efficient in neutralizing the B. atrox venom major toxins. Thus, we suggest that it is possible to obtain pan-specific effective antivenoms for Bothrops envenomations through immunization with venoms from only a few species of snakes, if these venoms contain protein classes that are representative of all species to which the antivenom is targeted. PMID

  14. Epidemiological, clinical and therapeutic aspects of Bothrops asper bites.

    Science.gov (United States)

    Otero-Patiño, Rafael

    2009-12-01

    Bothrops asper inflicts the majority of snakebites in Central America and in the northern regions of South America, mostly affecting young agricultural workers in rural settings. This species is capable of provoking severe envenomings associated with local and systemic manifestations. The main clinical features are: local edema, ecchymoses, blisters, dermonecrosis, myonecrosis, defibrinogenation, thrombocytopenia, systemic bleeding, hypotension and renal alterations. In addition, soft-tissue infection, acute renal failure, compartmental syndrome, central nervous system hemorrhage and, in pregnant women, abortion, fetal wastage and abruptio placentae have been described as complications. Intravenous administration of antivenom constitutes the mainstay in the therapy. Antivenoms composed of either whole IgG or F(ab')(2) fragments, manufactured in Brazil, Colombia, Costa Rica and Mexico, have been tested in controlled clinical trials, and rational protocols for antivenom administration have been developed. In addition to antivenom therapy, a number of ancillary interventions are recommended in the treatment of B. asper bites.

  15. Neuromuscular activity of Bothrops fonsecai snake venom in vertebrate preparations

    Science.gov (United States)

    Fernandes, Carla T; Giaretta, Vânia MA; Prudêncio, Luiz S; Toledo, Edvana O; da Silva, Igor RF; Collaço, Rita CO; Barbosa, Ana M; Hyslop, Stephen; Rodrigues-Simioni, Léa; Cogo, José C

    2014-01-01

    The neuromuscular activity of venom from Bothrops fonsecai, a lancehead endemic to southeastern Brazil, was investigated. Chick biventer cervicis (CBC) and mouse phrenic nerve-diaphragm (PND) preparations were used for myographic recordings and mouse diaphragm muscle was used for membrane resting potential (RP) and miniature end-plate potential (MEPP) recordings. Creatine kinase release and muscle damage were also assessed. In CBC, venom (40, 80 and 160μg/ml) produced concentration- and time-dependent neuromuscular blockade (50% blockade in 85±9 min and 73±8 min with 80 and 160μg/ml, respectively) and attenuated the contractures to 110μM ACh (78–100% inhibition) and 40mM KCl (45–90% inhibition). The venom-induced decrease in twitch-tension in curarized, directly-stimulated preparations was similar to that in indirectly stimulated preparations. Venom (100 and 200μg/ml) also caused blockade in PND preparations (50% blockade in 94±13 min and 49±8 min with 100 and 200μg/ml, respectively) but did not alter the RP or MEPP amplitude. In CBC, venom caused creatine kinase release and myonecrosis. The venom-induced decrease in twitch-tension and in the contractures to ACh and K+ were abolished by preincubating venom with commercial antivenom. These findings indicate that Bothrops fonsecai venom interferes with neuromuscular transmission essentially through postsynaptic muscle damage that affects responses to ACh and KCl. These actions are effectively prevented by commercial antivenom. PMID:25028603

  16. Biochemical and immunological alterations of {sup 60} Co irradiated Bothrops jararacussu venom; Alteracoes bioquimicas e imunologicas do veneno de Bothrops jararacussu irradiado com {sup 60} Co

    Energy Technology Data Exchange (ETDEWEB)

    Spencer, Patrick J.

    1995-12-31

    Proteins irradiation leads to structural alterations resulting in activity and function loss. This process has been useful to detoxify animal venoms and toxins, resulting in low toxicity products which increased immunogenicity. The Bothrops jararacussu venom behaves as a weak immunogen and its lethal activity in not neutralized by either autologous, heterologous or bothropic polyvalent antisera. This venom is markedly myotoxic and and the commercial bothropic antiserum does not neutralize this activity, because of this low immunogenicity of the myotoxins. This present work was done in order to evaluate the possibility of irradiating Bothrops jararacussu, intending to increase the immunogenicity of the myotoxic components, leading to productions of myotoxins neutralizing antibodies. Bothrops jararacussu venom samples were irradiated with 500, 1000 and 2000 Gy of {sup 60} Co gamma rays. A 2.3 folds decrease of toxicity was observed for the 1000 Gy irradiated samples while the 2000 Gy irradiated sample was at least 3.7 folds attenuated. On the other hand, the 500 Gy did not promote any detoxification. Electrophoresis and HPLC data indicate that the irradiation lead to the formation of high molecular weight products (aggregates). The proteolytic and phospholipasic activities decreased in a dose dependent manner, the phospholipases being more resistant than the proteases. Both the animals (rabbit) immunized with either native or 2000 Gy irradiated venom produced native venom binding antibodies, a slightly higher titer being obtained in the serum of the rabbit immunized with the irradiated samples. Western blot data indicate that the anti-irradiated venom Ig Gs recognised a greater amount of either autologous or heterologous venom bands, both sera behaving as genus specific. The anti-native serum did not neutralize the myotoxic activity of native venom, while the anti-irradiated one was able to neutralize this activity. (author). 56 refs., 7 figs.

  17. Isolation and Functional Characterization of an Acidic Myotoxic Phospholipase A2 from Colombian Bothrops asper Venom

    OpenAIRE

    Posada Arias, Silvia; Rey-Suárez, Paola; Pereáñez J, Andrés; Acosta, Cristian; Rojas, Mauricio; Delazari dos Santos, Lucilene; Ferreira Jr, Rui Seabra; Núñez, Vitelbina

    2017-01-01

    Myotoxic phospholipases A2 (PLA2) are responsible for many clinical manifestations in envenomation by Bothrops snakes. A new myotoxic acidic Asp49 PLA2 (BaCol PLA2) was isolated from Colombian Bothrops asper venom using reverse-phase high performance liquid chromatography (RP-HPLC). BaCol PLA2 had a molecular mass of 14,180.69 Da (by mass spectrometry) and an isoelectric point of 4.4. The complete amino acid sequence was obtained by cDNA cloning (GenBank accession No. MF319968) and revealed a...

  18. Determination of inorganic elements in blood of mice immunized with Bothrops Snake venom using XRF and NAA

    International Nuclear Information System (INIS)

    Da Silva, L F F Lopes; Zamboni, C B; Bahovschi, V; Metairon, S; Suzuki, M F; Sant' Anna, O A; Rizzutto, M A

    2015-01-01

    In this work, mice genetically modified [H III line] were immunized against different Bothrops snake venoms to produce anti-Bothrops serum (antivenom). The Neutron Activation Analysis (NAA) and Energy Dispersive X-Ray Fluorescence (EDXRF) techniques were used to evaluate Ca and Fe concentrations in blood of these immunized mice in order to establish a potential correlation between both phenotypes: antibody response and blood constituents after Bothrops venom administration. The results were compared with the control group (mice not immunized) and with human being estimative. These data are important for clinical screening of patients submitted to immunological therapy as well as the understanding of the envenoming mechanisms. (paper)

  19. Determination of inorganic elements in blood of mice immunized with Bothrops Snake venom using XRF and NAA

    Science.gov (United States)

    Lopes da Silva, L. F. F.; Zamboni, C. B.; Bahovschi, V.; Metairon, S.; Suzuki, M. F.; Sant'Anna, O. A.; Rizzutto, M. A.

    2015-07-01

    In this work, mice genetically modified [HIII line] were immunized against different Bothrops snake venoms to produce anti-Bothrops serum (antivenom). The Neutron Activation Analysis (NAA) and Energy Dispersive X-Ray Fluorescence (EDXRF) techniques were used to evaluate Ca and Fe concentrations in blood of these immunized mice in order to establish a potential correlation between both phenotypes: antibody response and blood constituents after Bothrops venom administration. The results were compared with the control group (mice not immunized) and with human being estimative. These data are important for clinical screening of patients submitted to immunological therapy as well as the understanding of the envenoming mechanisms.

  20. Reproductive Biology of Bothrops erythromelas from the Brazilian Caatinga

    Directory of Open Access Journals (Sweden)

    Verônica Alberto Barros

    2014-01-01

    Full Text Available The reproductive biology of Bothrops erythromelas, a small pit viper from the Caatinga, a semiarid biome in Brazil, is described based on analysis of individuals deposited in zoological collections. Males are smaller and also attain sexual maturity at a smaller size than females. Female reproductive cycle is seasonal with an extended period of secondary vitellogenesis and births occurring in a restricted period from late spring to early summer. Sperm storage in females may probably occur in infundibular tubular glands and uterine muscular twisting (UMT, which is a polymorphic condition within B. erythromelas. Seasonal spermatogenesis in males is variable with some intraspecific variation regarding the male reproductive stage per season. Most males are reproductively active during spring/summer and reproductively quiescent during autumn/winter, although some individuals vary (e.g., show testicular spermatogenesis and active sexual segment of the kidneys (SSK during winter. The SSK could be identified in every male. Most males showed highly hypertrophied SSK in spring/summer and moderately hypertrophied SSK in autumn/winter. The ampulla ductus deferentis was observed and histochemical reactions were conducted. We discuss the probable influence of the unique environmental conditions of the Caatinga region and phylogenetic inertia in the reproductive patterns of this snake species.

  1. Phylogeography of the Central American lancehead Bothrops asper (SERPENTES: VIPERIDAE.

    Directory of Open Access Journals (Sweden)

    Mónica Saldarriaga-Córdoba

    Full Text Available The uplift and final connection of the Central American land bridge is considered the major event that allowed biotic exchange between vertebrate lineages of northern and southern origin in the New World. However, given the complex tectonics that shaped Middle America, there is still substantial controversy over details of this geographical reconnection, and its role in determining biogeographic patterns in the region. Here, we examine the phylogeography of Bothrops asper, a widely distributed pitviper in Middle America and northwestern South America, in an attempt to evaluate how the final Isthmian uplift and other biogeographical boundaries in the region influenced genealogical lineage divergence in this species. We examined sequence data from two mitochondrial genes (MT-CYB and MT-ND4 from 111 specimens of B. asper, representing 70 localities throughout the species' distribution. We reconstructed phylogeographic patterns using maximum likelihood and Bayesian methods and estimated divergence time using the Bayesian relaxed clock method. Within the nominal species, an early split led to two divergent lineages of B. asper: one includes five phylogroups distributed in Caribbean Middle America and southwestern Ecuador, and the other comprises five other groups scattered in the Pacific slope of Isthmian Central America and northwestern South America. Our results provide evidence of a complex transition that involves at least two dispersal events into Middle America during the final closure of the Isthmus.

  2. Phylogeography of the Central American lancehead Bothrops asper (SERPENTES: VIPERIDAE).

    Science.gov (United States)

    Saldarriaga-Córdoba, Mónica; Parkinson, Christopher L; Daza, Juan M; Wüster, Wolfgang; Sasa, Mahmood

    2017-01-01

    The uplift and final connection of the Central American land bridge is considered the major event that allowed biotic exchange between vertebrate lineages of northern and southern origin in the New World. However, given the complex tectonics that shaped Middle America, there is still substantial controversy over details of this geographical reconnection, and its role in determining biogeographic patterns in the region. Here, we examine the phylogeography of Bothrops asper, a widely distributed pitviper in Middle America and northwestern South America, in an attempt to evaluate how the final Isthmian uplift and other biogeographical boundaries in the region influenced genealogical lineage divergence in this species. We examined sequence data from two mitochondrial genes (MT-CYB and MT-ND4) from 111 specimens of B. asper, representing 70 localities throughout the species' distribution. We reconstructed phylogeographic patterns using maximum likelihood and Bayesian methods and estimated divergence time using the Bayesian relaxed clock method. Within the nominal species, an early split led to two divergent lineages of B. asper: one includes five phylogroups distributed in Caribbean Middle America and southwestern Ecuador, and the other comprises five other groups scattered in the Pacific slope of Isthmian Central America and northwestern South America. Our results provide evidence of a complex transition that involves at least two dispersal events into Middle America during the final closure of the Isthmus.

  3. Fatal bothropic snakebite in a horse: a case report

    Directory of Open Access Journals (Sweden)

    NS Silva

    2011-01-01

    Full Text Available The present study reports a snakebite in a horse in the state of Pará, Brazil. At initial evaluation the animal was reluctant to walk and had tachycardia, tachypnea, severe lameness, bleeding on the pastern and swelling around the left hind leg. Blood samples from the bleeding sites, took on the first day, showed leukocytosis and neutrophilia, whereas biochemical values of urea and creatinine were significantly increased. The chosen treatment was snake antivenom, fluid therapy, antibiotics, anti-inflammatory agents and diuretic drugs. On the fourth day of therapy, the hematological values were within normal parameters. There was improvement related to the clinical lameness and swelling of the limb. However, a decrease in water intake and oliguria were observed. On the seventh day the animal died. Necropsy revealed areas of hemorrhagic edema in the left hind limb and ventral abdomen; the kidneys presented equimosis in the capsule, and when cut they were wet. Moreover, the cortex was pale, slightly yellow and the medullary striae had the same aspect. Based on these data, we concluded that the snakebite in the present study was caused by Bothrops spp. and that renal failure contributed to death.

  4. Phylogeography of the Central American lancehead Bothrops asper (SERPENTES: VIPERIDAE)

    Science.gov (United States)

    Parkinson, Christopher L.; Daza, Juan M.; Wüster, Wolfgang

    2017-01-01

    The uplift and final connection of the Central American land bridge is considered the major event that allowed biotic exchange between vertebrate lineages of northern and southern origin in the New World. However, given the complex tectonics that shaped Middle America, there is still substantial controversy over details of this geographical reconnection, and its role in determining biogeographic patterns in the region. Here, we examine the phylogeography of Bothrops asper, a widely distributed pitviper in Middle America and northwestern South America, in an attempt to evaluate how the final Isthmian uplift and other biogeographical boundaries in the region influenced genealogical lineage divergence in this species. We examined sequence data from two mitochondrial genes (MT-CYB and MT-ND4) from 111 specimens of B. asper, representing 70 localities throughout the species’ distribution. We reconstructed phylogeographic patterns using maximum likelihood and Bayesian methods and estimated divergence time using the Bayesian relaxed clock method. Within the nominal species, an early split led to two divergent lineages of B. asper: one includes five phylogroups distributed in Caribbean Middle America and southwestern Ecuador, and the other comprises five other groups scattered in the Pacific slope of Isthmian Central America and northwestern South America. Our results provide evidence of a complex transition that involves at least two dispersal events into Middle America during the final closure of the Isthmus. PMID:29176806

  5. Circadian pattern of Bothrops moojeni in captivity (Serpentes: Viperidae

    Directory of Open Access Journals (Sweden)

    DR Stuginski

    2012-01-01

    Full Text Available Members of the subfamily Crotalinae are considered to be essentially nocturnal and most of the data about these snakes have been collected from the field. Information on how nutritional status affects the movement rate and activity patterns is a key point to elucidating the ecophysiology of snakes. In this study, we distributed 28 lancehead Bothrops moojeni into three groups under distinct feeding regimens after a month of fasting. Groups were divided as follows: ingestion of meals weighing (A 40%, (B 20%, or (C 10% of the snake body mass. Groups were monitored for five days before and after food intake and the activity periods and movement rates were recorded. Our results show that B. moojeni is prevalently nocturnal, and the activity peak occurs in the first three hours of the scotophase. After feeding, a significant decrease in activity levels in groups A and B was detected. The current results corroborate previous field data that describe B. moojeni as a nocturnal species with low movement rates. The relationship between motion and the amount of food consumed by the snake may be associated with its hunting strategy.

  6. Inhibitory properties of the anti-bothropic complex from Didelphis albiventris serum on toxic and pharmacological actions of metalloproteases and myotoxins from Bothrops asper venom.

    Science.gov (United States)

    Trento, E P; Garcia, O S; Rucavado, A; França, S C; Batalini, C; Arantes, E C; Giglio, J R; Soares, A M

    2001-12-01

    Anti-bothropic complex (ABC) was isolated from the serum of the South American opossum (Didelphis albiventris) by single-step affinity chromatography using a Sepharose-immobilized metalloprotease (BaP1) from Bothrops asper as the binding protein. Biochemical characterization of ABC showed the presence of two glycosylated subunits of 43 and 45 kDa, respectively, with an isoelectric point asper venom. In addition to the anti-hemorrhagic and anti-proteolytic activities, ABC also showed anti-myotoxic, anti-lethal, and anti-edematogenic effects against myotoxic phospholipases A(2) isolated from the same venom. Moreover, it had inhibitory effects on the phospholipase A(2) activity of the crude venom as well as the isolated venom phospholipases A(2).

  7. Activity evaluation from different native or irradiated with 60 Co gamma rays snake venoms and their inhibitory effect on Leishmania (Leishmania) amazonensis

    International Nuclear Information System (INIS)

    Lourenco, Cecilia de Oliveira

    2000-01-01

    Cutaneous leishmaniasis is a disease, caused by Leishmania parasites, that occurs frequently in tropical and sub-tropical regions of the world. Skin lesions that could results in disfiguring aspect characterize it. The treatment is based on few drugs as antimony salts or pentamidine that are toxic with increasing resistance by the parasite. Alternative forms of disease treatment are in constant search, including natural components as snake venoms. Previous studies demonstrate that some components of snake venoms have an inhibitory effect against those parasites, including Leishmania species. Although snake venoms presented high toxicity, several methods have been described to detoxify most or some of their toxic components, with favorable results by the use of gamma irradiation. In this report we tested several native and irradiated snake venoms for inhibitory effect against Leishmania (Leishmania) amazonensis parasite and LLCMK 2 mammalian cells, with enzymatic tests and electrophoresis. There are significant activity in Acanthophis antarcticus, Agkistrodon bilineatus, Bothrops moojeni, Bothrops jararaca, Hoplocephalus stephensi, Naja melanoleuca, Naja mossambica, Pseudechis australis, Pseudechis colletti, Pseudechis guttatus and Pseudechis porphyriacus, venom being inactive Pseudonaja textilis, Notechis ater niger, Notechis scutatus. Oxyuranus microlepidotus and Oxyuranus scutellatus venoms. After 2 KGy of 60 Co irradiation most venom loses significantly their activity. Venoms with antileishmanial activity presented L-amino acid oxidase (L-AO) activity and showed common protein with a molecular weight about 60kDa in SDS-PAGE. These results indicate that L-AO activity in those venoms are probably related with antileishmanial effect. (author)

  8. Activity evaluation from different native or irradiated with {sup 60} Co gamma rays snake venoms and their inhibitory effect on Leishmania (Leishmania) amazonensis; Avaliacao da atividade de diferentes venenos de serpentes, nativos ou irradiados, com radiacao gama de {sup 60} Co, quanto ao poder inibitorio do crescimento de Leishmania (Leishmania) amazonensis

    Energy Technology Data Exchange (ETDEWEB)

    Lourenco, Cecilia de Oliveira

    2000-07-01

    Cutaneous leishmaniasis is a disease, caused by Leishmania parasites, that occurs frequently in tropical and sub-tropical regions of the world. Skin lesions that could results in disfiguring aspect characterize it. The treatment is based on few drugs as antimony salts or pentamidine that are toxic with increasing resistance by the parasite. Alternative forms of disease treatment are in constant search, including natural components as snake venoms. Previous studies demonstrate that some components of snake venoms have an inhibitory effect against those parasites, including Leishmania species. Although snake venoms presented high toxicity, several methods have been described to detoxify most or some of their toxic components, with favorable results by the use of gamma irradiation. In this report we tested several native and irradiated snake venoms for inhibitory effect against Leishmania (Leishmania) amazonensis parasite and LLCMK{sub 2} mammalian cells, with enzymatic tests and electrophoresis. There are significant activity in Acanthophis antarcticus, Agkistrodon bilineatus, Bothrops moojeni, Bothrops jararaca, Hoplocephalus stephensi, Naja melanoleuca, Naja mossambica, Pseudechis australis, Pseudechis colletti, Pseudechis guttatus and Pseudechis porphyriacus, venom being inactive Pseudonaja textilis, Notechis ater niger, Notechis scutatus. Oxyuranus microlepidotus and Oxyuranus scutellatus venoms. After 2 KGy of {sup 60}Co irradiation most venom loses significantly their activity. Venoms with antileishmanial activity presented L-amino acid oxidase (L-AO) activity and showed common protein with a molecular weight about 60kDa in SDS-PAGE. These results indicate that L-AO activity in those venoms are probably related with antileishmanial effect. (author)

  9. Toxicity of Bothrops sp snake venoms from Ecuador and preclinical assessment of the neutralizing efficacy of a polyspecific antivenom from Costa Rica.

    Science.gov (United States)

    Laines, Johana; Segura, Álvaro; Villalta, Mauren; Herrera, María; Vargas, Mariángela; Alvarez, Gladys; Gutiérrez, José María; León, Guillermo

    2014-09-01

    The toxicological profile of the venoms of the snakes Bothrops asper and Bothrops atrox from Ecuador was investigated, together with the venom of a population of B. asper formerly classified as 'Bothrops xanthogrammus'. The three venoms exerted lethal, hemorrhagic, myotoxic, coagulant and defibrinogenating effects, in agreement with the characteristic toxicological profile of Bothrops sp venoms. A polyspecific antivenom (bothropic-crotalic-lachesic) manufactured in Costa Rica was assessed for its preclinical efficacy against the toxic activities of these Ecuadorian venoms. Antivenom was effective in the neutralization of the five activities tested in the three venoms. These observations are in agreement with previous reports on the extensive cross-reactivity and paraspecific neutralization of antivenoms manufactured in Latin America against the venoms of Bothrops sp snakes. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. Biological and immunological characteristics of the poison of Bothrops cotiara (Serpentes: Viperidae)

    International Nuclear Information System (INIS)

    Roodt, Adolfo Rafael de; Dolab, Jorge Adrian; Manzanelli, Marcelo Victor; Pineiro, Nicolas; Estevez, Judith; Paniagua, Jorge Francisco; Urs Vogt, Alejandro

    2006-01-01

    Bothrops cotiara is a venomous snake sporadically found in the province of Misiones in Argentina, South of Brazil and Paraguay. Data on the clinics of the poisoning produced by its bite and on its venom are scarce. There is no information on the neutralizing capacity of the antivenoms available. In this study, the lethal potency, hemorrhagic, necrotizing, coagulant and thrombin-like, defibrinogenasing, indirect hemolytic and fibrinolytic activities of the venom of B. cotiara specimens from the province of Misiones were determined. The toxic activities were within the range of those described for the other Bothrops species from Argentina, and the electrophoretic and chromatographic studies showed similarities with those described for the other bothropic venoms. The immunochemical reactivity of six South American anti Viper antivenoms (ELISA) have a strong reactivity with all the antivenoms studied. The neutralizing capacity of three of these therapeutic antivenoms against the lethal potency and hemorrhagic, necrotizing, coagulant, thrombin-like and hemolytic activities showed a very close neutralizing capacity. Our data strongly suggest that the antivenoms for therapeutic use available in this area of South America are useful to neutralize the toxic and enzymatic activities of the venom of this uncommon specie of Bothrops. (author) [es

  11. Naa Technique for Clinical Investigation of Mice Immunized with BOTHROP Venom

    Science.gov (United States)

    Zamboni, C. B.; Aguiar, R. O.; Kovacs, L.; Suzuki, M.; Sant'Anna, O. A.

    2009-06-01

    In the present study Neutron Activation Analysis (NAA) technique was used to determine sodium concentration in whole blood of mice immunized with Bothrops venom. With this value it was possible to perform clinical investigation in this animal model using whole blood.

  12. Biochemical and immunological alterations of 60 Co irradiated Bothrops jararacussu venom

    International Nuclear Information System (INIS)

    Spencer, Patrick J.

    1995-01-01

    Proteins irradiation leads to structural alterations resulting in activity and function loss. This process has been useful to detoxify animal venoms and toxins, resulting in low toxicity products which increased immunogenicity. The Bothrops jararacussu venom behaves as a weak immunogen and its lethal activity in not neutralized by either autologous, heterologous or bothropic polyvalent antisera. This venom is markedly myotoxic and and the commercial bothropic antiserum does not neutralize this activity, because of this low immunogenicity of the myotoxins. This present work was done in order to evaluate the possibility of irradiating Bothrops jararacussu, intending to increase the immunogenicity of the myotoxic components, leading to productions of myotoxins neutralizing antibodies. Bothrops jararacussu venom samples were irradiated with 500, 1000 and 2000 Gy of 60 Co gamma rays. A 2.3 folds decrease of toxicity was observed for the 1000 Gy irradiated samples while the 2000 Gy irradiated sample was at least 3.7 folds attenuated. On the other hand, the 500 Gy did not promote any detoxification. Electrophoresis and HPLC data indicate that the irradiation lead to the formation of high molecular weight products (aggregates). The proteolytic and phospholipasic activities decreased in a dose dependent manner, the phospholipases being more resistant than the proteases. Both the animals (rabbit) immunized with either native or 2000 Gy irradiated venom produced native venom binding antibodies, a slightly higher titer being obtained in the serum of the rabbit immunized with the irradiated samples. Western blot data indicate that the anti-irradiated venom Ig Gs recognised a greater amount of either autologous or heterologous venom bands, both sera behaving as genus specific. The anti-native serum did not neutralize the myotoxic activity of native venom, while the anti-irradiated one was able to neutralize this activity. (author)

  13. The distribution and elimination of Bothrops erythromelas venom labeled with {sup 131} I after intravenous injection in mice

    Energy Technology Data Exchange (ETDEWEB)

    Rocha, M.L. [Pernambuco Univ., Recife, PE (Brazil). Dept. de Zoologia]. E-mail: rocha@cascavel.uefs.br

    1999-07-01

    Pharmacokinetic studies can be used to study the systemic effects of snake venoms and to develop standard serotherapy protocols for envenomation. Bothrops erythromelas is probably responsible for most of the snakebite in Pernambuco. The objective of this study was to investigate the pharmacokinetics of B. erythromelas venom (BeV) in mice, and to evaluate the efficacy of bothropic antivenom. BeV showed bicompartmental distribution in the blood of the experimental animals. (author)

  14. The distribution and elimination of Bothrops erythromelas venom labeled with 131 I after intravenous injection in mice

    International Nuclear Information System (INIS)

    Rocha, M.L.

    1999-01-01

    Pharmacokinetic studies can be used to study the systemic effects of snake venoms and to develop standard serotherapy protocols for envenomation. Bothrops erythromelas is probably responsible for most of the snakebite in Pernambuco. The objective of this study was to investigate the pharmacokinetics of B. erythromelas venom (BeV) in mice, and to evaluate the efficacy of bothropic antivenom. BeV showed bicompartmental distribution in the blood of the experimental animals. (author)

  15. Acerca de la localidad tipo de Bothrops ammodytoides Leybold (Serpentes: viperidae) y Pseudotomodon trigonatus (Leybold) (Serpentes: colubridae)

    OpenAIRE

    Scrocchi, Gustavo

    1997-01-01

    De las especies del género Bothrops, Bothrops ammodytoides es una de las más típicas, ya que su característica "nariz" la hace inconfundible. Probablemente sea debido a esto que la sinonimia de esta especie sea bastante corta, ya que parece haber sido conocida claramente desde el momento de su descripción original.

  16. Protective effect of Dracontium dubium against Bothrops asper venom.

    Science.gov (United States)

    Caro, Daneiva; Ocampo, Yanet; Castro, Jenny; Barrios, Lia; Salas, Rubén; Franco, Luis A

    2017-05-01

    In Colombia, Bothrops asper is responsible for 70-90% of ophidians accidents reported annually. Envenoming occurs mainly in rural areas where both antivenom and health centers are scarce. Thus, patients are frequently treated by local healers that employ medicinal herbs; including several species belonging to Dracontium genus. In this work, we evaluated the neutralizing activity of Dracontium dubium Kunth against the lethal, inflammatory, coagulant and hemolytic effects produced by B. asper venom. Mice treated with D. dubium extract (500 and 1000μg/g, ip), survived to the administration of lethal doses of venom, with remarkable recovery of macroscopic and histology damage. Furthermore, D. dubium exerted a significant inhibition of inflammatory damage promoted by paw injection of B. asper venom. Such activity might be related to the inhibition of macrophage activation and NO production, as demonstrated using LPS-stimulated RAW 264.7 cells. Moreover, the extract of D. dubium remarkably diminished the indirect hemolytic effect of snake venom. On the other hand, no substantial differences were observed in clotting time of plasma incubated with venom when compared to extract treated plasma. Noteworthy, D. dubium extract did not alter the electrophoretic pattern of venom before the assays. Phytochemistry screening revealed the presence of phenolic compounds, flavonoids, tannins and steroids/triterpenoids, which might explain the bioactivity of the extract. Our results, provides strong evidence that support the employment of D. dubium in folk medicine. Further studies are needed to isolate and identify the metabolites responsible for the activity, in order to provide a useful and accessible treatment for snakebite envenoming in low-income rural areas. Copyright © 2017. Published by Elsevier Masson SAS.

  17. Dieta e uso de habitat da jararaca-do-brejo, Mastigodryas bifossatus Raddi (Serpentes, Colubridade) em domínio subtropical do Brasil Diet and habitat use of swamp racer snake, Mastigodryas bifossatus Raddi (Serpentes, Colubridae) in subtropical domains of Brazil

    OpenAIRE

    Pedro T. Leite; Simone de F. Nunes; Sonia Z. Cechin

    2007-01-01

    A serpente Mastigodryas bifossatus Raddi, 1820 é um grande colubrídeo neotropical que habita áreas abertas na América do Sul. Sua dieta é composta principalmente por anfíbios, mas inclui outros itens como mamíferos e lagartos. A dissecção de 224 espécimes desta serpente, provenientes dos estados do Rio Grande do Sul, Santa Catarina e Paraná, preservados em coleções herpetológicas do Brasil e um estudo de campo entre 1996 e 1998, fornecem informações sobre os hábitos alimentares e uso de habit...

  18. Dieta e uso de habitat da jararaca-do-brejo, Mastigodryas bifossatus Raddi (Serpentes, Colubridade em domínio subtropical do Brasil Diet and habitat use of swamp racer snake, Mastigodryas bifossatus Raddi (Serpentes, Colubridae in subtropical domains of Brazil

    Directory of Open Access Journals (Sweden)

    Pedro T. Leite

    2007-01-01

    Full Text Available A serpente Mastigodryas bifossatus Raddi, 1820 é um grande colubrídeo neotropical que habita áreas abertas na América do Sul. Sua dieta é composta principalmente por anfíbios, mas inclui outros itens como mamíferos e lagartos. A dissecção de 224 espécimes desta serpente, provenientes dos estados do Rio Grande do Sul, Santa Catarina e Paraná, preservados em coleções herpetológicas do Brasil e um estudo de campo entre 1996 e 1998, fornecem informações sobre os hábitos alimentares e uso de habitat dessa serpente em domínio subtropical no Brasil. Essa serpente alimenta-se de anfíbios (80%, em sua maioria da família Leptodactylidae, mamíferos (10% e lagartos (2%. Foi verificada mudança ontogenética na dieta de M. bifossatus, o tamanho das presas ingeridas aumenta com o aumento de tamanho da serpente. Essa espécie ocorre principalmente em áreas abertas, algumas vezes perto de áreas antrópicas em domínio subtropical no Brasil.Mastigodryas bifossatus Raddi, 1820 is a large neotropical colubrid snake that inhabits open areas in South America. The diet is mainly composed by frogs, but it includes other items like mammals and lizards. The dissection of 224 specimens of this snake, proceeding from the states of Rio Grande do Sul, Santa Catarina and Paraná, stored in herpetological collections in Brazil and a field study between 1996 and 1998, provided information on dietary habits and habitat use of this snake in subtropical domains in Brazil. This snake eats mainly amphibians of the family Leptodactylidae (80%, mammals (10% and lizards (2%. There is ontogenetic diet shift in M. bifossatus, as the snake grows, the range of preys grows as well. M. bifossatus occur in open areas.

  19. Distribution of 131 I- labeled Bothrops erythromelas venom in mice

    International Nuclear Information System (INIS)

    Vasconcelos, C.M.L.; Valenca, R.C.; Araujo, E.A.; Modesto, J.C.A.; Pontes, M.M.; Guarnieri, M.C.; Brazil, T.K.

    1998-01-01

    Bothrops erythromelas is responsible for many snake bites in northeastern Brazil. In the present study we determined the in vivo distribution of the venom following its subcutaneous injection into mice. B. erythromelas venom and albumin were labeled individually with 131 I by the chloramine T method, and separated in a Sephacryl S-200 column. The efficiency of labeling was 68%.Male Swiss mice (40-45 g), which had been provided with drinking water containing 0.05% KI over a period of 10 days prior to the experiment, were inoculated dorsally (sc) with 0.3 ml (2.35 x 10 5 cpm/mouse) of 131 I-venom (N = 42), 131 -albumin or 131 I (controls, N = 28 each). Thirty minutes and 1,3, 6, 12, 18 and 24 h after inoculation, the animals were perfused with 0.85% Na Cl and skin and various organs were collected in order to determine radioactivity content. There was a high rate of venom absorption int he skin (51%) within the first 30 min compared to albumin (20.1%) and free iodine (8.2%). Up to the third hour after injection there was a tendency for venom and albumin to concentrate in the stomach ( 3 rd h),small intestine (3 rd h) and large intestine (6th h). Both control groups had more radioactivity in the digestive tract, especially in the stomach, but these levels decreased essentially to baseline by 12-18 h postinjection. In the kidneys, the distribution profiles of venom, albumin and iodine were similar. Counts at 30 min postinjection were low in all three groups (1.37, 1.86 and 0.77, respectively), and diminished to essentially 0% by 12-18 h. Albumin tended to concentrate in muscle until the 3 rd h postinjection (1.98%).There was a low binding of labeled venom in the liver (B. erythromelas venom does not specifically target most internal organs. That is, the systemic effects of envenomation ar mainly due to an indirect action. (author)

  20. Inhibition of the toxic effects of Bothrops asper venom by pinostrobin, a flavanone isolated from Renealmia alpinia (Rottb.) MAAS.

    Science.gov (United States)

    Gómez-Betancur, Isabel; Benjumea, Dora; Patiño, Arley; Jiménez, Nora; Osorio, Edison

    2014-09-29

    Renealmia alpinia has been traditionally used to treat snakebites by indigenous Embera-Katíos tribes belonging to the regions of Antioquia and Chocó, Colombia, and it has been shown to inhibit the enzymatic and biological activities of Bothrops venoms and their purified phospholipase A2 (PLA2) toxins. In addition to its common local usage against snakebites, Renealmia alpinia is commonly used to treat pain. To evaluate the inhibitory ability of pinostrobin, the main compound in the dichloromethane extract of Renealmia alpinia, on the toxic effects of Bothrops asper venom through in vitro and in vivo models and to evaluate its activity against pain and edema. Pinostrobin was isolated from the dichloromethane extract of Renealmia alpinia leaves. The protective properties of the extract and of pinostrobin against the indirect hemolytic, coagulant and proteolytic effects of Bothrops asper venom were evaluated in vitro, and the anti-hemorrhagic and anti-inflammatory activity were evaluated in vivo. Renealmia alpinia extract significantly inhibited the proteolytic activity and indirect hemolytic activity of Bothrops asper venom at a venom:extract ratio of 1:20. Moreover, the present data demonstrate that pinostrobin may mitigate some venom-induced local tissue damage due to hemorrhagic effects, and the compound is also responsible for the analgesic and anti-inflammatory activity of the extract from Renealmia alpinia. This is the first report to describe pinostrobin in the species Renealmia alpinia and its properties in vitro against Bothrops asper venom. Our studies of the activity of Renealmia alpinia against the venom of Bothrops asper have confirmed that this species possesses inhibitory effects against Bothrops asper venom in both in vitro and in vivo models and that these effects may be due to pinostrobin, supporting the traditional usage of the plant. Additionally, pinostrobin may be responsible for the anti-hemorrhagic and analgesic activity (peripheral

  1. Snake Venomics and Antivenomics of Bothrops diporus, a Medically Important Pitviper in Northeastern Argentina

    Science.gov (United States)

    Gay, Carolina; Sanz, Libia; Calvete, Juan J.; Pla, Davinia

    2015-01-01

    Snake species within genus Bothrops are responsible for more than 80% of the snakebites occurring in South America. The species that cause most envenomings in Argentina, B. diporus, is widely distributed throughout the country, but principally found in the Northeast, the region with the highest rates of snakebites. The venom proteome of this medically relevant snake was unveiled using a venomic approach. It comprises toxins belonging to fourteen protein families, being dominated by PI- and PIII-SVMPs, PLA2 molecules, BPP-like peptides, L-amino acid oxidase and serine proteinases. This toxin profile largely explains the characteristic pathophysiological effects of bothropic snakebites observed in patients envenomed by B. diporus. Antivenomic analysis of the SAB antivenom (Instituto Vital Brazil) against the venom of B. diporus showed that this pentabothropic antivenom efficiently recognized all the venom proteins and exhibited poor affinity towards the small peptide (BPPs and tripeptide inhibitors of PIII-SVMPs) components of the venom. PMID:26712790

  2. Inactivation and fragmentation of lectin from Bothrops leucurus snake venom by gamma irradiation

    International Nuclear Information System (INIS)

    Nunes, E.S.; Souza, M.A.A.; Vaz, A.F.M.; Coelho, L.C.B.B.; Aguiar, J.S.; Silva, T.G.; Guarnieri, M.C.; Melo, A.M.M.A.; Oliva, M.L.V.; Correia, M.T.S.

    2012-01-01

    Gamma radiation alters the molecular structure of biomolecules and is able to mitigate the action of snake venoms and their isolated toxins. The effect of γ-radiation on the folding of Bothrops lecurus venom lectin was measured by a hemagglutinating assay, intrinsic and bis-ANS fluorescence. Intrinsic and bis-ANS fluorescence analyses indicated that irradiation caused unfolding followed by aggregation of the lectin. Our results suggest that irradiation can lead to significant changes in the protein structure, which may promote the loss of its binding property and toxic action. - Highlights: ► Gamma radiation alters the molecular structure of biomolecules. ► The radiation has been able to mitigate snake venoms and its isolated toxins. ► Our aim was to evaluate the effects of radiation in Bothrops lecurus venom lectin. ► The irradiation acts as a detoxification strategy in snake venoms.

  3. Snake venomics and antivenomics of Bothrops colombiensis, a medically important pitviper of the Bothrops atrox-asper complex endemic to Venezuela: Contributing to its taxonomy and snakebite management.

    Science.gov (United States)

    Calvete, Juan J; Borges, Adolfo; Segura, Alvaro; Flores-Díaz, Marietta; Alape-Girón, Alberto; Gutiérrez, José María; Diez, Nardy; De Sousa, Leonardo; Kiriakos, Demetrio; Sánchez, Eladio; Faks, José G; Escolano, José; Sanz, Libia

    2009-03-06

    The taxonomic status of the medically important pitviper of the Bothrops atrox-asper complex endemic to Venezuela, which has been classified as Bothrops colombiensis, remains incertae cedis. To help resolving this question, the venom proteome of B. colombiensis was characterized by reverse-phase HPLC fractionation followed by analysis of each chromatographic fraction by SDS-PAGE, N-terminal sequencing, MALDI-TOF mass fingerprinting, and collision-induced dissociation tandem mass spectrometry of tryptic peptides. The venom contained proteins belonging to 8 types of families. PI Zn(2+)-metalloproteinases and K49 PLA(2) molecules comprise over 65% of the venom proteins. Other venom protein families comprised PIII Zn(2+)-metalloproteinases (11.3%), D49 PLA(2)s (10.2%), l-amino acid oxidase (5.7%), the medium-sized disintegrin colombistatin (5.6%), serine proteinases (1%), bradykinin-potentiating peptides (0.8%), a DC-fragment (0.5%), and a CRISP protein (0.1%). A comparison of the venom proteomes of B. colombiensis and B. atrox did not support the suggested synonymy between these two species. The closest homologues to B. colombiensis venom proteins appeared to be toxins from B. asper. A rough estimation of the similarity between the venoms of B. colombiensis and B. asper indicated that these species share approximately 65-70% of their venom proteomes. The close kinship of B. colombiensis and B. asper points at the ancestor of B. colombiensis as the founding Central American B. asper ancestor. This finding may be relevant for reconstructing the natural history and cladogenesis of Bothrops. Further, the virtually indistinguishable immunological crossreactivity of a Venezuelan ABC antiserum (raised against a mixture of B. colombiensis and Crotalus durissus cumanensis venoms) and the Costa Rican ICP polyvalent antivenom (generated against a mixture of B. asper, Crotalus simus, and Lachesis stenophrys venoms) towards the venoms of B. colombiensis and B. asper, supports this

  4. Isolation from a polyvalent antivenom of antibodies to a myotoxin in Bothrops asper snake venom

    OpenAIRE

    Lomonte, Bruno; Mata, Eugenia; Gutiérrez, José María

    1985-01-01

    Antibodies against Bothrops asper rmyotoxin were purified from a polyvalent antivenom by affinity chromatography . These antibodies neutralized most of the myotoxic activity of crude B. asper venom, as judged by histologic evaluation of skeletal muscle and determination of plasma creatine kinase levels in mice. When tested by immunoelectrophoresis, purified antibodies formed two superimposed bands of precipitation against an homogenous by SDS-PAGE analysis) preparation of B. asper myotoxin...

  5. Reversible posterior leukoencephalopathy in a venomous snake (Bothrops asper) bite victim.

    Science.gov (United States)

    Delgado, Miguel E; Del Brutto, Oscar H

    2012-03-01

    An 18-year-old man developed posterior reversible leukoencephalopaty after being bitten by a venomous snake (Bothrops asper). It is possible that this previously unrecognized neurological complication of snake bite envenoming occurred as the result of endothelial dysfunction induced by the venom of the offending snake. This pathogenetic mechanism has also been implicated as the cause of cerebral infarctions in snake bite victims. Alternatively, the leukoencephalopathy might have been a complication of antivenom therapy.

  6. Reversible Posterior Leukoencephalopathy in a Venomous Snake (Bothrops asper) Bite Victim

    OpenAIRE

    Delgado, Miguel E.; Del Brutto, Oscar H.

    2012-01-01

    An 18-year-old man developed posterior reversible leukoencephalopaty after being bitten by a venomous snake (Bothrops asper). It is possible that this previously unrecognized neurological complication of snake bite envenoming occurred as the result of endothelial dysfunction induced by the venom of the offending snake. This pathogenetic mechanism has also been implicated as the cause of cerebral infarctions in snake bite victims. Alternatively, the leukoencephalopathy might have been a compli...

  7. Accidents caused by Bothrops and Bothropoides in the State of Paraiba: epidemiological and clinical aspects.

    Science.gov (United States)

    Oliveira, Fagner Neves; Brito, Monalisa Taveira; Morais, Isabel Cristina Oliveira de; Fook, Sayonara Maria Lia; Albuquerque, Helder Neves de

    2010-01-01

    Bothrops and Bothropoides snakes cause 70% of the ophidic accidents in Brazil. The species that cause ophidic accidents in State of Paraíba are Bothropoides erythromelas, Bothrops leucurus and Bothropoides neuwiedi. This is a prospective and transverse study, following a quantitative approach of accidents involving Bothrops and Bothropoides admitted to the Toxicological Assistance and Information Centers of Campina Grande and João Pessoa (Ceatox-CG and Ceatox-JP), aimed at identifying the epidemiological and clinical profile of such accidents. All of the patients admitted had medical diagnoses and were monitored at Ceatox-CG or Ceatox-JP. The genera Bothrops and Bothropoides caused 91.7% of the ophidic accidents reported. Snake bites were frequent in men (75.1%), rural workers (65.1%), literate individuals (69%) between 11 and 20 years-old (21.7%), and toes the most common area attacked (52.7%). Most (86.6%) patients were admitted within 6 hours after the accident/bite, with a predominance of mild cases (64.6%). The annual occurrence in Paraíba was 5.5 accidents/100,000 inhabitants and lethality was 0.2%. Positive changes in the profiles of these accidents were verified, such as the non-application of inadequate solutions, including the use of tourniquet, coffee grounds, garlic, suction and/or cutting the bitten area. Moreover, the Itinerant Laboratory project, linked to Paraíba State University in partnership with Ceatox-CG, has contributed positively, providing several cities of the state with information regarding the prevention of accidents involving venomous animals. The local press has also contributed, reporting the educational work developed by the centers.

  8. Isolation and Functional Characterization of an Acidic Myotoxic Phospholipase A2 from Colombian Bothrops asper Venom

    Science.gov (United States)

    Posada Arias, Silvia; Rey-Suárez, Paola; Pereáñez J, Andrés; Acosta, Cristian; Rojas, Mauricio; Ferreira Jr, Rui Seabra; Núñez, Vitelbina

    2017-01-01

    Myotoxic phospholipases A2 (PLA2) are responsible for many clinical manifestations in envenomation by Bothrops snakes. A new myotoxic acidic Asp49 PLA2 (BaCol PLA2) was isolated from Colombian Bothrops asper venom using reverse-phase high performance liquid chromatography (RP-HPLC). BaCol PLA2 had a molecular mass of 14,180.69 Da (by mass spectrometry) and an isoelectric point of 4.4. The complete amino acid sequence was obtained by cDNA cloning (GenBank accession No. MF319968) and revealed a mature product of 124 amino acids with Asp at position 49. BaCol PLA2 showed structural homology with other acidic PLA2 isolated from Bothrops venoms, including a non-myotoxic PLA2 from Costa Rican B. asper. In vitro studies showed cell membrane damage without exposure of phosphatidylserine, an early apoptosis hallmark. BaCol PLA2 had high indirect hemolytic activity and moderate anticoagulant action. In mice, BaCol PLA2 caused marked edema and myotoxicity, the latter seen as an increase in plasma creatine kinase and histological damage to gastrocnemius muscle fibers that included vacuolization and hyalinization necrosis of the sarcoplasm. PMID:29072602

  9. Isolation and Functional Characterization of an Acidic Myotoxic Phospholipase A₂ from Colombian Bothrops asper Venom.

    Science.gov (United States)

    Posada Arias, Silvia; Rey-Suárez, Paola; Pereáñez J, Andrés; Acosta, Cristian; Rojas, Mauricio; Delazari Dos Santos, Lucilene; Ferreira, Rui Seabra; Núñez, Vitelbina

    2017-10-26

    Myotoxic phospholipases A₂ (PLA₂) are responsible for many clinical manifestations in envenomation by Bothrops snakes. A new myotoxic acidic Asp49 PLA₂ (BaCol PLA₂) was isolated from Colombian Bothrops asper venom using reverse-phase high performance liquid chromatography (RP-HPLC). BaCol PLA₂ had a molecular mass of 14,180.69 Da (by mass spectrometry) and an isoelectric point of 4.4. The complete amino acid sequence was obtained by cDNA cloning (GenBank accession No. MF319968) and revealed a mature product of 124 amino acids with Asp at position 49. BaCol PLA₂ showed structural homology with other acidic PLA₂ isolated from Bothrops venoms, including a non-myotoxic PLA₂ from Costa Rican B. asper . In vitro studies showed cell membrane damage without exposure of phosphatidylserine, an early apoptosis hallmark. BaCol PLA₂ had high indirect hemolytic activity and moderate anticoagulant action. In mice, BaCol PLA₂ caused marked edema and myotoxicity, the latter seen as an increase in plasma creatine kinase and histological damage to gastrocnemius muscle fibers that included vacuolization and hyalinization necrosis of the sarcoplasm.

  10. Isolation and Functional Characterization of an Acidic Myotoxic Phospholipase A2 from Colombian Bothrops asper Venom

    Directory of Open Access Journals (Sweden)

    Silvia Posada Arias

    2017-10-01

    Full Text Available Myotoxic phospholipases A2 (PLA2 are responsible for many clinical manifestations in envenomation by Bothrops snakes. A new myotoxic acidic Asp49 PLA2 (BaCol PLA2 was isolated from Colombian Bothrops asper venom using reverse-phase high performance liquid chromatography (RP-HPLC. BaCol PLA2 had a molecular mass of 14,180.69 Da (by mass spectrometry and an isoelectric point of 4.4. The complete amino acid sequence was obtained by cDNA cloning (GenBank accession No. MF319968 and revealed a mature product of 124 amino acids with Asp at position 49. BaCol PLA2 showed structural homology with other acidic PLA2 isolated from Bothrops venoms, including a non-myotoxic PLA2 from Costa Rican B. asper. In vitro studies showed cell membrane damage without exposure of phosphatidylserine, an early apoptosis hallmark. BaCol PLA2 had high indirect hemolytic activity and moderate anticoagulant action. In mice, BaCol PLA2 caused marked edema and myotoxicity, the latter seen as an increase in plasma creatine kinase and histological damage to gastrocnemius muscle fibers that included vacuolization and hyalinization necrosis of the sarcoplasm.

  11. Methemoglobinemia associated with loxoscelism Meta-hemoglobinemia associada ao loxoscelismo

    Directory of Open Access Journals (Sweden)

    Orlando C. de O. Barreto

    1990-02-01

    Full Text Available In twenty five patients who presented the cutaneous form of loxoscelism, serum haptoglobin and lactic dehydrogenase, erythrocyte glucose-6-phosphate dehydrogenase, glutathione reductase, glutathione peroxidase, methemoglobin, bilirubin and reticulocytes were investigated after bite. No hemolysis was detected but an increase in methemoglobin was found in 54% of the cases; in 7% it was between 1.1% and 2%, in 27% it ranged from 2.1% to 4%, and in 20% from 4.1% to 8%. Blood samples of a normal, blood group 0 individual and of a patient who exhibited methemoglobinemia after Loxosceles bite were incubated separately with antisera against Loxosceles gaucho, Crotalus terrificus, Bothrops jararaca, with Loxosceles gaucho venom and 0.3% phenol. No methemoglobin was found after 1, 4,8 and 15 days in both sets of samples. At the 25th day all the samples, including the controls, exhibited similar methemoglobin reductase decrease. The data suggest that the methemoglobinemia which occurs in 50% of the patients probably arises from in vivo venom metabolism, inasmuch as the crude venom does not induce methemoglobinemia.Vinte e cinco pacientes que apresentaram a forma cutânea do loxoscelismo foram estudados após a picada, determinando-se a glicose-6-fosfato desidrogenase, glutationa redutase e glutationa peroxidase eritrocitárias, haptoglobina e latico desidrogenase séricas, bilirrubina, reticulócitos e meta-hemoglobina. Não foi observada hemólise aumentada, mas foi detectado aumento da meta-hemoglobina em 54% dos casos: em 7% entre 1,1% e 2%, em 27% variou de 2,1% a 4%, e em 20% de 4,1 a 8%. Amostras de sangue de um indivíduo normal do grupo 0 de uma paciente que exibiu meta-hemoglobina após picada por Loxosceles foram incubadas separadamente com anti-soros contra Loxosceles gancho, Crotalus terrificus e Bothrops jararaca, com veneno de Loxosceles gaucho e fenol a 3%, e não se detectou aumento de meta-hemoglobina depois de 1, 4, 8 e 15 dias em todas

  12. A transcriptomic analysis of gene expression in the venom gland of the snake Bothrops alternatus (urutu

    Directory of Open Access Journals (Sweden)

    Menossi Marcelo

    2010-10-01

    Full Text Available Abstract Background The genus Bothrops is widespread throughout Central and South America and is the principal cause of snakebite in these regions. Transcriptomic and proteomic studies have examined the venom composition of several species in this genus, but many others remain to be studied. In this work, we used a transcriptomic approach to examine the venom gland genes of Bothrops alternatus, a clinically important species found in southeastern and southern Brazil, Uruguay, northern Argentina and eastern Paraguay. Results A cDNA library of 5,350 expressed sequence tags (ESTs was produced and assembled into 838 contigs and 4512 singletons. BLAST searches of relevant databases showed 30% hits and 70% no-hits, with toxin-related transcripts accounting for 23% and 78% of the total transcripts and hits, respectively. Gene ontology analysis identified non-toxin genes related to general metabolism, transcription and translation, processing and sorting, (polypeptide degradation, structural functions and cell regulation. The major groups of toxin transcripts identified were metalloproteinases (81%, bradykinin-potentiating peptides/C-type natriuretic peptides (8.8%, phospholipases A2 (5.6%, serine proteinases (1.9% and C-type lectins (1.5%. Metalloproteinases were almost exclusively type PIII proteins, with few type PII and no type PI proteins. Phospholipases A2 were essentially acidic; no basic PLA2 were detected. Minor toxin transcripts were related to L-amino acid oxidase, cysteine-rich secretory proteins, dipeptidylpeptidase IV, hyaluronidase, three-finger toxins and ohanin. Two non-toxic proteins, thioredoxin and double-specificity phosphatase Dusp6, showed high sequence identity to similar proteins from other snakes. In addition to the above features, single-nucleotide polymorphisms, microsatellites, transposable elements and inverted repeats that could contribute to toxin diversity were observed. Conclusions Bothrops alternatus venom gland

  13. Active immunization of cattle with a bothropic toxoid does not abrogate envenomation by Bothrops asper venom, but increases the likelihood of survival.

    Science.gov (United States)

    Herrera, María; González, Katherine; Rodríguez, Carlos; Gómez, Aarón; Segura, Álvaro; Vargas, Mariángela; Villalta, Mauren; Estrada, Ricardo; León, Guillermo

    2017-03-01

    This study assessed the protective effect of active immunization of cattle to prevent the envenomation induced by B. asper venom. Two groups of oxen were immunized with a bothropic toxoid and challenged by an intramuscular injection of either 10 or 50 mg B. asper venom, to induce moderate or severe envenomations, respectively. Non-immunized oxen were used as controls. It was found that immunized oxen developed local edema similar to those observed in non-immunized animals. However, systemic effects were totally prevented in immunized oxen challenged with 10 mg venom, and therefore antivenom treatment was not required. When immunized oxen were challenged with 50 mg venom, coagulopathy was manifested 3-16 h later than in non-immunized oxen, demonstrating a delay in the onset of systemic envenomation. In these animals, active immunization did not eliminate the need for antivenom treatment, but increased the time lapse in which antivenom administration is still effective. All experimentally envenomed oxen completely recovered after a week following venom injection. Our results suggest that immunization of cattle with a bothropic toxoid prevents the development of systemic effects in moderate envenomations by B. asper, but does not abrogate these effects in severe envenomation. Copyright © 2016 International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.

  14. Proteomic and functional profiling of the venom of Bothrops ayerbei from Cauca, Colombia, reveals striking interspecific variation with Bothrops asper venom.

    Science.gov (United States)

    Mora-Obando, Diana; Guerrero-Vargas, Jimmy Alexander; Prieto-Sánchez, Rodrigo; Beltrán, José; Rucavado, Alexandra; Sasa, Mahmood; Gutiérrez, José María; Ayerbe, Santiago; Lomonte, Bruno

    2014-01-16

    Bothrops ayerbei, a pitviper inhabiting the Patía River's basin (Valle Alto del Río Patía) in the Southwestern Department of Cauca, Colombia, was considered as a variant form of Bothrops asper prior to being proposed as a new species in 2010, on the basis of subtle morphological differences. This study reports the proteomic and functional profiling of B. ayerbei venom. Its most striking feature is an almost complete absence (0.7%) of phospholipases A2 (PLA2), which is in contrast to the high proportion of these enzymes (25.3%) in the venom of B. asper from Cauca, as well as in other species of Bothrops. The predominant proteins in B. ayerbei venom are metalloproteinases (53.7%), in agreement with its higher hemorrhagic and lethal activities compared to B. asper venom. Moreover, the negligible content of PLA2s in B. ayerbei venom correlates with its weaker myotoxic effect, in contrast to B. asper venom, here shown to contain abundant Asp49- and Lys49-type PLA2s responsible for its strong myotoxic activity. Other components identified in B. ayerbei venom include bradykinin-potentiating-like peptides and proteins belonging to the C-type lectin/lectin-like, serine proteinase, l-amino acid oxidase, disintegrin, cysteine-rich secretory protein, nerve growth factor, and phosphodiesterase families. The venom composition of B. ayerbei resembles that of neonate specimens of B. asper, which shows a predominance of metalloproteinases, with only low amounts of PLA2s. Therefore, the present findings suggest that the expression of venom proteins in B. ayerbei, in contrast to B. asper, might retain a marked 'paedomorphic' condition. Altogether, the proteomic and toxicological characterization of the venom of B. ayerbei here reported argues in favor of its taxonomical separation from B. asper in Cauca, Colombia. B. ayerbei, a pitviper found in Cauca, Colombia, had been considered as a variant form of B. asper, but was recently described as a new species on the basis of subtle

  15. A eficácia do antiveneno botrópico-crotálico na neutralização das principais atividades do veneno de Bothrops jararacussu The efficacy of the bothropic/crotalic antivenom on the neutralization of the main Bothrops jararacussu venom activities

    Directory of Open Access Journals (Sweden)

    Maria Cristina dos-Santos

    1992-04-01

    Full Text Available A mionecrose é um dos efeitos causados pelo veneno de Bothrops jararacussu. Uma miotoxina com homologia estrutural à fosfolipase A2 (PLA2, mas sem atividade enzimática, foi isolada desse veneno. O veneno de Crotalus durissus terrificus apresenta também atividade miotóxica, que vem sendo atribuída à crotoxina e à PLA2 (crotoxina B, o componente básico do complexo crotoxina. O veneno de Bothrops jararacussu apresenta três proteínas, que têm identidade imunológica com a PLA2 da crotoxina. O presente trabalho comparou a eficiência dos antivenenos polivalentes comerciais produzidos pelo Instituto Butantan - o antiveneno botrópico (AB e o antiveneno botrópico-crotálico (AB/C - na neutralização das atividades letal, hemorrágica, coagulante e miotóxica do veneno de B. jararacussu. Os dois antivenenos neutralizaram de maneira semelhante a atividade hemorrágica, mas o AB/C foi três vezes mais potente que o AB em neutralizar a ação miotóxica e duas vezes mais potente na neutralização da letalidade e na ação coagulante do veneno de B. jararacussu. Os dados sugerem que a utilização do AB/C pode ser vantajosa no tratamento de pacientes picados por serpentes dessa espécieMyonecrosis is one of the effects of Bothrops jararacussu venom, from which a myotoxin was isolated showing structural homology to phospholipase A2 (PLA2, but without enzimatic activity. Such myotoxic activity is also present in the Crotalus durissus terrificus venom, and is atributed to crotoxin and to PLA2 (crotoxin B, the basic component of the crotoxin complex. The Bothrops jararacussu venom showed three proteins with immunologic identity to PLA2 from crotoxin. The bothropic (AB and the bothropic/crotalic (AB/C antivenoms, two commercial polyvalent antivenoms produced at Instituto Butantan, were compared in order to assess their capacity for neutralization of the lethal, hemorrhagic, coagulant and myotoxic activities of Bothrops jararacussu venom. Both

  16. Design of Fab-based chimeric antibodies against Bothrops asper toxins

    DEFF Research Database (Denmark)

    M. Haack, Aleksander; B. Hallgren, Malte; U. W. Friis, Rasmus

    Snakebite is one of the world’s most neglected tropical diseases, with an estimated 5 million bites per year, resulting in about 125.000 deaths. The only current treatment for snakebite envenoming is antiserum derived from the blood of immunized mammals(typically horses). These antisera are expen...... are expensive to produce and carry a high risk of causing hyper-allergic reactions in human recipients due to their heterologous origin. Here we report the discovery of chimeric scFvs against Bothrops asper toxins....

  17. Variação entre filhotes de representantes do complexo Bothrops newied (Serpentes, Viperidae, Crotalinae

    Directory of Open Access Journals (Sweden)

    Vinícius Xavier

    2002-01-01

    Full Text Available External morphological characters of 141 young specimens (69 males and 72 femalesof the Bothrops newied complex were analyzed. Regression analysis was used in the study of morphometric characters and principal components analysis was used in the study of meristic and qualitative characters. Sexual dimorphism was confirmed in the meristic and morphometric characters. Males showed higher counts of subcaudals and longer tails. Females showed eventually higher number of ventrals and dorsal rows, and larger heads. Six different drawing patterns were diagnosed and can indicate the existence of different species. Ontogenetic variation was described.

  18. Inactivation and fragmentation of lectin from Bothrops leucurus snake venom by gamma irradiation

    Science.gov (United States)

    Nunes, E. S.; Souza, M. A. A.; Vaz, A. F. M.; Coelho, L. C. B. B.; Aguiar, J. S.; Silva, T. G.; Guarnieri, M. C.; Melo, A. M. M. A.; Oliva, M. L. V.; Correia, M. T. S.

    2012-04-01

    Gamma radiation alters the molecular structure of biomolecules and is able to mitigate the action of snake venoms and their isolated toxins. The effect of γ-radiation on the folding of Bothrops lecurus venom lectin was measured by a hemagglutinating assay, intrinsic and bis-ANS fluorescence. Intrinsic and bis-ANS fluorescence analyses indicated that irradiation caused unfolding followed by aggregation of the lectin. Our results suggest that irradiation can lead to significant changes in the protein structure, which may promote the loss of its binding property and toxic action.

  19. The analgesics morphine and tramadol do not alter the acute toxicity induced by Bothrops asper snake venom in mice

    OpenAIRE

    Gutiérrez, José María; Herrera Arias, Cristina

    2014-01-01

    Analgesics tramadol and morphine were used in mice injected with Bothrops asper venom. Analgesics did not alter the acute toxicity of B. asper venom in mice. Precautionary (prophylactic) analgesia should be introduced routinely in Toxinology. UCR::Investigación::Unidades de Investigación::Ciencias de la Salud::Instituto Clodomiro Picado (ICP)

  20. Fatal stroke after Bothrops snakebite in the Amazonas state, Brazil: A case report.

    Science.gov (United States)

    Silva de Oliveira, Sâmella; Freitas-de-Sousa, Luciana Aparecida; Alves, Eliane Campos; de Lima Ferreira, Luiz Carlos; da Silva, Iran Mendonça; de Lacerda, Marcus Vinícius Guimarães; Fan, Hui Wen; Moura-da-Silva, Ana Maria; Monteiro, Wuelton Marcelo

    2017-11-01

    Bothrops atrox is the snake responsible for the majority of snakebites in the Brazilian Amazon. Patients generally evolve to local manifestations such as edema, pain and ecchymoses. Systemic effects of B. atrox venom are usually restricted to blood incoagulability and spontaneous bleeding. However, in a few cases, bleeding in the central nervous system may occur, which can lead to sequels and deaths. Here, we report a case of a 59 year-old woman who presented edema, pain and ecchymoses on the right foot, headache, nausea, diarrhea, hypertension and blood incoagulability after the bite by Bothrops snake in the Brazilian Amazon. This case evolved with stroke resulting in death despite the antivenom and conservative therapy employed. In addition, we were able to identify the presence of venom in the patient's brain tissue after death. Direct action of toxins present in the snake's venom in the induction of systemic hemorrhage allied to blood incoagulability and hypertension presented by the patient could be involved in the mechanism of stroke in this case. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. The Triterpenoid Betulin Protects against the Neuromuscular Effects of Bothrops jararacussu Snake Venom In Vivo

    Science.gov (United States)

    Ferraz, Miriéle Cristina; de Oliveira, Jhones Luiz; de Oliveira Junior, Joel Reis; Cogo, José Carlos; dos Santos, Márcio Galdino; Franco, Luiz Madaleno; Puebla, Pilar; Ferraz, Helena Onishi; Ferraz, Humberto Gomes; da Rocha, Marisa Maria Teixeira; Hyslop, Stephen

    2015-01-01

    We confirmed the ability of the triterpenoid betulin to protect against neurotoxicity caused by Bothrops jararacussu snake venom in vitro in mouse isolated phrenic nerve-diaphragm (PND) preparations and examined its capability of in vivo protection using the rat external popliteal/sciatic nerve-tibialis anterior (EPSTA) preparation. Venom caused complete, irreversible blockade in PND (40 μg/mL), but only partial blockade (~30%) in EPSTA (3.6 mg/kg, i.m.) after 120 min. In PND, preincubation of venom with commercial bothropic antivenom (CBA) attenuated the venom-induced blockade, and, in EPSTA, CBA given i.v. 15 min after venom also attenuated the blockade (by ~70% in both preparations). Preincubation of venom with betulin (200 μg/mL) markedly attenuated the venom-induced blockade in PND; similarly, a single dose of betulin (20 mg, i.p., 15 min after venom) virtually abolished the venom-induced decrease in contractility. Plasma creatine kinase activity was significantly elevated 120 min after venom injection in the EPSTA but was attenuated by CBA and betulin. These results indicate that betulin given i.p. has a similar efficacy as CBA given i.v. in attenuating the neuromuscular effects of B. jararacussu venom in vivo and could be a useful complementary measure to antivenom therapy for treating snakebite. PMID:26633987

  2. New findings from the first transcriptome of the Bothrops moojeni snake venom gland.

    Science.gov (United States)

    Amorim, Fernanda Gobbi; Morandi-Filho, Romualdo; Fujimura, Patricia Tieme; Ueira-Vieira, Carlos; Sampaio, Suely Vilela

    2017-12-15

    Snakebites are a serious health problem in tropical countries. In Brazil, the genus Bothrops (Viperidae family) causes most of the ophidic accidents, characterized by proteolysis and haemorrhage. Snake venoms are rich sources of toxins with great therapeutic and biotechnological potential and omics approaches is a valuable tool for identification of new bioactive components in the venom. In this study, we described the first transcriptome of the venom gland of Bothrops moojeni snake, using the next-generation sequencing with the Illumina platform. We identified: (i) 20 venom components classes, among which metalloproteases were the most expressed ones, followed by serine proteases and phospholipases; and (ii) the 33 full-length amino acid sequences of toxins that have never been reported before in B. moojeni venom, such as one cysteine-rich secretory protein (Moojin), two hyaluronidases (BmooHyal-1 and BmooHyal-2), and one three-finger toxin (Bmoo-3FTx). Altogether, the transcripts identified herein represent a starting point for the analysis of structure-function relationships of toxins, which shall help develop novel biological tools and therapeutic drugs. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. The Triterpenoid Betulin Protects against the Neuromuscular Effects of Bothrops jararacussu Snake Venom In Vivo

    Directory of Open Access Journals (Sweden)

    Miriéle Cristina Ferraz

    2015-01-01

    Full Text Available We confirmed the ability of the triterpenoid betulin to protect against neurotoxicity caused by Bothrops jararacussu snake venom in vitro in mouse isolated phrenic nerve-diaphragm (PND preparations and examined its capability of in vivo protection using the rat external popliteal/sciatic nerve-tibialis anterior (EPSTA preparation. Venom caused complete, irreversible blockade in PND (40 μg/mL, but only partial blockade (~30% in EPSTA (3.6 mg/kg, i.m. after 120 min. In PND, preincubation of venom with commercial bothropic antivenom (CBA attenuated the venom-induced blockade, and, in EPSTA, CBA given i.v. 15 min after venom also attenuated the blockade (by ~70% in both preparations. Preincubation of venom with betulin (200 μg/mL markedly attenuated the venom-induced blockade in PND; similarly, a single dose of betulin (20 mg, i.p., 15 min after venom virtually abolished the venom-induced decrease in contractility. Plasma creatine kinase activity was significantly elevated 120 min after venom injection in the EPSTA but was attenuated by CBA and betulin. These results indicate that betulin given i.p. has a similar efficacy as CBA given i.v. in attenuating the neuromuscular effects of B. jararacussu venom in vivo and could be a useful complementary measure to antivenom therapy for treating snakebite.

  4. Effects of Co60 gamma radiation on the immunogenic and antigenic properties of Bothrops jararacussu venom

    International Nuclear Information System (INIS)

    Spencer, Patrick J.; Nascimento, Nanci do; Rogero, Jose R.

    1997-01-01

    Ionizing radiation has been successfully employed to attenuate animals toxins and venoms for immunizing antisera producing animals. However, the radiation effects on antigenicity and immunogenecity have not yet been elucidated. In the present work, we investigated the effects of gamma rays on the antigenic and immunogenicity have not yet been elucidated. In the present work, we investigated the effects of gamma rays on the antigenic and immunogenic behaviour of Bothrops jararacussu venon. Venom samples (2mg/ml in 150 mM NaCl) were irradiated with 500, 1000 and 2000 Gy of 60 Co gamma rays. These samples were submitted to antigen capture ELISA on plates coated with commercial bothropic antiserum. Results suggest a loss of reactivity of the 1000 and 2000 Gy irradiated samples. Antibodies against native and 2000 Gy irradiated venoms were produced in rabbits. Both sera able to bind native venom with a slightly higher titer for anti-irradiated serum. These data suggest that radiation promoted structural modification on the antigen molecules. However since the antibodies produced against irradiated antivenom were able to recognize native venom, there must have been preservation of some antigenic determinants. It has already been demosntrated that irradiation of proteins leads to structural modifications and unfolding of the molecules. Our data suggest that irradiation led to conformational epitopes destruction with preservation of linear epitopes and that the response against irradiated venom may be attributed to these linear antigenic determinants. (author). 8 refs., 3 figs

  5. In vitro hemolytic activity of Bothrops lanceolatus (fer-de-lance venom

    Directory of Open Access Journals (Sweden)

    LJ Martins

    2009-01-01

    Full Text Available Bothrops lanceolatus venom contains a variety of enzymatic and biological activities. The present work investigated the hemolytic activity of this venom and its phospholipase A2 (PLA2. Bothrops lanceolatus venom (6.7 µg/mL caused indirect hemolysis of cow, horse, rat and sheep erythrocytes, with horse erythrocytes being the most sensitive; no direct hemolysis was observed. Hemolysis in sheep erythrocytes was concentration-dependent (5-11.7 µg/mL and markedly attenuated by heating the venom for 30 minutes at ≥ 40°C and by the PLA2 inhibitor p-bromophenacyl bromide. An acidic PLA2 (5 µg/mL purified from B. lanceolatus venom also caused hemolysis. This PLA2 showed immunoprecipitin lines with antivenom against B. lanceolatus, which suggests that the enzymatic and hemolytic activities of this enzyme may be neutralized during antivenom therapy. These results indicate that B. lanceolatus venom and its PLA2 can cause hemolysis in vitro.

  6. Screening of Bothrops snake venoms for L-amino acid oxidase activity

    Energy Technology Data Exchange (ETDEWEB)

    Pessati, M.L.; Fontana, J.D.; Guimaraes, M.F. [Federal Univ. of Parana, Curitiba (Brazil)

    1995-12-31

    Toxins, enzymes, and biologically active peptides are the main components of snake venoms from the genus Bothrops. Following the venom inoculation, the local effects are hemorrhage, edema, and myonecrosis. Nineteen different species of Brazilian Bothrops were screened for protein content and L-amino acid oxidase activity. B. cotiara, formerly found in the South of Brazil, is now threatened with extinction. Its venom contains a highly hemorrhagic fraction and, as expected from the deep yellow color of the corresponding lyophilized powder, a high L-amino acid oxidase (LAO) activity was also characterized. Flavin adenine dinucleotide (FAD) is its associate coenzyme. B. cotiara venom LAO catalyzed the oxidative deamination of several L-amino acids, and the best substrates were methionine, leucine, tryptophan, and phenylalanine, hence, its potential application for the use in biosensors for aspartame determination and for the removal of amino acids from plasma. High levels for LAO were also found in other species than B. cotiara. In addition, the technique of isoelectric focusing (IEF) was employed as a powerful tool to study the iso- or multi-enzyme distribution for LAO activity in the B. cotiara snake venom.

  7. Local inflammation, lethality and cytokine release in mice injected with Bothrops atrox venom

    Directory of Open Access Journals (Sweden)

    S. F. Barros

    1998-01-01

    Full Text Available We have provided evidence that: (a lethality of mice to crude Bothrops venom varies according the isogenic strain (A/J > C57Bl/6 > A/Sn > BALB/c > C3H/ HePas > DBA/2 > C3H/He; (bBALB/c mice (LD50=100.0 μg were injected i.p. with 50 μg of venom produced IL-6, IL-10, INF-γ, TNF-α and NO in the serum. In vitro the cells from the mice injected and challenged with the venom only released IL-10 while peritoneal macrophages released IL-10, INF-γ and less amounts of IL-6; (c establishment of local inflammation and necrosis induced by the venom, coincides with the peaks of TNF-α, IFN-γ and NO and the damage was neutralized when the venom was incubated with a monoclonal antibody against a 60 kDa haemorrhagic factor. These results suggest that susceptibility to Bothrops a trox venom is genetically dependent but MHC independent; that IL-6, IL10, TNF-α, IFN-γ and NO can be involved in the mediation of tissue damage; and that the major venom component inducers of the lesions are haemorrhagins.

  8. Ecology of ticks in a taxocenosis of snakes from the Serra do Mendanha, Rio de Janeiro, Brazil, with new host records

    Directory of Open Access Journals (Sweden)

    Jorge A. L. Pontes

    2009-06-01

    Full Text Available We studied the ecology of ticks found in different species of a taxocenosis of snakes from the Serra do Mendanha, an area of Atlantic Rainforest located in the state of Rio de Janeiro, southeastern Brazil. Snakes were sampled monthly in the field during a period of 48 months. The specific identity of the hosts and their parasites, the number of parasites, and and snout-vent length and body mass of each host were recorded. A total of 25% of the species of snakes in the area were parasitized by ticks (larvae, nymphs and adult females Amblyomma rotundatum Koch, 1844 was the predominant parasite species. The infestation parameters varied among the species of snakes sampled, with the highest prevalence of A. rotundatum being observed in the viperid Bothrops jararaca (Wied, 1824 (71.4%, followed by the colubrids Xenodon neuwiedii Günther, 1820 (33%, Chironius laevicollis (Wied, 1824 and Spilotes pullatus (Linnaeus, 1758 (both with 22%. The latter three species also showed the highest rates of infestation by A. rotundatum. The results of the present study suggest that a combination of skin shedding, habitat of the host, type of scale and pattern of scale distribution on the body of the host can influence the degree to which a given species is parasitized by ticks

  9. Parasitological and immunological diagnoses from feces of captive-bred snakes at Vital Brazil Institute

    Directory of Open Access Journals (Sweden)

    Janaína Lima de Souza

    Full Text Available Fecal samples from 56 snakes at the Vital Brazil Institute, in the city of Niterói, Rio de Janeiro, were tested using the sedimentation and flotation techniques to investigate the evolutionary forms of parasites such as helminths and protozoa, and using enzyme immunoassay techniques to detect antigens of Cryptosporidium sp. and Giardiasp. Among the animals tested, 80.3% were positive for parasites. Out of these, there were 16 Bothrops jararaca, 16 B. jararacussu and 13 Crotalus durissus. The prevalence of parasitic nematodes was 41.1%, and nematodes were found in all three snake species. Among these, the most frequent finding was eggs of Kalicephalus sp., which were diagnosed in 25% of the snakes. The positivity for protozoa detected using parasite concentration techniques was 75%, including oocysts of Caryospora sp. in 75%, cysts with morphology similar to Giardia sp. 3.6%, amoeboid cysts in 41.1% and unsporulated coccidia oocysts in 8.9%. Immunoassays for Cryptosporidium sp. antigens produced positive findings in 60.7%. Pseudoparasites were detected in 64.3%. These results show that there is a need to improve the sanitary handling of captive-bred snakes, and also for the animal house that supplies rodents to feed them. The results also highlight that diagnostic tests should be performed periodically on stool specimens from captive-bred snakes.

  10. Snake Venom Peptides and Low Mass Proteins: Molecular Tools and Therapeutic Agents.

    Science.gov (United States)

    Almeida, J R; Resende, L M; Watanabe, R K; Carregari, V C; Huancahuire-Vega, S; da S Caldeira, C A; Coutinho-Neto, A; Soares, A M; Vale, N; de C Gomes, P A; Marangoni, S; de A Calderon, L; Da Silva, S L

    2017-01-01

    Snake venoms are natural sources of biologically active molecules that are able to act selectively and specifically on different cellular targets, modulating physiological functions. Thus, these mixtures, composed mainly of proteins and peptides, provide ample and challenging opportunities and a diversified molecular architecture to design and develop tools and agents of scientific and therapeutic interest. Among these components, peptides and small proteins play diverse roles in numerous physiological processes, exerting a wide range of pharmacological activities, such as antimicrobial, antihypertensive, analgesic, antitumor, analgesic, among others. The pharmaceutical and cosmetic industries have recognized the huge potential of these privileged frameworks and believe them to be a promising alternative to contemporary drugs. A number of natural or synthetic peptides from snake venoms have already found preclinical or clinical applications for the treatment of pain, hypertension, cardiovascular diseases and aging skin. A well-known example is captopril, whose natural peptide precursor was isolated from Bothrops jararaca snake venom, which is a peptide-based drug that inhibits the angiotensin-converting enzyme, producing an anti-hypertensive effect. The present review looks at the main peptides (natriuretic peptides, bradykinin-potentiating peptides and sarafotoxins) and low mass proteins (crotamine, disintegrins and three-Finger toxins) from snake venoms, as well as synthetic peptides inspired by them, describing their biochemical, structural and physiological features, as well as their applications as research tools and therapeutic agents. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  11. Snake Venom: From Deadly Toxins to Life-saving Therapeutics.

    Science.gov (United States)

    Waheed, Humera; Moin, Syed F; Choudhary, M I

    2017-01-01

    Snakes are fascinating creatures and have been residents of this planet well before ancient humans dwelled the earth. Venomous snakes have been a figure of fear, and cause notable mortality throughout the world. The venom constitutes families of proteins and peptides with various isoforms that make it a cocktail of diverse molecules. These biomolecules are responsible for the disturbance in fundamental physiological systems of the envenomed victim, leading to morbidity which can lead to death if left untreated. Researchers have turned these life-threatening toxins into life-saving therapeutics via technological advancements. Since the development of captopril, the first drug that was derived from bradykininpotentiating peptide of Bothrops jararaca, to the disintegrins that have potent activity against certain types of cancers, snake venom components have shown great potential for the development of lead compounds for new drugs. There is a continuous development of new drugs from snake venom for coagulopathy and hemostasis to anti-cancer agents. In this review, we have focused on different snake venom proteins / peptides derived drugs that are in clinical use or in developmental stages till to date. Also, some commonly used snake venom derived diagnostic tools along with the recent updates in this exciting field are discussed. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  12. Interaction between bradykinin potentiating nonapeptide (BPP9a) and {beta}-cyclodextrin: A structural and thermodynamic study

    Energy Technology Data Exchange (ETDEWEB)

    Lula, Ivana; De Sousa, Frederico B. [Departamento de Quimica, Instituto de Ciencias Exatas, Universidade Federal de Minas Gerais, Av. Antonio Carlos, 6627, 31270-901, Belo Horizonte, MG (Brazil); Denadai, Angelo M.L. [Centro Federal de Educacao Tecnologica de Minas Gerais, CEFET-MG, Campus VII, 35.183-006, Timoteo, MG (Brazil); Ferreira de Lima, Guilherme; Duarte, Helio Anderson [Departamento de Quimica, Instituto de Ciencias Exatas, Universidade Federal de Minas Gerais, Av. Antonio Carlos, 6627, 31270-901, Belo Horizonte, MG (Brazil); Mares Guia, Thiago R. dos [Departamento de Bioquimica e Imunologia, ICB, Universidade Federal de Minas Gerais, 31270-901, Belo Horizonte, MG (Brazil); Faljoni-Alario, Adelaide [Departamento de Bioquimica, Instituto de Quimica, Universidade de Sao Paulo, 05508-900, Sao Paulo, SP (Brazil); Santoro, Marcelo M. [Departamento de Bioquimica e Imunologia, ICB, Universidade Federal de Minas Gerais, 31270-901, Belo Horizonte, MG (Brazil); Camargo, Antonio C.M. de [Center for Applied Toxinology CAT-CEPID, Laboratorio Especial de Toxicologia Aplicada, Instituto Butantan, 05503-900, Sao Paulo, SP (Brazil); Santos, Robson A.S. dos [Departamento de Fisiologia e Biofisica, ICB, Universidade Federal de Minas Gerais, 31270-901, Belo Horizonte, MG (Brazil); and others

    2012-02-01

    Herein, we demonstrate the physical and chemical characterizations of the supramolecular complex formed between {beta}-cyclodextrin ({beta}CD) and bradykinin potentiating nonapeptide (BPP9a), an endogenous toxin found in Bothrops jararaca. Circular dichroism results indicate a conformational change in the BPP9a secondary structure upon its complexation with {beta}CD. Nuclear magnetic resonance results, mainly from NOESY experiments, and theoretical calculations showed a favorable interaction between the tryptophan residue of BPP9a and the {beta}CD cavity. Thermodynamic inclusion parameters were investigated by isothermal titration calorimetry, demonstrating that {beta}CD/BPP9a complex formation is an exothermic process that results in a reduction in entropy. Additionally, in vitro degradation study of BPP9a against trypsin (37 Degree-Sign C, pH 7.2) showed higher stability of peptide in presence of {beta}CD. This {beta}CD/BPP9a complex, which presents new chemical properties arising from the peptide inclusion process, may be useful as an antihypertensive drug in oral pharmaceutical formulations. Highlights: Black-Right-Pointing-Pointer Cd and NMR showed evidences for the existence of more than one structure in solution. Black-Right-Pointing-Pointer Complexation with {beta}CD reduces the conformational rigidity of the peptide. Black-Right-Pointing-Pointer {beta}CD cavity recognize Trp and/or Pro segments of BPP9a. Black-Right-Pointing-Pointer Interactions involving disaggregation of BPP9a assemblies and binding with {beta}CD.

  13. Parasitological and immunological diagnoses from feces of captive-bred snakes at Vital Brazil Institute.

    Science.gov (United States)

    Souza, Janaína Lima de; Barbosa, Alynne da Silva; Vazon, Adriana Prado; Uchôa, Claudia Maria Antunes; Nunes, Beatriz Coronato; Cortez, Myrian Bandeira Vianna; Silva, Valmir Laurentino da; Más, Leonora Brazil; Melgarejo, Aníbal Rafael; Bastos, Otilio Machado Pereira

    2014-01-01

    Fecal samples from 56 snakes at the Vital Brazil Institute, in the city of Niterói, Rio de Janeiro, were tested using the sedimentation and flotation techniques to investigate the evolutionary forms of parasites such as helminths and protozoa, and using enzyme immunoassay techniques to detect antigens of Cryptosporidium sp. and Giardia sp. Among the animals tested, 80.3% were positive for parasites. Out of these, there were 16 Bothrops jararaca, 16 B. jararacussu and 13 Crotalus durissus. The prevalence of parasitic nematodes was 41.1%, and nematodes were found in all three snake species. Among these, the most frequent finding was eggs of Kalicephalus sp., which were diagnosed in 25% of the snakes. The positivity for protozoa detected using parasite concentration techniques was 75%, including oocysts of Caryospora sp. in 75%, cysts with morphology similar to Giardia sp. 3.6%, amoeboid cysts in 41.1% and unsporulated coccidia oocysts in 8.9%. Immunoassays for Cryptosporidium sp. antigens produced positive findings in 60.7%. Pseudoparasites were detected in 64.3%. These results show that there is a need to improve the sanitary handling of captive-bred snakes, and also for the animal house that supplies rodents to feed them. The results also highlight that diagnostic tests should be performed periodically on stool specimens from captive-bred snakes.

  14. Serpentes encontradas mortas em rodovias do estado de Santa Catarina, Brasil

    Directory of Open Access Journals (Sweden)

    Tobias Saraiva Kunz

    2009-08-01

    Full Text Available http://dx.doi.org/10.5007/2175-7925.2009v22n2p91 Estradas causam diversos impactos aos ecossistemas, incluindo perda e fragmentação de hábitat, alteração da paisagem natural e morte de animais, sendo que pouco se sabe sobre o impacto destas mortes sobre as populações. A fauna de serpentes do Estado de Santa Catarina é a menos estudada do Sul do Brasil, não havendo estudos abordando serpentes atropeladas. Neste trabalho apresentamos dados sobre serpentes encontradas mortas em diversas estradas do Estado entre os anos de 2003 e 2008. Foram registrados 165 exemplares pertencentes a 38 espécies que correspondem a 50% das serpentes com ocorrência esperada para o Estado de Santa Catarina. As quatro espécies mais freqüentes foram Philodryas patagoniensis (N= 22; 13,33%, Liophis miliaris (n = 21; 12,72%, P. aestiva (n = 13; 7,87% e Bothrops jararaca (n = 12; 7,27%, que somadas representam cerca de 41% das serpentes encontradas mortas. Amplia-se ao sul a distribuição de Imantodes cenchoa em cerca de 60km.

  15. Snake venom serine proteinases specificity mapping by proteomic identification of cleavage sites.

    Science.gov (United States)

    Zelanis, André; Huesgen, Pitter F; Oliveira, Ana Karina; Tashima, Alexandre K; Serrano, Solange M T; Overall, Christopher M

    2015-01-15

    Many snake venom toxins are serine proteases but their specific in vivo targets are mostly unknown. Various act on components of the coagulation cascade, and fibrinolytic and kallikrein-kinin systems to trigger various pathological effects observed in the envenomation. Despite showing high similarity in terms of primary structure snake venom serine proteinases (SVSPs) show exquisite specificity towards macromolecular substrates. Therefore, the characterization of their peptide bond specificity is important for understanding the active site preference associated with effective proteolysis as well as for the design of peptide substrates and inhibitors. Bothrops jararaca contains various SVSPs among which Bothrops protease A is a specific fibrinogenolytic agent and PA-BJ is a platelet-activating enzyme. In this study we used proteome derived peptide libraries in the Proteomic Identification of protease Cleavage Sites (PICS) approach to explore the peptide bond specificity of Bothrops protease A and PA-BJ in order to determine their individual peptide cleavage sequences. A total of 371 cleavage sites (208 for Bothrops protease A and 163 for PA-BJ) were detected and both proteinases displayed a clear preference for arginine at the P1 position. Moreover, the analysis of the specificity profiles of Bothrops protease A and PA-BJ revealed subtle differences in the preferences along P6-P6', despite a common yet unusual preference for Pro at P2. Taken together, these results map the subsite specificity of both SVSPs and shed light in the functional differences between these proteinases. Proteolysis is key to various pathological effects observed upon envenomation by viperid snakes. The use of the Proteomic Identification of protease Cleavage Sites (PICS) approach for the easy mapping of proteinase subsite preferences at both the prime- and non-prime sides concurrently gives rise to a fresh understanding of the interaction of the snake venom serine proteinases with peptide and

  16. Bothrops snake myotoxins induce a large efflux of ATP and potassium with spreading of cell damage and pain.

    Science.gov (United States)

    Cintra-Francischinelli, Mariana; Caccin, Paola; Chiavegato, Angela; Pizzo, Paola; Carmignoto, Giorgio; Angulo, Yamileth; Lomonte, Bruno; Gutiérrez, José María; Montecucco, Cesare

    2010-08-10

    Myotoxins play a major role in the pathogenesis of the envenomations caused by snake bites in large parts of the world where this is a very relevant public health problem. We show here that two myotoxins that are major constituents of the venom of Bothrops asper, a deadly snake present in Latin America, induce the release of large amounts of K(+) and ATP from skeletal muscle. We also show that the released ATP amplifies the effect of the myotoxins, acting as a "danger signal," which spreads and causes further damage by acting on purinergic receptors. In addition, the release of ATP and K(+) well accounts for the pain reaction characteristic of these envenomations. As Bothrops asper myotoxins are representative of a large family of snake myotoxins with phospholipase A(2) structure, these findings are expected to be of general significance for snake bite envenomation. Moreover, they suggest potential therapeutic approaches for limiting the extent of muscle tissue damage based on antipurinergic drugs.

  17. Características biológicas e inmunológicas del veneno de Bothrops cotiara (Serpentes: Viperidae

    Directory of Open Access Journals (Sweden)

    Adolfo Rafael de Roodt

    2006-09-01

    Full Text Available Bothrops cotiara es una serpiente que se encuentra en la provincia de Misiones (Argentina, el Sur de Brasil y Paraguay. La información sobre las características clínicas de los accidentes por esta serpiente es muy escasa y existen pocos datos sobre su veneno y la capacidad neutralizante de las actividades tóxicas del mismo por antivenenos terapéuticos. En este trabajo se estudiaron características bioquímicas, actividades tóxicas y la reactividad inmunoquímica del veneno de B. cotiara. Seis antivenenos anti Viperinos Sudamericanos fueron estudiados frente a este veneno por el método ELISA y se probó la capacidad neutralizante de tres de estos frente a las actividades hemorrágica, necrotizante, procoagulante, trombina-símil, hemolítica indirecta y la potencia letal de veneno de ejemplares de B. cotiara de la provincia de Misiones. Los patrones cromatográficos y electroforéticos mostraron características similares a los de otros venenos de Bothrops. Las actividades tóxicas estuvieron dentro de los ámbitos descritos para los venenos botrópicos. Los seis antivenenos mostraron gran reactividad inmunoquímica por ELISA y las potencias neutralizantes de los tres estudiados fueron muy próximas para las actividades letal, hemorrágica, necrotizante, hemolítica indirecta, coagulante y trombina-símil. Los resultados de los estudios de neutralización indicarían que ante la mordedura de esta poco común especie de Bothrops, pueden usarse los diferentes tipos de antivenenos botrópicos o botrópico-crotálicos para uso terapéutico disponibles en esa región.Biological and immunological characteristics of the poison of Bothrops cotiara (Serpentes: Viperidae. Bothrops cotiara is a venomous snake sporadically found in the province of Misiones in Argentina, South of Brazil and Paraguay. Data on the clinics of the envenomation produced by its bite and on its venom are scarce. There is no information on the neutralizing capacity of the

  18. In vitro and in vivo genotoxic evaluation of Bothrops moojeni snake venom.

    Science.gov (United States)

    Novak Zobiole, Nathalia; Caon, Thiago; Wildgrube Bertol, Jéssica; Pereira, Cintia Alves de Souza; Okubo, Brunna Mary; Moreno, Susana Elisa; Cardozo, Francielle Tramontini Gomes de Sousa

    2015-06-01

    Bothrops moojeni Hoge (Viperidae) venom is a complex mixture of compounds with therapeutic potential that has been included in the research and development of new drugs. Along with the biological activity, the pharmaceutical applicability of this venom depends on its toxicological profile. This study evaluates the cytotoxicity and genotoxicity of the Bothrops moojeni venom (BMV). The in vitro cytotoxicity and genotoxicity of a pooled sample of BMV was assessed by the MTT and Comet assay, respectively. Genotoxicity was also evaluated in vivo through the micronucleus assay. BMV displayed a 50% cytotoxic concentration (CC50) on Vero cells of 4.09 µg/mL. Vero cells treated with 4 µg/mL for 90 min and 6 h presented significant (p < 0.05, ANOVA/Newman-Keuls test) higher DNA damage than the negative control in the Comet assay. The lower DNA damage found after 6 h compared with the 90 min treatment suggests a DNA repair effect. Mice intraperitoneally treated with BMV at 10, 30, or 80 µg/animal presented significant genotoxicity (p < 0.05, ANOVA/Newman-Keuls test) in relation to the negative control after 24 h of treatment. Contrary to the in vitro results, no DNA repair seemed to occur in vivo up to 96 h post-venom inoculation at a dose of 30 µg/animal. The results show that BMV presents cyto- and genotoxicity depending on the concentration/dose used. These findings emphasize the importance of toxicological studies, including assessment of genotoxicity, in the biological activity research of BMV and/or in the development of BMV-derived products.

  19. Molecular Cloning and Pharmacological Properties of an Acidic PLA2 from Bothrops pauloensis Snake Venom

    Science.gov (United States)

    Ferreira, Francis Barbosa; Gomes, Mário Sérgio Rocha; Naves de Souza, Dayane Lorena; Gimenes, Sarah Natalie Cirilo; Castanheira, Letícia Eulalio; Borges, Márcia Helena; Rodrigues, Renata Santos; Yoneyama, Kelly Aparecida Geraldo; Homsi Brandeburgo, Maria Inês; Rodrigues, Veridiana M.

    2013-01-01

    In this work, we describe the molecular cloning and pharmacological properties of an acidic phospholipase A2 (PLA2) isolated from Bothrops pauloensis snake venom. This enzyme, denominated BpPLA2-TXI, was purified by four chromatographic steps and represents 2.4% of the total snake venom protein content. BpPLA2-TXI is a monomeric protein with a molecular mass of 13.6 kDa, as demonstrated by Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) analysis and its theoretical isoelectric point was 4.98. BpPLA2-TXI was catalytically active and showed some pharmacological effects such as inhibition of platelet aggregation induced by collagen or ADP and also induced edema and myotoxicity. BpPLA2-TXI displayed low cytotoxicity on TG-180 (CCRF S 180 II) and Ovarian Carcinoma (OVCAR-3), whereas no cytotoxicity was found in regard to MEF (Mouse Embryonic Fibroblast) and Sarcoma 180 (TIB-66). The N-terminal sequence of forty-eight amino acid residues was determined by Edman degradation. In addition, the complete primary structure of 122 amino acids was deduced by cDNA from the total RNA of the venom gland using specific primers, and it was significantly similar to other acidic D49 PLA2s. The phylogenetic analyses showed that BpPLA2-TXI forms a group with other acidic D49 PLA2s from the gender Bothrops, which are characterized by a catalytic activity associated with anti-platelet effects. PMID:24304676

  20. Molecular Cloning and Pharmacological Properties of an Acidic PLA2 from Bothrops pauloensis Snake Venom

    Directory of Open Access Journals (Sweden)

    Francis Barbosa Ferreira

    2013-12-01

    Full Text Available In this work, we describe the molecular cloning and pharmacological properties of an acidic phospholipase A2 (PLA2 isolated from Bothrops pauloensis snake venom. This enzyme, denominated BpPLA2-TXI, was purified by four chromatographic steps and represents 2.4% of the total snake venom protein content. BpPLA2-TXI is a monomeric protein with a molecular mass of 13.6 kDa, as demonstrated by Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF analysis and its theoretical isoelectric point was 4.98. BpPLA2-TXI was catalytically active and showed some pharmacological effects such as inhibition of platelet aggregation induced by collagen or ADP and also induced edema and myotoxicity. BpPLA2-TXI displayed low cytotoxicity on TG-180 (CCRF S 180 II and Ovarian Carcinoma (OVCAR-3, whereas no cytotoxicity was found in regard to MEF (Mouse Embryonic Fibroblast and Sarcoma 180 (TIB-66. The N-terminal sequence of forty-eight amino acid residues was determined by Edman degradation. In addition, the complete primary structure of 122 amino acids was deduced by cDNA from the total RNA of the venom gland using specific primers, and it was significantly similar to other acidic D49 PLA2s. The phylogenetic analyses showed that BpPLA2-TXI forms a group with other acidic D49 PLA2s from the gender Bothrops, which are characterized by a catalytic activity associated with anti-platelet effects.

  1. Effect of Bothrops alternatus snake venom on macrophage phagocytosis and superoxide production: participation of protein kinase C

    Directory of Open Access Journals (Sweden)

    SS Setubal

    2011-01-01

    Full Text Available Envenomations caused by different species of Bothrops snakes result in severe local tissue damage, hemorrhage, pain, myonecrosis, and inflammation with a significant leukocyte accumulation at the bite site. However, the activation state of leukocytes is still unclear. According to clinical cases and experimental work, the local effects observed in envenenomation by Bothrops alternatus are mainly the appearance of edema, hemorrhage, and necrosis. In this study we investigated the ability of Bothrops alternatus crude venom to induce macrophage activation. At 6 to 100 ¼g/mL, BaV is not toxic to thioglycollate-elicited macrophages; at 3 and 6 ¼g/mL, it did not interfere in macrophage adhesion or detachment. Moreover, at concentrations of 1.5, 3, and 6 ¼g/mL the venom induced an increase in phagocytosis via complement receptor one hour after incubation. Pharmacological treatment of thioglycollate-elicited macrophages with staurosporine, a protein kinase (PKC inhibitor, abolished phagocytosis, suggesting that PKC may be involved in the increase of serum-opsonized zymosan phagocytosis induced by BaV. Moreover, BaV also induced the production of anion superoxide (O2_ by thioglycollate-elicited macrophages. This BaV stimulated superoxide production was abolished after treating the cells with staurosporine, indicating that PKC is an important signaling pathway for the production of this radical. Based on these results, we suggest that phagocytosis and reactive oxygen species are involved in the pathogenesis of local tissue damage characteristic of Bothrops spp. envenomations.

  2. Unmasking Snake Venom of Bothrops leucurus: Purification and Pharmacological and Structural Characterization of New PLA2 Bleu TX-III

    Science.gov (United States)

    Marangoni, Fábio André; Ponce-Soto, Luis Alberto; Marangoni, Sergio; Landucci, Elen Cristina Teizem

    2013-01-01

    Bleu TX-III was isolated from Bothrops leucurus snake venom on one-step analytical chromatography reverse phase HPLC, was homogeneous on SDS-PAGE, and was confirmed by Q-Tof Ultima API ESI/MS (TOF MS mode) mass spectrometry in 14243.8 Da. Multiple alignments of Bleu TX-III show high degree of homology with basic PLA2 myotoxins from other Bothrops venoms. Our studies on local and systemic myotoxicity “in vivo” reveal that Bleu TX-III is myotoxin with local but not systemic action due to the decrease in the plasmatic CK levels when Bleu TX-III is administrated by intravenous route in mice (dose 1 and 5 μg). And at a dose of 20 μg myotoxin behaves like a local and systemic action. Bleu TX-III induced moderate marked paw edema, evidencing the local increase in vascular permeability. The inflammatory events induced in the mice (I.M.) were investigated. The increase in the levels of IL-1, IL-6, and TNF-α was observed in the plasma. It is concluded that Bleu TX-III induces inflammatory events in this model. The enzymatic phospholipid hydrolysis may be relevant to these phenomena. Bothrops leucurus venom is still not extensively explored, and the knowledge of its toxins separately through the study of structure/function will contribute for a better understanding of its action mechanism. PMID:23509815

  3. ENVENENAMIENTO OFIDICO POR EL GENERO Bothrops COMPLICADO CON MIOCARDIOPATIA TÓXICA: A PROPOSITO DE UN CASO.

    Directory of Open Access Journals (Sweden)

    Sajar Abusaid Palomo

    2014-12-01

    Full Text Available El envenenamiento ofídico en Venezuela es un problema de salud pública, afectando a agricultores y mineros, implicada con mayor frecuencia la familia Viperidae, genero Bothrops. Presenta clínica variable, desde manifestaciones locales hasta sistémicas. Paciente femenina de 40 años, agricultora, consulta por dolor y aumento de volumen en cara lateral de pie izquierdo por envenenamiento ofídico. Acude a ambulatorio de su localidad, 24 horas después es trasladada al Hospital Universitario Ruiz y Páez, ingresando con diagnóstico de Envenenamiento Ofídico por Bothrops. Evoluciona tórpidamente presentando Insuficiencia Renal Aguda en terapia de hemodiálisis, Sepsis punto de partida respiratorio, Insuficiencia Respiratoria Aguda y Miocardiopatía Tóxica, por lo que ingresa a Unidad de Cuidados Intensivos y Cuidados Coronarios. Luego de marcada mejoría clínica es dada de alta. El envenenamiento ofídico por Bothrops corresponde 80% de los accidentes por mordeduras de serpientes, su pronóstico depende de la especie, atención medica-hospitalaria, el intervalo de tiempo ocurrido y el uso de la terapia antiofídica especifica.

  4. Biological characterization of a myotoxin phosphoplipase A2 homologue purified from the venom of the snake Bothrops moojeni

    Directory of Open Access Journals (Sweden)

    MR Queiroz

    2011-01-01

    Full Text Available A myotoxin phospholipase A2 homologue, BmooMtx, was isolated from the venom of Bothrops moojeni by a combination of ion-exchange chromatography on DEAE-Sephacel column and gel filtration on Sephadex G-75. SDS-PAGE showed the enzyme to be a monomer with a molecular weight of 16,500. BmooMtx induced release of creatine kinase and morphological analyses indicated that it provoked an intense myonecrosis, with visible leukocyte infiltrate and damaged muscle cells 24 hours after injection. Anti-BmooMTx antibodies partially neutralized the myotoxic activity of BmooMtx and crude B. moojeni venom, as judged by determination of plasma creatine kinase levels and histological evaluation of skeletal muscle in mice. Anti-BmooMTx antibodies were effective in reducing the plasma creatine kinase levels of crude B. alternatus and B. leucurus venoms, evidencing immunological cross-reactivity between BmooMTx and other bothropic venoms. Intraplantar (i.pl. injection of BmooMtx (1 to 15 μg/animal caused a dose- and time-dependent hyperalgesia and edematogenic responses. Dexamethasone (0.4 mg/kg, meloxicam (2 mg/kg and promethazine (5 mg/kg markedly reduced the hyperalgesia. Our data suggest that these drugs may likely serve as complementary therapies in cases of accidents with Bothrops moojeni, provided that such pharmacological treatments are administered immediately after the incident.

  5. Natural history of the terciopelo Bothrops asper (Serpentes: Viperidae) in Costa Rica.

    Science.gov (United States)

    Sasa, Mahmood; Wasko, Dennis K; Lamar, William W

    2009-12-01

    The terciopelo Bothrops asper is the only lancehead species widely distributed in the humid lowlands of Middle America and northwestern South America. Its large body size, relative abundance and cryptic habits contribute to the high incidence of snakebites induced by this species throughout its distribution. The terciopelo plays an important role in ecosystems, both as prey and as a generalist predator. Diet comprises a great variety of prey items, including some species that are considered nuisances. B. asper, as other lancehead species, exhibits a notable ontogenetic shift in diet, consuming ectotherms (mainly frogs and lizards) when young, and increasingly incorporating birds, rodents, and other small mammals with maturity. Adult terciopelos also consume large anurans, especially when endothermic prey availability is low. Using radiotelemetry we determined home range and movement patterns from 28 individual B. asper at La Selva Biological Station, Costa Rica. Overall home range estimates are relatively small compared with other pitvipers, averaging between 3.71ha and 5.95ha; home range size did not differ between males and females. Movement patterns are largely aseasonal and consist of short (50 m) movements to new foraging areas. Habitat use is related to prey availability and therefore to foraging strategy. Our data support a strong preference for areas near swamps by both sexes. Reproduction in B. asper is highly seasonal, and--apparently--biannual. Reproductive cycles in Costa Rica are tightly related to rainfall patterns. Therefore, the timing of breeding differs between populations in the Caribbean and Pacific lowlands. Bothrops asper is adapted to areas with low levels of disturbance along the agricultural frontier, and consequently it is not rare to find it in or near human dwellings. However, despite popular belief, no evidence supports a purported increase in population density of this species in Costa Rica. Despite human persecution and substantial

  6. Functional proteomic analyses of Bothrops atrox venom reveals phenotypes associated with habitat variation in the Amazon.

    Science.gov (United States)

    Sousa, Leijiane F; Portes-Junior, José A; Nicolau, Carolina A; Bernardoni, Juliana L; Nishiyama, Milton Y; Amazonas, Diana R; Freitas-de-Sousa, Luciana A; Mourão, Rosa Hv; Chalkidis, Hipócrates M; Valente, Richard H; Moura-da-Silva, Ana M

    2017-04-21

    Venom variability is commonly reported for venomous snakes including Bothrops atrox. Here, we compared the composition of venoms from B. atrox snakes collected at Amazonian conserved habitats (terra-firme upland forest and várzea) and human modified areas (pasture and degraded areas). Venom samples were submitted to shotgun proteomic analysis as a whole or compared after fractionation by reversed-phase chromatography. Whole venom proteomes revealed a similar composition among the venoms with predominance of SVMPs, CTLs, and SVSPs and intermediate amounts of PLA 2 s and LAAOs. However, when distribution of particular isoforms was analyzed by either method, the venom from várzea snakes showed a decrease in hemorrhagic SVMPs and an increase in SVSPs, and procoagulant SVMPs and PLA 2 s. These differences were validated by experimental approaches including both enzymatic and in vivo assays, and indicated restrictions in respect to antivenom efficacy to variable components. Thus, proteomic analysis at the isoform level combined to in silico prediction of functional properties may indicate venom biological activity. These results also suggest that the prevalence of functionally distinct isoforms contributes to the variability of the venoms and could reflect the adaptation of B. atrox to distinct prey communities in different Amazon habitats. In this report, we compared isoforms present in venoms from snakes collected at different Amazonian habitats. By means of a species venom gland transcriptome and the in silico functional prediction of each isoform, we were able to predict the principal venom activities in vitro and in animal models. We also showed remarkable differences in the venom pools from snakes collected at the floodplain (várzea habitat) compared to other habitats. Not only was this venom less hemorrhagic and more procoagulant, when compared to the venom pools from the other three habitats studied, but also this enhanced procoagulant activity was not

  7. Treatment of Bothrops alternatus envenomation by Curcuma longa and Calendula officinalis extracts and ar-turmerone Tratamento local do envenenamento por Bothrops alternatus com extrato de Curcuma longa e Calendula officinalis e ar-turmerone

    Directory of Open Access Journals (Sweden)

    M.M. Melo

    2005-02-01

    Full Text Available It was investigated the efficiency of two extracts of plants and one fraction of their properties against the local effects of bothropic envenomation. Bothrops alternatus venom (1.25µg diluted in 100µl of sterile saline solution was inoculated (intradermally into the shaved dorsal back skin of 30 New Zealand rabbits. The animals were divided in six groups receiving the following treatments: group I: subcutaneous application of Curcuma longa extract (1.0ml; group II: topic treatment of Curcuma longa hydroalcoholic extract (1.0ml; group III: topic application of ar-turmerone in vaseline (1.0g; group IV: topic application of Curcuma longa methanolic extract (1.0ml; group V: topic application of Calendula officinalis ointment (1.0g; group VI: topic application of saline (1.0ml. These treatments were done at 30 minutes, and at 2, 4, 24 and 72 hours after venom inoculation. Intensity of local edema, hemorrhagic halo and necrosis were evaluated until 168h after that. Additionally, seven days after the Bothrops venom inoculation, blood was collected from heart with and without EDTA (10% for hemogram and biochemical parameters (total protein, blood urea nitrogen, creatinine, and fibrinogen and all the animals were anesthetized, sacrificed by ether inhalation and submitted to necropsy. Fragments of tissues were taken for histopathological evaluation. The most efficient treatment for inhibition of edema, necrosis and local hemorrhage after Bothrops alternatus venom was the topic application of ar-turmerone.Investigou-se a eficácia do extrato de plantas no tratamento local do envenenamento botrópico. Veneno de serpentes Bothrops alternatus (1,25µg diluído em 100µl de solução salina estéril foi inoculado (via intradérmica entre as escápulas de 30 coelhos. Os animais foram divididos em seis grupos (tratamentos: grupo I: tratamento subcutâneo com extrato de Curcuma longa; grupo II: tratamento tópico com extrato hidroalcoólico de Curcuma longa

  8. A New Platelet-Aggregation-Inhibiting Factor Isolated from Bothrops moojeni Snake Venom

    Directory of Open Access Journals (Sweden)

    Bruna Barbosa de Sousa

    2017-01-01

    Full Text Available This work reports the purification and functional characterization of BmooPAi, a platelet-aggregation-inhibiting factor from Bothrops moojeni snake venom. The toxin was purified by a combination of three chromatographic steps (ion-exchange on DEAE-Sephacel, molecular exclusion on Sephadex G-75, and affinity chromatography on HiTrap™ Heparin HP. BmooPAi was found to be a single-chain protein with an apparent molecular mass of 32 kDa on 14% SDS-PAGE, under reducing conditions. Sequencing of BmooPAi by Edman degradation revealed the amino acid sequence LGPDIVPPNELLEVM. The toxin was devoid of proteolytic, haemorrhagic, defibrinating, or coagulant activities and induced no significant oedema or hyperalgesia. BmooPAi showed a rather specific inhibitory effect on ristocetin-induced platelet aggregation in human platelet-rich plasma, whereas it had little or no effect on platelet aggregation induced by collagen and adenosine diphosphate. The results presented in this work suggest that BmooPAi is a toxin comprised of disintegrin-like and cysteine-rich domains, originating from autolysis/proteolysis of PIII SVMPs from B. moojeni snake venom. This toxin may be of medical interest because it is a platelet aggregation inhibitor, which could potentially be developed as a novel therapeutic agent to prevent and/or treat patients with thrombotic disorders.

  9. Pulsed ultrasound therapy accelerates the recovery of skeletal muscle damage induced by Bothrops jararacussu venom

    Directory of Open Access Journals (Sweden)

    J. Saturnino-Oliveira

    2012-06-01

    Full Text Available We studied the effect of pulsed ultrasound therapy (UST and antibothropic polyvalent antivenom (PAV on the regeneration of mouse extensor digitorum longus muscle following damage by Bothrops jararacussu venom. Animals (Swiss male and female mice weighing 25.0 ± 5.0 g; 5 animals per group received a perimuscular injection of venom (1 mg/kg and treatment with UST was started 1 h later (1 min/day, 3 MHz, 0.3 W/cm², pulsed mode. Three and 28 days after injection, muscles were dissected and processed for light microscopy. The venom caused complete degeneration of muscle fibers. UST alone and combined with PAV (1.0 mL/kg partially protected these fibers, whereas muscles receiving no treatment showed disorganized fascicules and fibers with reduced diameter. Treatment with UST and PAV decreased the effects of the venom on creatine kinase content and motor activity (approximately 75 and 48%, respectively. Sonication of the venom solution immediately before application decreased the in vivo and ex vivo myotoxic activities (approximately 60 and 50%, respectively. The present data show that UST counteracts some effects of B. jararacussu venom, causing structural and functional improvement of the regenerated muscle after venom injury.

  10. The effects of low-level laser on muscle damage caused by Bothrops neuwiedi venom

    Energy Technology Data Exchange (ETDEWEB)

    Dourado, D.M.; Matias, R.; Almeida, M.F.; Paula, K.R. de; Carvalho, P.T.C. [University for the Development of the State and of the Region of Pantanal (UNIDERP), Campo Grande, MS (Brazil). Lab. of Experimental Histopathology]. E-mail: ccfi@uniderp.br; Vieira, R.P. [University of Sao Paulo (USP), SP (Brazil). School of Medicine. Dept. of Pathology and Physical Therapy; Oliveira, L.V.F. [Nove de Julho University (UNINOVE), Sao Paulo, SP (Brazil). Masters Program in Rehabilitation Sciences

    2008-07-01

    The present study aimed to assess the effects of low-level laser (660 nm) on myonecrosis caused by the insertion of Bothrops neuwiedi venom in the gastrocnemius muscle of rats. Male Wistar rats were divided into three groups (n = 24 each): Group S (0.9% saline solution); Group V (venom) and Group VLLL (venom plus low-level laser). These categories were subdivided into four additional groups (n = 6) based on the euthanasia timing (3 hours, 24 hours, 3 days and 7 days). The groups V and VLLL were inoculated with 100 {mu}L of concentrated venom (40 {mu}g/mL) in the gastrocnemius muscle. The muscle was irradiated using a gallium-aluminum-arsenide laser (GaAlAs) at 35 mW power and 4 J/cm{sup 2} energy density for 3 hours, 24 hours, 3 days or 7 days after venom inoculation. To evaluate the myotoxic activity of the venom, CK activity was measured and the muscle was histologically analyzed. The low-level laser reduced venom-induced CK activity in the groups euthanized at 3 hours, 24 hours and 3 days (p < 0.0001). Histological analysis revealed that low-level laser reduced neutrophilic inflammation as well as myofibrillar edema, hemorrhage and myonecrosis following B. neuwiedi envenomation. These results suggest that low-level laser can be useful as an adjunct therapy following B. neuwiedi envenomation. (author)

  11. The effects of low-level laser on muscle damage caused by Bothrops neuwiedi venom

    International Nuclear Information System (INIS)

    Dourado, D.M.; Matias, R.; Almeida, M.F.; Paula, K.R. de; Carvalho, P.T.C.; Vieira, R.P.; Oliveira, L.V.F.

    2008-01-01

    The present study aimed to assess the effects of low-level laser (660 nm) on myonecrosis caused by the insertion of Bothrops neuwiedi venom in the gastrocnemius muscle of rats. Male Wistar rats were divided into three groups (n = 24 each): Group S (0.9% saline solution); Group V (venom) and Group VLLL (venom plus low-level laser). These categories were subdivided into four additional groups (n = 6) based on the euthanasia timing (3 hours, 24 hours, 3 days and 7 days). The groups V and VLLL were inoculated with 100 μL of concentrated venom (40 μg/mL) in the gastrocnemius muscle. The muscle was irradiated using a gallium-aluminum-arsenide laser (GaAlAs) at 35 mW power and 4 J/cm 2 energy density for 3 hours, 24 hours, 3 days or 7 days after venom inoculation. To evaluate the myotoxic activity of the venom, CK activity was measured and the muscle was histologically analyzed. The low-level laser reduced venom-induced CK activity in the groups euthanized at 3 hours, 24 hours and 3 days (p < 0.0001). Histological analysis revealed that low-level laser reduced neutrophilic inflammation as well as myofibrillar edema, hemorrhage and myonecrosis following B. neuwiedi envenomation. These results suggest that low-level laser can be useful as an adjunct therapy following B. neuwiedi envenomation. (author)

  12. The effects of low-level laser on muscle damage caused by Bothrops neuwiedi venom

    Directory of Open Access Journals (Sweden)

    DM Dourado

    2008-01-01

    Full Text Available The present study aimed to assess the effects of low-level laser (660 nm on myonecrosis caused by the insertion of Bothrops neuwiedi venom in the gastrocnemius muscle of rats. Male Wistar rats were divided into three groups (n = 24 each: Group S (0.9% saline solution; Group V (venom and Group VLLL (venom plus low-level laser. These categories were subdivided into four additional groups (n = 6 based on the euthanasia timing (3 hours, 24 hours, 3 days and 7 days. The groups V and VLLL were inoculated with 100 µL of concentrated venom (40 µg/mL in the gastrocnemius muscle. The muscle was irradiated using a gallium-aluminum-arsenide laser (GaAlAs at 35 mW power and 4 J/cm² energy density for 3 hours, 24 hours, 3 days or 7 days after venom inoculation. To evaluate the myotoxic activity of the venom, CK activity was measured and the muscle was histologically analyzed. The low-level laser reduced venom-induced CK activity in the groups euthanized at 3 hours, 24 hours and 3 days (p < 0.0001. Histological analysis revealed that low-level laser reduced neutrophilic inflammation as well as myofibrillar edema, hemorrhage and myonecrosis following B. neuwiedi envenomation. These results suggest that low-level laser can be useful as an adjunct therapy following B. neuwiedi envenomation.

  13. Envenomations by Bothrops and Crotalus snakes induce the release of mitochondrial alarmins.

    Science.gov (United States)

    Zornetta, Irene; Caccin, Paola; Fernandez, Julián; Lomonte, Bruno; Gutierrez, José María; Montecucco, Cesare

    2012-01-01

    Skeletal muscle necrosis is a common manifestation of viperid snakebite envenomations. Venoms from snakes of the genus Bothrops, such as that of B. asper, induce muscle tissue damage at the site of venom injection, provoking severe local pathology which often results in permanent sequelae. In contrast, the venom of the South American rattlesnake Crotalus durissus terrificus, induces a clinical picture of systemic myotoxicity, i.e., rhabdomyolysis, together with neurotoxicity. It is known that molecules released from damaged muscle might act as 'danger' signals. These are known as 'alarmins', and contribute to the inflammatory reaction by activating the innate immune system. Here we show that the venoms of B. asper and C. d. terrificus release the mitochondrial markers mtDNA (from the matrix) and cytochrome c (Cyt c) from the intermembrane space, from ex vivo mouse tibialis anterior muscles. Cyt c was released to a similar extent by the two venoms whereas B. asper venom induced the release of higher amounts of mtDNA, thus reflecting hitherto some differences in their pathological action on muscle mitochondria. At variance, injection of these venoms in mice resulted in a different time-course of mtDNA release, with B. asper venom inducing an early onset increment in plasma levels and C. d. terrificus venom provoking a delayed release. We suggest that the release of mitochondrial 'alarmins' might contribute to the local and systemic inflammatory events characteristic of snakebite envenomations.

  14. Experimental pathology of local tissue damage induced by Bothrops asper snake venom.

    Science.gov (United States)

    Gutiérrez, José María; Rucavado, Alexandra; Chaves, Fernando; Díaz, Cecilia; Escalante, Teresa

    2009-12-01

    Envenomations by Bothrops asper are often associated with complex and severe local pathological manifestations, including edema, blistering, dermonecrosis, myonecrosis and hemorrhage. The pathogenesis of these alterations has been investigated at the experimental level. These effects are mostly the consequence of the direct action of zinc-dependent metalloproteinases (SVMPs) and myotoxic phospholipases A(2) (PLA(2)s). SVMPs induce hemorrhage, blistering, dermonecrosis and general extracellular matrix degradation, whereas PLA(2)s induce myonecrosis and also affect lymphatic vessels. In addition, the prominent vascular alterations leading to hemorrhage and edema may contribute to ischemia and further tissue necrosis. The mechanisms of action of SVMPs and PLA(2)s are discussed in detail in this review. Venom-induced tissue damage plays also a role in promoting bacterial infection. A prominent inflammatory reaction develops as a consequence of these local pathological alterations, with the synthesis and release of abundant mediators, resulting in edema and pain. However, whether inflammatory cells and mediators contribute to further tissue damage is not clear at present. Muscle tissue regeneration after venom-induced pathological effects is often impaired, thus resulting in permanent tissue loss and dysfunction. SVMP-induced microvessel damage is likely to be responsible of this poor regenerative outcome. Antivenoms are only partially effective in the neutralization of B. asper-induced local effects, and the search for novel toxin inhibitors represents a potential avenue for improving the treatment of this serious aspect of snakebite envenomation.

  15. Biochemistry and toxicology of toxins purified from the venom of the snake Bothrops asper.

    Science.gov (United States)

    Angulo, Yamileth; Lomonte, Bruno

    2009-12-01

    The isolation and study of individual snake venom components paves the way for a deeper understanding of the pathophysiology of envenomings--thus potentially contributing to improved therapeutic modalities in the clinical setting--and also opens possibilities for the discovery of novel toxins that might be useful as tools for dissecting cellular and molecular processes of biomedical importance. This review provides a summary of the different toxins that have been isolated and characterized from the venom of Bothrops asper, the snake species causing the majority of human envenomings in Central America. This venom contains proteins belonging to at least eight families: metalloproteinase, serine proteinase, C-type lectin-like, L-amino acid oxidase, disintegrin, DC-fragment, cystein-rich secretory protein, and phospholipase A(2). Some 25 venom proteins within these families have been isolated and characterized. Their main biochemical properties and toxic actions are described, including, in some cases, their possible relationships to the pathologic effects induced by B. asper venom.

  16. Inhibition of myotoxic activity of Bothrops asper myotoxin II by the anti-trypanosomal drug suramin.

    Science.gov (United States)

    Murakami, Mário T; Arruda, Emerson Z; Melo, Paulo A; Martinez, Ana B; Calil-Eliás, Sabrińa; Tomaz, Marcelo A; Lomonte, Bruno; Gutiérrez, José M; Arni, Raghuvir K

    2005-07-15

    Suramin, a synthetic polysulfonated compound, developed initially for the treatment of African trypanosomiasis and onchocerciasis, is currently used for the treatment of several medically relevant disorders. Suramin, heparin, and other polyanions inhibit the myotoxic activity of Lys49 phospholipase A2 analogues both in vitro and in vivo, and are thus of potential importance as therapeutic agents in the treatment of viperid snake bites. Due to its conformational flexibility around the single bonds that link the central phenyl rings to the secondary amide backbone, the symmetrical suramin molecule binds by an induced-fit mechanism complementing the hydrophobic surfaces of the dimer and adopts a novel conformation that lacks C2 symmetry in the dimeric crystal structure of the suramin-Bothrops asper myotoxin II complex. The simultaneous binding of suramin at the surfaces of the two monomers partially restricts access to the nominal active sites and significantly changes the overall charge of the interfacial recognition face of the protein, resulting in the inhibition of myotoxicity.

  17. Experimental pathophysiology of systemic alterations induced by Bothrops asper snake venom.

    Science.gov (United States)

    Gutiérrez, José María; Escalante, Teresa; Rucavado, Alexandra

    2009-12-01

    Moderate and severe envenomations by the snake Bothrops asper provoke systemic alterations, such as systemic bleeding, coagulopathy, hypovolemia, hemodynamic instability and shock, and acute renal failure. Systemic hemorrhage is a typical finding of these envenomations, and is primarily caused by the action of P-III snake venom metalloproteinases (SVMPs). This venom also contains a thrombin-like serine proteinase and a prothrombin-activating P-III SVMP, both of which cause defibrin(ogen)ation. Thrombocytopenia, predominantly induced by a C-type lectin-like protein, and platelet hypoaggregation, caused by the two defibrin(ogen)ating enzymes, also contribute to hemostatic disturbances, which potentiate the systemic bleeding induced by hemorrhagic SVMPs. Cardiovascular disturbances leading to shock are due to the combined effects of hemorrhagic toxins, other venom components that increase vascular permeability, the action of hypotensive agents in the venom and of endogenous mediators, and the potential cardiotoxic effect of some toxins. Renal alterations are likely to be caused by direct cytotoxicity of venom components in the kidney, and by renal ischemia resultant from hypovolemia and hypoperfusion. Lethality induced by B. asper venom is the consequence of several combined effects among which the action of P-III SVMPs is especially relevant.

  18. Evaluation of anti-Bothrops asper venom activity of ethanolic extract of Brownea rosademonte leaves.

    Science.gov (United States)

    Salazar, Marcos; Chérigo, Lilia; Acosta, Hildaura; Otero, Rafael; Martínez-Luis, Sergio

    2014-12-01

    Significant inhibition of the coagulant and hemorrhagic effects of Bothrops asper venom was demonstrated by ethanolic extract prepared from the leaves of Brownea rosademonte. In vitro experiments preincubating 5.5 mg of extract kg-1 b.m. for 30 min with a minimum hemorrhagic dose of venom (273.8 ± 16.1 μg of venom kg-1 b.m.) lowered the hemorrhagic activity of the venom alone in CD-1 mice by 51.5 ± 2.6 %. Additionally, 1.7 mg extract L-1 plasma prolonged 5.1 times the plasma coagulation time. Fractionation of the extract led to the isolation of two compounds: ononitol (1) and quercetrin (2). The structure of compounds 1 and 2 was established by spectroscopic analyses, including APCI-HRMS and NMR (1H, 13C, HSQC, HMBC and COSY). A quercetrin concentration of 0.11 μmol L-1 prolonged the plasma coagulation time 2.6 times demonstrating that this compound was one of the active constituents of the Brownea rosademonte extract.

  19. Studies on the venom proteome of Bothrops asper: perspectives and applications.

    Science.gov (United States)

    Alape-Girón, Alberto; Flores-Díaz, Marietta; Sanz, Libia; Madrigal, Marvin; Escolano, José; Sasa, Mahmood; Calvete, Juan J

    2009-12-01

    Bothrops asper is responsible for the vast majority of snakebite accidents in Central America and several studies have demonstrated that specific toxic and enzymatic activities of its venom vary with the geographic origin and age of the specimens. Variability in venom proteins and enzymes between specimens from the Caribbean and the Pacific versants of Costa Rica has been reported since 1964. Recently, we performed a comparative proteomic characterization of the venoms from one population of each versant. Proteins belonging to several families, including disintegrin, phospholipases A(2), serine proteinases, C-type lectins, CRISP, l-amino acid oxidase, and Zn(2+)-dependent metalloproteinases show a variable degree of relative occurrence in the venoms of both populations. The occurrence of prominent differences in the protein profile between venoms from adults and newborns, and among venom samples from individual specimens of the same region or developmental stage, further demonstrated the existence of geographic, ontogenetic and individual variability in the venom proteome of this species. These findings provide new insights towards understanding the biology of B. asper, contribute to a deeper understanding of the pathology induced by its venom and underscore the importance of the use of venoms pooled from specimens from both regions for producing antivenom exhibiting the broadest cross-reactivity. Furthermore, knowledge of the protein composition of B. asper venom paves the way for detailed future structure-function studies of individual toxins as well as for the development of new protocols to study the reactivity of therapeutic antivenoms.

  20. Abarema cochliacarpos Extract Decreases the Inflammatory Process and Skeletal Muscle Injury Induced by Bothrops leucurus Venom

    Science.gov (United States)

    Saturnino-Oliveira, Jeison; Santos, Daiana Do Carmo; Guimarães, Adriana Gibara; Santos Dias, Antônio; Tomaz, Marcelo Amorim; Monteiro-Machado, Marcos; Estevam, Charles Santos; Lucca Júnior, Waldecy De; Maria, Durvanei Augusto; Melo, Paulo A.; Araújo, Adriano Antunes de Souza; Santos, Márcio Roberto Viana; Almeida, Jackson Roberto Guedes da Silva; Oliveira, Rita de Cássia Meneses; Pereira de Oliveira, Aldeidia; Quintans Júnior, Lucindo José

    2014-01-01

    Snakebites are a public health problem, especially in tropical countries. However, treatment with antivenom has limited effectiveness against venoms' local effects. Here, we investigated the ability of Abarema cochliacarpos hydroethanolic extract (EAc) to protect mice against injection of Bothrops leucurus venom. Swiss mice received perimuscular venom injection and were subsequently treated orally with EAc in different doses. Treatment with EAc 100, 200, and 400 mg/kg reduced the edema induced by B. leucurus in 1%, 13%, and 39%, respectively. Although lower doses showed no antihypernociceptive effect in the Von Frey test, the higher dose significantly reduced hyperalgesia induced by the venom. Antimyotoxic activity of EAc was also observed by microscopy assessment, with treated muscles presenting preserved structures, decreased edema, and inflammatory infiltrate as compared to untreated ones. Finally, on the rotarod test, the treated mice showed better motor function, once muscle fibers were preserved and there were less edema and pain. Treated mice could stand four times more time on the rotating rod than untreated ones. Our results have shown that EAc presented relevant activities against injection of B. leucurus venom in mice, suggesting that it can be considered as an adjuvant in the treatment of envenomation. PMID:25136627

  1. ACIDENTES POR SERPENTE (BOTHROPS SPP. E CROTALLUS SPP. EM CRIANÇAS: RELATO DE DOIS CASOS

    Directory of Open Access Journals (Sweden)

    Beatriz Ferreira Martins

    2012-01-01

    Full Text Available Los accidentes provocados por serpientes poseen importancia sanitaria debido a la frecuencia y gravedad. La estandarización de la atención es imprescindible para auxiliar a los equipos de salud en el diagnóstico precoz y en el tratamiento específico a las víctimas. Pero, la evolución clínica del caso presenta particularidades, principalmente en accidentes infantiles y en ancianos. El objetivo fue describir y analizar dos casos clínicos de accidentes infantiles con serpientes de los géneros Bothrops spp. y Crotallus spp., notificados a un centro de información y asistencia toxicológica del Noroeste del Paraná/Brasil. Se encontraron aspectos positivos en la atención, como precocidad del acceso a la unidad de salud, identificación de síntomas y seroterapia precoz, y no ocurrencia de reacciones adversas a la seroterapia antivenenosa, y como aspectos negativos, la ausencia de la solicitud de exámenes bioquímicos preconizados y alta hospitalaria precoz pos seroterapia, considerando la posibilidad de reacciones adversas.

  2. Irradiation of the Crude Venom of Bothrops jararacussu to Obtain Toxoid

    International Nuclear Information System (INIS)

    Ferreira, Camila G.; Avalloni, Tania M.; Oshima-Franco, Yoko; Oliveira, Sara de J; Oliveira, Jose M. Jr. de; Cogo, Jose C.

    2011-01-01

    The aim of this work was to reduce the toxicity of Bothrops jararacussu venom using gamma-rays of low-energy coming from a source of Americium-241 (E = 59.6 keV and 3.7x10 9 Bq of activity) in order to obtain a toxoid. The radiation dose that each sample received was controlled by exposure time of the venom to the radiation beam. Mouse nerve phrenic-diaphragm preparation was used for testing the loss of venom toxicity, since the venom causes an irreversible neuromuscular blockade. In this condition, the several samples of irradiated venom, when assayed in neuromuscular preparation showed that with a dose of 0.051 Gy the paralysis caused by the irradiated venom was of 91%, at 0.360 Gy was of 79%, at 1.662 Gy was of 50% and at 2.448 Gy was of 42%. Therefore, it can be concluded that the irradiation model was able to induce a progressive loss of the venom toxicity.

  3. Irradiation of the Crude Venom of Bothrops jararacussu to Obtain Toxoid

    Science.gov (United States)

    Ferreira, Camila G.; Avalloni, Tânia M.; Oshima-Franco, Yoko; de J. Oliveira, Sara; de Oliveira, José M.; Cogo, José C.

    2011-08-01

    The aim of this work was to reduce the toxicity of Bothrops jararacussu venom using gamma-rays of low-energy coming from a source of Americium-241 (E = 59.6 keV and 3.7×109 Bq of activity) in order to obtain a toxoid. The radiation dose that each sample received was controlled by exposure time of the venom to the radiation beam. Mouse nerve phrenic-diaphragm preparation was used for testing the loss of venom toxicity, since the venom causes an irreversible neuromuscular blockade. In this condition, the several samples of irradiated venom, when assayed in neuromuscular preparation showed that with a dose of 0.051 Gy the paralysis caused by the irradiated venom was of 91%, at 0.360 Gy was of 79%, at 1.662 Gy was of 50% and at 2.448 Gy was of 42%. Therefore, it can be concluded that the irradiation model was able to induce a progressive loss of the venom toxicity.

  4. Envenomations by Bothrops and Crotalus snakes induce the release of mitochondrial alarmins.

    Directory of Open Access Journals (Sweden)

    Irene Zornetta

    Full Text Available Skeletal muscle necrosis is a common manifestation of viperid snakebite envenomations. Venoms from snakes of the genus Bothrops, such as that of B. asper, induce muscle tissue damage at the site of venom injection, provoking severe local pathology which often results in permanent sequelae. In contrast, the venom of the South American rattlesnake Crotalus durissus terrificus, induces a clinical picture of systemic myotoxicity, i.e., rhabdomyolysis, together with neurotoxicity. It is known that molecules released from damaged muscle might act as 'danger' signals. These are known as 'alarmins', and contribute to the inflammatory reaction by activating the innate immune system. Here we show that the venoms of B. asper and C. d. terrificus release the mitochondrial markers mtDNA (from the matrix and cytochrome c (Cyt c from the intermembrane space, from ex vivo mouse tibialis anterior muscles. Cyt c was released to a similar extent by the two venoms whereas B. asper venom induced the release of higher amounts of mtDNA, thus reflecting hitherto some differences in their pathological action on muscle mitochondria. At variance, injection of these venoms in mice resulted in a different time-course of mtDNA release, with B. asper venom inducing an early onset increment in plasma levels and C. d. terrificus venom provoking a delayed release. We suggest that the release of mitochondrial 'alarmins' might contribute to the local and systemic inflammatory events characteristic of snakebite envenomations.

  5. Evaluation of anti-Bothrops asper venom activity of ethanolic extract of Brownea rosademonte leaves

    Directory of Open Access Journals (Sweden)

    Salazar Marcos

    2014-12-01

    Full Text Available Significant inhibition of the coagulant and hemorrhagic effects of Bothrops asper venom was demonstrated by ethanolic extract prepared from the leaves of Brownea rosademonte. In vitro experiments preincubating 5.5 mg of extract kg-1 b.m. for 30 min with a minimum hemorrhagic dose of venom (273.8 ± 16.1 μg of venom kg-1 b.m. lowered the hemorrhagic activity of the venom alone in CD-1 mice by 51.5 ± 2.6 %. Additionally, 1.7 mg extract L-1 plasma prolonged 5.1 times the plasma coagulation time. Fractionation of the extract led to the isolation of two compounds: ononitol (1 and quercetrin (2. The structure of compounds 1 and 2 was established by spectroscopic analyses, including APCI-HRMS and NMR (1H, 13C, HSQC, HMBC and COSY. A quercetrin concentration of 0.11 μmol L-1 prolonged the plasma coagulation time 2.6 times demonstrating that this compound was one of the active constituents of the Brownea rosademonte extract.

  6. Acción de la miotoxina del veneno de Bothrops brazili Hoge, 1953 (Ophidia: Viperidae

    Directory of Open Access Journals (Sweden)

    Carmen Pantigoso

    2013-06-01

    Full Text Available Se ha estudiado el modo de acción de la miotoxina aislada del veneno de la serpiente Bothrops brazili. La inoculación de la miotoxina en el músculo gastrocnemius de ratones albinos produce durante la primera hora de acción la liberación de creatina kinasa y lactato deshidrogenasa, mientras que por PAGE-SDS, se revela que la incubación de la miotoxina con músculo gastrocnemius aislado, produce además la liberación de otras proteínas musculares. Asimismo, la miotoxina produce hipercontracción, lesiones delta e incrementa los niveles de calcio intramuscular, tanto in vivo como in vitro, lo cual no depende del ingreso de calcio extracelular vía receptores de dihidropiridina. Este incremento de calcio explicaría la hipercontracción observada y podría generar la activación de proteasas y lipasas endógenas dependientes de calcio, que conducirían a la necrosis muscular.

  7. Muscle phospholipid hydrolysis by Bothrops asper Asp49 and Lys49 phospholipase A₂ myotoxins--distinct mechanisms of action.

    Science.gov (United States)

    Fernández, Julián; Caccin, Paola; Koster, Grielof; Lomonte, Bruno; Gutiérrez, José M; Montecucco, Cesare; Postle, Anthony D

    2013-08-01

    Bothrops snakes are the major cause of ophidian envenomings in Latin America. Their venom contains myotoxins that cause prominent muscle damage, which may lead to permanent disability. These toxins include myotoxins Mt-I and Mt-II, which share the phospholipase A₂ (PLA₂) fold, but Mt-II lacks enzymatic activity because the essential active site Asp49 is replaced by Lys. Both myotoxins cause sarcolemma alterations, with Ca²⁺ entry and loss of ATP and K⁺ from muscle cells, but the molecular lesions at the basis of their cellular action are not known, particularly the role of phospholipid hydrolysis. Here we tested their PLA₂ activity in vivo, and evaluated the hypothesis that Ca²⁺-activated endogenous PLA₂s may be involved in the action of Mt-II. The time course of phospholipid hydrolysis by Mt-I and Mt-II in myotubes in culture and in tibialis anterior mouse muscles was determined. Mt-I rapidly hydrolyzed phosphatidylcholine and phosphatidylethanolamine but not phosphatidylserine, but no phospho-lipids were hydrolyzed in the presence of Mt-II. Whole Bothrops asper venom induced a higher extent of phospholipid hydrolysis than Mt-I alone. These results demonstrate in vivo PLA₂ activity of Mt-I for the first time, and indicate that it acts only on the external monolayer of the sarcolemma. They also exclude activation of endogenous PLA₂s in the action of Mt-II, implying that plasma membrane disruption by this toxin does not depend on phospholipid hydrolysis. Therefore, both Bothrops myotoxins induce Ca²⁺ entry and release of ATP and cause myonecrosis, but through different biochemical mechanisms. © 2013 FEBS.

  8. ENVENENAMIENTO OFIDICO POR EL GENERO Bothrops COMPLICADO CON MIOCARDIOPATIA TÓXICA: A PROPOSITO DE UN CASO.

    OpenAIRE

    Sajar Abusaid Palomo; Carlos Muñoz Medina; Milagros Mota Carvajal; Marisol Sandoval

    2014-01-01

    El envenenamiento ofídico en Venezuela es un problema de salud pública, afectando a agricultores y mineros, implicada con mayor frecuencia la familia Viperidae, genero Bothrops. Presenta clínica variable, desde manifestaciones locales hasta sistémicas. Paciente femenina de 40 años, agricultora, consulta por dolor y aumento de volumen en cara lateral de pie izquierdo por envenenamiento ofídico. Acude a ambulatorio de su localidad, 24 horas después es trasladada al Hospital Universitario Ruiz y...

  9. First record of Porocephalus cf. clavatus (Pentastomida: Porocephalida as a parasite on Bothrops asper (Squamata: Viperidae in Costa Rica

    Directory of Open Access Journals (Sweden)

    G. Alvarado

    Full Text Available Abstract Pentastomids are parasites that infect respiratory cavities of vertebrates, they are pretty common but poorly known in wildlife veterinary. A Bothrops asper snake (Garman, 1884 was captured in the Caribbean region of Costa Rica and had its lung infested with pentastomids, identified as ca Porocephalus clavatus (Wyman, 1845. This represents the first record of Porocephalus (Humboldt, 1812 on B. asper as well as P. cf. clavatus in Costa Rica. Further studies are needed to clarify their taxonomic position, images and scanning electron microscopy photographs (SEM of the specimens are given.

  10. First record of Porocephalus cf. clavatus (Pentastomida: Porocephalida) as a parasite on Bothrops asper (Squamata: Viperidae) in Costa Rica.

    Science.gov (United States)

    Alvarado, G; Sánchez-Monge, A

    2015-11-01

    Pentastomids are parasites that infect respiratory cavities of vertebrates, they are pretty common but poorly known in wildlife veterinary. A Bothrops asper snake (Garman, 1884) was captured in the Caribbean region of Costa Rica and had its lung infested with pentastomids, identified as ca Porocephalus clavatus (Wyman, 1845). This represents the first record of Porocephalus (Humboldt, 1812) on B. asper as well as P. cf. clavatus in Costa Rica. Further studies are needed to clarify their taxonomic position, images and scanning electron microscopy photographs (SEM) of the specimens are given.

  11. Eficacia experimental de anticuerpos IgY producidos en huevos, contra el veneno de la serpiente peruana Bothrops atrox Experimental efficacy of IgY antibodies produced in eggs against the venom of the Peruvian snake Bothrops atrox

    Directory of Open Access Journals (Sweden)

    Julio C. Mendoza

    2012-03-01

    Full Text Available Objetivos. Desarrollar un protocolo de inmunización para producir inmunoglobulinas IgY de origen aviar contra el veneno de la serpiente peruana Bothrops atrox y evaluar la capacidad neutralizante. Materiales y métodos. Se inmunizaron seis gallinas de postura de la raza hy line brown con 500 μg/dosis de veneno de B. atrox en un periodo de dos meses. Cada semana, los huevos fueron colectados para el aislamiento de inmunoglobulinas IgY a partir de la yema, usando dos pasos consecutivos con αcido caprνlico y sulfato de amonio. La detecciσn de anticuerpos se realizσ por inmunodifusiσn doble mientras que el tνtulo y reactividad cruzada se determinaron por las técnicas de ELISA y Western blot. El cálculo de DL50 y de la DE50 del antiveneno IgY producido se realizó utilizando el método de Probits. Resultados. La masa de anticuerpos aislados fue de 8,5 ± 1,35 mg de IgY/mL de yema. Asimismo, la DE50 del antiveneno aviar fue calculada en 575 μL de antiveneno/mg de veneno. Adicionalmente, los ensayos de reactividad cruzada mostraron que el veneno de B. atrox comparte mas epνtopes comunes con el veneno de B. brazili (47% que con otros veneno del mismo género, en tanto que los venenos de Lachesis muta (19% y Crotalus durissus (12% mostraron una baja reactividad cruzada. Conclusiones. Se ha obtenido IgY purificada contra el veneno de B. atrox con capacidad neutralizante y se ha demostrado su utilidad como herramienta inmunoanalítica para evaluar la reactividad cruzada con venenos de otras especies.Objectives. To develop an immunization protocol in order to produce avian IgY immunoglobulins against Bothrops atrox Peruvian snake venom and to evaluate its neutralizing capacity. Materials and methods. Six Hy Line Brown hens were immunized each two weeks using 500μg/doses of B. atrox venom in a period of two months. Each week, eggs were collected for IgY isolation from yolk using two consecutive steps with caprilic acid and ammonium sulfate

  12. Inflammation induced by Bothrops asper venom: release of proinflammatory cytokines and eicosanoids, and role of adhesion molecules in leukocyte infiltration.

    Science.gov (United States)

    Zamuner, Stella Regina; Zuliani, Juliana Pavan; Fernandes, Cristina Maria; Gutiérrez, José Maria; de Fátima Pereira Teixeira, Catarina

    2005-12-01

    Bothrops asper venom (BaV) causes systemic and local effects characterized by an acute inflammatory reaction with accumulation of leukocytes and release of endogenous mediators. In this study, the effects of BaV on the release of the cytokines IL-1, IL-6 and TNF-alpha and the eicosanoids LTB4 and TXA2 in the peritoneal cavity of mice were analyzed. We also investigated the participation of beta2 integrin chain, l-selectin, LFA-1, ICAM-1 and PECAM-1 adhesion molecules in the BaV-induced leukocyte accumulation. Levels of proinflammatory cytokines IL-6 and TNF-alpha, as well as eicosanoids LTB4 and TXA2 were significantly increased after BaV injection (250 microg/kg), whereas no increment in IL-1 was observed. Anti-mouse l-selectin, LFA-1, ICAM-1, PECAM-1 and beta2 integrin chain monoclonal antibodies resulted in a reduction of neutrophil accumulation induced by BaV injection compared with isotype-matched control injected animals. These data suggest that BaV is able to induce the activation of leukocytes and endothelium to express adhesion molecules involved in the recruitment of neutrophils into the inflammed site. Furthermore, these results showed that BaV induces the release of cytokines and eicosanoids in the local of the venom injection; these inflammatory mediators may be important for the initiation and amplification of the inflammatory reaction characteristic from Bothrops sp envenomation.

  13. Evaluation of platelet number and function and fibrinogen level in patients bitten by snakes of the Bothrops genus

    Directory of Open Access Journals (Sweden)

    Fábio Cardoso Luan

    1995-03-01

    Full Text Available Platelet function and plasma fibrinogen levels were evaluated in 14 patients, 10 males and 4females, aged 13-59years bitten by Bothrops genus snakes. There was a statistical difference (p Foram avaliadas a função plaquetária e os níveis séricos de fibrinogênio em 14 doentes picados por serpentes do gênero Bothrops, sendo 10 do sexo masculino e 4 do sexo feminino, com idades compreendidas entre 13 e 59 anos. Houve diferença estatística (p < 0,05 entre os níveis séricos defibrinogênio avaliados 24 e 48 horas após o acidente. Houve tendência à normalização após 48 horas do tratamento. A plaquetopenia foi evidente nas avaliações de 24 e 48 horas. Houve tendência à nomalização no 8o dia após o tratamento (p <0,05. Os níveis de produtos de degradação defibrina (PDF mostraram-se alterados em 83,33 % dos pacientes avaliados. Os autores sugerem que a hipoagregação esteja relacionada com níveis baixos de fibrinogênio e elevados de PDF.

  14. ACUTE KIDNEY INJURY CAUSED BY Crotalus AND Bothrops SNAKE VENOM: A REVIEW OF EPIDEMIOLOGY, CLINICAL MANIFESTATIONS AND TREATMENT

    Directory of Open Access Journals (Sweden)

    Polianna L.M.M. Albuquerque

    2013-09-01

    Full Text Available SUMMARY Ophidic accidents are an important public health problem due to their incidence, morbidity and mortality. An increasing number of cases have been registered in Brazil in the last few years. Several studies point to the importance of knowing the clinical complications and adequate approach in these accidents. However, knowledge about the risk factors is not enough and there are an increasing number of deaths due to these accidents in Brazil. In this context, acute kidney injury (AKI appears as one of the main causes of death and consequences for these victims, which are mainly young males working in rural areas. Snakes of the Bothrops and Crotalus genera are the main responsible for renal involvement in ophidic accidents in South America. The present study is a literature review of AKI caused by Bothrops and Crotalus snake venom regarding diverse characteristics, emphasizing the most appropriate therapeutic approach for these cases. Recent studies have been carried out searching for complementary therapies for the treatment of ophidic accidents, including the use of lipoic acid, simvastatin and allopurinol. Some plants, such as Apocynaceae, Lamiaceae and Rubiaceae seem to have a beneficial role in the treatment of this type of envenomation. Future studies will certainly find new therapeutic measures for ophidic accidents.

  15. Purification and Biochemical Characterization of Three Myotoxins from Bothrops mattogrossensis Snake Venom with Toxicity against Leishmania and Tumor Cells

    Directory of Open Access Journals (Sweden)

    Andréa A. de Moura

    2014-01-01

    Full Text Available Bothrops mattogrossensis snake is widely distributed throughout eastern South America and is responsible for snakebites in this region. This paper reports the purification and biochemical characterization of three new phospholipases A2 (PLA2s, one of which is presumably an enzymatically active Asp49 and two are very likely enzymatically inactive Lys49 PLA2 homologues. The purification was obtained after two chromatographic steps on ion exchange and reverse phase column. The 2D SDS-PAGE analysis revealed that the proteins have pI values around 10, are each made of a single chain, and have molecular masses near 13 kDa, which was confirmed by MALDI-TOF mass spectrometry. The N-terminal similarity analysis of the sequences showed that the proteins are highly homologous with other Lys49 and Asp49 PLA2s from Bothrops species. The PLA2s isolated were named BmatTX-I (Lys49 PLA2-like, BmatTX-II (Lys49 PLA2-like, and BmatTX-III (Asp49 PLA2. The PLA2s induced cytokine release from mouse neutrophils and showed cytotoxicity towards JURKAT (leukemia T and SK-BR-3 (breast adenocarcinoma cell lines and promastigote forms of Leishmania amazonensis. The structural and functional elucidation of snake venoms components may contribute to a better understanding of the mechanism of action of these proteins during envenomation and their potential pharmacological and therapeutic applications.

  16. ACUTE KIDNEY INJURY CAUSED BY Crotalus AND Bothrops SNAKE VENOM: A REVIEW OF EPIDEMIOLOGY, CLINICAL MANIFESTATIONS AND TREATMENT

    Science.gov (United States)

    Albuquerque, Polianna L.M.M.; Jacinto, Camilla N.; Silva, Geraldo B.; Lima, Juliana B.; Veras, Maria do Socorro B.; Daher, Elizabeth F.

    2013-01-01

    SUMMARY Ophidic accidents are an important public health problem due to their incidence, morbidity and mortality. An increasing number of cases have been registered in Brazil in the last few years. Several studies point to the importance of knowing the clinical complications and adequate approach in these accidents. However, knowledge about the risk factors is not enough and there are an increasing number of deaths due to these accidents in Brazil. In this context, acute kidney injury (AKI) appears as one of the main causes of death and consequences for these victims, which are mainly young males working in rural areas. Snakes of the Bothrops and Crotalus genera are the main responsible for renal involvement in ophidic accidents in South America. The present study is a literature review of AKI caused by Bothrops and Crotalus snake venom regarding diverse characteristics, emphasizing the most appropriate therapeutic approach for these cases. Recent studies have been carried out searching for complementary therapies for the treatment of ophidic accidents, including the use of lipoic acid, simvastatin and allopurinol. Some plants, such as Apocynaceae, Lamiaceae and Rubiaceae seem to have a beneficial role in the treatment of this type of envenomation. Future studies will certainly find new therapeutic measures for ophidic accidents. PMID:24037282

  17. Isolation and Biochemical Characterization of a New Thrombin-Like Serine Protease from Bothrops pirajai Snake Venom

    Directory of Open Access Journals (Sweden)

    Kayena D. Zaqueo

    2014-01-01

    Full Text Available This paper presents a novel serine protease (SP isolated from Bothrops pirajai, a venomous snake found solely in Brazil that belongs to the Viperidae family. The identified SP, named BpirSP-39, was isolated by three chromatographic steps (size exclusion, bioaffinity, and reverse phase chromatographies. The molecular mass of BpirSP-39 was estimated by SDS-PAGE and confirmed by mass spectrometry (39,408.32 Da. The protein was able to form fibrin networks, which was not observed in the presence of serine protease inhibitors, such as phenylmethylsulfonyl fluoride (PMSF. Furthermore, BpirSP-39 presented considerable thermal stability and was apparently able to activate factor XIII of the blood coagulation cascade, unlike most serine proteases. BpirSP-39 was capable of hydrolyzing different chromogenic substrates tested (S-2222, S-2302, and S-2238 while Cu2+ significantly diminished BspirSP-39 activity on the three tested substrates. The enzyme promoted platelet aggregation and also exhibited fibrinogenolytic, fibrinolytic, gelatinolytic, and amidolytic activities. The multiple alignment showed high sequence similarity to other thrombin-like enzymes from snake venoms. These results allow us to conclude that a new SP was isolated from Bothrops pirajai snake venom.

  18. PROVENANCE SNAKES OF GENUS Bothrops AND Crotalus (VIPERIDAE CATALOGED IN SERPENTARIUM ZOO UNIVERSITY OF CAXIAS DO SUL (UCS

    Directory of Open Access Journals (Sweden)

    Ezequiel Capeletti

    2016-09-01

    Full Text Available Snakes are reptiles that have large environmental adaptations, which favored it's distribution among the various ecosystems. In Brazil, there are found 392 species of snakes, while in Rio Grande do Sul (RS, this research field, 79 species have been described, of which there is the Viperidae family. The objective of this work is to verify the origin of snakes of genus Bothrops and Crotalus, according to the records of the last 15 years of the serpentarium the University of Caxias do Sul, representing the first survey in captivity of the northeastern region of RS. By extension, we seek to investigate whether there is a relationship between the amount of snakes records and abiotic factors: precipitation, temperature and humidity during the period. The research was developed at UCS's serpentarium, by documental analysis of notebook records describing serpents entries between the years 2000 and 2014, in addition to literature review that helped to understand the distribution of the species and gather of the environmental parameters at the Instituto Nacional de Metereologia for further analysis. It was found that the received species are distributed in 26 cities in the RS state, of which the highlights are the municities of Nova Petrópolis and Caxias do Sul, with 148 and 42 individuals respectively. Further, it was found that there was no relationship between the amount of animals and registered temperature, precipitation and humidity. Keywords: Geographic distribution; Bothrops; Crotalus; abiotic factors; northeastern RS.

  19. Effects of irradiated Bothropstoxin-1 and Bothrops jararacussu crude venom on the immune system

    International Nuclear Information System (INIS)

    Caproni, Priscila

    2009-01-01

    Ionizing radiation has been successfully employed to modify the immunological properties of biomolecules and has been proven to be a powerful tool to attenuate snake venoms toxicity without affecting and even increasing their immunogenic properties. Very promising results were obtained when crude animal venoms, as well as isolated toxins, were treated with 60 Co gamma rays, yielding toxoids with good immunogenicity, however, little is known about the modifications that irradiated molecules undergo and even less about the immunological response that such antigens elicit. At the present work, we have evaluated the effects on immune system of B10.PL and BALB/c mice of Bothrops jararacussu crude venom and isolated bothropstoxin-1 (Bthx-1), before and after gamma radiation exposition. According to our data, irradiation process promoted structural modifications on both isolated toxin and crude venom, characterized by higher molecular weight protein (aggregates and oligomers) formation. Irradiated samples were immunogenic and the antibodies elicited by them were able to recognize the native toxin in ELISA. These results indicate that irradiation of toxic proteins can promote significant modifications in their structures, but still retain many of the original antigenic and immunological properties. Also, our data indicate that the irradiated protein induced higher titers of IgG2b, suggesting that Th1 cells were predominantly involved. Results from Western blot assay showed that antibodies raised against irradiated bothropstoxin-1 recognize both native isolated toxin or crude venom. Cytotoxicity assay showed that irradiated toxin and crude venom were less toxic than their native counterpart. Thus, the viability of the macrophages cultured in the presence of irradiated Bthx-1 or crude venom was higher if compared with their native forms. LDH Assay showed that irradiated Bthx-1 promotes less muscular damage than the native form. Our data confirm a potential use of ionizing

  20. A C-Type Lectin from Bothrops jararacussu Venom Disrupts Staphylococcal Biofilms

    Science.gov (United States)

    Klein, Raphael Contelli; Fabres-Klein, Mary Hellen; de Oliveira, Leandro Licursi; Feio, Renato Neves; Malouin, François; Ribon, Andréa de Oliveira Barros

    2015-01-01

    Bovine mastitis is a major threat to animal health and the dairy industry. Staphylococcus aureus is a contagious pathogen that is usually associated with persistent intramammary infections, and biofilm formation is a relevant aspect of the outcome of these infections. Several biological activities have been described for snake venoms, which led us to screen secretions of Bothrops jararacussu for antibiofilm activity against S. aureus NRS155. Crude venom was fractionated by size-exclusion chromatography, and the fractions were tested against S. aureus. Biofilm growth, but not bacterial growth, was affected by several fractions. Two fractions (15 and 16) showed the best activities and were also assayed against S. epidermidis NRS101. Fraction 15 was identified by TripleTOF mass spectrometry as a galactose-binding C-type lectin with a molecular weight of 15 kDa. The lectin was purified from the crude venom by D-galactose affinity chromatography, and only one peak was observed. This pure lectin was able to inhibit 75% and 80% of S. aureus and S. epidermidis biofilms, respectively, without affecting bacterial cell viability. The lectin also exhibited a dose-dependent inhibitory effect on both bacterial biofilms. The antibiofilm activity was confirmed using scanning electron microscopy. A pre-formed S. epidermidis biofilm was significantly disrupted by the C-type lectin in a time-dependent manner. Additionally, the lectin demonstrated the ability to inhibit biofilm formation by several mastitis pathogens, including different field strains of S. aureus, S. hyicus, S. chromogenes, Streptococcus agalactiae, and Escherichia coli. These findings reveal a new activity for C-type lectins. Studies are underway to evaluate the biological activity of these lectins in a mouse mastitis model. PMID:25811661

  1. A C-type lectin from Bothrops jararacussu venom disrupts Staphylococcal biofilms.

    Directory of Open Access Journals (Sweden)

    Raphael Contelli Klein

    Full Text Available Bovine mastitis is a major threat to animal health and the dairy industry. Staphylococcus aureus is a contagious pathogen that is usually associated with persistent intramammary infections, and biofilm formation is a relevant aspect of the outcome of these infections. Several biological activities have been described for snake venoms, which led us to screen secretions of Bothrops jararacussu for antibiofilm activity against S. aureus NRS155. Crude venom was fractionated by size-exclusion chromatography, and the fractions were tested against S. aureus. Biofilm growth, but not bacterial growth, was affected by several fractions. Two fractions (15 and 16 showed the best activities and were also assayed against S. epidermidis NRS101. Fraction 15 was identified by TripleTOF mass spectrometry as a galactose-binding C-type lectin with a molecular weight of 15 kDa. The lectin was purified from the crude venom by D-galactose affinity chromatography, and only one peak was observed. This pure lectin was able to inhibit 75% and 80% of S. aureus and S. epidermidis biofilms, respectively, without affecting bacterial cell viability. The lectin also exhibited a dose-dependent inhibitory effect on both bacterial biofilms. The antibiofilm activity was confirmed using scanning electron microscopy. A pre-formed S. epidermidis biofilm was significantly disrupted by the C-type lectin in a time-dependent manner. Additionally, the lectin demonstrated the ability to inhibit biofilm formation by several mastitis pathogens, including different field strains of S. aureus, S. hyicus, S. chromogenes, Streptococcus agalactiae, and Escherichia coli. These findings reveal a new activity for C-type lectins. Studies are underway to evaluate the biological activity of these lectins in a mouse mastitis model.

  2. Effects of Bothrops asper snake venom on lymphatic vessels: insights into a hidden aspect of envenomation.

    Science.gov (United States)

    Mora, Javier; Mora, Rodrigo; Lomonte, Bruno; Gutiérrez, José María

    2008-01-01

    Envenomations by the snake Bothrops asper represent a serious medical problem in Central America and parts of South America. These envenomations concur with drastic local tissue pathology, including a prominent edema. Since lymph flow plays a role in the maintenance of tissue fluid balance, the effect of B. asper venom on collecting lymphatic vessels was studied. B. asper venom was applied to mouse mesentery, and the effects were studied using an intravital microscopy methodology coupled with an image analysis program. B. asper venom induced a dose-dependent contraction of collecting lymphatic vessels, resulting in a reduction of their lumen and in a halting of lymph flow. The effect was reproduced by a myotoxic phospholipase A(2) (PLA(2)) homologue isolated from this venom, but not by a hemorrhagic metalloproteinase or a coagulant thrombin-like serine proteinase. In agreement with this, treatment of the venom with fucoidan, a myotoxin inhibitor, abrogated the effect, whereas no inhibition was observed after incubation with the peptidomimetic metalloproteinase inhibitor Batimastat. Moreover, fucoidan significantly reduced venom-induced footpad edema. The myotoxic PLA(2) homologue, known to induce skeletal muscle necrosis, was able to induce cytotoxicity in smooth muscle cells in culture and to promote an increment in the permeability to propidium iodide in these cells. Our observations indicate that B. asper venom affects collecting lymphatic vessels through the action of myotoxic PLA(2)s on the smooth muscle of these vessels, inducing cell contraction and irreversible cell damage. This activity may play an important role in the pathogenesis of the pronounced local edema characteristic of viperid snakebite envenomation, as well as in the systemic biodistribution of the venom, thus representing a potential therapeutical target in these envenomations.

  3. Thrombelastographic characterization of the thrombin-like activity of Crotalus simus and Bothrops asper venoms.

    Science.gov (United States)

    Nielsen, Vance G; Boyer, Leslie V; Redford, Daniel T; Ford, Paul

    2017-04-01

    : Annually, thousands suffer venomous snake-bite from Crotalus simus and Bothrops asper vipers in central and South America. The goals of the present study were to generally characterize the thrombin-like effects of venom from these snakes in human plasma with viscoelastic methods. Human plasma was exposed to the venom of three different C. simus subspecies and venoms obtained from B. asper vipers located in three different locations in Mexico. To characterize the factor X-activating and thrombin-like activity of these venoms, plasma (normal or factor XIII deficient) was pretreated with a variety of additives (e.g., heparin) in the absence or presence of calcium prior to exposure to 2.0 μg/ml of each viper's venom. These profiles were compared with plasma without venom that had contact activation of coagulation. Coagulation kinetics were determined with thrombelastography. All venoms had thrombin-like activity, with C. s. simus creating a slow growing, weak clot that was likely mediated by metalloproteinases. In contrast, B. asper venoms had rapid onset of coagulation and a high velocity of thrombus growth. Further, B. asper venom activity was calcium-independent, activated prothrombin, activated factor XIII, and independently polymerized fibrinogen. The viscoelastic methods used were able to differentiate subspecies of C. simus and specimens of B. asper, and provide insight into the mechanisms by which the venoms acted on plasma. These methods may be useful in the profiling of similar venoms and perhaps can assist in the assessment of interventions designed to treat envenomation (e.g., antivenom).

  4. Effects of Bothrops asper snake venom on lymphatic vessels: insights into a hidden aspect of envenomation.

    Directory of Open Access Journals (Sweden)

    Javier Mora

    Full Text Available Envenomations by the snake Bothrops asper represent a serious medical problem in Central America and parts of South America. These envenomations concur with drastic local tissue pathology, including a prominent edema. Since lymph flow plays a role in the maintenance of tissue fluid balance, the effect of B. asper venom on collecting lymphatic vessels was studied.B. asper venom was applied to mouse mesentery, and the effects were studied using an intravital microscopy methodology coupled with an image analysis program. B. asper venom induced a dose-dependent contraction of collecting lymphatic vessels, resulting in a reduction of their lumen and in a halting of lymph flow. The effect was reproduced by a myotoxic phospholipase A(2 (PLA(2 homologue isolated from this venom, but not by a hemorrhagic metalloproteinase or a coagulant thrombin-like serine proteinase. In agreement with this, treatment of the venom with fucoidan, a myotoxin inhibitor, abrogated the effect, whereas no inhibition was observed after incubation with the peptidomimetic metalloproteinase inhibitor Batimastat. Moreover, fucoidan significantly reduced venom-induced footpad edema. The myotoxic PLA(2 homologue, known to induce skeletal muscle necrosis, was able to induce cytotoxicity in smooth muscle cells in culture and to promote an increment in the permeability to propidium iodide in these cells.Our observations indicate that B. asper venom affects collecting lymphatic vessels through the action of myotoxic PLA(2s on the smooth muscle of these vessels, inducing cell contraction and irreversible cell damage. This activity may play an important role in the pathogenesis of the pronounced local edema characteristic of viperid snakebite envenomation, as well as in the systemic biodistribution of the venom, thus representing a potential therapeutical target in these envenomations.

  5. Radioiodination and biodistribution of Leucurolysin-B isolated from Bothrops Leucurus in mice bearing Ehrlich

    International Nuclear Information System (INIS)

    Gabriel, L.M.; Soares, M.A.; Bicalho, M.S.; Santos, R.G.; Sanchez, E.O.F.; Silva, S.G.

    2009-01-01

    Integrins are family of heterodimeric cell surface adhesion receptors able to recognize and bind to proteins in the extracellular matrix (ECM). This recognition is mainly through the RGD domain present in both the cell surface as the protein in the ECM. Various integrins have been identified as regulators of tumor progression. The RGD domain is also found in some snake venoms named disintegrins. Disintegrins inhibit cell-matrix and a cell-cell interaction mediated by integrin and has been shown that these proteins are able to inhibit metastasis in processes dependent on integrin. The disintegrin-like (ECD), as well as RGD-disintegrin are also able to bind to cell surface integrins and inhibit their adherence to the natural ligands. Leucurolysin-B (Leuc-B) is a metalloproteinase class P-III isolated from Bothrops leucurus (BLV) and possesses a disintegrin-like domain (ECD). The goals of this work were to synthesize a radioactive probe analog to Leuc-B using radioiodine 125 I and evaluate the interaction of 125 I-Leuc-B in tumor cells through the study of biodistribution in animals bearing Ehrlich tumor.125I-Leuc-B was synthesized using lactoperoxidase with high yield (90%) and specific activity of 1.2x10-7Bq/mmol. It was observed that 125 I-Leuc-B had very fast clearance from the blood stream (T1/2= 0.01 h). Tumor uptake of 125I-Leuc-B gradually increased up to (2 min) and remained for a quite long period. The tumor/normal tissue uptake ratios of 125 I-Leuc-B were 1.77 (tumor/normal paw) and 8.44 tumor/skeletal muscle. The results suggest that 125 I-Leuc- B may constitute a good template for development of a tool for detection of solid tumors. (author)

  6. Crystal structure of myotoxin-II: a myotoxic phospholipase A2 - homologue from Bothrops moojeni venom

    International Nuclear Information System (INIS)

    Azevedo, W.F.; Ward, R.J.; Lombardi, F.R.; Arni, R.K.; Soares, A.M.; Giglio, J.R.; Fontes, M.R.M.

    1997-01-01

    Full text. Phospho lipases A2 (PLA 2 ; E C 3.1.1.4, phosphatides s n-2 acyl hydrolases) hydrolysis the s n-2 ester bond of phospholipids showing enhanced activity at lamellar or membrane surfaces. Intracellular PLA 2 s are involved at phospholipid metabolism and signal transduction, whereas extracellular PLA 2 s are found in mammalian pancreatic juices, the venoms of snakes, lizards and insects. Based on their high primary sequence similarity, extracellular PLA 2 s are separated into Classes I, II and III. Class II PLA 2 s are found in snake venoms of Crotalidae an Viperidae species, and include the sub-family of Lys PLA 2 s homologue. he coordination of the Ca 2+ ion in the PLA 2 calcium-binding loop includes and aspartate at position 49. In the catalytically active PLA 2 s, this calcium ion plays a critical role in the stabilization of the tetrahedral transition state intermediate in the catalytic mechanism. The conservative substitution Asp49-Lys results in a decreased calcium affinity with a concomitant loss of catalytic activity, and naturally occurring PLA 2 s-homologues showing the same substitution are catalytically inactive. However, the Lys PLA 2 s possess cytolytic and myotoxic activities and furthermore retain the ability to disrupt the integrity of both plasma membranes and model lipid layers by a ca 2+ -independent mechanism for which there is no evidence of lipid hydrolysis. Lys 49 PLA 2 homologues have been isolated from several Bothrops spp. venoms including B. moojeni. Therefore, in order to improve our understanding of the molecular basis of the myotoxic and Ca 2+ independent membrane damaging activities we have determined the crystal structure of MjTX-II, a Lys 49 homologue from the venom of B. moojeni. The model presented has been determined at 2.0 A resolution and refined to a crystallographic residual of 19.7% (R f ree=28.1%). (author)

  7. Neuromuscular activity of Bothrops neuwiedi pauloensis snake venom in mouse nerve-muscle preparations

    Directory of Open Access Journals (Sweden)

    A. M. Durigon

    2005-03-01

    Full Text Available The pharmacological effects of Bothrops neuwiedi pauloensis venom on mouse phrenic nerve-diaphragm (PND preparations were studied. Venom (20 mug/ml irreversibly inhibited indirectly evoked twitches in PND preparations (60 ± 10% inhibition, mean ± SEM; p<0.05; n=6. At 50 mug/ml, the venom blocked indirectly and directly (curarized preparations evoked twitches in mouse hemidiaphragms. In the absence of Ca2+, venom (50 mug/ml, produced partial blockade only after an 80 min incubation, which reached 40.3 ± 7.8% (p<0.05; n=3 after 120 min. Venom (20 mug/ml increased (25 ± 2%, p< 0.05 the frequency of giant miniature end-plate potentials in 9 of 10 end-plates after 30 min and the number of miniature end-plate potentials which was maximum (562 ± 3%, p<0.05 after 120 min. During the same period, the resting membrane potential decreased from - 81 ± 1.4 mV to - 41.3 ± 3.6 mV 24 fibers; p<0.01; n=4 in the end-plate region and from - 77.4 ± 1.4 to -44.6 ± 3.9 mV (24 fibers; p<0.01; n=4 in regions distant from the end-plate. These results indicate that B. n. pauloensis venom acts primarily at presynaptic sites. They also suggest that enzymatic activity may be involved in this pharmacological action.

  8. Biochemical and functional characterization of Bothropoidin: the first haemorrhagic metalloproteinase from Bothrops pauloensis snake venom.

    Science.gov (United States)

    Gomes, Mário Sérgio R; Naves de Souza, Dayane L; Guimarães, Denise O; Lopes, Daiana S; Mamede, Carla C N; Gimenes, Sarah Natalie C; Achê, David C; Rodrigues, Renata S; Yoneyama, Kelly A G; Borges, Márcia H; de Oliveira, Fábio; Rodrigues, Veridiana M

    2015-03-01

    We present the biochemical and functional characterization of Bothropoidin, the first haemorrhagic metalloproteinase isolated from Bothrops pauloensis snake venom. This protein was purified after three chromatographic steps on cation exchange CM-Sepharose fast flow, size-exclusion column Sephacryl S-300 and anion exchange Capto Q. Bothropoidin was homogeneous by SDS-PAGE under reducing and non-reducing conditions, and comprised a single chain of 49,558 Da according to MALDI TOF analysis. The protein presented an isoelectric point of 3.76, and the sequence of six fragments obtained by MS (MALDI TOF\\TOF) showed a significant score when compared with other PIII Snake venom metalloproteinases (SVMPs). Bothropoidin showed proteolytic activity on azocasein, Aα-chain of fibrinogen, fibrin, collagen and fibronectin. The enzyme was stable at pH 6-9 and at lower temperatures when assayed on azocasein. Moreover, its activity was inhibited by EDTA, 1.10-phenanthroline and β-mercaptoethanol. Bothropoidin induced haemorrhage [minimum haemorrhagic dose (MHD) = 0.75 µg], inhibited platelet aggregation induced by collagen and ADP, and interfered with viability and cell adhesion when incubated with endothelial cells in a dose and time-dependent manner. Our results showed that Bothropoidin is a haemorrhagic metalloproteinase that can play an important role in the toxicity of B. pauloensis envenomation and might be used as a tool for studying the effects of SVMPs on haemostatic disorders and tumour metastasis. © The Authors 2014. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.

  9. Ontogenetic variations in the venom proteome of the Amazonian snake Bothrops atrox

    Directory of Open Access Journals (Sweden)

    Sousa Marcelo V

    2006-05-01

    Full Text Available Abstract Background Bothrops atrox is responsible for the majority of snakebite accidents in the Brazilian Amazon region. Previous studies have demonstrated that the biological and pharmacological activities of B. atrox venom alter with the age of the animal. Here, we present a comparative proteome analysis of B. atrox venom collected from specimens of three different stages of maturation: juveniles, sub-adults and adults. Results Optimized conditions for two-dimensional gel electrophoresis (2-DE of pooled venom samples were achieved using immobilized pH gradient (IPG gels of non-linear 3–10 pH range during the isoelectric focusing step and 10–20% gradient polyacrylamide gels in the second dimension. Software-assisted analysis of the 2-DE gels images demonstrated differences in the number and intensity of spots in juvenile, sub-adult and adult venoms. Although peptide mass fingerprinting (PMF failed to identify even a minor fraction of spots, it allowed us to group spots that displayed similar peptide maps. The spots were subjected to a combination of tandem mass spectrometry and Mascot and MS BLAST database searches that identified several classes of proteins, including metalloproteinases, serine proteinases, lectins, phospholipases A2, L-amino oxidases, nerve growth factors, vascular endothelial growth factors and cysteine-rich secretory proteins. Conclusion The analysis of B. atrox samples from specimens of different ages by 2-DE and mass spectrometry suggested that venom proteome alters upon ontogenetic development. We identified stage specific and differentially expressed polypeptides that may be responsible for the activities of the venom in each developmental stage. The results provide insight into the molecular basis of the relation between symptomatology of snakebite accidents in humans and the venom composition. Our findings underscore the importance of the use of venoms from individual specimen at various stages of maturation for

  10. Preparación toxoide a partir de la fracción hemorrágica del veneno de Bothrops asper (serpiente de América Central y del Sur) (Toxoid preparation from hemorrhagic fraction of the venom from Bothrops asper (snake from Central and South America).

    Science.gov (United States)

    Rodríguez-Acosta, A; Aguilar, I; Girón, M E

    1993-01-01

    A technique is described for preparing a toxoid from the hemorrhagic fraction of the Bothrops asper venom. This method conserves a high degree of immunogenicity although it eliminates lethal effects. None of the animals vaccinated with the toxoid from this fraction had hemorrhagic lesions after they were injected the venom from the hemorrhagic fraction.

  11. Immunogenicity of Bothrops atrox (Ophidia: Viperidae venom and its evaluation by immunoenzymatic methods Inmunogenicidad del veneno de Bothrops atrox (Ophidia: Viperidae y su evaluación por métodos inmunoenzimáticos

    Directory of Open Access Journals (Sweden)

    Gustavo A. Sandoval

    2012-02-01

    Full Text Available The immunogenicity of Bothrops atrox, “jergón”, venom was studied using ELISA and Western Blot methods, as well as cross-reactivity patterns against venoms of Bothrops brazili, Lachesis muta and Crotalus durissus. For this purpose, New Zealand white rabbits (2 kg aprox were immunized with four 500 μg doses of B. atrox venom in a period of 90 days. Antibody production was followed using ELISA technique, and title of hiper-immune serum was determined at the end of immunization protocol. Additionally, electrophoretic patterns of venoms were analyzed by SDS-PAGE and venom reactivity against obtained serum by ELISA and Western Blot. Immunization schedule allowed a pronounced antibody production since day 20 of protocol. At the end of process, serum title was 256000, which demonstrated both efficacy and usefulness of the developed procedure. On the other hand, studied venoms showed a heterogenic protein composition according to their electrophoretic patterns, whereas cross-reactivity values of 23,7%, 4,0% and 1,8% were obtained between B. atrox venom and B. brazili, L. muta and C. durissus venoms, respectively, using immunoenzymatic methods. According to our results, this procedure constitutes an initial step for further assays directed to optimization in immunoserum production for envenoming treatment and development of kits for diagnosis and species identification of snakes.Se estudió la inmunogenicidad del veneno de la serpiente Bothrops atrox, “jergón”, utilizando los métodos inmunoenzimáticos de ELISA y Western Blot, así como los patrones de reactividad cruzada empleando los venenos de las serpientes Bothrops brazili, Lachesis muta y Crotalus durissus. Para este fin se inmunizaron conejos albinos Nueva Zelanda (2 kg aprox con cuatro dosis de 500 μg del veneno de B. atrox en un periodo de 90 días. La producción de anticuerpos fue monitoreada mediante la técnica de ELISA, determinándose el título del suero hiperinmune obtenido

  12. Snake venomics of the lancehead pitviper Bothrops asper: geographic, individual, and ontogenetic variations.

    Science.gov (United States)

    Alape-Girón, Alberto; Sanz, Libia; Escolano, José; Flores-Díaz, Marietta; Madrigal, Marvin; Sasa, Mahmood; Calvete, Juan J

    2008-08-01

    We report the comparative proteomic characterization of the venoms of adult and newborn specimens of the lancehead pitviper Bothrops asper from two geographically isolated populations from the Caribbean and the Pacific versants of Costa Rica. The crude venoms were fractionated by reverse-phase HPLC, followed by analysis of each chromatographic fraction by SDS-PAGE, N-terminal sequencing, MALDI-TOF mass fingerprinting, and collision-induced dissociation tandem mass spectrometry of tryptic peptides. The two B. asper populations, separated since the late Miocene or early Pliocene (8-5 mya) by the Guanacaste Mountain Range, Central Mountain Range, and Talamanca Mountain Range, contain both identical and different (iso)enzymes from the PLA 2, serine proteinase, and SVMP families. Using a similarity coefficient, we estimate that the similarity of venom proteins between the two B. asper populations may be around 52%. Compositional differences between venoms among different geographic regions may be due to evolutionary environmental pressure acting on isolated populations. To investigate venom variability among specimens from the two B. asper populations, the reverse-phase HPLC protein profiles of 15 venoms from Caribbean specimens and 11 venoms from snakes from Pacific regions were compared. Within each B. asper geographic populations, all major venom protein families appeared to be subjected to individual variations. The occurrence of intraspecific individual allopatric variability highlights the concept that a species, B. asper in our case, should be considered as a group of metapopulations. Analysis of pooled venoms of neonate specimens from Caribbean and Pacific regions with those of adult snakes from the same geographical habitat revealed prominent ontogenetic changes in both geographical populations. Major ontogenetic changes appear to be a shift from a PIII-SVMP-rich to a PI-SVMP-rich venom and the secretion in adults of a distinct set of PLA 2 molecules than in

  13. Intraspecies geographical variability in the South American tigra mariposa (Bothrops venezuelensis Sandner 1952) snake venom activities.

    Science.gov (United States)

    Girón, María E; Padrón, Vanessa; Ramos, María I; Sánchez, Elda E; Guerrero, Belsy; García, Alberto; Uzcátegui, Néstor L; Navarrete, Luis F; Rodríguez-Acosta, Alexis

    2018-03-15

    Bothrops venezuelensis snake venoms, from five localities in the North-Central Venezuelan regions, showed biochemical and haemostatic differences. In this study, bioactivities of B. venezuelensis venoms from different regions (Aragua state; Waraira Repano (Capital District); Baruta, La Boyera and Lagunetica (Miranda state)) were compared using both natural and synthetic substrates. The protein contents of these venoms were Lagunetica 89%, La Boyera 79%, Baruta 71%, Waraira Repano 68% and Aragua 64%. Toxic activities effects were: Intraperitoneal LD 50 s: Aragua-14 mg/kg; Waraira Repano-6.4 mg/kg; Baruta: 8.3 mg/kg; La Boyera-4.4 mg/kg; Lagunetica-16.2 mg/kg. The MHD results: Aragua-21.4 μg/mouse; Waraira Repano-2.5 μg/mouse; Baruta-1.2 μg/mouse; La Boyera-1.4 μg/mouse and Lagunetica-12 μg/mouse. The hide powder azure results: Aragua-1.24 U/mg; La Boyera-2.26 U/mg; Baruta-2.83 U/mg; Lagunetica-3.28 U/mg and Waraira Repano-5.77 U/mg. Esterase specific activity on BAEE results: Waraira Repano-666.66 U/mg; La Boyera-805.5 U/mg; Baruta-900.00 U/mg; Lagunetica-922.19 U/mg and Aragua-1960.67 U/mg. Casein zymography showed digestion bands in the molecular weight above 100 and at 66.2 and 21.5 kDa. Analysis of casein degradation by SDS-PAGE showed two different degradation patterns. Fibrinolytic activity (mm 2 /μg) on fibrin plates results: Aragua-6.07; Lagunetica-27.6; Waraira Repano-35.7; La Boyera-44.27 and Baruta-45.63. In the fibrinogenolytic assay, the five venoms completely degraded the α chain after 1 min of incubation. None of the venoms completely degraded the β and γ chains after 24 h incubation. The research indicated that venoms of B. venezuelensis of different geographic areas in Venezuela exhibit variances in composition and component concentrations; except the Aragua venom, all of them had high proteolytic activities. Copyright © 2018 Elsevier Ltd. All rights reserved.

  14. Distribution of {sup 131} I- labeled Bothrops erythromelas venom in mice

    Energy Technology Data Exchange (ETDEWEB)

    Vasconcelos, C.M.L.; Valenca, R.C.; Araujo, E.A. [Pernambuco Univ., Recife, PE (Brazil). Dept. de Biofisica e Radiobiologia; Modesto, J.C.A.; Pontes, M.M.; Guarnieri, M.C. [Pernambuco Univ., Recife, PE (Brazil). Dept. de Zoologia; Brazil, T.K. [Bahia Univ., Salvador, BA (Brazil). Inst. de Biologia

    1998-03-01

    Bothrops erythromelas is responsible for many snake bites in northeastern Brazil. In the present study we determined the in vivo distribution of the venom following its subcutaneous injection into mice. B. erythromelas venom and albumin were labeled individually with {sup 131} I by the chloramine T method, and separated in a Sephacryl S-200 column. The efficiency of labeling was 68%.Male Swiss mice (40-45 g), which had been provided with drinking water containing 0.05% KI over a period of 10 days prior to the experiment, were inoculated dorsally (sc) with 0.3 ml (2.35 x 10{sup 5} cpm/mouse) of {sup 131} I-venom (N = 42), {sup 131} -albumin or {sup 131} I (controls, N = 28 each). Thirty minutes and 1,3, 6, 12, 18 and 24 h after inoculation, the animals were perfused with 0.85% Na Cl and skin and various organs were collected in order to determine radioactivity content. There was a high rate of venom absorption int he skin (51%) within the first 30 min compared to albumin (20.1%) and free iodine (8.2%). Up to the third hour after injection there was a tendency for venom and albumin to concentrate in the stomach ( 3 rd h),small intestine (3 rd h) and large intestine (6th h). Both control groups had more radioactivity in the digestive tract, especially in the stomach, but these levels decreased essentially to baseline by 12-18 h postinjection. In the kidneys, the distribution profiles of venom, albumin and iodine were similar. Counts at 30 min postinjection were low in all three groups (1.37, 1.86 and 0.77, respectively), and diminished to essentially 0% by 12-18 h. Albumin tended to concentrate in muscle until the 3 rd h postinjection (1.98%).There was a low binding of labeled venom in the liver (B. erythromelas venom does not specifically target most internal organs). That is, the systemic effects of envenomation ar mainly due to an indirect action. (author) 14 refs., 2 figs.

  15. Ecology of a snake assemblage in the Atlantic Forest of southeastern Brazil

    Directory of Open Access Journals (Sweden)

    Paulo A. Hartmann

    2009-01-01

    Full Text Available The main objective of this study was to examine the natural history and the ecology of the species that constitute a snake assemblage in the Atlantic Rainforest, at Núcleo Picinguaba, Parque Estadual da Serra do Mar, located on the northern coast of the state of São Paulo, southeastern Brazil. The main aspects studied were: richness, relative abundance, daily and seasonal activity, and substrate use. We also provide additional information on natural history of the snakes. A total of 282 snakes, distributed over 24 species, belonging to 16 genera and four families, has been found within the area of the Núcleo Picinguaba. Species sampled more frequently were Bothrops jararaca and B. jararacussu. The methods that yielded the best results were time constrained search and opportunistic encounters. Among the abiotic factors analyzed, minimum temperature, followed by the mean temperature and the rainfall are apparently the most important in determining snake abundance. Most species presented a diet concentrated on one prey category or restricted to a few kinds of food items. The large number of species that feed on frogs points out the importance of this kind of prey as an important food resource for snakes in the Atlantic Rainforest. Our results indicate that the structure of the Picinguaba snake assemblage reflects mainly the phylogenetic constraints of each of its lineages.O principal objetivo deste estudo foi obter informações sobre a história natural e a ecologia das espécies que compõem uma taxocenoses de serpentes da Mata Atlântica, no Núcleo Picinguaba do Parque Estadual da Serra do Mar, localizado no litoral norte do estado no Estado de São Paulo, sudeste do Brasil. Os principais aspectos estudados foram: riqueza, abundância relativa de espécies, padrões de atividade diária e sazonal, utilização do ambiente e dieta. Um total de 282 serpentes, distribuídas em 24 espécies, pertencentes a 16 gêneros e quatro famílias, foi

  16. Protistas "incerta sedis" de ofídios do Brasil: Toddia França, 1912 e Pirhemocyton Chatton & Blanc, 1914: estudo comparativo "Incerta sedis" Portists from Brazilian snakes: Toddia França, 1912 and Pirhemocyton Chatton & Blanc, 1914: comparative study

    Directory of Open Access Journals (Sweden)

    Maria Auxiliadora de Sousa

    1973-01-01

    Full Text Available Dentre 683 serpentes procedentes de diferentes pontos do Brasil, 16 (2,4% apresentaram em seu sangue (em glóbulos vermelhos maturos e imaturos Toddia ou Pirhemocyton. Denominamos provisoriamente Toddia os parasitos encontrados em Bothrops moojeni, B. pradoi, B. jararaca e Chironius flavolineatus e Pirhemocyton os encontrados em B. alternatus; fornecemos a primeira referência nestes hospedeiros. Estes parasitos foram estudados comparativamente com o objetivo de estabelecermos a extensão de suas afinidades. Com Toddia de B. moojeni realizamos técnica citoquímica para caracterização de ácidos nucleicos e encontramos positividade apenas para DNA. As maiores afinidades entre os parasitos estudados residiam nas características dos corpúsculos cromáticos e alterações celulares que determinam; observarmos quase total identidade nas maneiras como os parasitos da mesma espécie de serpente se apresentaram. Constatamos a coexistência de inclusões globóides, cristalóides e formas intermediárias associada aos parasitos de B. pradoi e B. alternatus; em C. flavolineatus encotnramos além dos cristais, raríssimas formas intermediárias entre estes e globóides. Afinidades entre Toddia e Pirhemocyton abrangendo o tipo de inclusão contrariam as descrições originais, tornando pouco válida esta separação genérica. Discutimos também os fundamentos da criação das espécies e a possível natureza virótica destes parasitos.Among 683 snakes from several localities of Brazil, 16 (2,4% presented in their blood (in mature or immature red cells Toddia or Pirhemocyton. We named provisorily Toddia the parasites found in Bothrops moojeni, B. pradoi, B. jararaca and Chironius flavolineatus and Pirhemocyton those found in B. alternatus; we give the first reference in these hosts. These parasites were comparatively studied in view to establish the range of their relationship. With Toddia from B. moojeni we attempted nucleic acids by cytochemical

  17. Hemograma de cães envenenados experimentalmente com Bothrops alternatus após diferentes tratamentos.

    OpenAIRE

    D. O. Jacome; M. M. Melo; M. M. B. Santos; K. M. Ferreira; R. M. Sabaini

    2005-01-01

    RESUMO: O objetivo deste trabalho foi verificar a influência de diferentes tratamentos (soro antibotrópico, flunixina meglumina e extrato aquoso de C. longa) sobre o hemograma de cães envenenados experimentalmente com Bothrops alternatus. Foram utilizados 12 cães adultos, sem raça definida, inoculados com 0,3mg/kg do veneno via IM, tratados duas horas após o envenenamento, divididos em três grupos de 4 animais: Grupo I- soro antibotrópico diluído em solução fisiológica 0,9%, dose ...

  18. Isolation and characterization of a myotoxin from Bothrops brazili Hoge, 1953 Hoge, 1953 snake venom (Ophidia: Viperidae.

    Directory of Open Access Journals (Sweden)

    Carmen Pantigoso

    2014-06-01

    Full Text Available A myotoxin from the venom of the snake Bothrops brazili has been purified by ion-exchange chromatography on CM-Sephadex C-50 with 0,05 M ammonium acetate buffer pH 7. The homogeneity was evaluated by PAGE with and without SDS, immunodiffusion and immunoelectrophoresis. The myotoxin is a basic protein with 15,6% of Lys+Arg; it is not a glicoprotein, has not enzymatic activity, and corresponds to 25% of the whole venom protein. The molecular weight of the myotoxin was determined by PAGE-SDS and gel filtration chromatography. The myotoxin has 30 KDa of molecular weight and two polypeptide chains of 15 KDa each. Myotoxin produces a severe necrosis on the gastrocnemius muscle of white mice. The myotoxin does not have hemolytic nor anticoagulant activity. However, produces edema with a DEM of 32,6 mg of protein.

  19. Assessment of metalloproteinase inhibitors clodronate and doxycycline in the neutralization of hemorrhage and coagulopathy induced by Bothrops asper snake venom.

    Science.gov (United States)

    Rucavado, Alexandra; Henríquez, Mónica; García, Jonielle; Gutiérrez, José María

    2008-12-01

    Snake venom metalloproteinases (SVMPs) play a prominent role in the local and systemic manifestations of viperid snakebite envenomations. Thus, the possibility of using metalloproteinase inhibitors in the treatment of these envenomations is a promising therapeutic alternative. This study assessed the ability of two metalloproteinase inhibitors, the biphosphonate clodronate and the tetracycline doxycycline, to inhibit proteolytic, hemorrhagic, coagulant and defibrinogenating effects of Bothrops asper venom. Both compounds were able to inhibit these activities, at concentrations in the mM range, when incubated with venom prior to testing. However, when inhibition of hemorrhage was assessed in assays involving independent injection of venom and drug, inhibition was poor, even when these compounds were injected immediately after envenomation. These findings support the concept that the effectiveness of compounds, such as clodronate and doxycycline, whose inhibitory action on SVMPs is based on zinc chelation alone, is limited, and stress the view that more specific molecules are required for an effective inhibition of SVMPs in vivo.

  20. Edema induced by Bothrops asper (Squamata: Viperidae) snake venom and its inhibition by Costa Rican plant extracts.

    Science.gov (United States)

    Badilla, Beatriz; Chaves, Fernando; Mora, Gerardo; Poveda, Luis J

    2006-06-01

    We tested the capacity of leaf (Urera baccifera, Loasa speciosa, Urtica leptuphylla, Chaptalia nutans, and Satureja viminea) and root (Uncaria tomentosa) extracts to inhibit edema induced by Bothrops asper snake venom. Edema-forming activity was studied plethysmographically in the rat hind paw model. Groups of rats were injected intraperitoneally with various doses of each extract and, one hour later, venom was injected subcutaneously in the right hind paw. Edema was assessed at various time intervals. The edematogenic activity was inhibited in those animals that received an injection U. tomentosa, C. nutans or L. speciosa extract. The extract of U. baccifera showed a slight inhibition of the venom effect. Extract from S. viminea and, to a lesser extent that of U. leptuphylla, induced a pro-inflammatory effect, increasing the edema at doses of 250 mg/kg at one and two hours.

  1. Aislamiento y caracterización parcial de una miotoxina del veneno de la serpiente Bothrops atrox (Ophidia: Viperidae

    Directory of Open Access Journals (Sweden)

    Sergio Huatuco

    2013-06-01

    Full Text Available Se ha purificado una miotoxina del veneno de la serpiente Bothrops atrox, empleando una columna de intercambio catiónico de CM-Sephadex C-50 equilibrada con buffer acetato de amonio 0,05M a pH 7. La miotoxina es una proteína básica, y por cromatografía de filtración y PAGE-SDS se ha determinado que tiene un peso molecular de 27 kDa, estando formada por dos cadenas polipeptídicas de 14 kDa cada una. La inoculación de la miotoxina en el músculo gastrocnemius de ratones albinos produce la liberación de creatina kinasa así como la necrosis del tejido. La miotoxina tiene actividad de fosfolipasa, anticoagulante y edemática, pero no hemolítica.

  2. Description of Serpentirhabdias atroxi n. sp. (Nematoda: Rhabdiasidae), a parasite of Bothrops atrox (Linnaeus) (Reptilia: Serpentes: Viperidae) in Brazilian Amazonia.

    Science.gov (United States)

    Kuzmin, Yuriy; Giese, Elane Guerreiro; Melo, Francisco Tiago de Vasconcelos; da Costa, Paulo André Ferreira Borges; Maschio, Gleomar Fabiano; dos Santos, Jeannie Nascimento

    2016-01-01

    A new lung-dwelling nematode species is described from the common lancehead Bothrops atrox (Linnaeus) in the Brazilian Amazon Region. The species is assigned to the genus Serpentirhabdias Tkach, Kuzmin & Snyder, 2014 based on the presence of six lips arranged in two lateral groups, the absence of prominent cuticular inflations, and lung parasitism in snakes. Serpentirhabdias atroxi n. sp. differs from other species of the genus mainly by details of the morphology of the anterior end: cuticularised ring surrounding the anterior part of the buccal cavity and six minute onchia present in the oesophastome. Serpentirhabdias atroxi n. sp. is the seventh species of the genus known from the Neotropical Realm and the second species described from viperid snakes.

  3. Biochemical characterization, action on macrophages, and superoxide anion production of four basic phospholipases A2 from Panamanian Bothrops asper snake venom.

    Science.gov (United States)

    Rueda, Aristides Quintero; Rodríguez, Isela González; Arantes, Eliane C; Setúbal, Sulamita S; Calderon, Leonardo de A; Zuliani, Juliana P; Stábeli, Rodrigo G; Soares, Andreimar M

    2013-01-01

    Bothrops asper (Squamata: Viperidae) is the most important venomous snake in Central America, being responsible for the majority of snakebite accidents. Four basic PLA2s (pMTX-I to -IV) were purified from crude venom by a single-step chromatography using a CM-Sepharose ion-exchange column (1.5 × 15 cm). Analysis of the N-terminal sequence demonstrated that pMTX-I and III belong to the catalytically active Asp49 phospholipase A2 subclass, whereas pMTX-II and IV belong to the enzymatically inactive Lys49 PLA2s-like subclass. The PLA2s isolated from Panama Bothrops asper venom (pMTX-I, II, III, and IV) are able to induce myotoxic activity, inflammatory reaction mainly leukocyte migration to the muscle, and induce J774A.1 macrophages activation to start phagocytic activity and superoxide production.

  4. ESI-MS/MS Identification of a Bradykinin-Potentiating Peptide from Amazon Bothrops atrox Snake Venom Using a Hybrid Qq-oaTOF Mass Spectrometer

    Science.gov (United States)

    Coutinho-Neto, Antonio; Caldeira, Cleópatra A. S.; Souza, Gustavo H. M. F.; Zaqueo, Kayena D.; Kayano, Anderson M.; Silva, Rodrigo S.; Zuliani, Juliana P.; Soares, Andreimar M.; Stábeli, Rodrigo G.; Calderon, Leonardo A.

    2013-01-01

    A bradykinin-potentiating peptide (BPP) from Amazon Bothrops atrox venom with m/z 1384.7386 was identified and characterized by collision induced dissociation (CID) using an ESI-MS/MS spectra obtained in positive ion mode on a hybrid Qq-oaTOF mass spectrometer, Xevo G2 QTof MS (Waters, Manchester, UK). De novo peptide sequence analysis of the CID fragmentation spectra showed the amino acid sequence ZKWPRPGPEIPP, with a pyroglutamic acid and theoretical monoisotopic m/z 1384.7378, which is similar to experimental data, showing a mass accuracy of 0.6 ppm. The peptide is homologous to other BPP from Bothrops moojeni and was named as BPP-BAX12. PMID:23430539

  5. Biochemical Characterization, Action on Macrophages, and Superoxide Anion Production of Four Basic Phospholipases A2 from Panamanian Bothrops asper Snake Venom

    Directory of Open Access Journals (Sweden)

    Aristides Quintero Rueda

    2013-01-01

    Full Text Available Bothrops asper (Squamata: Viperidae is the most important venomous snake in Central America, being responsible for the majority of snakebite accidents. Four basic PLA2s (pMTX-I to -IV were purified from crude venom by a single-step chromatography using a CM-Sepharose ion-exchange column (1.5 × 15 cm. Analysis of the N-terminal sequence demonstrated that pMTX-I and III belong to the catalytically active Asp49 phospholipase A2 subclass, whereas pMTX-II and IV belong to the enzymatically inactive Lys49 PLA2s-like subclass. The PLA2s isolated from Panama Bothrops asper venom (pMTX-I, II, III, and IV are able to induce myotoxic activity, inflammatory reaction mainly leukocyte migration to the muscle, and induce J774A.1 macrophages activation to start phagocytic activity and superoxide production.

  6. Edema induced by Bothrops asper (Squamata: Viperidae snake venom and its inhibition by Costa Rican plant extracts

    Directory of Open Access Journals (Sweden)

    Beatriz Badilla

    2006-06-01

    Full Text Available We tested the capacity of leaf (Urera baccifera, Loasa speciosa, Urtica leptuphylla, Chaptalia nutans, and Satureja viminea and root (Uncaria tomentosa extracts to inhibit edema induced by Bothrops asper snake venom. Edema-forming activity was studied plethysmographically in the rat hind paw model. Groups of rats were injected intraperitoneally with various doses of each extract and, one hour later, venom was injected subcutaneously in the right hind paw. Edema was assessed at various time intervals. The edematogenic activity was inhibited in those animals that received an injection U. tomentosa, C. nutans or L. speciosa extract. The extract of U. baccifera showed a slight inhibition of the venom effect. Extract from S. viminea and, to a lesser extent that of U. leptuphylla, induced a pro-inflammatory effect, increasing the edema at doses of 250 mg/kg at one and two hours. Rev. Biol. Trop. 54(2: 245-252. Epub 2006 Jun 01.Se investigó la capacidad de los extractos de las hojas de Urera baccifera, Loasa speciosa, Urtica leptuphylla, Chaptalia nutans, Satureja viminea y de la raíz de Uncaria tomentosa para inhibir el edema inducido por el veneno de Bothrops asper por métodos pletismométricos. Los grupos de ratas fueron inyectados intraperitonealmente con varias dosis de cada extracto y una hora mas tarde se inyectó veneno por vía subcutánea en la pata trasera derecha de la rata. Se evaluó el edema en distintos intervalos de tiempo. Los resultados muestran que la actividad edematogénica fue inhibida en los animales que recibieron los extractos de raíz de U. tomentosa, hojas de C. nutans y L. speciosa. Los extractos de hojas de U. baccifera mostraron leve inhibición del efecto del veneno. El extracto de hojas de S. viminea y en menor grado el de U. leptuphylla indujeron un efecto pro inflamatorio.

  7. Bothrops asper snake venom and its metalloproteinase BaP-1 activate the complement system. Role in leucocyte recruitment.

    Science.gov (United States)

    Farsky, S H; Gonçalves, L R; Gutiérrez, J M; Correa, A P; Rucavado, A; Gasque, P; Tambourgi, D V

    2000-01-01

    The venom of the snake Bothrops asper, the most important poisonous snake in Central America, evokes an inflammatory response, the mechanisms of which are not well characterized. The objectives of this study were to investigate whether B. asper venom and its purified toxins--phospholipases and metalloproteinase--activate the complement system and the contribution of the effect on leucocyte recruitment. In vitro chemotaxis assays were performed using Boyden's chamber model to investigate the ability of serum incubated with venom and its purified toxins to induce neutrophil migration. The complement consumption by the venom was evaluated using an in vitro haemolytic assay. The importance of complement activation by the venom on neutrophil migration was investigated in vivo by injecting the venom into the peritoneal cavity of C5-deficient mice. Data obtained demonstrated that serum incubated with crude venom and its purified metalloproteinase BaP-1 are able to induce rat neutrophil chemotaxis, probably mediated by agent(s) derived from the complement system. This hypothesis was corroborated by the capacity of the venom to activate this system in vitro. The involvement of C5a in neutrophil chemotaxis induced by venom-activated serum was demonstrated by abolishing migration when neutrophils were pre-incubated with antirat C5a receptor antibody. The relevance of the complement system in in vivo leucocyte mobilization was further demonstrated by the drastic decrease of this response in C5-deficient mice. Pre-incubation of serum with the soluble human recombinant complement receptor type 1 (sCR 1) did not prevent the response induced by the venom, but abolished the migration evoked by metalloproteinase-activated serum. These data show the role of the complement system in bothropic envenomation and the participation of metalloproteinase in the effect. Also, they suggest that the venom may contain other component(s) which can cause direct activation of C5a. PMID:11200361

  8. Biochemical, biological and molecular characterization of an L-Amino acid oxidase (LAAO) purified from Bothrops pictus Peruvian snake venom.

    Science.gov (United States)

    Lazo, Fanny; Vivas-Ruiz, Dan E; Sandoval, Gustavo A; Rodríguez, Edith F; Kozlova, Edgar E G; Costal-Oliveira, F; Chávez-Olórtegui, Carlos; Severino, Ruperto; Yarlequé, Armando; Sanchez, Eladio F

    2017-12-01

    An L-amino acid oxidase from Peruvian Bothrops pictus (Bpic-LAAO) snake venom was purified using a combination of size-exclusion and ion-exchange chromatography. Bpic-LAAO is a homodimeric glycosylated flavoprotein with molecular mass of ∼65 kDa under reducing conditions and ∼132 kDa in its native form as analyzed by SDS-PAGE and gel filtration chromatography, respectively. N-terminal amino acid sequencing showed highly conserved residues in a glutamine-rich motif related to binding substrate. The enzyme exhibited optimal activity towards L-Leu at pH 8.5, and like other reported SV-LAAOs, it is stable until 55 °C. Kinetic studies showed that the cations Ca 2+ , Mg 2+ and Mn 2+ did not alter Bpic-LAAO activity; however, Zn 2+ is an inhibitor. Some reagents such as β-mercaptoethanol, glutathione and iodoacetate had inhibitory effect on Bpic-LAAO activity, but PMSF, EDTA and glutamic acid did not affect its activity. Regarding the biological activities of Bpic-LAAO, this enzyme induced edema in mice (MED = 7.8 μg), and inhibited human platelet aggregation induced by ADP in a dose-dependent manner and showed antibacterial activity on Gram (+) and Gram (-) bacteria. Bpic-LAAO cDNA of 1494 bp codified a mature protein with 487 amino acid residues comprising a signal peptide of 11 amino acids. Finally, the phylogenetic tree obtained with other sequences of LAAOs, evidenced its similarity to other homologous enzymes, showing two well-established monophyletic groups in Viperidae and Elapidae families. Bpic-LAAO is evolutively close related to LAAOs from B. jararacussu, B. moojeni and B. atrox, and together with the LAAO from B. pauloensis, form a well-defined cluster of the Bothrops genus. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. Bothrops asper snake venom and its metalloproteinase BaP–1 activate the complement system. Role in leucocyte recruitment

    Directory of Open Access Journals (Sweden)

    Sandra H. P. Farsky

    2000-01-01

    Full Text Available The venom of the snake Bothrops asper, the most important poisonous snake in Central America, evokes an inflammatory response, the mechanisms of which are not well characterized. The objectives of this study were to investigate whether B. asper venom and its purified toxins – phospholipases and metalloproteinase – activate the complement system and the contribution of the effect on leucocyte recruitment. In vitro chemotaxis assays were performed using Boyden's chamber model to investigate the ability of serum incubated with venom and its purified toxins to induce neutrophil migration. The complement consumption by the venom was evaluated using an in vitro haemolytic assay. The importance of complement activation by the venom on neutrophil migration was investigated in vivo by injecting the venom into the peritoneal cavity of C5-deficient mice. Data obtained demonstrated that serum incubated with crude venom and its purified metalloproteinase BaP–1 are able to induce rat neutrophil chemotaxis, probably mediated by agent(s derived from the complement system. This hypothesis was corroborated by the capacity of the venom to activate this system in vitro. The involvement of C5a in neutrophil chemotaxis induced by venom-activated serum was demonstrated by abolishing migration when neutrophils were pre-incubated with antirat C5a receptor antibody. The relevance of the complement system in in vivo leucocyte mobilization was further demonstrated by the drastic decrease of this response in C5-deficient mice. Pre-incubation of serum with the soluble human recombinant complement receptor type 1 (sCR 1 did not prevent the response induced by the venom, but abolished the migration evoked by metalloproteinase-activated serum. These data show the role of the complement system in bothropic envenomation and the participation of metalloproteinase in the effect. Also, they suggest that the venom may contain other component(s which can cause direct activation

  10. In Vitro Antiplasmodial Activity of Phospholipases A2 and a Phospholipase Homologue Isolated from the Venom of the Snake Bothrops asper

    OpenAIRE

    Castillo, Juan; Vargas, Leidy; Segura, Cesar; Gutiérrez, José; Pérez, Juan

    2012-01-01

    The antimicrobial and antiparasite activity of phospholipase A2 (PLA2) from snakes and bees has been extensively explored. We studied the antiplasmodial effect of the whole venom of the snake Bothrops asper and of two fractions purified by ion-exchange chromatography: one containing catalytically-active phospholipases A2 (PLA2) (fraction V) and another containing a PLA2 homologue devoid of enzymatic activity (fraction VI). The antiplasmodial effect was assessed on in vitro cultures of Plasmod...

  11. Immunoglobulin G and F(ab')2 polyvalent antivenoms do not differ in their ability to neutralize hemorrhage, edema and myonecrosis induced by Bothrops asper (terciopelo) snake venom

    OpenAIRE

    León Montero, Guillermo; Rojas Céspedes, Gustavo; Lomonte, Bruno; Gutiérrez, José María

    1997-01-01

    The ability of whole immunoglobulin G (IgG) and F(ab')2 polyvalent (Crotalinae) antivenoms to neutralize the hemorrhagic, edema-forming and myotoxic activities of Bothrops asper venom was studied. Both antivenoms were adjusted to the same neutralizing potency against lethal and hemorrhagic activities in experiments where venom and antivenoms were incubated before injection. Thus, in these experimental conditions, differences in the neutralizing ability in experiments involving independent inj...

  12. Avaliação da uniformidade de conteúdo do captopril, em cápsulas e comprimidos comercializados em farmácias e drogarias de Votuporanga, SP, Brasil

    Directory of Open Access Journals (Sweden)

    Gabriela Oliveira Rodrigues NASCIMENTO

    2014-01-01

    Full Text Available O captopril é um fármaco indicado para o tratamento de hipertensão arterial sistêmica (HAS e insuficiência cardíaca (IC, sendo o primeiro fármaco desenvolvido a partir da síntese do veneno da cobra Bothrops jararaca. É um protótipo para outras substâncias usadas no controle da hipertensão arterial. No Sistema Único de Saúde (SUS, o captopril está inserido na Relação Nacional de Medicamentos Essenciais (Rename; também está citado em quase todas as Relações Municipais de Medicamentos Essenciais (Remumes. O objetivo desse trabalho foi avaliar a uniformidade de conteúdo de comprimidos e cápsulas contendo captopril, na concentração de 25 mg, produzidos por indústrias, classificados, de acordo com registro, em: referência, genéricos e similares, bem como os manipulados em farmácias magistrais do município de Votuporanga, SP, com base no conceito da qualidade, aplicado à obediência à legislação vigente. Os resultados obtidos apresentaram variações nas concentrações do fármaco, em relação ao produto de referência, onde foi confirmada a massa declarada de 25 mg do fármaco, sendo que nas demais apresentações foram observadas, em valores percentuais, de 69 % para menor concentração e a maior em 11 2% do valor declarado. Dessa forma, foi evidenciada a falta de controle da qualidade na uniformidade de conteúdo, o que pode comprometer a eficácia e segurança do medicamento para o paciente.

  13. Microbiological evaluation of different strategies for management of snakes in captivity.

    Science.gov (United States)

    Campagner, M V; Bosco, S M G; Bagagli, E; Cunha, M L R S; Jeronimo, B C; Saad, E; Biscola, N P; Ferreira, R S; Barraviera, B

    2012-01-01

    Keeping snakes in captivity to produce venom for scientific research and production of inputs is now a worldwide practice. Maintaining snakes in captivity involves capture, infrastructure investments, management techniques, and appropriate qualified personnel. Further, the success of the project requires knowledge of habitat, nutrition, and reproduction, and control of opportunistic infections. This study evaluated the management of snakes in three types of captivity (quarantine, intensive, and semiextensive) and diagnosed bacterial and fungal contaminants. A bacteriological profile was obtained by swabbing the oral and cloacal cavities, scales, and venoms of healthy adult snakes from Bothrops jararaca (Bj) and Crotalus durissus terrificus (Cdt). There was predominance of Enterobacteriaceae, especially non-fermenting Gram-negative bacilli excluding Pseudomonas spp and Gram- positive bacteria. Statistically, intensive captivity resulted in the highest number of bacterial isolates, followed by recent capture (quarantine) and by semiextensive captivity. No statistical difference was found between Bj and Cdt bacterial frequency. In vitro bacterial susceptibility testing found the highest resistance against the semisynthetic penicillins (amoxicillin and ampicillin) and highest sensitivity to amicacin and tobramycin aminoglycosides. To evaluate mycological profile of snakes from intensive captivity, samples were obtained from two healthy Bj and one B. moojeni, one B. pauloensis, and one Cdt showing whitish lesions on the scales suggestive of ringworm. Using conventional methods and DNA-based molecular procedures, five samples of Trichosporon asahii were identified. Despite the traditional role of intense captivity in ophidian venom production, semiextensive captivity was more effective in the present study by virtue of presenting superior control of bacterial and fungal transmission, easier management, lowest cost, and decreased rate of mortality; therefore, it should be

  14. Lachesis stenophrys venom reduces the equine antibody response towards Bothrops asper venom used as co-immunogen in the production of polyspecific snake antivenom.

    Science.gov (United States)

    Arroyo, Cynthia; Solano, Sergio; Herrera, María; Segura, Álvaro; Estrada, Ricardo; Vargas, Mariángela; Villalta, Mauren; Gutiérrez, José María; León, Guillermo

    2015-09-01

    The anti-bothropic activity of an antivenom prepared from the plasma of horses immunized with Bothrops asper venom (anti-B antivenom) was compared with a similar formulation produced from the plasma of horses immunized with a mixture of B. asper and Lachesis stenophrys venoms (anti-BL antivenom). Likewise, a comparison between the anti-lachesic activity of the anti-BL antivenom and a similar formulation prepared from horses immunized only with L. stenophrys venom (anti-L antivenom) was performed. The anti-BL antivenom had lower concentration of anti-bothropic antibodies than the anti-B antivenom. This difference was associated to a lower response towards all components of B. asper venom, but particularly towards some D49-phospholipases A2 (PLA2s) and PIII-metalloproteinases. Consequently, the anti-BL antivenom was less effective neutralizing lethal, coagulant, defibrinogenating, PLA2, and myotoxic activities of B. asper venom. On the other hand, anti-BL and anti-L antivenoms showed similar concentration of anti-lachesic antibodies, and similar capacity to recognize the HPLC fractions of L. stenophrys venom and to neutralize lethal, coagulant, proteolytic, hemorrhagic, PLA2 and myotoxic activities induced by this venom. It is concluded that, when used as co-immunogens, the venom of L. stenophrys reduces the antibody response towards B. asper venom, whereas the latter does not affect the anti-lachesic response. Copyright © 2015 Elsevier Ltd. All rights reserved.

  15. Clinical poisoning in bovine the venom of Bothrops atrox the municipality of Oriximiná-Pará, Central Amazonia, Brazil - Case report

    Directory of Open Access Journals (Sweden)

    Ubaldo de Almeida Farias Junior

    2015-10-01

    Full Text Available ABSTRACT. Farias Junior U. de A. & Chalkidis H.M. [Clinical poisoning in bovine the venom of Bothrops atrox the municipality of Oriximiná-Pará, Central Amazonia, Brazil - Case report.] Envenenamento clínico de bovino por peçonha de Bothrops atrox no município de Oriximiná-Pará, Amazô- nia Central, Brasil - Relato de caso. Revista Brasileira de Medicina Veterinária, 37(3:264-268, 2015. Laboratório de Pesquisas Zoológicas, Faculdades Integradas do Tapajós, Rua Rosa Vermelha, 335, Aeroporto Velho, Santarém, PA 68010-200, Brasil. E-mail: chalkidis@hotmail.com It explains a case of poisoning bovine by Bothrops atrox, abundant snake family Viperidae, prevalent in northern Brazil, assigned as the etiological agent of over 90% of cases of snakebite in the State of Pará. Report the examination semiological and the clinical symptoms observed due to its evolution as well. Clinical signs are confronted with the findings conferred in similar cases reported by veterinarians and ranchers in the region. The treatment in this particular case was not proceeded in order to examine symptoms presented by the accuracy of these reports.

  16. Influence of phospholipasic inhibition on neuromuscular activity of Bothrops fonsecai snake venom.

    Science.gov (United States)

    Schezaro-Ramos, Raphael; Collaço, Rita de Cássia O; Randazzo-Moura, Priscila; Rocha, Thalita; Cogo, José Carlos; Rodrigues-Simioni, Léa

    2017-05-01

    Bothrops fonsecai (B. fonsecai), a pitviper endemic to southeastern Brazil, has a venom mainly composed by snake venom phospholipases (PLA 2 ) and metalloproteases, compounds that could interfere with neuromuscular junction in vitro. In this work, we investigated the role of PLA 2 in the myotoxicity and neuromuscular blockade caused by B. fonsecai venom using different procedures frequently associated with PLA 2 activity inhibition: 24 °C bath temperature, Ca 2+ - Sr 2+ replacement and chemical modification with p-bromophenacyl bromide (p-BPB). Mice extensor digitorum longus preparations (EDL) were incubated with usual or modified Tyrode solution (prepared with Ca 2+ or Sr 2+ respectively) at 24 °C or 37 °C (as controls) and in addition of B. fonsecai venom (100 μg/mL) alone or after its incubation with buffer (24 h, 23 °C) on the absence (alkylation control) and presence of p-BPB; all muscle were processed for histological analysis. The PLA 2 , proteolytic and amidolytic activities under the same conditions (24 °C or 37 °C, Ca 2+ - Sr 2+ replacement, absence or presence p-BPB) were also assessed. The B. fonsecai venom caused total neuromuscular blockade after 100 min of incubation, in Ca 2+ Tyrode solution at 37 °C (usual conditions); on Sr 2+ Tyrode solution (37 °C) the twitch height were 31.7 ± 7.4% of basal, and at 24 °C (Ca 2+ Tyrode solution) were 53.6 ± 7.0% of basal. The alkylation of PLA 2 with p-BPB promoted a great blockade decrease at 100 min of incubation (88.7 ± 5.7% of basal), but it was also observed on alkylation control preparations (66.2 ± 6.6%). The venom produced 50% of blockade at 40.5 ± 5.9 min, in Ca 2+ Tyrode solution at 37 °C. The protocols delayed the time for 50% blockade: 105.7 ± 7.1 min (at 24 °C, in Ca 2+ Tyrode solution) and 71.1 ± 9.0 min (at 37 °C, in Sr 2+ Tyrode solution). Regarding p-BPB incubation and alkylation control preparations, 50% of blockade was not reached

  17. Daño renal en ratas inducido por veneno de Bothrops neuwiedii diporus de Argentina Renal injury in rats poisoned by venom of Bothrops neuwiedii diporus from Argentina

    Directory of Open Access Journals (Sweden)

    Patricia Koscinczuk

    2004-08-01

    Full Text Available La insuficiencia renal aguda es una de las complicaciones sistémicas más frecuentes después de un accidente ofídico. En este estudio se evalúan los efectos que el veneno de Bothrops neuwiedii diporus produce en el riñón de ratas machos de la cepa Wistar. La histopatología permitió comprobar el desarrollo de necrosis tubular aguda; las lesiones iniciales se observaron a las 3 horas de la inoculación de una dosis de 700 µg del veneno, observándose en corteza renal congestión y degeneración granulohialina de las células epiteliales tubulares, acompañadas de dilatación y cilindros hialinos en la luz tubular. A las 24 horas se presentó necrosis tubular aguda en una superficie extensa de la corteza sin daño de la membrana basal tubular. Las lesiones de degeneración turbia de células epiteliales tubulares, dilatación de la luz tubular y cilindros hialinos se mantuvieron presentes hasta las 4 semanas post-inoculación. Si bien los parámetros de la bioquímica clínica asociados con insuficiencia renal aguda aumentaron a las 6 horas de la administración del veneno (urea: 1.10±0.22 g/dl; creatinina: 19.60±1.51mg/dl, a la semana descendieron a valores normales. Las densidades urinarias, en cambio, a la semana se mantuvieron más bajas que lo normal, 1.005 ± 0.001 (pAcute renal failure is one of the systemic complications that can be found in bothropic accidents. In this study the effects on male Wistar rats induced by the venom of Bothrops neuwiedii diporus were evaluated. The histopathology revealed acute tubular necrosis, lesions firstly were observed 3 hours post inoculation of 700 µg of venom. Cortical kidney congestion and granulohialin degeneration of tubular epithelial cells were observed, these lesions achieved a maximum at 24 hours after inoculation. Tubular epithelial hidropic degeneration and dilatation of tubular lumen with hyalin casts were present inclusive up to 4 weeks after inoculation. Biochemical parameter

  18. Hemograma de cães envenenados experimentalmente com Bothrops alternatus após diferentes tratamentos.

    Directory of Open Access Journals (Sweden)

    D. O. Jacome

    2005-03-01

    Full Text Available RESUMO: O objetivo deste trabalho foi verificar a influência de diferentes tratamentos (soro antibotrópico, flunixina meglumina e extrato aquoso de C. longa sobre o hemograma de cães envenenados experimentalmente com Bothrops alternatus. Foram utilizados 12 cães adultos, sem raça definida, inoculados com 0,3mg/kg do veneno via IM, tratados duas horas após o envenenamento, divididos em três grupos de 4 animais: Grupo I- soro antibotrópico diluído em solução fisiológica 0,9%, dose única suficiente para neutralizar 0,3mg/kg do veneno. Grupo II- flunixina meglumina 1,1mg/kg via IM, uma vez ao dia, por 5 dias. Grupo III- extrato aquoso de Curcuma longa (10%, tópica no local da injeção do veneno, 3 vezes ao dia, por cinco dias. A análise dos resultados demonstrou uma queda significativa (p<0,05 nos valores médios da contagem total de eritrócitos, na concentração de hemoglobina e no hematócrito, a partir do momento controle em todos os grupos. A análise do leucograma revelou aumento significativo (p<0,05 para a contagem total de leucócitos e neutrófilos segmentados em todos os grupos 24 horas após a inoculação do veneno, caracterizando uma leucocitose por neutrofilia. Os valores médios dos bastonetes mantiveram-se dentro da normalidade, exceto 30 horas após a inoculação no grupo II e 24 e 168 horas após a inoculação no grupo III, encontraram-se acima dos valores de referência. Houve uma queda significativa (p<0,05 nos valores médios de eosinófilos nos três grupos entre seis horas e 168 horas após a inoculação do veneno. Com relação aos linfócitos, observou-se diferença significativa (p<0,05 entre os grupos 30 horas após a inoculação para o grupo I e grupo II e 336 horas após a inoculação para o grupo I e grupo III. Um aumento dos monócitos foi observado em todos os grupos 336 horas após a

  19. Análisis inmunoenzimático (ELISA para determinar niveles de IgG anti Bothrops atrox en accidente ofídico Immunoenzymatic determination of IgG anti bothrops atrox levels after snake bites

    Directory of Open Access Journals (Sweden)

    John J. Estrada

    1993-01-01

    Full Text Available Se desarrolló un método de inmunización de conejos con veneno de Bothrops atrox con el fin de preparar antisueros y estandarizar un inmunoanálisis (ELISA para medir niveles de IgG en pacientes con accidente ofídico. La respuesta Inmune de los conejos se siguió por inmunodifusión en doble dimensión (Ouchterlony e inmunoelectroforesis, demostrando la presencia de bandas nítidas desde el día 60 y en todas las sangrias posteriores; se comprobó que hay variabilidad individual en su respuesta inmune. El ELISA para detección de IgG humana anti B. atrox en los indígenas del Chocó fue una prueba simple y sensible (83.3% pero inespecífica por las reacciones cruzadas en individuos que habían sufrido accidentes por B. nasutus. La técnica para detectar IgG equina anti B. atrox en pacientes tratados con antiveneno fue tambIén simple y muy sensible. We developed an immunization method for the production of rabbit antisera against Bothrops atroxvenoms. An enzyme-Ilnked assay (ELISA was standardized in order to measure IgG levels after snake bites. The immune response of rabbits, as determined by Ouchterlony and immunoelectrophoresis techniques, revealed bands of precipitation from the sixtieth day on. Individual variability in the immune response of rabbits was demonstrated. For the measurement of IgG levels In Indians from the Department of Choco (Colombia, ELISA proved to be a sensitive (83.3% and simple but not an specific procedure, since there were cross-reactions in those previously bitten by B. nasutus. ELISA was also simple and sensitive (100% for the determination of equine anti B. atrox IgG antibodies in patients treated with antivenom

  20. Crystal structure of a Bothrops toxin I, a Myotoxin K 49 like Phospholipase A{sub 2}

    Energy Technology Data Exchange (ETDEWEB)

    Giotto, M.T.S.; Azevedo, W.F.; Horjales, E.; Oliva, G.; Mascarenhas, Y.P.; Garratt, R.C. [Sao Paulo Univ., Sao Carlos, SP (Brazil). Inst. de Fisica]|[Sao Paulo Univ., Ribeirao Preto, SP (Brazil). Faculdade de Medicina

    1996-12-31

    Full text. Bothrops toxin-I (BthTX-I) is a mytoxin isolated from the Brazilian snake Bothrops jararacussu which is a member of the phospholipase A{sub 2} but presents no catalytic activity due to a D49K substitution. Protein was provided by Prof. Dr. J.R. Giglio and Prof{sup a}. Dr{sup a}. A.C.O. Cintra from the Departamento de Medicina de Ribeirao Preto and used in crystallization experiments which were performed using the vapor diffusion technique in hanging drops at 18{sup 0}C. The BthTX-I crystallized in 0.1 M HEPES, pH ranging from 7.0 to 7.6. The precipitant was (NH{sub 4})SO{sub 4} in concentrations ranging from 7.0 to 7.6. The precipitant was (NH{sub 4})SO{sub 4} in concentrations ranging from 57 to 62%. The data collection was initially performed using the automatic difractometer R-AXIS IIC from the Rigaku Co. at the Laboratorio de Cristalografia of IFESC-USP. Subsequently a second data set was collected at SERC Daresbury Laboratory in England using synchrotron radiation. BthTX-I crystallizes in space group P3{sub 1} 21 with a=b=57.58 A, c=131.27 A, {alpha}={beta}=900{sup 0}C and {gamma}=120{sup 0}C. Processing of the data was performed with the MOSFLM program yielding an R{sub merge}= 6.3% and completeness of 99.6% at 2.1 A of resolution. The structure was solved by Molecular Replacement using the package AMoRe with the Agksitrodon piscivorus piscivorus enyme as search model and refined using the refinement program X-PLOR to a final R=18.7% and R{sub free}=27.4%. The assymetric unit contains two monomers, which can be chosen such that they present similar interactions to those described for the homogeneous myotoxin II from B. asper. The interface is, however, surprisingly small when compared to other dimeric structures and is less complementary than expected. A theoretical model for phospholipid building to BthTX-I suggests that no direct interactions between the sn-2 ester bond and K49 would be expected, thus explaining the lack of catalytic activity.

  1. Effect of Echinacea purpurea (Asteraceae aqueous extract on antibody response to Bothrops asper venom and immune cell response

    Directory of Open Access Journals (Sweden)

    Fernando Chaves

    2007-03-01

    Full Text Available The effect of aqueous extract of Echinacea purpurea roots on the murine antibody response to Bothrops asper snake venom in vivo was studied. Three groups were used. Group #1, baseline control, was treated with snake venom plus PBS. Group #2 was treated with snake venom plus sodium alginate as adjuvant (routine method used at Instituto Clodomiro Picado, and group #3 or experimental group, was treated with snake venom plus aqueous extract of E. purpurea root as adjuvant. In all groups, the first inoculation was done with Freund’s complete adjuvant (FCA. By the time of the second bleeding, mice in group #3 showed a remarkable increment in the level of anti-venom antibodies compared with those in groups #1 or #2. In vitro immune cell proliferation as a response to aqueous extract of E. purpurea root was studied using human lymphocytes activated with different lectins (Con A, PHA and PWM. In all cases, increase in percentage of lymphoproliferation was greater when E. purpurea root extract was used in addition to individual lectins. Rev. Biol. Trop. 55 (1: 113-119. Epub 2007 March. 31.Se estudió in vivo, el efecto del extracto acuoso de las raíces de Echinacea purpurea en la respuesta de los anticuerpos murinos al veneno de la serpiente Bothrops asper. El grupo 1 control, fue tratado con el veneno y PBS. El grupo 2 con veneno y alginato de sodio (método utilizado en el Instituto Clodomiro Picado, y el grupo 3 o experimental, con veneno y extracto acuoso de las raíces de E. purpurea. En todos los grupos, la primera inmunización fue hecha con FCA (Freund’s Complete Adjuvant. En las muestras correspondientes a la segunda sangría, los ratones del grupo 3 mostraron un marcado incremento en el nivel de anticuerpos, en comparación con los ratones de los otros grupos. También se determinó la proliferación de células inmunes in vitro, como respuesta al extracto acuoso de la raíz de E. purpurea, utilizando linfocitos humanos activados con

  2. Could mesenchymal stem cell therapy help in the treatment of muscle damage caused by Bothrops alternatus venom?

    Directory of Open Access Journals (Sweden)

    Thalita da Costa Telles

    2018-03-01

    Full Text Available ABSTRACT: The aim of this study was to evaluate the use of mesenchymal stem cells (MSC in the treatment of myonecrosis induced by Bothrops alternatus venom in rats. Seventy-five male adult Wistar rats were divided into three experimental groups. G1 and G2 were injected in the gastrocnemius muscle with 120μg of B. alternatus venom, while G3 received 200μL of PBS only. Three days after the venom injection, 12 rats from G1 were treated with 5.0 x 106 MSC in PBS, whereas G2 and G3 rats received PBS. Every three days, blood and muscle samples of five animals from each group were taken for serum biochemical and pathological analyses. Histological examinations showed more intense muscle lesions following MSC treatment, characterized by disorganization and loss of muscle fibers, with focal necrosis and inflammatory infiltration by mononuclear cells. In conclusion, the use of MSC for the treatment of local damage caused by inoculation of B. alternatus venom impaired muscle regeneration and interfered in the healing process.

  3. Crystallization and preliminary X-ray diffraction analysis of three myotoxic phospholipases A2 from Bothrops brazili venom

    International Nuclear Information System (INIS)

    Fernandes, Carlos A. H.; Gartuzo, Elaine C. G.; Pagotto, Ivan; Comparetti, Edson J.; Huancahuire-Vega, Salomón; Ponce-Soto, Luis Alberto; Costa, Tássia R.; Marangoni, Sergio; Soares, Andreimar M.; Fontes, Marcos R. M.

    2012-01-01

    Two myotoxic and noncatalytic Lys49-phospholipases A 2 (braziliantoxin-II and MT-II) and a myotoxic and catalytic phospholipase A 2 (braziliantoxin-III) from B. brazili were crystallized. X-ray diffraction data sets were collected and molecular-replacement solutions were obtained. Two myotoxic and noncatalytic Lys49-phospholipases A 2 (braziliantoxin-II and MT-II) and a myotoxic and catalytic phospholipase A 2 (braziliantoxin-III) from the venom of the Amazonian snake Bothrops brazili were crystallized. The crystals diffracted to resolutions in the range 2.56–2.05 Å and belonged to space groups P3 1 21 (braziliantoxin-II), P6 5 22 (braziliantoxin-III) and P2 1 (MT-II). The structures were solved by molecular-replacement techniques. Both of the Lys49-phospholipases A 2 (braziliantoxin-II and MT-II) contained a dimer in the asymmetric unit, while the Asp49-phospholipase A 2 braziliantoxin-III contained a monomer in its asymmetric unit. Analysis of the quaternary assemblies of the braziliantoxin-II and MT-II structures using the PISA program indicated that both models have a dimeric conformation in solution. The same analysis of the braziliantoxin-III structure indicated that this protein does not dimerize in solution and probably acts as a monomer in vivo, similar to other snake-venom Asp49-phospholipases A 2

  4. A phospholipase A₂ from Bothrops asper snake venom activates neutrophils in culture: expression of cyclooxygenase-2 and PGE₂ biosynthesis.

    Science.gov (United States)

    Moreira, Vanessa; Gutiérrez, José María; Amaral, Rafaela Bacci; Lomonte, Bruno; Purgatto, Eduardo; Teixeira, Catarina

    2011-02-01

    In this study, the production of prostaglandin E₂ (PGE₂) and up-regulation in cyclooxygenase (COX) pathway induced by a phospholipase A₂ (PLA₂), myotoxin-III (MT-III), purified from Bothrops asper snake venom, in isolated neutrophils were investigated. The arachidonic acid (AA) production and the participation of intracellular PLA₂s (cytosolic PLA₂ and Ca(2+)-independent PLA₂) in these events were also evaluated. MT-III induced COX-2, but not COX-1 gene and protein expression in neutrophils and increased PGE₂ levels. Pretreatment of neutrophils with COX-2 and COX-1 inhibitors reduced PGE₂ production induced by MT-III. Arachidonyl trifluoromethyl ketone (AACOCF₃), an intracellular PLA₂ inhibitor, but not bromoenol lactone (BEL), an iPLA₂ inhibitor, suppressed the MT-III-induced AA and PGE₂ release. In conclusion, MT-III directly stimulates neutrophils inducing COX-2 mRNA and protein expression followed by production of PGE₂. COX-2 isoform is preeminent over COX-1 for production of PGE₂ stimulated by MT-III. PGE₂ and AA release by MT-III probably is related to cPLA₂ activation. Copyright © 2010 Elsevier Ltd. All rights reserved.

  5. Venom of Bothrops asper from Mexico and Costa Rica: intraspecific variation and cross-neutralization by antivenoms.

    Science.gov (United States)

    Segura, Alvaro; Herrera, María; Villalta, Mauren; Vargas, Mariángela; Uscanga-Reynell, Alfredo; de León-Rosales, Samuel Ponce; Jiménez-Corona, María Eugenia; Reta-Mares, José Francisco; Gutiérrez, José María; León, Guillermo

    2012-01-01

    Bothrops asper is the species that induces the highest incidence of snakebite envenomation in southern Mexico, Central America and parts of northern South America. The intraspecies variability in HPLC profile and toxicological activities between the venoms from specimens collected in Mexico (Veracruz) and Costa Rica (Caribbean and Pacific populations) was investigated, as well as the cross-neutralization by antivenoms manufactured in these countries. Venoms differ in their HPLC profiles and in their toxicity, since venom from Mexican population showed higher lethal and defibrinogenating activities, whereas those from Costa Rica showed higher hemorrhagic and in vitro coagulant activities. In general, antivenoms were more effective in the neutralization of homologous venoms. Overall, both antivenoms effectively neutralized the various toxic effects of venoms from the two populations of B. asper. However, antivenom raised against venom from Costa Rican specimens showed a higher efficacy in the neutralization of defibrinogenating and coagulant activities, thus highlighting immunochemical differences in the toxins responsible for these effects associated with hemostatic disturbances in snakebite envenoming. These observations illustrate how intraspecies venom variation may influence antivenom neutralizing profile. Copyright © 2011 Elsevier Ltd. All rights reserved.

  6. Isolation and characterization of a serine proteinase with thrombin-like activity from the venom of the snake Bothrops asper

    Directory of Open Access Journals (Sweden)

    A.V Pérez

    2008-01-01

    Full Text Available A serine proteinase with thrombin-like activity was isolated from the venom of the Central American pit viper Bothrops asper. Isolation was performed by a combination of affinity chromatography on aminobenzamidine-Sepharose and ion-exchange chromatography on DEAE-Sepharose. The enzyme accounts for approximately 0.13% of the venom dry weight and has a molecular mass of 32 kDa as determined by SDS-PAGE, and of 27 kDa as determined by MALDI-TOF mass spectrometry. Its partial amino acid sequence shows high identity with snake venom serine proteinases and a complete identity with a cDNA clone previously sequenced from this species. The N-terminal sequence of the enzyme is VIGGDECNINEHRSLVVLFXSSGFL CAGTLVQDEWVLTAANCDSKNFQ. The enzyme induces clotting of plasma (minimum coagulant dose = 4.1 µg and fibrinogen (minimum coagulant dose = 4.2 µg in vitro, and promotes defibrin(ogenation in vivo (minimum defibrin(ogenating dose = 1.0 µg. In addition, when injected intravenously in mice at doses of 5 and 10 µg, it induces a series of behavioral changes, i.e., loss of the righting reflex, opisthotonus, and intermittent rotations over the long axis of the body, which closely resemble the `gyroxin-like' effect induced by other thrombin-like enzymes from snake venoms.

  7. Effect of Echinacea purpurea (Asteraceae) aqueous extract on antibody response to Bothrops asper venom and immune cell response.

    Science.gov (United States)

    Chaves, Fernando; Chacón, Mauricio; Badilla, Beatriz; Arévalo, Carolina

    2007-03-01

    The effect of aqueous extract of Echinacea purpurea roots on the murine antibody response to Bothrops asper snake venom in vivo was studied. Three groups were used. Group #1, baseline control, was treated with snake venom plus PBS. Group #2 was treated with snake venom plus sodium alginate as adjuvant (routine method used at Instituto Clodomiro Picado), and group #3 or experimental group, was treated with snake venom plus aqueous extract ofE. purpurea root as adjuvant. In all groups, the first inoculation was done with Freund's complete adjuvant (FCA). By the time of the second bleeding, mice in group #3 showed a remarkable increment in the level of anti-venom antibodies compared with those in groups #1 or #2. In vitro immune cell proliferation as a response to aqueous extract of E. purpurea root was studied using human lymphocytes activated with different lectins (Con A, PHA and PWM). In all cases, increase in percentage of lymphoproliferation was greater when E. purpurea root extract was used in addition to individual lectins.

  8. Impact of regional variation in Bothrops asper snake venom on the design of antivenoms: integrating antivenomics and neutralization approaches.

    Science.gov (United States)

    Gutiérrez, José María; Sanz, Libia; Flores-Díaz, Marietta; Figueroa, Lucía; Madrigal, Marvin; Herrera, María; Villalta, Mauren; León, Guillermo; Estrada, Ricardo; Borges, Adolfo; Alape-Girón, Alberto; Calvete, Juan J

    2010-01-01

    Intraspecific snake venom variations have implications in the preparation of venom pools for the generation of antivenoms. The impact of such variation in the cross-reactivity of antivenoms against Bothrops asper venom was assessed by comparing two commercial and four experimental antivenoms. All antivenoms showed similar immunorecognition pattern toward the venoms from adult and neonate specimens. They completely immunodepleted most P-III snake venom metalloproteinases (SVMPs), l-amino acid oxidases, serine proteinases, DC fragments, cysteine-rich secretory proteins (CRISPs), and C-type lectin-like proteins, and partially immunodepleted medium-sized disintegrins, phospholipases A(2) (PLA(2)s), some serine proteinases, and P-I SVMPs. Although all antivenoms abrogated the lethal, hemorrhagic, coagulant, proteinase, and PLA(2) venoms activities, monospecific experimental antivenoms were more effective than the polyspecific experimental antivenom. In addition, the commercial antivenoms, produced in horses subjected to repeated immunization cycles, showed higher neutralization than experimental polyspecific antivenom, produced by a single round of immunization. Overall, a conspicuous pattern of cross-neutralization was evident for all effects by all antivenoms, and monospecific antivenoms raised against venom from the Caribbean population were effective against venom from the Pacific population, indicating that geographic variations in venom proteomes of B. asper from Costa Rica do not result in overt variations in immunological cross-reactivity between antivenoms.

  9. Role of nitric oxide in the local and systemic pathophysiological effects induced by Bothrops asper snake venom in mice.

    Science.gov (United States)

    Chaves, F; Teixeira, C F P; Gutiérrez, J M

    2006-06-01

    To assess the role of nitric oxide in the most relevant local and systemic manifestations in mice injected with the venom of the snake Bothrops asper. Mice were pretreated with nitric oxide synthase inhibitors, and the modifications of the pathological effects induced by the venom were tested. Inhibition of NO synthesis did not affect acute local myonecrosis and hemorrhage in muscle tissue upon intramuscular injection of venom. Local footpad edema was reduced in mice pretreated with the NO synthase inhibitor L-NAME, and a reduction in the extent of inflammatory infiltrate in muscle tissue was observed after envenomation in mice pretreated with L-NAME and aminoguanidine. The most pronounced effect of NOS inhibition by L-NAME was an increment in the lethal activity of the venom, when injected by the intraperitoneal route. Nitric oxide does not seem to play a significant role in the local acute pathological alterations (hemorrhage and myonecrosis) induced by B. asper venom in mice, although it contributes to edema and inflammatory infiltrate. Nitric oxide exerts a protective role in the systemic pathophysiological manifestations leading to lethality.

  10. Isolation and characterization of a serine proteinase with thrombin-like activity from the venom of the snake Bothrops asper.

    Science.gov (United States)

    Pérez, A V; Rucavado, A; Sanz, L; Calvete, J J; Gutiérrez, J M

    2008-01-01

    A serine proteinase with thrombin-like activity was isolated from the venom of the Central American pit viper Bothrops asper. Isolation was performed by a combination of affinity chromatography on aminobenzamidine-Sepharose and ion-exchange chromatography on DEAE-Sepharose. The enzyme accounts for approximately 0.13% of the venom dry weight and has a molecular mass of 32 kDa as determined by SDS-PAGE, and of 27 kDa as determined by MALDI-TOF mass spectrometry. Its partial amino acid sequence shows high identity with snake venom serine proteinases and a complete identity with a cDNA clone previously sequenced from this species. The N-terminal sequence of the enzyme is VIGGDECNINEHRSLVVLFXSSGFL CAGTLVQDEWVLTAANCDSKNFQ. The enzyme induces clotting of plasma (minimum coagulant dose = 4.1 microg) and fibrinogen (minimum coagulant dose = 4.2 microg) in vitro, and promotes defibrin(ogen)ation in vivo (minimum defibrin(ogen)ating dose = 1.0 microg). In addition, when injected intravenously in mice at doses of 5 and 10 microg, it induces a series of behavioral changes, i.e., loss of the righting reflex, opisthotonus, and intermittent rotations over the long axis of the body, which closely resemble the ;gyroxin-like' effect induced by other thrombin-like enzymes from snake venoms.

  11. Biochemical characterization and pharmacological properties of a phospholipase A2 myotoxin inhibitor from the plasma of the snake Bothrops asper.

    Science.gov (United States)

    Lizano, S; Lomonte, B; Fox, J W; Gutiérrez, J M

    1997-01-01

    A protein that neutralizes the biological activities of basic phospholipase A2 (PLA2) myotoxin isoforms from the venom of the snake Bothrops asper was isolated from its blood by affinity chromatography with Sepharose-immobilized myotoxins. Biochemical characterization of this B. asper myotoxin inhibitor protein (BaMIP) indicated a subunit molecular mass of 23-25 kDa, an isoelectric point of 4, and glycosylation. Gel-filtration studies revealed a molecular mass of 120 kDa, suggesting that BaMIP possesses an oligomeric structure composed of five 23-25 kDa subunits. Functional studies indicated that BaMIP inhibits the PLA2 activity of B. asper basic myotoxins I and III, as well as the myotoxicity and edema-forming activity in vivo and cytolytic activity in vitro towards cultured endothelial cells, of all four myotoxin isoforms (I-IV) tested. Sequence analysis of the first 63 amino acid residues from the N-terminus of BaMIP indicated more than 65% sequence similarity to the PLA2 inhibitors isolated from the blood of the crotalid snakes Trimeresurus flavoviridis and Agkistrodon blomhoffii siniticus. These inhibitors also share sequences similar to the carbohydrate-recognition domains of human and rabbit cellular PLA2 receptors, suggesting a common domain evolution among snake plasma PLA2 inhibitors and mammalian PLA2 receptors. Despite this similarity, this is the first description of a natural anti-myotoxic factor from snake blood. PMID:9307037

  12. Phenotypic differences in a cryptic predator: factors influencing morphological variation in the terciopelo Bothrops asper (Garman, 1884; Serpentes: Viperidae).

    Science.gov (United States)

    Saldarriaga-Córdoba, Mónica María; Sasa, Mahmood; Pardo, Rodrigo; Méndez, Marco Antonio

    2009-12-01

    The terciopelo Bothrops asper, is a cryptic lancehead pitviper widely distributed in humid environments of Middle America and the north-western portion of South America. Throughout its extensive distribution range, the terciopelo exhibits great morphological variation in external characters, a situation that has complicated its proper separation from other related species. In this paper, we analyzed the phenotypic variation of B. asper based in a sample of 514 specimens from nine distinct physiographic regions. Univariate and multivariate analyses demonstrated great phenotypic differentiation among most pre-established groups, and the pattern is fairly congruent between sexes. However, no correspondence was observed between morphological variation and molecular divergence, inferred from mDNA sequences, between individuals representing the physiographical regions under study. Geographic variation in the number of interrictals, ventral scales, subcaudal scales and dorsal blotches was positively correlated with latitude and number of dry months, but inversely related with precipitation. However, other variables do not exhibit such an effect. The observed relationships between scale counts and environmental variables are explained in terms of selective pressures to improve water balance along the distributional range of the species.

  13. Isolation of bothrasperin, a disintegrin with potent platelet aggregation inhibitory activity, from the venom of the snake Bothrops asper.

    Science.gov (United States)

    Pinto, Adrián; Angulo, Yamileth; Jiménez, Rafael; Lomonte, Bruno

    2003-03-01

    The venom of Bothrops asper induces severe coagulation disturbances in accidentally envenomed humans. However, only few studies have been conducted to identify components that interact with the hemostatic system in this venom. In the present work, we fractionated B. asper venom in order to investigate the possible presence of inhibitors of platelet aggregation. Using a combination of gel filtration, anion-exchange chromatography, and reverse-phase high performance liquid chromatography, we isolated an acidic protein which shows a single chain composition, with a molecular mass of approximately 8 kDa, estimated by SDS-polyacrylamide gel electrophoresis. Its N-terminal sequence has high similarity to disintegrins isolated from different snake venoms, which are known to bind to cellular integrins such as the GPIIb/IIIa fibrinogen receptor on platelets. The purified protein exerted potent aggregation inhibitory activity on ADP-stimulated human platelets in vitro, with an estimated IC50 of 50 nM. This biological activity, together with the biochemical characteristics observed, demonstrate that the protein isolated from B. asper venom is a disintegrin, hereby named "bothrasperin". This is the first disintegrin isolated from Central American viperid snake species.

  14. Neutralization of Bothrops asper venom by antibodies, natural products and synthetic drugs: contributions to understanding snakebite envenomings and their treatment.

    Science.gov (United States)

    Lomonte, Bruno; León, Guillermo; Angulo, Yamileth; Rucavado, Alexandra; Núñez, Vitelbina

    2009-12-01

    Interest in studies on the neutralization of snake venoms and toxins by diverse types of inhibitors is two-fold. From an applied perspective, results enclose the potential to be translated into useful therapeutic products or procedures, to benefit patients suffering from envenomings. From a basic point of view, on the other hand, neutralizing agents may be used as powerful dissecting tools to determine the relative role of toxins within the context of the overall pathology induced by a venom, or to increase our understanding on the molecular mechanisms by which toxins exert their harmful actions upon particular targets. The venom of the snake Bothrops asper has been the subject of a number of experimental studies addressing its neutralization by antibodies, as well as by non-immunologic inhibitors, including natural products derived from plants or animals, or synthetic drugs. As summarized in the present review, neutralization studies on this venom and some of its isolated toxins have contributed to a better understanding of envenomings by this species, and their treatment. In addition, such studies have provided valuable knowledge on the mechanisms of action and the relative functional importance of particular toxins of this venom, especially in the case of its myotoxic phospholipases A(2) and hemorrhagic metalloproteinases.

  15. Bothrops asper envenoming in cattle: Clinical features and management using equine-derived whole IgG antivenom.

    Science.gov (United States)

    Rodríguez, C; Estrada, R; Herrera, M; Gómez, A; Segura, Á; Vargas, M; Villalta, M; León, G

    2016-01-01

    Snakebite envenoming is an important problem in the livestock industry in Costa Rica. Of the 22 species of venomous snakes in the country, Bothrops asper is involved in most animal envenomings. Envenomation is typically characterised by swelling and bleeding at the bite site, coagulopathy, systemic haemorrhage, and, in some cases, death. The aims of the present study were to describe the clinical manifestations of B. asper envenomation in cattle and to evaluate the treatment efficacy of antivenom administration. The clinical effects of naturally occurring envenomation were reproduced experimentally in cattle by giving an intramuscular injection of either 10 mg or 50 mg venom to replicate mild and severe envenomings, respectively. Intravenous antivenom given 6 h after experimental venom injection controlled the symptoms; a dose of 120 mL was found to be appropriate for moderate and 200 mL for severe naturally occurring envenomings. Although administration of antivenom within the first 6 h following a snakebite prevented systemic effects, it did not reduce the extent of swelling at the bite site. Delayed administration of antivenom was not effective in saving naturally envenomed animals. The results indicate that, when promptly administered, antivenom constitutes an effective treatment for B. asper snakebite envenomation in cattle.

  16. Human neutrophil migration and activation by BJcuL, a galactose binding lectin purified from Bothrops jararacussu venom

    Directory of Open Access Journals (Sweden)

    Fernandes Luiz

    2011-01-01

    Full Text Available Abstract Background Neutrophil migration to an inflamed site constitutes the first line of the innate immune response against invading microorganisms. Given the crucial role of endogenous lectins in neutrophil mobilization and activation, lectins from exogenous sources have often been considered as putative modulators of leukocyte function. Lectins purified from snake venom have been described as galactoside ligands that induce erythrocyte agglutination and platelet aggregation. This study evaluated human neutrophil migration and activation by C-type lectin BJcuL purified from Bothrops jararacussu venom. Results Utilizing fluorescence microscopy, we observed that biotinylated-BJcuL was evenly distributed on the neutrophil surface, selectively inhibited by D-galactose. Lectin was able to induce modification in the neutrophil morphology in a spherical shape for a polarized observed by optical microscopy and exposure to BJcuL in a Boyden chamber assay resulted in cell migration. After 30 minutes of incubation with BJcuL we found enhanced neutrophil functions, such as respiratory burst, zymozan phagocytosis and an increase in lissosomal volume. In addition, BJcuL delays late apoptosis neutrophils. Conclusion These results demonstrate that BJcuL can be implicated in a wide variety of immunological functions including first-line defense against pathogens, cell trafficking and induction of the innate immune response since lectin was capable of inducing potent neutrophil activation.

  17. Effect of perimuscular injection of Bothrops jararacussu venom on plasma creatine kinase levels in mice: influence of dose and volume

    Directory of Open Access Journals (Sweden)

    Calil-Elias S.

    2002-01-01

    Full Text Available The effect of dose and volume of a perimuscular injection of Bothrops jararacussu venom on myonecrosis of skeletal muscle was studied in mice. An increase of the venom dose (0.25 to 2.0 µg/g at a given volume (50 µl resulted in an increase in plasma creatine kinase (CK levels 2 h after injection. Plasma CK activity increased from the basal level of 129.27 ± 11.83 (N = 20 to 2392.80 ± 709.43 IU/l (N = 4 for the 1.0 µg/g dose. Histological analysis of extensor digitorum longus muscle 4 h after injection showed lesion of peripheral muscle fibers, disorganization of the bundles or the complete degeneration of muscle fibers. These lesions were more extensive when higher doses were injected. Furthermore, an increase in volume (12.5 to 100 µl by dilution of a given dose (0.5 µg/g also increased plasma CK levels from 482.31 ± 122.79 to 919.07 ± 133.33 IU/l (N = 4, respectively. These results indicate that care should be taken to standardize volumes and sites of venom injections.

  18. Crystal structure of pira toxin-I: a calcium-independent, myotoxic phospholipase A2 - homologue from Bothrops pirajai venom

    International Nuclear Information System (INIS)

    Canduri, R.J.; Ward, R.J.; Azevedo Junior, G.W.F. de; Arni, R.K.; Soares, A.M.; Giglio, J.R.

    1997-01-01

    Full text. Phospho lipases A2 (PLA 2 ) are small enzymes that specifically hydrolysed the sn-2 ester bond of phospholipids, preferentially in lamellar or micellar aggregates at membrane surfaces. These enzymes are widely distributed in nature and have been extensively studied. Toxic proteins from venoms from Bothrops species include catalytically active PLA 2 s and calcium independent PLA 2L ys 49 homologues. The substitution of Asp49 by Lys greatly diminishes the ability of these PLA 2 to bind calcium, an ion that plays a critical role in the stabilization of the tetrahedral transition state intermediate in the catalytic mechanism. The Lys 49 PLA 2 homologues and therefore catalytically inactive yet maintain cytolytic and myotoxic activities and furthermore retain the ability to disrupt the integrity of both plasma membranes and model lipid bilayers by a poorly understood Ca 2+ independente mechanism. Lys49 PLA 2 homologues demonstrate a specific toxic activity against skeletal muscle, affecting only muscle fibers and leaving other tissue structure such as connective tissue, nerves and vessels essentially unharmed. In order to improve our understanding of the molecular basis of the myotoxic and Ca 2+ -independent membrane damaging activities, we have determined the crystal structure of Pr TX-I, a Lys49 variant from the venom of B. pirajai. The model presented has been determined at 2.8 angstrom resolution and refined to a crystallographic residual of 19.7% (R free =29.7%). (author)

  19. Purification and Characterization of BmooAi: A New Toxin from Bothrops moojeni Snake Venom That Inhibits Platelet Aggregation

    Directory of Open Access Journals (Sweden)

    Mayara Ribeiro de Queiroz

    2014-01-01

    Full Text Available In this paper, we describe the purification/characterization of BmooAi, a new toxin from Bothrops moojeni that inhibits platelet aggregation. The purification of BmooAi was carried out through three chromatographic steps (ion-exchange on a DEAE-Sephacel column, molecular exclusion on a Sephadex G-75 column, and reverse-phase HPLC chromatography on a C2/C18 column. BmooAi was homogeneous by SDS-PAGE and shown to be a single-chain protein of 15,000 Da. BmooAi was analysed by MALDI-TOF Spectrometry and revealed two major components with molecular masses 7824.4 and 7409.2 as well as a trace of protein with a molecular mass of 15,237.4 Da. Sequencing of BmooAi by Edman degradation showed two amino acid sequences: IRDFDPLTNAPENTA and ETEEGAEEGTQ, which revealed no homology to any known toxin from snake venom. BmooAi showed a rather specific inhibitory effect on platelet aggregation induced by collagen, adenosine diphosphate, or epinephrine in human platelet-rich plasma in a dose-dependent manner, whereas it had little or no effect on platelet aggregation induced by ristocetin. The effect on platelet aggregation induced by BmooAi remained active even when heated to 100°C. BmooAi could be of medical interest as a new tool for the development of novel therapeutic agents for the prevention and treatment of thrombotic disorders.

  20. Comparison of the adjuvant activity of aluminum hydroxide and calcium phosphate on the antibody response towards Bothrops asper snake venom.

    Science.gov (United States)

    Olmedo, Hidekel; Herrera, María; Rojas, Leonardo; Villalta, Mauren; Vargas, Mariángela; Leiguez, Elbio; Teixeira, Catarina; Estrada, Ricardo; Gutiérrez, José María; León, Guillermo; Montero, Mavis L

    2014-01-01

    The adjuvanticity of aluminum hydroxide and calcium phosphate on the antibody response in mice towards the venom of the snake Bothrops asper was studied. It was found that, in vitro, most of the venom proteins are similarly adsorbed by both mineral salts, with the exception of some basic phospholipases A2, which are better adsorbed by calcium phosphate. After injection, the adjuvants promoted a slow release of the venom, as judged by the lack of acute toxicity when lethal doses of venom were administered to mice. Leukocyte recruitment induced by the venom was enhanced when it was adsorbed on both mineral salts; however, venom adsorbed on calcium phosphate induced a higher antibody response towards all tested HPLC fractions of the venom. On the other hand, co-precipitation of venom with calcium phosphate was the best strategy for increasing: (1) the capacity of the salt to couple venom proteins in vitro; (2) the venom ability to induce leukocyte recruitment; (3) phagocytosis by macrophages; and (4) a host antibody response. These findings suggest that the chemical nature is not the only one determining factor of the adjuvant activity of mineral salts.

  1. Isolation of bothrasperin, a disintegrin with potent platelet aggregation inhibitory activity, from the venom of the snake Bothrops asper

    International Nuclear Information System (INIS)

    Pinto, A.; Angulo, Y.; Jimenez, R.; Lomonte, B.

    2003-01-01

    The venom of Bothrops asper induces severe coagulation disturbances in accidentally envenomed humans. However, only few studies have been conducted to identify components that interact with the hemostatic system in this venom. In the present work, we fractionated B. asper venom in order to investigate the possible presence of inhibitors of platelet aggregation. Using a combination of gel filtration, anion-exchange chromatography, and reverse-phase high performance liquid chromatography, we isolated an acidic protein which shows a single chain composition, with a molecular mass of ∼8 kDa, estimated by SDS-polyacrylamide gel electrophoresis. Its N-terminal sequence has high similarity to disintegrins isolated from different snake venoms, which are known to bind to cellular integrins such as the GPIIb/IIIa fibrinogen receptor on platelets. The purified protein exerted potent aggregation inhibitory activity on ADP-stimulated human platelets in vitro, with an estimated IC 50 of 50 nM. This biological activity, together with the biochemical characteristics observed, demonstrate that the protein isolated from B. asper venom is a disintegrin, hereby named bothrasperin. This is the first disintegrin isolated from Central American viperid snake species. (Author)

  2. Biology and conservation status of Piraja’s Lancehead Snake Bothrops piraña Amaral, 1923 (Serpentes: Viperidae, Brazil

    Directory of Open Access Journals (Sweden)

    M.A.D. Freitas

    2014-09-01

    Full Text Available The venomous snake Bothrops pirajai (Amaral, 1923 is endemic to Brazil. Despite being described almost a century ago, very little is known about this species, which has never been studied in situ. Here, we present new data on the biology and natural history of B. pirajai based on a review of existing museum specimens and a field study (1504 man hours carried out throughout the species range. The distribution of B. pirajai was found to be restricted to the eastern Atlantic forest of the state of Bahia, Brazil, between Todosos Santos Bay, Itabuna and Ilhéus (12050’S-14050’S, 88-835 m. We recommend the species be uplisted to Endangered in the International Union for the Conservation of Nature’s global Red List of Threatened Species as its estimated extent of occurrence is under 5000km2. The effective conservation of B. pirajai will rely on addressing two key issues: improving our knowledge of the species and successfully mitigating habitat loss and fragmentation.

  3. Effect of calcineurin inhibitors on myotoxic activity of crotoxin and Bothrops asper phospholipase A2 myotoxins in vivo and in vitro.

    Science.gov (United States)

    Miyabara, E H; Baptista, I L; Lomonte, B; Selistre-de-Araújo, H S; Gutiérrez, J M; Moriscot, A S

    2006-07-01

    Previous studies have shown that calcineurin activity plays a critical role in the myotoxic activity induced by crotoxin (CTX), a group II phospholipase A(2) (PLA(2)) with neurotoxic and myotoxic actions. In order to address whether calcineurin is also important for the activity of non-neurotoxic group II PLA(2) myotoxins we have compared the effects of calcineurin inhibition on the myotoxic capacity of CTX and the non-neurotoxic PLA(2)s, myotoxin II (Mt II) and myotoxin III (Mt III) from Bothrops asper venom. Rats were treated with cyclosporin A (CsA) or FK506, calcineurin inhibitors, and received an intramuscular injection of either CTX, Mt II or Mt III into the tibialis anterior. Animals were killed 24 h after injection of toxins. Tibialis anterior was removed and stored in liquid nitrogen. Myofibers in culture were also treated with CsA or FK506 and exposed to CTX, Mt II and Mt III. It was observed that, in contrast to CTX, CsA and FK506 do not attenuate myotoxic effects induced by both Mt II and Mt III in vivo and in vitro. The results of the present study suggest that calcineurin is not essential for the myotoxic activity of Mt II and Mt III, indicating that distinct intracellular pathways might be involved in myonecrosis induced by neurotoxic CTX and non-neurotoxic Bothrops sp. PLA(2) myotoxins. Alternatively, calcineurin dependent fast fiber type shift might render the muscle resistant to the action of CTX, without affecting its susceptibility to Bothrops sp. myotoxins.

  4. Inhibition of the Myotoxicity Induced by Bothrops jararacussu Venom and Isolated Phospholipases A2 by Specific Camelid Single-Domain Antibody Fragments.

    Directory of Open Access Journals (Sweden)

    Nidiane D R Prado

    Full Text Available Antivenoms, produced using animal hyperimmune plasma, remains the standard therapy for snakebites. Although effective against systemic damages, conventional antivenoms have limited efficacy against local tissue damage. Additionally, the hypersensitivity reactions, often elicited by antivenoms, the high costs for animal maintenance, the difficulty of producing homogeneous lots, and the instability of biological products instigate the search for innovative products for antivenom therapy. In this study, camelid antibody fragments (VHH with specificity to Bothropstoxin I and II (BthTX-I and BthTX-II, two myotoxic phospholipases from Bothrops jararacussu venom, were selected from an immune VHH phage display library. After biopanning, 28 and 6 clones recognized BthTX-I and BthTX-II by ELISA, respectively. Complementarity determining regions (CDRs and immunoglobulin frameworks (FRs of 13 VHH-deduced amino acid sequences were identified, as well as the camelid hallmark amino acid substitutions in FR2. Three VHH clones (KF498607, KF498608, and KC329718 were capable of recognizing BthTX-I by Western blot and showed affinity constants in the nanomolar range against both toxins. VHHs inhibited the BthTX-II phospholipase A2 activity, and when tested for cross-reactivity, presented specificity to the Bothrops genus in ELISA. Furthermore, two clones (KC329718 and KF498607 neutralized the myotoxic effects induced by B. jararacussu venom, BthTX-I, BthTX-II, and by a myotoxin from Bothrops brazili venom (MTX-I in mice. Molecular docking revealed that VHH CDRs are expected to bind the C-terminal of both toxins, essential for myotoxic activity, and to epitopes in the BthTX-II enzymatic cleft. Identified VHHs could be a biotechnological tool to improve the treatment for snake envenomation, an important and neglected world public health problem.

  5. Crystallization and preliminary X-ray diffraction analysis of suramin, a highly charged polysulfonated napthylurea, complexed with a myotoxic PLA2 from Bothrops asper venom.

    Science.gov (United States)

    Murakami, Mário T; Gava, Lisandra M; Zela, Sandro P; Arruda, Emerson Z; Melo, Paulo A; Gutierrez, José M; Arni, Raghuvir K

    2004-12-01

    Suramin is a highly charged polysulfonated napthylurea that interferes in a number of physiologically relevant processes such as myotoxicity, blood coagulation and several kinds of cancers. This synthetic compound was complexed with a myotoxic Lys49 PLA(2) from Bothrops asper venom and crystallized by the hanging-drop vapor diffusion method at 18 degrees C. The crystals belong to the orthorhombic space group P2(1)2(1)2(1), with unit cell parameters a=49.05, b=63.84 and c=85.67 angstroms. Diffraction data was collected to 1.78 angstroms.

  6. Biochemical Characterization, Action on Macrophages, and Superoxide Anion Production of Four Basic Phospholipases A 2 from Panamanian Bothrops asper Snake Venom

    OpenAIRE

    Rueda, Aristides Quintero; Rodr?guez, Isela Gonz?lez; Arantes, Eliane C.; Set?bal, Sulamita S.; Calderon, Leonardo de A.; Zuliani, Juliana P.; St?beli, Rodrigo G.; Soares, Andreimar M.

    2012-01-01

    Bothrops asper (Squamata: Viperidae) is the most important venomous snake in Central America, being responsible for the majority of snakebite accidents. Four basic PLA2s (pMTX-I to -IV) were purified from crude venom by a single-step chromatography using a CM-Sepharose ion-exchange column (1.5 × 15 cm). Analysis of the N-terminal sequence demonstrated that pMTX-I and III belong to the catalytically active Asp49 phospholipase A2 subclass, whereas pMTX-II and IV belong to the enzymatically inac...

  7. Poor efficacy of preemptive amoxicillin clavulanate for preventing secondary infection from Bothrops snakebites in the Brazilian Amazon: A randomized controlled clinical trial.

    Science.gov (United States)

    Sachett, Jacqueline A G; da Silva, Iran Mendonça; Alves, Eliane Campos; Oliveira, Sâmella S; Sampaio, Vanderson S; do Vale, Fábio Francesconi; Romero, Gustavo Adolfo Sierra; Dos Santos, Marcelo Cordeiro; Marques, Hedylamar Oliveira; Colombini, Mônica; da Silva, Ana Maria Moura; Wen, Fan Hui; Lacerda, Marcus V G; Monteiro, Wuelton M; Ferreira, Luiz C L

    2017-07-01

    Secondary bacterial infections from snakebites contribute to the high complication rates that can lead to permanent function loss and disabilities. Although common in endemic areas, routine empirical prophylactic use of antibiotics aiming to prevent secondary infection lacks a clearly defined policy. The aim of this work was to estimate the efficacy of amoxicillin clavulanate for reducing the secondary infection incidence in patients bitten by Bothrops snakes, and, secondarily, identify risk factors for secondary infections from snakebites in the Western Brazilian Amazon. This was an open-label, two-arm individually randomized superiority trial to prevent secondary infection from Bothrops snakebites. The antibiotic chosen for this clinical trial was oral amoxicillin clavulanate per seven days compared to no intervention. A total of 345 patients were assessed for eligibility in the study period. From this total, 187 accomplished the inclusion criteria and were randomized, 93 in the interventional group and 94 in the untreated control group. All randomized participants completed the 7 days follow-up period. Enzyme immunoassay confirmed Bothrops envenoming diagnosis in all participants. Primary outcome was defined as secondary infection (abscess and/or cellulitis) until day 7 after admission. Secondary infection incidence until 7 days after admission was 35.5% in the intervention group and 44.1% in the control group [RR = 0.80 (95%CI = 0.56 to 1.15; p = 0.235)]. Survival analysis demonstrated that the time from patient admission to the onset of secondary infection was not different between amoxicillin clavulanate treated and control group (Log-rank = 2.23; p = 0.789).Secondary infections incidence in 7 days of follow-up was independently associated to fibrinogen >400 mg/dL [AOR = 4.78 (95%CI = 2.17 to 10.55; p44 IU/L [AOR = 2.52 (95%CI = 1.06 to 5.98; p = 0.037)], C-reactive protein >6.5 mg/L [AOR = 2.98 (95%CI = 1.40 to 6.35; p = 0.005)], moderate pain [AOR = 24

  8. Efeito do extrato das raÃzes de Bredemeyera floribunda Willd sobre a aÃÃo local do veneno de Bothrops jararacussu em camundongos

    OpenAIRE

    Natacha Teresa Queiroz Alves

    2013-01-01

    Acidentes ofÃdicos representam considerÃvel problema de saÃde pÃblica. No Brasil, a maior parte desses acidentes à causada por serpentes do gÃnero Bothrops, que podem levar ao desenvolvimento de dano tecidual local, incluindo edema e hemorragia. A soroterapia tradicional atua na neutralizaÃÃo de efeitos sistÃmicos, mas tem eficÃcia limitada contra efeitos locais. Compostos isolados de plantas usadas popularmente para tratar envenenamentos ofÃdicos mostram-se uma boa alternativa a fim de se co...

  9. Effects of irradiated Bothropstoxin-1 and Bothrops jararacussu crude venom on the immune system; Acao da Bothropstoxina-1 e do veneno total de Bothrops jararacussu irradiados sobre o sistema imune

    Energy Technology Data Exchange (ETDEWEB)

    Caproni, Priscila

    2009-07-01

    Ionizing radiation has been successfully employed to modify the immunological properties of biomolecules and has been proven to be a powerful tool to attenuate snake venoms toxicity without affecting and even increasing their immunogenic properties. Very promising results were obtained when crude animal venoms, as well as isolated toxins, were treated with {sup 60}Co gamma rays, yielding toxoids with good immunogenicity, however, little is known about the modifications that irradiated molecules undergo and even less about the immunological response that such antigens elicit. At the present work, we have evaluated the effects on immune system of B10.PL and BALB/c mice of Bothrops jararacussu crude venom and isolated bothropstoxin-1 (Bthx-1), before and after gamma radiation exposition. According to our data, irradiation process promoted structural modifications on both isolated toxin and crude venom, characterized by higher molecular weight protein (aggregates and oligomers) formation. Irradiated samples were immunogenic and the antibodies elicited by them were able to recognize the native toxin in ELISA. These results indicate that irradiation of toxic proteins can promote significant modifications in their structures, but still retain many of the original antigenic and immunological properties. Also, our data indicate that the irradiated protein induced higher titers of IgG2b, suggesting that Th1 cells were predominantly involved. Results from Western blot assay showed that antibodies raised against irradiated bothropstoxin-1 recognize both native isolated toxin or crude venom. Cytotoxicity assay showed that irradiated toxin and crude venom were less toxic than their native counterpart. Thus, the viability of the macrophages cultured in the presence of irradiated Bthx-1 or crude venom was higher if compared with their native forms. LDH Assay showed that irradiated Bthx-1 promotes less muscular damage than the native form. Our data confirm a potential use of

  10. Blister formation and skin damage induced by BaP1, a haemorrhagic metalloproteinase from the venom of the snake Bothrops asper.

    Science.gov (United States)

    Rucavado, A; Núñez, J; Gutiérrez, J M

    1998-01-01

    Blister formation and skin damage can be induced by BaP1, a haemorrhagic metalloproteinase from the venom of the snake Bothrops asper. Pathological changes in the skin were investigated after intramuscular injections of Bothrops asper haemorrhagic metalloproteinase BaP1. Blisters developed within the first hour, with separation of epidermis from the dermal-epidermal junction, whereas acantholysis of epithelial cells was not observed. After the third hour there was ulceration with formation of a proteinaceous scab and inflammatory infiltrate. By 7 to 14 days there was evidence of a regenerative process in dermis and epidermis. Haemorrhage occurred in both dermis and hypodermis as a consequence of BaP1 injection, together with damage of sebaceous glands and an inflammatory reaction in which enlarged macrophages were the predominant cell type. Zymography assays showed the presence of several endogenous metalloproteinases in the exudate, skin homogenates and plasma. In addition, BaP1 was detected in exudates and plasma by immunoblotting. This technique also demonstrated the presence of components immunologically related to laminin and collagen type IV in exudates. It is suggested that BaP1, and probably endogenous matrix metalloproteinases, degrade some protein components at the dermal-epidermal junction, inducing the formation of blisters. PMID:9797720

  11. Myotoxic phospholipases A(2) isolated from Bothrops brazili snake venom and synthetic peptides derived from their C-terminal region: cytotoxic effect on microorganism and tumor cells.

    Science.gov (United States)

    Costa, Tassia R; Menaldo, Danilo L; Oliveira, Clayton Z; Santos-Filho, Norival A; Teixeira, Sabrina S; Nomizo, Auro; Fuly, André L; Monteiro, Marta C; de Souza, Bibiana M; Palma, Mário S; Stábeli, Rodrigo G; Sampaio, Suely V; Soares, Andreimar M

    2008-10-01

    This paper reports the purification and biochemical/pharmacological characterization of two myotoxic phospholipases A(2) (PLA(2)s) from Bothrops brazili venom, a native snake from Brazil. Both myotoxins (MTX-I and II) were purified by a single chromatographic step on a CM-Sepharose ion-exchange column up to a high purity level, showing M(r) approximately 14,000 for the monomer and 28,000Da for the dimer. The N-terminal and internal peptide amino acid sequences showed similarity with other myotoxic PLA(2)s from snake venoms, MTX-I belonging to Asp49 PLA(2) class, enzymatically active, and MTX-II to Lys49 PLA(2)s, catalytically inactive. Treatment of MTX-I with BPB and EDTA reduced drastically its PLA(2) and anticoagulant activities, corroborating the importance of residue His48 and Ca(2+) ions for the enzymatic catalysis. Both PLA(2)s induced myotoxic activity and dose-time dependent edema similar to other isolated snake venom toxins from Bothrops and Crotalus genus. The results also demonstrated that MTXs and cationic synthetic peptides derived from their 115-129 C-terminal region displayed cytotoxic activity on human T-cell leukemia (JURKAT) lines and microbicidal effects against Escherichia coli, Candida albicans and Leishmania sp. Thus, these PLA(2) proteins and C-terminal synthetic peptides present multifunctional properties that might be of interest in the development of therapeutic strategies against parasites, bacteria and cancer.

  12. A new l-amino acid oxidase from Bothrops jararacussu snake venom: Isolation, partial characterization, and assessment of pro-apoptotic and antiprotozoal activities.

    Science.gov (United States)

    Carone, Sante E I; Costa, Tássia R; Burin, Sandra M; Cintra, Adélia C O; Zoccal, Karina F; Bianchini, Francine J; Tucci, Luiz F F; Franco, João J; Torqueti, Maria R; Faccioli, Lúcia H; Albuquerque, Sérgio de; Castro, Fabíola A de; Sampaio, Suely V

    2017-10-01

    A new l-amino acid oxidase (LAAO) from Bothrops jararacussu venom (BjussuLAAO-II) was isolated by using a three-step chromatographic procedure based on molecular exclusion, hydrophobicity, and affinity. BjussuLAAO-II is an acidic enzyme with pI=3.9 and molecular mass=60.36kDa that represents 0.3% of the venom proteins and exhibits high enzymatic activity (4884.53U/mg/mim). We determined part of the primary sequence of BjussuLAAO-II by identifying 96 amino acids, from which 34 compose the N-terminal of the enzyme (ADDRNPLEECFRETDYEEFLEIARNGLSDTDNPK). Multiple alignment of the partial BjussuLAAO-II sequence with LAAOs deposited in the NCBI database revealed high similarity (95-97%) with other LAAOs isolated from Bothrops snake venoms. BjussuLAAO-II exerted a strong antiprotozoal effect against Leishmania amazonensis (IC 50 =4.56μg/mL) and Trypanosoma cruzi (IC 50 =4.85μg/mL). This toxin also induced cytotoxicity (IC 50 =1.80μg/mL) and apoptosis in MCF7 cells (a human breast adenocarcinoma cell line) by activating the intrinsic and extrinsic apoptosis pathways, but were not cytotoxic towards MCF10A cells (a non-tumorigenic human breast epithelial cell line). The results reported herein add important knowledge to the field of Toxinology, especially for the development of new therapeutic agents. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Structural and biophysical studies with the MjTX-I, a Lys49-phospholipase A2 homologue from Bothrops moojeni venom

    International Nuclear Information System (INIS)

    Salvador, G.H.M.; Fernandes, C.A.H.; Fernandez, R.M.; Fontes, M.R.M.; Marchi-Salvador, D.P.; Soares, A.M.; Oliveira, C.L.P

    2012-01-01

    Full text: Phospholipases A 2 (PLA 2 ) are small proteins found in a great diversity of organisms and belong to a superfamily of proteins involved in many important pharmacological processes, such as neurotoxicity, myotoxicity, platelet aggregation, and anticoagulant activity. Ophidic accidents caused by snakes from Bothrops genus are not efficiently neutralized by conventional serum therapy, and then detailed studies with this class of proteins may be very important to supplement this conventional therapy. Miotoxin-I (MjTX-I) is a basic Lys49-PLA 2 , isolated from Bothrops moojeni snake venom, which induces a drastic local myonecrosis. Crystal structure of MjTX-I shows four molecules in the asymmetric unit, an unusually oligomeric conformation for snake venom Lys49-PLA 2 s. However, bioinformatics techniques indicate a dimer as the biological oligomeric conformation. To get additional information of its biological conformation, we also performed Dynamic Light Scattering, Size Exclusion Chromatography and Small Angle X-ray Scattering experiments. These techniques showed a monomer as the most probable biological conformation in water; however small changes in pH and ionic strength result in different oligomeric assemblies. These novel information for Lys49-PLA 2 s may result in important conclusions for this intriguing class of toxins. (author)

  14. Neutralization of pharmacological and toxic activities of Bothrops jararacussu snake venom and isolated myotoxins by Serjania erecta methanolic extract and its fractions

    Directory of Open Access Journals (Sweden)

    RS Fernandes

    2011-01-01

    Full Text Available Most of the snakebites recorded in Brazil are caused by the Bothrops genus. Given that the local tissue damage caused by this genus cannot be treated by antivenom therapy, numerous studies are focusing on supplementary alternatives, such as the use of medicinal plants. Serjania erecta has already demonstrated anti-inflammatory, antiseptic and healing properties. In the current study, the aerial parts of S. erecta were extracted with methanol, then submitted to chromatographic fractionation on a Sephadex LH20 column and eluted with methanol, which resulted in four main fractions. The crude extract and fractions neutralized the toxic activities of Bothrops jararacussu snake venom and isolated myotoxins (BthTX-I and II. Results showed that phospholipase A2, fibrinogenolytic, myotoxic and hemorrhagic activities were inhibited by the extract. Moreover, the myotoxic and edematous activities induced by BthTX-I, and phospholipase A2 activity induced by BthTX-II, were inhibited by the extract of S. erecta and its fraction. The clotting time on bovine plasma was significantly prolonged by the inhibitory action of fractions SF3 and SF4. This extract is a promising source of natural inhibitors, such as flavonoids and tannins, which act by forming complexes with metal ions and proteins, inhibiting the action of serineproteases, metalloproteases and phospholipases A2.

  15. Structural and biophysical studies with the MjTX-I, a Lys49-phospholipase A{sub 2} homologue from Bothrops moojeni venom

    Energy Technology Data Exchange (ETDEWEB)

    Salvador, G.H.M.; Fernandes, C.A.H.; Fernandez, R.M.; Fontes, M.R.M. [UNESP, Universidade Estadual Paulista, Botucatu, SP (Brazil); Marchi-Salvador, D.P. [Universidade Federal da Paraiba (UFPB), Joao Pessoa, PB (Brazil); Soares, A.M. [Universidade de Sao Paulo (USP-RP), Ribeirao Preto, SP (Brazil); Oliveira, C.L.P [Universidade de Sao Paulo (USP), SP (Brazil)

    2012-07-01

    Full text: Phospholipases A{sub 2} (PLA{sub 2}) are small proteins found in a great diversity of organisms and belong to a superfamily of proteins involved in many important pharmacological processes, such as neurotoxicity, myotoxicity, platelet aggregation, and anticoagulant activity. Ophidic accidents caused by snakes from Bothrops genus are not efficiently neutralized by conventional serum therapy, and then detailed studies with this class of proteins may be very important to supplement this conventional therapy. Miotoxin-I (MjTX-I) is a basic Lys49-PLA{sub 2}, isolated from Bothrops moojeni snake venom, which induces a drastic local myonecrosis. Crystal structure of MjTX-I shows four molecules in the asymmetric unit, an unusually oligomeric conformation for snake venom Lys49-PLA{sub 2}s. However, bioinformatics techniques indicate a dimer as the biological oligomeric conformation. To get additional information of its biological conformation, we also performed Dynamic Light Scattering, Size Exclusion Chromatography and Small Angle X-ray Scattering experiments. These techniques showed a monomer as the most probable biological conformation in water; however small changes in pH and ionic strength result in different oligomeric assemblies. These novel information for Lys49-PLA{sub 2}s may result in important conclusions for this intriguing class of toxins. (author)

  16. Acidentes ofidicos causados por Bothrops moojeni: correlação do quadro clínico com o tamanho da serpente

    Directory of Open Access Journals (Sweden)

    João Aris Kouyoumdjian

    1989-04-01

    Full Text Available Durante um período de 19 meses (março 1986 a setembro 1987 foram estudados 22 casos de acidentes ofídicos causados por Bothrops moojeni na região de São José do Rio Preto, São Paulo, nos quais o tamanho da serpente foi sistematicamente medido. Foram constituídos dois grupos de pacientes de acordo com o tamanho da serpente: grupo I - 9 casos de serpentes pequenas (30 a 53 cm e grupo II - 13 casos de serpentes grandes (80 a 147 cm. Os resultados mostraram: 1. efeitos locais iniciais - dor e edema - mais leves no grupo I; 2. tempo de coagulação prolongado/incoagulável levemente mais freqüente no grupo I; 3. complicações locais - necrose, infecção e síndrome compartimentai - exclusivamente, e em mais da metade dos casos do grupo II, apesar da terapia com antiveneno ter sido mais rápida e em doses maiores nesse grupo. Conclui-se que as serpentes Bothrops moojeni maiores apresentam grande incremento nas suas ações locais - edema, necrose e infecção secundária - e leve perda em sua ação coagulante.

  17. Avaliação das lesões locais de cães envenenados experimentalmente com Bothrops alternatus após diferentes tratamentos

    Directory of Open Access Journals (Sweden)

    Santos M.M.B.

    2003-01-01

    Full Text Available The aim of this study was to evaluate the therapeutic action of flunixin meglumine and aqueous extract of Curcuma longa (10% on local lesions caused by bothropic venom, comparing with the specific antivenom. Twelve adult dogs were inoculated in the middle third of the lateral face of the thigh with 0.3mg/kg of venom. The dogs were divided into three groups of four animals and the treatment was done two hours after venom inoculation as follows: Group I- specific antivenom diluted in saline (one dosage sufficient to neutralize the venom, Group II- flunixin meglumine (1.1mg/kg, IM, once a day for five days, Group III- topical application of aqueous extract of C. longa (10% (three times a day for five days. Necrosis was observed in all dogs of groups I and II, and in two dogs of group III. Edema and hemorrhagic halo were observed in all dogs. Better recovery of lesions was observed in dogs of group I and III. The results denote a satisfactory effect of C. longa on treatment of local lesions caused by bothropic venom in dogs.

  18. Isolation of bothrasperin, a disintegrin with potent platelet aggregation inhibitory activity, from the venom of the snake Bothrops asper

    Directory of Open Access Journals (Sweden)

    Adrián Pinto

    2003-03-01

    Full Text Available The venom of Bothrops asper induces severe coagulation disturbances in accidentally envenomed humans. However, only few studies have been conducted to identify components that interact with the hemostatic system in this venom. In the present work, we fractionated B. asper venom in order to investigate the possible presence of inhibitors of platelet aggregation. Using a combination of gel filtration, anion-exchange chromatography, and reverse-phase high performance liquid chromatography, we isolated an acidic protein which shows a single chain composition, with a molecular mass of ~8 kDa, estimated by SDS-polyacrylamide gel electrophoresis. Its N-terminal sequence has high similarity to disintegrins isolated from different snake venoms, which are known to bind to cellular integrins such as the GPIIb/IIIa fibrinogen receptor on platelets. The purified protein exerted potent aggregation inhibitory activity on ADP-stimulated human platelets in vitro, with an estimated IC50 of 50 nM. This biological activity, together with the biochemical characteristics observed, demonstrate that the protein isolated from B. asper venom is a disintegrin, hereby named "bothrasperin". This is the first disintegrin isolated from Central American viperid snake species.El veneno de la serpiente Bothrops asper induce graves alteraciones de la coagulación en los humanos accidentalmente envenenados. Sin embargo, se han realizado pocos estudios para identificar los componentes del veneno que interactúan con el sistema hemostático. En el presente trabajo, fraccionamos el veneno de B. asper para investigar la posible presencia de inhibidores de la agregación plaquetaria. Empleando una combinación de técnicas cromatográficas (filtración en gel, intercambio aniónico y cromatografía líquida de alto desempeño en fase reversa, aislamos una proteína acídica de cadena simple, con una masa molecular de ~8 kDa, estimada mediante electroforesis en gel de poliacrilamida con

  19. Effects of two serine proteases from Bothrops pirajai snake venom on the complement system and the inflammatory response.

    Science.gov (United States)

    Menaldo, Danilo L; Bernardes, Carolina P; Pereira, Juliana C; Silveira, Denise S C; Mamede, Carla C N; Stanziola, Leonilda; Oliveira, Fábio de; Pereira-Crott, Luciana S; Faccioli, Lúcia H; Sampaio, Suely V

    2013-04-01

    The present study aimed to evaluate the effects of two serine proteases from Bothrops pirajai snake venom, named BpirSP27 and BpirSP41, on the complement system and the inflammatory response. The effects of these enzymes on the human complement system were assessed by kinetic hemolytic assays, evaluating the hemolysis promoted by the classical/lectin (CP/LP) and alternative (AP) pathways after incubation of normal human serum with the serine proteases. The results suggested that these enzymes were able to induce modulation of CP/LP and AP at different levels: BpirSP41 showed higher inhibitory effects on the hemolytic activity of CP/LP than BpirSP27, with inhibition values close to 40% and 20%, respectively, for the highest concentration assayed. Regarding AP, both enzymes showed percentages of inhibition of the hemolytic activity around 20% for the highest concentrations tested, indicating similar effects on this complement pathway. The proinflammatory effects of B. pirajai serine proteases were evaluated regarding their ability to induce paw edema, variations in the pain threshold and leukocyte recruitment at the site of injection. Both showed mild effects on these inflammatory processes, leading to low levels of increase of paw volumes and decrease in pain thresholds in rats up to 6 h after injection, and inducing neutrophil recruitment without significant increases in the total number of leukocytes in the inflammatory exudates after 6 and 24 h of administration into mice peritoneal cavity. These results suggest that serine proteases must present a minor role in the inflammation caused by B. pirajai snake venom. Copyright © 2013 Elsevier B.V. All rights reserved.

  20. Intramuscular administration of antivenoms in experimental envenomation by Bothrops asper: comparison between Fab and IgG.

    Science.gov (United States)

    Chaves, Fernando; Loría, Gilbert D; Salazar, Adriana; Gutiérrez, José María

    2003-02-01

    The efficacy of intramuscular (im) administration of sheep Fab and IgG antivenoms was assessed in a mouse experimental model of envenomation by Bothrops asper, in order to test if the more rapid absorption of Fab improves neutralization. Both antivenoms were adjusted to have a similar neutralizing potency in assays involving preincubation of venom and antivenom. Neither antivenom was effective in neutralizing lethality, nor in prolonging the time of death, in mice injected with either 3, 2 or 1.5 LD(50)s of venom by the intraperitoneal (ip) route, in experiments in which antivenoms were administered im immediately after envenomation. Antivenoms were effective in the neutralization of defibrinating activity, even if treatment was performed 30 min after envenomation, with no differences between IgG and Fab. Regarding neutralization of local effects, i.e. myonecrosis and hemorrhage, im administration of antivenoms at a site distant from the venom-injection site was completely ineffective in reducing the extent of local tissue damage. However, partial neutralization of these effects was achieved if antivenoms were administered im at the same site of venom injection, provided treatment was performed immediately after envenomation. Fab antivenom was slightly more effective than IgG antivenom in the neutralization of myotoxicity under these conditions, although a similar efficacy was observed between these antivenoms regarding neutralization of hemorrhagic effect. Our observations do not evidence major differences in the neutralizing ability of Fab and IgG antivenoms when applied by the im route, and do not support the hypothesis that im administration of Fab antivenoms constitutes an effective alternative to treat B. asper envenomations.

  1. Muscle Tissue Damage Induced by the Venom of Bothrops asper: Identification of Early and Late Pathological Events through Proteomic Analysis.

    Directory of Open Access Journals (Sweden)

    Cristina Herrera

    2016-04-01

    Full Text Available The time-course of the pathological effects induced by the venom of the snake Bothrops asper in muscle tissue was investigated by a combination of histology, proteomic analysis of exudates collected in the vicinity of damaged muscle, and immunodetection of extracellular matrix proteins in exudates. Proteomic assay of exudates has become an excellent new methodological tool to detect key biomarkers of tissue alterations for a more integrative perspective of snake venom-induced pathology. The time-course analysis of the intracellular proteins showed an early presence of cytosolic and mitochondrial proteins in exudates, while cytoskeletal proteins increased later on. This underscores the rapid cytotoxic effect of venom, especially in muscle fibers, due to the action of myotoxic phospholipases A2, followed by the action of proteinases in the cytoskeleton of damaged muscle fibers. Similarly, the early presence of basement membrane (BM and other extracellular matrix (ECM proteins in exudates reflects the rapid microvascular damage and hemorrhage induced by snake venom metalloproteinases. The presence of fragments of type IV collagen and perlecan one hour after envenoming suggests that hydrolysis of these mechanically/structurally-relevant BM components plays a key role in the genesis of hemorrhage. On the other hand, the increment of some ECM proteins in the exudate at later time intervals is likely a consequence of the action of endogenous matrix metalloproteinases (MMPs or of de novo synthesis of ECM proteins during tissue remodeling as part of the inflammatory reaction. Our results offer relevant insights for a more integrative and systematic understanding of the time-course dynamics of muscle tissue damage induced by B. asper venom and possibly other viperid venoms.

  2. Poor regenerative outcome after skeletal muscle necrosis induced by Bothrops asper venom: alterations in microvasculature and nerves.

    Science.gov (United States)

    Hernández, Rosario; Cabalceta, Carmen; Saravia-Otten, Patricia; Chaves, Alessandra; Gutiérrez, José María; Rucavado, Alexandra

    2011-01-01

    Viperid snakebite envenoming is characterized by prominent local tissue damage, including muscle necrosis. A frequent outcome of such local pathology is deficient skeletal muscle regeneration, which causes muscle dysfunction, muscle loss and fibrosis, thus provoking permanent sequelae that greatly affect the quality of life of patients. The causes of such poor regenerative outcome of skeletal muscle after viperid snakebites are not fully understood. A murine model of muscle necrosis and regeneration was adapted to study the effects of the venom and isolated toxins of Bothrops asper, the medically most important snake in Central America. Gastrocnemius muscle was injected with either B. asper venom, a myotoxic phospholipase A(2) (Mtx), a hemorrhagic metalloproteinase (SVMP), or saline solution. At various time intervals, during one month, tissue samples were collected and analyzed by histology, and by immunocytochemical and immunohistochemical techniques aimed at detecting muscle fibers, collagen, endothelial cells, myoblasts, myotubes, macrophages, TUNEL-positive nuclei, and axons. A successful regenerative response was observed in muscle injected with Mtx, which induces myonecrosis but does not affect the microvasculature. In contrast, poor regeneration, with fibrosis and atrophic fibers, occurred when muscle was injected with venom or SVMP, both of which provoke necrosis, microvascular damage leading to hemorrhage, and poor axonal regeneration. The deficient skeletal muscle regeneration after injection of B. asper venom is likely to depend on the widespread damage to the microvasculature, which affects the removal of necrotic debris by phagocytes, and the provision of nutrients and oxygen required for regeneration. In addition, deficient axonal regeneration is likely to contribute to the poor regenerative outcome in this model.

  3. Immunochemical and biological characterization of monoclonal antibodies against BaP1, a metalloproteinase from Bothrops asper snake venom.

    Science.gov (United States)

    Fernandes, I; Assumpção, G G; Silveira, C R F; Faquim-Mauro, E L; Tanjoni, I; Carmona, A K; Alves, M F M; Takehara, H A; Rucavado, A; Ramos, O H P; Moura-da-Silva, A M; Gutiérrez, J M

    2010-11-01

    BaP1 is a P-I class of Snake Venom Metalloproteinase (SVMP) relevant in the local tissue damage associated with envenomations by Bothrops asper, a medically-important species in Central America and parts of South America. Six monoclonal antibodies (MoAb) against BaP1 (MABaP1) were produced and characterized regarding their isotype, dissociation constant (K(d)), specificity and ability to neutralize BaP1-induced hemorrhagic and proteolytic activity. Two MABaP1 are IgM, three are IgG1 and one is IgG2b. The K(d)s of IgG MoAbs were in the nM range. All IgG MoAbs recognized conformational epitopes of BaP1 and B. asper venom components but failed to recognize venoms from 27 species of Viperidae, Colubridae and Elapidae families. Clone 7 cross-reacted with three P-I SVMPs tested (moojeni protease, insularinase and neuwiedase). BaP1-induced hemorrhage was totally neutralized by clones 3, 6 and 8 but not by clone 7. Inhibition of BaP1 enzymatic activity on a synthetic substrate by MABaP1 was totally achieved by clones 3 and 6, and partially by clone 8, but not by clone 7. In conclusion, these neutralizing MoAbs against BaP1 may become important tools to understand structure-function relationships of BaP1 and the role of P-I class SVMP in snakebite envenomation. (c) 2010 Elsevier Ltd. All rights reserved.

  4. Muscle Tissue Damage Induced by the Venom of Bothrops asper: Identification of Early and Late Pathological Events through Proteomic Analysis.

    Science.gov (United States)

    Herrera, Cristina; Macêdo, Jéssica Kele A; Feoli, Andrés; Escalante, Teresa; Rucavado, Alexandra; Gutiérrez, José María; Fox, Jay W

    2016-04-01

    The time-course of the pathological effects induced by the venom of the snake Bothrops asper in muscle tissue was investigated by a combination of histology, proteomic analysis of exudates collected in the vicinity of damaged muscle, and immunodetection of extracellular matrix proteins in exudates. Proteomic assay of exudates has become an excellent new methodological tool to detect key biomarkers of tissue alterations for a more integrative perspective of snake venom-induced pathology. The time-course analysis of the intracellular proteins showed an early presence of cytosolic and mitochondrial proteins in exudates, while cytoskeletal proteins increased later on. This underscores the rapid cytotoxic effect of venom, especially in muscle fibers, due to the action of myotoxic phospholipases A2, followed by the action of proteinases in the cytoskeleton of damaged muscle fibers. Similarly, the early presence of basement membrane (BM) and other extracellular matrix (ECM) proteins in exudates reflects the rapid microvascular damage and hemorrhage induced by snake venom metalloproteinases. The presence of fragments of type IV collagen and perlecan one hour after envenoming suggests that hydrolysis of these mechanically/structurally-relevant BM components plays a key role in the genesis of hemorrhage. On the other hand, the increment of some ECM proteins in the exudate at later time intervals is likely a consequence of the action of endogenous matrix metalloproteinases (MMPs) or of de novo synthesis of ECM proteins during tissue remodeling as part of the inflammatory reaction. Our results offer relevant insights for a more integrative and systematic understanding of the time-course dynamics of muscle tissue damage induced by B. asper venom and possibly other viperid venoms.

  5. Poor regenerative outcome after skeletal muscle necrosis induced by Bothrops asper venom: alterations in microvasculature and nerves.

    Directory of Open Access Journals (Sweden)

    Rosario Hernández

    Full Text Available BACKGROUND: Viperid snakebite envenoming is characterized by prominent local tissue damage, including muscle necrosis. A frequent outcome of such local pathology is deficient skeletal muscle regeneration, which causes muscle dysfunction, muscle loss and fibrosis, thus provoking permanent sequelae that greatly affect the quality of life of patients. The causes of such poor regenerative outcome of skeletal muscle after viperid snakebites are not fully understood. METHODOLOGY/PRINCIPAL FINDINGS: A murine model of muscle necrosis and regeneration was adapted to study the effects of the venom and isolated toxins of Bothrops asper, the medically most important snake in Central America. Gastrocnemius muscle was injected with either B. asper venom, a myotoxic phospholipase A(2 (Mtx, a hemorrhagic metalloproteinase (SVMP, or saline solution. At various time intervals, during one month, tissue samples were collected and analyzed by histology, and by immunocytochemical and immunohistochemical techniques aimed at detecting muscle fibers, collagen, endothelial cells, myoblasts, myotubes, macrophages, TUNEL-positive nuclei, and axons. A successful regenerative response was observed in muscle injected with Mtx, which induces myonecrosis but does not affect the microvasculature. In contrast, poor regeneration, with fibrosis and atrophic fibers, occurred when muscle was injected with venom or SVMP, both of which provoke necrosis, microvascular damage leading to hemorrhage, and poor axonal regeneration. CONCLUSIONS/SIGNIFICANCE: The deficient skeletal muscle regeneration after injection of B. asper venom is likely to depend on the widespread damage to the microvasculature, which affects the removal of necrotic debris by phagocytes, and the provision of nutrients and oxygen required for regeneration. In addition, deficient axonal regeneration is likely to contribute to the poor regenerative outcome in this model.

  6. N-terminal domain of Bothrops asper Myotoxin II Enhances the Activity of Endothelin Converting Enzyme-1 and Neprilysin.

    Science.gov (United States)

    Smith, A Ian; Rajapakse, Niwanthi W; Kleifeld, Oded; Lomonte, Bruno; Sikanyika, Nkumbu L; Spicer, Alexander J; Hodgson, Wayne C; Conroy, Paul J; Small, David H; Kaye, David M; Parkington, Helena C; Whisstock, James C; Kuruppu, Sanjaya

    2016-03-02

    Neprilysin (NEP) and endothelin converting enzyme-1 (ECE-1) are two enzymes that degrade amyloid beta in the brain. Currently there are no molecules to stimulate the activity of these enzymes. Here we report, the discovery and characterisation of a peptide referred to as K49-P1-20, from the venom of Bothrops asper which directly enhances the activity of both ECE-1 and NEP. This is evidenced by a 2- and 5-fold increase in the Vmax of ECE-1 and NEP respectively. The K49-P1-20 concentration required to achieve 50% of maximal stimulation (AC50) of ECE-1 and NEP was 1.92 ± 0.07 and 1.33 ± 0.12 μM respectively. Using BLITZ biolayer interferometry we have shown that K49-P1-20 interacts directly with each enzyme. Intrinsic fluorescence of the enzymes change in the presence of K49-P1-20 suggesting a change in conformation. ECE-1 mediated reduction in the level of endogenous soluble amyloid beta 42 in cerebrospinal fluid is significantly higher in the presence of K49-P1-20 (31 ± 4% of initial) compared with enzyme alone (11 ± 5% of initial; N = 8, P = 0.005, unpaired t-test). K49-P1-20 could be an excellent research tool to study mechanism(s) of enzyme stimulation, and a potential novel drug lead in the fight against Alzheimer's disease.

  7. Snakebites and ethnobotany in the northwest region of Colombia. Part III: neutralization of the haemorrhagic effect of Bothrops atrox venom.

    Science.gov (United States)

    Otero, R; Núñez, V; Barona, J; Fonnegra, R; Jiménez, S L; Osorio, R G; Saldarriaga, M; Díaz, A

    2000-11-01

    Thirty-one of 75 extracts of plants used by traditional healers for snakebites, had moderate or high neutralizing ability against the haemorrhagic effect of Bothrops atrox venom from Antioquia and Chocó, north-western Colombia. After preincubation of several doses of every extract (7.8-4000 microg/mouse) with six minimum haemorrhagic doses (10 microg) of venom, 12 of them demonstrated 100% neutralizing capacity when the mixture was i.d. injected into mice (18-20 g). These were the stem barks of Brownea rosademonte (Caesalpiniaceae) and Tabebuia rosea (Bignoniaceae); the whole plants of Pleopeltis percussa (Polypodiaceae), Trichomanes elegans (Hymenophyllaceae) and Senna dariensis (Caesalpiniaceae); rhizomes of Heliconia curtispatha (Heliconiaceae); leaves and branches of Bixa orellana (Bixaceae), Philodendron tripartitum (Araceae), Struthanthus orbicularis (Loranthaceae) and Gonzalagunia panamensis (Rubiaceae); the ripe fruits of Citrus limon (Rutaceae); leaves, branches and stem of Ficus nymphaeifolia (Moraceae). Extracts of another 19 species showed moderate neutralization (21-72%) at doses up to 4 mg/mouse, e.g. the whole plants of Aristolochia grandiflora (Aristolochiaceae), Columnea kalbreyeriana (Gesneriaceae), Sida acuta (Malvaceae), Selaginella articulata (Selaginellaceae) and Pseudoelephantopus spicatus (Asteraceae); rhizomes of Renealmia alpinia (Zingiberaceae); the stem of Strychnos xinguensis (Loganiaceae); leaves, branches and stems of Hyptis capitata (Lamiaceae), Ipomoea cairica (Convolvulaceae), Neurolaena lobata (Asteraceae), Ocimum micranthum (Lamiaceae), Piper pulchrum (Piperaceae), Siparuna thecaphora (Monimiaceae), Castilla elastica (Moraceae) and Allamanda cathartica (Apocynaceae); the macerated ripe fruits of Capsicum frutescens (Solanaceae); the unripe fruits of Crescentia cujete (Bignoniaceae); leaves and branches of Piper arboreum (Piperaceae) and Passiflora quadrangularis (Passifloraceae). When the extracts were independently administered

  8. BaltDC: purification, characterization and infrared spectroscopy of an antiplatelet DC protein isolated from Bothrops alternatus snake venom.

    Science.gov (United States)

    Matias, Mariana Santos; de Sousa, Bruna Barbosa; da Cunha Pereira, Déborah Fernanda; Dias, Edigar Henrique Vaz; Mamede, Carla Cristine Neves; de Queiroz, Mayara Ribeiro; Silva, Anielle Christine Almeida; Dantas, Noelio Oliveira; Soares, Andreimar Martins; de Oliveira Costa, Júnia; de Oliveira, Fábio

    2017-01-01

    Snake venoms are a complex mixture of proteins, organic and inorganic compounds. Some of these proteins, enzymatic or non-enzymatic ones, are able to interact with platelet receptors, causing hemostatic disorders. The possible therapeutic potential of toxins with antiplatelet properties may arouse interest in the pharmacological areas. The present study aimed to purify and characterize an antiplatelet DC protein from Bothrops alternatus snake venom. The protein, called BaltDC (DC protein from B. alternatus snake venom), was purified by a combination of ion-exchange chromatography on DEAE-Sephacel column and gel filtration on Sephadex G-75. The molecular mass was estimated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE). The amino acid sequence of the N-terminal region was carried out by Edman degradation method. Platelet aggregation assays were performed in human platelet-rich plasma (PRP). Infrared (IR) spectroscopy was used in order to elucidate the interactions between BaltDC and platelet membrane. BaltDC ran as a single protein band on SDS-PAGE and showed apparent molecular mass of 32 kDa under reducing or non-reducing conditions. The N-terminal region of the purified protein revealed the amino acid sequence IISPPVCGNELLEVGEECDCGTPENCQNECCDA, which showed identity with other snake venom metalloproteinases (SVMPs). BaltDC was devoid of proteolytic, hemorrhagic, defibrinating or coagulant activities, but it showed a specific inhibitory effect on platelet aggregation induced by ristocetin and epinephrine in PRP. IR analysis spectra strongly suggests that PO 3 2- groups, present in BaltDC, form hydrogen bonds with the PO 2 - groups present in the non-lipid portion of the membrane platelets. BaltDC may be of medical interest since it was able to inhibit platelet aggregation.

  9. Angiogenenic effects of BpLec, a C-type lectin isolated from Bothrops pauloensis snake venom.

    Science.gov (United States)

    Castanheira, Letícia Eulalio; Lopes, Daiana Silva; Gimenes, Sarah Natalie Cirilo; Deconte, Simone Ramos; Ferreira, Bruno Antônio; Alves, Patricia Terra; Filho, Luiz Ricardo Goulart; Tomiosso, Tatiana Carla; Rodrigues, Renata Santos; Yoneyama, Kelly Aparecida Geraldo; Araújo, Fernanda de Assis; Rodrigues, Veridiana de Melo

    2017-09-01

    The present work reports the effects of a C-type lectin (BpLec) isolated from Bothrops pauloensis snake venom upon in vitro and in vivo angiogenesis models. Initially, we noted that BpLec was not cytotoxic to endothelial cells (tEnd) in doses up to 40μg/mL, but lower doses (2.5μg/mL, 5μg/mL, 10μg/mL and 20μg/mL) reduced tEnd cells adhesion to some extracellular matrix proteins and inhibited the in vitro vessel formation in Matrigel assay stimulated by bFGF. β-galactosides (d-lactose, N-acetyl-d-galactosamine and d-galactose) at 400mM reversed the effect of BpLec on tEnd cells adhesion, whereas d-galactose (400mM) partially reversed BpLec property of inhibiting vessel formation by tEnd cells in Matrigel. In vivo assays showed that BpLec increased hemoglobin content and capillary vessels number in polyether-polyurethane sponge discs subcutaneously implanted into dorsal skin mice. Additionally, BpLec also reduced collagen deposition and did not induce a pro-inflammatory response, as demonstrated by the decreased the secretion of some inflammatory cytokines, whereas myeloperoxidase (MPO) and N-acetylglucosaminidase (NAG) activities were not altered by BpLec. Taken together, our results indicate that BpLec might represent an interesting angiogenesis and inflammatory modulator that could also be used for searching possible therapeutic targets involved in these processes. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Structural and binding studies of a C-type galactose-binding lectin from Bothrops jararacussu snake venom.

    Science.gov (United States)

    Sartim, Marco A; Pinheiro, Matheus P; de Pádua, Ricardo A P; Sampaio, Suely V; Nonato, M Cristina

    2017-02-01

    BJcuL is a snake venom galactoside-binding lectin (SVgalL) isolated from Bothrops jararacussu and is involved in a wide variety of biological activities including triggering of pro-inflammatory response, disruption of microbial biofilm structure and induction of apoptosis. In the present work, we determined the crystallographic structure of BJcuL, the first holo structure of a SVgalL, and introduced the fluorescence-based thermal stability assay (Thermofluor) as a tool for screening and characterization of the binding mechanism of SVgalL ligands. BJcuL structure revealed the existence of a porous and flexible decameric arrangement composed of disulfide-linked dimers related by a five-fold symmetry. Each monomer contains the canonical carbohydrate recognition domain, a calcium ion required for BJcuL lectinic activity and a sodium ion required for protein stabilization. BJcuL thermostability was found to be induced by calcium ion and galactoside sugars which exhibit hyperbolic saturation profiles dependent on ligand concentration. Serendipitously, the gentamicin group of aminoglycoside antibiotics (gAGAs) was also identified as BJcuL ligands. On contrast, gAGAs exhibited a sigmoidal saturation profile compatible with a cooperative mechanism of binding. Thermofluor, hemagglutination inhibition assay and molecular docking strategies were used to identify a distinct binding site in BJcuL localized at the dimeric interface near the fully conserved intermolecular Cys86-Cys86 disulfide bond. The hybrid approach used in the present work provided novel insights into structural behavior and functional diversification of SVgaLs. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. Isotopic change in the tissues of Bothrops atrox in captivity collected from environments of the eastern Amazon

    Science.gov (United States)

    Martinez, M. G.; Chalkidis, H. D.; Amazonas, D. R.; da Silva, A. M.; De Oliveira, R., Jr.; Camargo, P. B.

    2013-12-01

    The Bothrops atrox is little studied because it is sympatric Amazonian animals, and very little is known about the ecology and natural history of this species. It has a generalist diet and the distribution of this species is very wide. The adult animals forage mostly on the ground, while the younger animals prefer to stay on the vegetation. They are easily find in the rainy months in areas near lakes and seasonally flooded and are difficult to find in the driest months, a period where there is less availability of preys in these environments. Due to its aggressiveness, is considered one of the most feared snakes in South America and in the eastern Amazon, being responsible for the largest number of snakebites in the region. Through stable isotope carbon-13 and nitrogen-15, is intended to characterize the variations of the feeding habits of these collected animals in different environments and also when they are kept in captivity, feeding the animal's bioterium. The serpents were collected in environments with different land uses, such as native forest, savannah, pasture and have been brought to the serpentarium Integrated College Tapajos (FIT), being retained in order to Samplings throughout the experiment with feeding mice's own bioterium. When these snakes came from different locations, samples were collected scales and blood (T0), before receiving the new supply (captive), and every time we fed the mice the vivarium, new tissue samples were collected, (T1, T2, T3) to exchange all the nature of food for the food captivity.Based on the results of δ13C and δ15N, the samples collected in the tissues of snakes of different environments (nature and captivity), it was observed that changes in food sources reflect changes in tissues (blood and scales), also reflecting the production of poison different periods of turnover of absorbed material in those tissues, contributing to the study of animal ecology and behavior in relation to habitat.

  12. Activation of J77A.1 Macrophages by Three Phospholipases A2 Isolated from Bothrops atrox Snake Venom

    Directory of Open Access Journals (Sweden)

    Juliana L. Furtado

    2014-01-01

    Full Text Available In the present study, we investigated the in vitro effects of two basic myotoxic phospholipases A2 (PLA2, BaTX-I, a catalytically inactive Lys-49 variant, and BaTX-II, a catalytically active Asp-49, and of one acidic myotoxic PLA2, BaPLA2, a catalytically active Asp-49, isolated from Bothrops atrox snake venom, on the activation of J774A.1 macrophages. At noncytotoxic concentrations, the toxins did not affect the adhesion of the macrophages, nor their ability to detach. The data obtained showed that only BaTX-I stimulated complement receptor-mediated phagocytosis. However, BaTX-I, BaTX-II, and BaPLA2 induced the release of the superoxide anion by J774A.1 macrophages. Additionally, only BaTX-I raised the lysosomal volume of macrophages after 15 min of incubation. After 30 min, all the phospholipases increased this parameter, which was not observed within 60 min. Moreover, BaTX-I, BaTX-II, and BaPLA2 increased the number of lipid bodies on macrophages submitted to phagocytosis and not submitted to phagocytosis. However, BaTX-II and BaPLA2 induced the release of TNF-α by J774A.1 macrophages. Taken together, the data show that, despite differences in enzymatic activity, the three toxins induced inflammatory events and whether the enzyme is acidic or basic does not seem to contribute to these effects.

  13. SISTEMA FIBRINOLÍTICO: MÉTODOS DE ESTUDIO Y HALLAZGOS EN VENENOS DE SERPIENTES DE LOS GENEROS Bothrops, Crotalus, Micrurus EN VENEZUELA | FIBRINOLYTIC SYSTEM: METHODS OF STUDY AND FINDINGS IN Bothrops, Crotalus, Micrurus SNAKE VENOMS IN VENEZUELA

    Directory of Open Access Journals (Sweden)

    Alexis Rodríguez-Acosta,

    2016-10-01

    Full Text Available En la presente revisión se muestran resultados relevantes de actividades hemostáticas, con especial énfasis a las relacionadas con la fibrinólisis, presentes en venenos de serpientes venezolanas de los géneros Bothrops, Crotalus y Micrurus, que existen en diversas localidades geográficas. Con esta descripción, se presenta además una revisión de diversos ensayos que se utilizan en investigación para estos estudios. Se han evidenciado importantes diferencias inter e intra-especies, e incluso entre ejemplares de distintos espacios geográficos, lo que debe influir en los mecanismos de acción de sus venenos y la sintomatología clínica. Estas diferencias en composición y concentración de componentes activos, deben estar asociadas a variaciones estacionales o geográficas, dieta, sexo, hábitat, edad, e inclusive variabilidades genéticas. Ello podría explicar las discrepancias en los cuadros clínicos de las víctimas y también ayudar a determinar mecanismos de acción que faciliten una mejor orientación a los médicos tratantes. Estas variables ayudarían al seguimiento y tratamiento de los pacientes, orientando a los especialistas en antivenenos, en la escogencia del inóculo, con una mezcla amplia de venenos de diferentes especies, géneros, edad, sexo y procedencia, así como con toxinas de relevancia, para la preparación y obtención de estos antivenenos con valores de gran eficacia y amplio espectro. Los componentes bioactivos aislados, como las colombienasas y la tenerplasminina-1, por sus características biológicas, representan asimismo moléculas con un uso potencial como trombolíticos y antifibrinolíticos, respectivamente.

  14. Biotechnological application of protein Leuc-B isolated from Bothrops leucurus venom as a prototype for antitumoral radiopharmaceutical;Aplicacao biotecnologica da proteina Leuc-B isolada da peconha de Bothrops leucurus como prototipo de radiofarmaco antitumoral

    Energy Technology Data Exchange (ETDEWEB)

    Gabriel, Lucilene Marcia

    2010-07-01

    According to the report of the International Agency for Research on Cancer, the growth of this disease implies the death of 17 million people a year by 2030. Although the knowledge on development of cancer is growing considerably, just a few advances in the diagnosis and therapy has been achieved. Faced with this scenario, it is clear the need for new substances more specifics with low toxicity to the patient, which can be used for diagnosis and treatment of cancer. Membrane receptors over expressed in tumor cells are promising target candidates for development of diagnostic and therapeutical tools. Integrins are a family of hetero dimeric cell surface adhesion receptors able to recognize and bind to proteins in the extracellular matrix (ECM). This recognition is mainly through the RGD domains presents in both the cell surface as in the protein from the ECM. Various integrins have been identified as regulators of tumor progression. The RGD domain is also found in some snake venoms named disintegrins. Disintegrins inhibit cell-matrix and a cell-cell interactions mediated by integrins and it has been shown that these proteins are able to inhibit metastasis in processes dependent on integrin. The disintegrin-like (ECD), as well as RGD-disintegrin are also able to bind to cell surface integrins and inhibit their adhesion to the natural ligands. In this work it was purified from Bothrops leucurus venom (VBL), a metalloproteinase-class P-III with disintegrin-like domain (ECD), Leucurolisina B (Leuc-B). This metalloproteinase and the crude venom were used to evaluate their applicability in the differential detection of tumors. In vitro results demonstrated that both VBL and Leuc-B have potent antitumoral effect on several cancer cell lines: U87, T98, RT2 (glioblastoma), MCF7 (breast), Ehrlich and UACC (melanoma) with IC{sub 50} values of approximately 0.6 muM. The morphological changes observed in these strains when treated with Leuc-B, and data from the DAPI staining

  15. Structural and Functional Studies of a Bothropic Myotoxin Complexed to Rosmarinic Acid: New Insights into Lys49-PLA2 Inhibition

    Science.gov (United States)

    dos Santos, Juliana I.; Cardoso, Fábio F.; Soares, Andreimar M.; dal Pai Silva, Maeli; Gallacci, Márcia; Fontes, Marcos R. M.

    2011-01-01

    Snakebite envenoming is an important public health problem in many tropical and subtropical countries, and is considered a neglected tropical disease by the World Health Organization. Most severe cases are inflicted by species of the families Elapidae and Viperidae, and lead to a number of systemic and local effects in the victim. One of the main problems regarding viperidic accidents is prominent local tissue damage whose pathogenesis is complex and involves the combined actions of a variety of venom components. Phospholipases A2 (PLA2s) are the most abundant muscle-damaging components of these venoms. Herein, we report functional and structural studies of PrTX-I, a Lys49-PLA2 from Bothops pirajai snake venom, and the influence of rosmarinic acid (RA) upon this toxin's activities. RA is a known active component of some plant extracts and has been reported as presenting anti-myotoxic properties related to bothopic envenomation. The myotoxic activity of Lys49-PLA2s is well established in the literature and although no in vivo neurotoxicity has been observed among these toxins, in vitro neuromuscular blockade has been reported for some of these proteins. Our in vitro studies show that RA drastically reduces both the muscle damage and the neuromuscular blockade exerted by PrTX-I on mice neuromuscular preparations (by ∼80% and ∼90%, respectively). These results support the hypothesis that the two effects are closely related and lead us to suggest that they are consequences of the muscle membrane-destabilizing activity of the Lys49-PLA2. Although the C-terminal region of these proteins has been reported to comprise the myotoxic site, we demonstrate by X-ray crystallographic studies that RA interacts with PrTX-I in a different region. Consequently, a new mode of Lys49-PLA2 inhibition is proposed. Comparison of our results with others in the literature suggests possible new ways to inhibit bothropic snake venom myotoxins and improve serum therapy. PMID:22205953

  16. Biotechnological application of protein Leuc-B isolated from Bothrops leucurus venom as a prototype for antitumoral radiopharmaceutical

    International Nuclear Information System (INIS)

    Gabriel, Lucilene Marcia

    2010-01-01

    According to the report of the International Agency for Research on Cancer, the growth of this disease implies the death of 17 million people a year by 2030. Although the knowledge on development of cancer is growing considerably, just a few advances in the diagnosis and therapy has been achieved. Faced with this scenario, it is clear the need for new substances more specifics with low toxicity to the patient, which can be used for diagnosis and treatment of cancer. Membrane receptors over expressed in tumor cells are promising target candidates for development of diagnostic and therapeutical tools. Integrins are a family of hetero dimeric cell surface adhesion receptors able to recognize and bind to proteins in the extracellular matrix (ECM). This recognition is mainly through the RGD domains presents in both the cell surface as in the protein from the ECM. Various integrins have been identified as regulators of tumor progression. The RGD domain is also found in some snake venoms named disintegrins. Disintegrins inhibit cell-matrix and a cell-cell interactions mediated by integrins and it has been shown that these proteins are able to inhibit metastasis in processes dependent on integrin. The disintegrin-like (ECD), as well as RGD-disintegrin are also able to bind to cell surface integrins and inhibit their adhesion to the natural ligands. In this work it was purified from Bothrops leucurus venom (VBL), a metalloproteinase-class P-III with disintegrin-like domain (ECD), Leucurolisina B (Leuc-B). This metalloproteinase and the crude venom were used to evaluate their applicability in the differential detection of tumors. In vitro results demonstrated that both VBL and Leuc-B have potent antitumoral effect on several cancer cell lines: U87, T98, RT2 (glioblastoma), MCF7 (breast), Ehrlich and UACC (melanoma) with IC 50 values of approximately 0.6 μM. The morphological changes observed in these strains when treated with Leuc-B, and data from the DAPI staining solution

  17. Crystal structure of a phospholipase A2from Bothrops asper venom: Insights into a new putative "myotoxic cluster".

    Science.gov (United States)

    Salvador, Guilherme H M; Dos Santos, Juliana I; Lomonte, Bruno; Fontes, Marcos R M

    2017-02-01

    Snake venoms from the Viperidae and Elapidae families often have several phospholipases A 2 (PLA 2 s), which may display different functions despite having a similar structural scaffold. These proteins are considered an important target for the development of drugs against local myotoxic damage because they are not efficiently neutralized by conventional serum therapy. PLA 2 s from these venoms are generally divided into two classes: (i) catalytic PLA 2 s (or Asp49-PLA 2 s) and (ii) non-catalytic PLA 2 -like toxins (or Lys49-PLA 2 s). In many Viperidae venoms, a subset of the basic Asp49-PLA 2 s displays some functional and structural characteristics of PLA 2 -like proteins and group within the same phylogenetic clade, but their myotoxic mechanism is still largely unknown. In the present study, we have crystallized and solved the structure of myotoxin I (MT-I), a basic myotoxic Asp49-PLA 2 isolated from Bothrops asper venom. The structure presents a dimeric conformation that is compatible with that of previous dimers found for basic myotoxic Asp49-PLA 2 s and Lys49-PLA 2 s and has been confirmed by other biophysical and bioinformatics techniques. This arrangement suggests a possible cooperative action between both monomers to exert myotoxicity via two different sites forming a putative membrane-docking site (MDoS) and a putative membrane disruption site (MDiS). This mechanism would resemble that proposed for Lys49-PLA 2 s, but the sites involved appear to be situated in a different region. Thus, as both sites are close to one another, they form a "myotoxic cluster", which is also found in two other basic myotoxic Asp49-PLA 2 s from Viperidae venoms. Such arrangement may represent a novel structural strategy for the mechanism of muscle damage exerted by the group of basic, Asp49-PLA 2 s found in viperid snake venoms. Copyright © 2016 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

  18. Diversity of metalloproteinases in Bothrops neuwiedi snake venom transcripts: evidences for recombination between different classes of SVMPs

    Directory of Open Access Journals (Sweden)

    Valente Richard H

    2011-11-01

    Full Text Available Abstract Background Snake venom metalloproteinases (SVMPs are widely distributed in snake venoms and are versatile toxins, targeting many important elements involved in hemostasis, such as basement membrane proteins, clotting proteins, platelets, endothelial and inflammatory cells. The functional diversity of SVMPs is in part due to the structural organization of different combinations of catalytic, disintegrin, disintegrin-like and cysteine-rich domains, which categorizes SVMPs in 3 classes of precursor molecules (PI, PII and PIII further divided in 11 subclasses, 6 of them belonging to PII group. This heterogeneity is currently correlated to genetic accelerated evolution and post-translational modifications. Results Thirty-one SVMP cDNAs were full length cloned from a single specimen of Bothrops neuwiedi snake, sequenced and grouped in eleven distinct sequences and further analyzed by cladistic analysis. Class P-I and class P-III sequences presented the expected tree topology for fibrinolytic and hemorrhagic SVMPs, respectively. In opposition, three distinct segregations were observed for class P-II sequences. P-IIb showed the typical segregation of class P-II SVMPs. However, P-IIa grouped with class P-I cDNAs presenting a 100% identity in the 365 bp at their 5' ends, suggesting post-transcription events for interclass recombination. In addition, catalytic domain of P-IIx sequences segregated with non-hemorrhagic class P-III SVMPs while their disintegrin domain grouped with other class P-II disintegrin domains suggesting independent evolution of catalytic and disintegrin domains. Complementary regions within cDNA sequences were noted and may participate in recombination either at DNA or RNA levels. Proteins predicted by these cDNAs show the main features of the correspondent classes of SVMP, but P-IIb and P-IIx included two additional cysteines cysteines at the C-termini of the disintegrin domains in positions not yet described. Conclusions In

  19. The inhibitory potential of the condensed-tannin-rich fraction of Plathymenia reticulata Benth. (Fabaceae) against Bothrops atrox envenomation.

    Science.gov (United States)

    de Moura, Valéria Mourão; da Silva, Wania Cristina Rodrigues; Raposo, Juliana D A; Freitas-de-Sousa, Luciana A; Dos-Santos, Maria Cristina; de Oliveira, Ricardo Bezerra; Veras Mourão, Rosa Helena

    2016-05-13

    Ethnobotanical studies have shown that Plathymenia reticulata Benth. (Fabaceae) has been widely used in cases of snake envenomation, particularly in Northern Brazil. In light of this, the aim of this study was to evaluate the inhibitory potential of the condensed-tannin-rich fraction obtained from the bark of P. reticulata against the main biological activities induced by Bothrops atrox venom (BaV). The chemical composition of the aqueous extract of P. reticulata (AEPr) was first investigated by thin-layer chromatography (TLC) and the extract was then fractionated by column chromatography on Sephadex LH-20. This yielded five main fractions (Pr1, Pr2, Pr3, Pr4 and Pr5), which were analyzed by colorimetry to determine their concentrations of total phenolics, total tannins and condensed tannins and to assess their potential for blocking the phospholipase activity of BaV. The Pr5 fraction was defined as the fraction rich in condensed tannins (CTPr), and its inhibitory potential against the activities of the venom was evaluated. CTPr was evaluated in different in vivo and in vitro experimental protocols. The in vivo protocols consisted of (1) pre-incubation (venom:CTPr, w/w), (2) pre-treatment (orally administered) and (3) post-treatment (orally administered) to evaluate the effect on the hemorrhagic and edematogenic activities of BaV; in the in vitro protocol the effect on phospholipase and coagulant activity using pre-incubation in both tests was evaluated. There was statistically significant inhibition (p<0.05) of hemorrhagic activity by CTPr when the pre-incubation protocol was used [55% (1:5, w/w) and 74% (1:10, w/w)] and when pre-treatment with doses of 50 and 100mg/kg was used (19% and 13%, respectively). However, for the concentrations tested, there was no statistically significant inhibition in the group subjected to post-treatment administered orally. CTPr blocked 100% of phospholipase activity and 63.3% (1:10, w/w) of coagulant activity when it was pre

  20. Snakebites by Bothrops spp in children in Campinas, São Paulo, Brazil Acidentes por serpentes do gênero Bothrops em crianças em Campinas, São Paulo, Brasil

    Directory of Open Access Journals (Sweden)

    Fábio BUCARETCHI

    2001-12-01

    Full Text Available From January, 1984 to March, 1999, 73 children under 15 y old (ages 1-14 y, median 9 y were admitted after being bitten by snakes of the genus Bothrops. Twenty-six percent of the children were classified as mild envenoming, 50.7% as moderate envenoming and 20.6% as severe envenoming. Two patients (2.7% showed no signs of envenoming. Most of the patients presented local manifestations, mainly edema (94.5%, pain (94.5% ecchymosis (73.9% and blisters (11%. Local and/or systemic bleeding was observed in 28.8% of the patients. Before antivenom (AV administration, blood coagulation disorders were observed in 60.7% (incoagulable blood in 39.3% of the 56 children that received AV only in our hospital. AV early reactions, most of which were considered mild, were observed in 44.6% of these cases (in 15/30 patients not pretreated and in 10/26 patients pretreated with hydrocortisone and histamine H1 and H2 antagonists. The main clinical complications observed were local infection (15.1%, compartment syndrome (4.1%, gangrene (1.4% and acute renal failure (1.4%. No deaths were recorded. There were no significant differences with regard to severity of envenoming versus the frequency of blood coagulation disorders among the three categories of envenoming (p = 0.75 or in the frequency of patients with AV early reactions between the groups that were and were not pretreated (p = 0.55. The frequency of local infection was significantly greater in severe cases (p De janeiro de 1984 a março de 1999, 73 crianças com menos de 15 anos de idade (1 a 14 anos, mediana = 9 anos foram admitidas após terem sido picadas por serpentes do gênero Bothrops. 26,0% das crianças foram classificadas como acidente leve, 50,7% como moderado e 20,6% como grave. Dois pacientes (2,7% não apresentaram sinais de envenenamento. A maioria dos pacientes apresentou manifestações locais, principalmente edema (94,5%, dor (94,5%, equimoses (73,9% e bolhas (11,0%. Sangramento local e/ou sist

  1. Bothrops asper metalloproteinase BaP1 is inhibited by alpha(2)-macroglobulin and mouse serum and does not induce systemic hemorrhage or coagulopathy.

    Science.gov (United States)

    Escalante, Teresa; Rucavado, Alexandra; Kamiguti, Aura S; Theakston, R David G; Gutiérrez, José María

    2004-02-01

    The ability of the P-I metalloproteinase BaP1, isolated from the venom of the snake Bothrops asper, to induce systemic bleeding, thrombocytopenia and defibrinogenation was assessed in an experimental mouse model. Intravenous administration of BaP1 caused neither systemic bleeding nor any evidence of pathology in lungs, kidneys, liver, heart and brain. Moreover, there were no alterations in the whole blood clotting time or in platelet numbers. In addition, BaP1 did not inhibit collagen-induced platelet aggregation in vitro. Proteolytic and hemorrhagic activities of BaP1 were readily inhibited by the plasma proteinase inhibitor, alpha(2)-macroglobulin, and normal mouse serum also inhibited hemorrhage. Such inhibition may explain why BaP1 induces multiple local tissue-damaging effects, but is largely devoid of systemic toxicity.

  2. Sarcophagidae and Calliphoridae related to Rhinella schneideri (Anura, Bufonidae, Bothrops moojeni (Reptilia, Serpentes and Mabuya frenata (Reptilia, Lacertilia carcasses in Brasília, Brazil

    Directory of Open Access Journals (Sweden)

    Roger Maia Dias Ledo

    2012-09-01

    Full Text Available Sarcophagidae and Calliphoridae related to Rhinella schneideri (Anura, Bufonidae, Bothrops moojeni (Reptilia, Serpentes and Mabuya frenata (Reptilia, Lacertilia carcasses in Brasília, Brazil. This paper presents a list of necrophagous insects associated with small size carrions of two reptiles and one amphibian, found in areas of riparian forests and Cerrado sensu stricto physiognomies in a Conservation Unit located in Brasilia, Distrito Federal. We found seven species of insects related to these carcasses, being five Sarcophagidae, one Calliphoridae and one Braconidae parasitoid wasp. Lucilia eximia and Peckia (Pattonella intermutans were the most abundant species in the study, corroborating with other studies that suggests that these species have specializations for colonization of small size animal carcasses.

  3. Inhibition of proteases and phospholipases A2 from Bothrops atrox and Crotalus durissus terrificus snake venoms by ascorbic acid, vitamin E, and B-complex vitamins.

    Science.gov (United States)

    Oliveira, Carlos H M; Simão, Anderson A; Trento, Marcus V C; César, Pedro H S; Marcussi, Silvana

    2016-01-01

    The enzyme inhibition by natural and/ or low-cost compounds may represent a valuable adjunct to traditional serotherapy performed in cases of snakebite, mainly with a view to mitigate the local effects of envenoming. The objective of this study was to evaluate possible interactions between vitamins and enzymes that comprise Bothrops atrox and Crotalus durissus terrificus venoms, in vitro. Proteolysis inhibition assays (substrates: azocasein, collagen, gelatin and fibrinogen), hemolysis, coagulation, hemagglutination were carried out using different proportions of vitamins in face of to inhibit minimum effective dose of each venom. The vitamins were responsible for reducing 100% of breaking azocasein by C.d.t. venom, thrombolysis induced by B. atrox and fibrinogenolysis induced by both venoms. It is suggested the presence of interactions between vitamin and the active site of enzymes, for example the interactions between hydrophobic regions present in the enzymes and vitamin E, as well as the inhibitions exercised by antioxidant mechanism.

  4. Study of the immune response by antibodies against the Bothrops asper venom for the elaboration of a antiophidic vaccine for bovines

    International Nuclear Information System (INIS)

    Gonzalez Rojas, Katherine

    2014-01-01

    Active immunization has determined against Bothrops asper snake venom (tested in murine and bovine models) a induced response by antibody able to prevent in immunized animals. A coagulopathy or death is developed after of be administered with adequate doses of poison. The amount of B. asper venom has defined the poisoning induced in bovine and murine models. The plasmatic concentration of equine antibodies against B. asper venom is specified to prevent coagulopathy and lethality induced by this venom in murine and bovine models. Murine and bovine models have verified the active immunization reached in a concentration of antibodies against B. asper venom equal or greater to the maximum concentration achieved by intravenous administration of antivenoms from equine origin. The concentration of antibodies induced by the active immunization is evaluated against B. asper venom to prevent the development of coagulopathy and lethality induced by the venom in murine and bovine models [es

  5. Digenetic trematodes infection in a Bothrops moojeni (Viperidae population from a fauna rescue in Porto Primavera, São Paulo State

    Directory of Open Access Journals (Sweden)

    Barrella T.H.

    2003-01-01

    Full Text Available No presente estudo é descrita a infecção por trematódeos digenéticos parasitas da cavidade oral e esôfago em uma população de serpentes Bothrops moojeni provenientes de resgate de fauna em Porto Primavera, Estado de São Paulo. Foi observada prevalência de infecção de 68%. O grau de infecção (número de trematódeos por serpente variou de 2 a 51 helmintos. Os trematódeos encontrados foram Ophisthogonimus spp. e Sticholecitha serpentis. A alta prevalência de infecção foi associada com a drástica alteração ambiental e o estresse multi-fatorial aos quais os animais foram submetidos, que poderiam ter favorecido o ciclo dos parasitas.

  6. Snake population venomics and antivenomics of Bothrops atrox: Paedomorphism along its transamazonian dispersal and implications of geographic venom variability on snakebite management.

    Science.gov (United States)

    Calvete, Juan J; Sanz, Libia; Pérez, Alicia; Borges, Adolfo; Vargas, Alba M; Lomonte, Bruno; Angulo, Yamileth; Gutiérrez, José María; Chalkidis, Hipócrates M; Mourão, Rosa H V; Furtado, M Fatima D; Moura-Da-Silva, Ana M

    2011-04-01

    We describe two geographically differentiated venom phenotypes across the wide distribution range of Bothrops atrox, from the Colombian Magdalena Medio Valley through Puerto Ayacucho and El Paují, in the Venezuelan States of Amazonas and Orinoquia, respectively, and São Bento in the Brazilian State of Maranhão. Colombian and Venezuelan venoms show an ontogenetic toxin profile phenotype whereas Brazilian venoms exhibit paedomorphic phenotypes. Venoms from each of the 16 localities sampled contain both population-specific toxins and proteins shared by neighboring B. atrox populations. Mapping the molecular similarity between conspecific populations onto a physical map of B. atrox range provides clues for tracing dispersal routes that account for the current biogeographic distribution of the species. The proteomic pattern is consistent with a model of southeast and southwest dispersal and allopatric fragmentation northern of the Amazon Basin, and trans-Amazonian expansion through the Andean Corridor and across the Amazon river between Monte Alegre and Santarém. An antivenomic approach applied to assess the efficacy towards B. atrox venoms of two antivenoms raised in Costa Rica and Brazil using Bothrops venoms different than B. atrox in the immunization mixtures showed that both antivenoms immunodepleted very efficiently the major toxins (PIII-SVMPs, serine proteinases, CRISP, LAO) of paedomorphic venoms from Puerto Ayacucho (Venezuelan Amazonia) through São Bento, but had impaired reactivity towards PLA(2) and P-I SVMP molecules abundantly present in ontogenetic venoms. The degree of immunodepletion achieved suggests that each of these antivenoms may be effective against envenomations by paedomorphic, and some ontogenetic, B. atrox venoms. Copyright © 2010 Elsevier B.V. All rights reserved.

  7. Isolation of an acidic phospholipase A2 from the venom of the snake Bothrops asper of Costa Rica: biochemical and toxicological characterization.

    Science.gov (United States)

    Fernández, Julián; Gutiérrez, José María; Angulo, Yamileth; Sanz, Libia; Juárez, Paula; Calvete, Juan J; Lomonte, Bruno

    2010-03-01

    Phospholipases A(2) (PLA(2)) are major components of snake venoms, exerting a variety of relevant toxic actions such as neurotoxicity and myotoxicity, among others. Since the majority of toxic PLA(2)s are basic proteins, acidic isoforms and their possible roles in venoms are less understood. In this study, an acidic enzyme (BaspPLA(2)-II) was isolated from the venom of Bothrops asper (Pacific region of Costa Rica) and characterized. BaspPLA(2)-II is monomeric, with a mass of 14,212 +/- 6 Da and a pI of 4.9. Its complete sequence of 124 amino acids was deduced through cDNA and protein sequencing, showing that it belongs to the Asp49 group of catalytically active enzymes. In vivo and in vitro assays demonstrated that BaspPLA(2)-II, in contrast to the basic Asp49 counterparts present in the same venom, lacks myotoxic, cytotoxic, and anticoagulant activities. BaspPLA(2)-II also differed from other acidic PLA(2)s described in Bothrops spp. venoms, as it did not show hypotensive and anti-platelet aggregation activities. Furthermore, this enzyme was not lethal to mice at intravenous doses up to 100 microg (5.9 microg/g), indicating its lack of neurotoxic activity. The only toxic effect recorded in vivo was a moderate induction of local edema. Therefore, the toxicological characteristics of BaspPLA(2)-II suggest that it does not play a key role in the pathophysiology of envenomings by B. asper, and that its purpose might be restricted to digestive functions. Immunochemical analyses using antibodies raised against BaspPLA(2)-II revealed that acidic and basic PLA(2)s form two different antigenic groups in B. asper venom. Copyright (c) 2009 Elsevier Masson SAS. All rights reserved.

  8. Neutralización del efecto hemorrágico inducido por veneno de Bothrops asper (Serpentes: Viperidae por extractos de plantas tropicales

    Directory of Open Access Journals (Sweden)

    Oscar Castro

    1999-09-01

    Full Text Available Se evaluó la capacidad de extractos orgánicos de 48 especies de plantas costarricenses para neutralizar la actividad hemorrágica del veneno de la serpiente Bothrops asper (terciopelo. Los extractos se evaluaron mediante un bioensayo basado en inyecciones intradérmicas de veneno en ratones, o de mezclas veneno-extracto, seguidas de la cuantificación macroscópica de la hemorragia. Se observó una inhibición total de la hemorragia con los extractos etanólico, de acetato de etilo y acuoso de Bursera simaruba, Clusia torresii, C. palmana, Croton draco, Persea americana, Phoebe brenesii, Pimenta dioica, Sapindus saponaria, Smilax cuculmeca y Virola koschnyi. El análisis químico de estos extractos permitió identificar catequinas, flavonas, antocianinas y taninos condensados, los cuales podrían jugar un papel en la inhibición del efecto hemorrágico debido a la capacidad de quelar el ion zinc requerido por las metaloproteinasas hemorrágicas para su acción.Organic extracts representing 48 species included in 30 families of Costa Rican tropical plants were evaluated for their ability to neutralize hemorrhagic activity induced by the venom of the snake Bothrops asper. A bioassay in mice was used, based on intradermal injection of either venom or venom-extract mixtures followed by the measurement of hemorrhagic areas. Total inhibition of hemorrhage was observed with the ethanolic, ethyl acetate and aquous extracts of Bursera simaruba, Clusia torresii, C. palmana, Croton draco, Persea americana, Phoebe brenesii, Pimenta dioica, Sapindus saponaria, Smilax cuculmeca and Virola koschnyi. Chemical analysis of these extracts identified catequines, flavones, anthocyanines and condensated tannins, which may be responsible for the inhibitory effect observed, probably owing to the chelation of the zinc required for the catalytic activity of venom’s hemorrhagic metalloproteinases.

  9. Comparison between IgG and F(ab′)2 polyvalent antivenoms: neutralization of systemic effects induced by Bothrops asper venom in mice, extravasation to muscle tissue, and potential for induction of adverse reactions

    OpenAIRE

    León Montero, Guillermo; Monge Monge, María; Rojas Umaña, Ermila; Lomonte, Bruno; Gutiérrez, José María

    2001-01-01

    Whole IgG and F(ab′)2 equine-derived polyvalent (Crotalinae) antivenoms, prepared from the same batch of hyperimmune plasma, were compared in terms of neutralization of the lethal and defibrinating activities induced by Bothrops asper venom, their ability to reach the muscle tissue compartment in envenomated mice, and their potential for the induction of adverse reactions. Both preparations were adjusted to the same potency against the lethal effect of B. asper venom in experiments involving ...

  10. Aspectos clínico-patológicos e laboratoriais do envenenamento experimental por Bothrops alternatus em bovinos Clinic and pathological and laboratory aspects of experimental poisoning by Bothrops alternatus venom in cattle

    Directory of Open Access Journals (Sweden)

    Saulo A. Caldas

    2008-06-01

    Full Text Available Esse estudo teve como objetivo determinar as alterações clínico-patológicas e os achados laboratoriais em bovinos inoculados com a peçonha de Bothrops alternatus, no intuito de fornecer subsídios para o estabelecimento do diagnóstico e do diagnóstico diferencial, bem como esclarecer pontos obscuros da literatura pertinente. O veneno liofilizado foi diluído em 1 ml de solução fisiológica e administrado a cinco bovinos, por via subcutânea, nas doses de 0,0625, 0,125 e 0,25 mg/kg e a dois outros, por via intramuscular, nas doses de 0,25 e 0,45 mg/kg. Seis bovinos foram a óbito e um que recebeu a dose de 0,0625mg/kg, por via subcutânea, recuperou-se. Os sinais clínicos tiveram início entre 25 minutos a 5 horas 30 minutos após a inoculação. O período de evolução variou de 7 horas 18 minutos a 66 horas 12 minutos. Um animal recuperou-se após 92 horas. O quadro clínico, independentemente das doses, caracterizou-se por aumento de volume (hemorragia/hematoma no local da inoculação, tempo de sangramento aumentado, mucosas hipocoradas e apatia. O exame laboratorial revelou progressiva anemia normocítica normocrômica, trombocitopenia, redução de fibrinogênio e proteínas plasmáticas totais, hematócrito e hemoglobina diminuídos, além de leve aumento dos níveis de creatinaquinase e desidrogenase lática. Á necropsia, havia, a partir do local da inoculação, extensos hematomas e áreas de hemorragia no tecido celular subcutâneo dos animais que receberam o veneno por via subcutânea; nos animais inoculados por via intramuscular, adicionalmente, havia hemorragia intramuscular. O endocárdio esquerdo apresentava extensas hemorragias e verificaram-se petéquias na serosa do rúmen e do omaso e na mucosa do abomaso e da vesícula biliar. Em cinco animais, o cólon, reto e região perirrenal estavam envoltos por coágulos de sangue. Ao exame histológico observou-se, além do quadro hemorragíparo, necrose muscular coagulativa

  11. Accidents caused by Bothrops and Bothropoides in the State of Paraiba: epidemiological and clinical aspects Acidentes causados por serpentes dos gêneros Bothrops e Bothropoides no Estado da Paraíba: aspectos clínicos e epidemiológicos

    Directory of Open Access Journals (Sweden)

    Fagner Neves Oliveira

    2010-12-01

    Full Text Available INTRODUCTION: Bothrops and Bothropoides snakes cause 70% of the ophidic accidents in Brazil. The species that cause ophidic accidents in State of Paraíba are Bothropoides erythromelas, Bothrops leucurus and Bothropoides neuwiedi. METHODS: This is a prospective and transverse study, following a quantitative approach of accidents involving Bothrops and Bothropoides admitted to the Toxicological Assistance and Information Centers of Campina Grande and João Pessoa (Ceatox-CG and Ceatox-JP, aimed at identifying the epidemiological and clinical profile of such accidents. All of the patients admitted had medical diagnoses and were monitored at Ceatox-CG or Ceatox-JP. RESULTS: The genera Bothrops and Bothropoides caused 91.7% of the ophidic accidents reported. Snake bites were frequent in men (75.1%, rural workers (65.1%, literate individuals (69% between 11 and 20 years-old (21.7%, and toes the most common area attacked (52.7%. Most (86.6% patients were admitted within 6 hours after the accident/bite, with a predominance of mild cases (64.6%. The annual occurrence in Paraíba was 5.5 accidents/100,000 inhabitants and lethality was 0.2%. CONCLUSIONS: Positive changes in the profiles of these accidents were verified, such as the non-application of inadequate solutions, including the use of tourniquet, coffee grounds, garlic, suction and/or cutting the bitten area. Moreover, the Itinerant Laboratory project, linked to Paraíba State University in partnership with Ceatox-CG, has contributed positively, providing several cities of the state with information regarding the prevention of accidents involving venomous animals. The local press has also contributed, reporting the educational work developed by the centers.INTRODUÇÃO: As serpentes Bothrops e Bothropoides são responsáveis por 70% dos acidentes ofídicos ocorridos no Brasil. As espécies causadoras de acidentes na Paraíba são Bothropoides erythromelas, Bothrops leucurus e Bothropoides neuwiedi

  12. Variaciones en las actividades enzimáticas del veneno de la serpiente Bothrops atrox "jergón", de tres zonas geográficas del Perú Variation of the enzymatic activity of Bothrops atrox "jergon" snake venom from three geographic regions, Peru

    Directory of Open Access Journals (Sweden)

    César Ortiz

    2012-06-01

    Full Text Available Objetivos. Estudiar la variabilidad en la composición y actividades enzimáticas entre venenos de ejemplares adultos de Bothrops atrox. Materiales y métodos. Se emplearon venenos de serpientes adultas procedentes de Amazonas, Junín y Ucayali. A cada una de las muestras se les realizó el análisis del contenido proteico y del número de bandas por PAGESDS, así como las actividades de fosfolipasa A2, hemolítica indirecta, amidolítica, coagulante, hemorrágica y proteolítica sobre caseína y mediante zimograma; además, se hicieron ensayos de inmunodifusión y neutralización in vitro con el suero antibotrópico polivalente del Instituto Nacional de Salud de Perú. Resultados. Las actividades amidolítica, coagulante, hemorrágica, proteolítica mediante zimograma, fosfolipasa A2 y hemolítica indirecta fueron variables, evidenciándose en las tres últimas una mayor actividad en los venenos de Amazonas, mientras que en la cantidad de proteína, bandas electroforéticas y actividad proteolítica sobre caseína no se observaron diferencias. Con respecto a las pruebas de neutralización, 0,5 dosis del antiveneno fueron suficientes para neutralizar con eficacia (más del 50% la actividad coagulante y fosfolipasa A2 de todas las muestras analizadas. Conclusiones. Algunas propiedades biológicas del veneno de ejemplares adultos de Bothrops atrox de Perú son variables, sin que ello afecte la neutralización in vitro por parte del suero antibotrópico polivalente sobre las actividades coagulante y fosfolipasa A2 del veneno.Objectives. To study the variability in the composition and enzymatic activity of venom from adult Bothrops atrox specimens. Materials and methods. We used venoms from adult snakes from Amazonas, Junín and Ucayali. Each of the venom samples underwent analysis for protein and number of bands by pagesds. Phospholipase A2, hemolytic, amidolytic, coagulant, hemorrhagic activity were analyzed, also and proteolytic activity on

  13. Sarcophagidae and Calliphoridae related to Rhinella schneideri (Anura, Bufonidae, Bothrops moojeni (Reptilia, Serpentes and Mabuya frenata (Reptilia, Lacertilia carcasses in Brasília, Brazil Sarcophagidae e Calliphoridae associados às carcaças de Rhinella schneideri (Anura, Bufonidae, Bothrops moojeni (Reptilia, Serpentes e Mabuya frenata (Reptilia, Lacertilia em Brasília, Distrito Federal, Brasil

    Directory of Open Access Journals (Sweden)

    Roger Maia Dias Ledo

    2012-09-01

    Full Text Available Sarcophagidae and Calliphoridae related to Rhinella schneideri (Anura, Bufonidae, Bothrops moojeni (Reptilia, Serpentes and Mabuya frenata (Reptilia, Lacertilia carcasses in Brasília, Brazil. This paper presents a list of necrophagous insects associated with small size carrions of two reptiles and one amphibian, found in areas of riparian forests and Cerrado sensu stricto physiognomies in a Conservation Unit located in Brasilia, Distrito Federal. We found seven species of insects related to these carcasses, being five Sarcophagidae, one Calliphoridae and one Braconidae parasitoid wasp. Lucilia eximia and Peckia (Pattonella intermutans were the most abundant species in the study, corroborating with other studies that suggests that these species have specializations for colonization of small size animal carcasses.Sarcophagidae e Calliphoridae associados às carcaças de Rhinella schneideri (Anura, Bufonidae, Bothrops moojeni (Reptilia, Serpentes e Mabuya frenata (Reptilia, Lacertilia em Brasília, Distrito Federal, Brasil. Este trabalho apresenta uma lista de insetos decompositores associados a carcaças de pequeno porte de dois répteis e de um anfíbio, encontrados em áreas de matas de galeria e de cerrado sensu stricto em unidades de conservação do Distrito Federal. Foram encontradas sete espécies de insetos associados a essas carcaças, sendo cinco sarcofagídeos, um califorídeo e uma vespa parasitóide Braconidae. Lucilia eximia e Peckia (Pattonella intermutans foram as espécies mais abundantes, corroborando com outros estudos que sugerem que estas espécies apresentam especializações para a colonização de carcaças menores.

  14. Construction of concept maps as tool for Biochemistry learning

    Directory of Open Access Journals (Sweden)

    Silvia Lopes de Menezes

    2006-07-01

    Full Text Available The use of concept maps on the teaching of sciences has been object of worldwide research with different purposes: to detect the previous knowledge of the students on certain topics or to evaluate learning, among others. Based on Ausubel´s cognitive psychology, concept maps assume that the learning is accomplished by assimilation of new concepts and propositions to the students´ cognitive structure, contributing to establish links between the previous and new knowledge. It is especially interesting on the approach of interdisciplinary issues, as many studied in Biochemistry.The relevance of the use of concept maps on biochemistry learning was evaluated on a thirty-hour undergraduation optional course, with interdisciplinary topics, which are not usually included on introductory Biochemistry courses. The course Biochemistry of Animal Venoms was structured in seven module where the biochemical action mechanisms of the venoms of Crotalus sp (south american rattlesnake, Bothrops sp (jararaca, Loxosceles sp (brown spider, Tityus sp (yellow scorpion, Phoneutria sp (armed spider, Apis mellifera (honey bee and Latrodectus sp (black widowwere discussed. The students worked in small groups and, at each module, there were (1 an oriented study, guided by questions, texts and schemes, supervised by the teachers, (2 the construction of individual concept maps, where the local and systemic effects of the venoms should be predicted by their biochemical composition and (3 the construction of a new map by the group, incorporating the information of the individual maps. The difficulty level of these tasks was gradually increased throughout the course, with lesser time to carry out the tasks, lesser assistance during the oriented study and even lesser information on the venom effects.The course assessment was given by the number, quality and correction of the concepts relationship present in the concept maps, through a questionnaire and by the

  15. Prevalência de Cryptosporidium serpentis em serpentes de cativeiro Prevalence of Cryptosporidium serpentis in captive snakes

    Directory of Open Access Journals (Sweden)

    Paulo Custório Ruggiero

    2011-11-01

    Full Text Available Cryptosporidium é um protozoário encontrado em uma grande variedade de espécies animais como responsável por casos de gastrite e enterite, porém com epidemiologia pouco conhecida em animais silvestres. A presente investigação teve como objetivo avaliar a prevalência de Cryptosporidium serpentis em lavado gástrico de serpentes mantidas em cativeiro no serpentário do Instituto Butantan (São Paulo, Brasil. A coleta foi realizada uma semana após alimentação, evitando, assim, a regurgitação devido à manipulação. Foram realizados esfregaços do sedimento do lavado gástrico, obtido por centrifugação, corados pela técnica de coloração de Kinyoun. Parte do sedimento foi submetido à técnica de RFLP-PCR para identificação da espécie de Cryptosporidium. O serpentário é dividido em três seções, por espécie - a primeira com oito jibóias (Boa constrictor amarali, a segunda com dez jararacas (Bothropoides jararaca e a última com sete cascavéis (Caudisona durissa. A prevalência de C. serpentis encontrada neste estudo para as serpentes C. durissa, B. jararaca e Boa c. amarali, foi de 57,14% (04/07, 40% (04/10 e 37,5% (03/08, respectivamente, revelando importante ocorrência desse protozoário em serpentes de cativeiro. Apesar da alta prevalência encontrada, apenas as jiboias apresentaram sintomas como perda de peso e regurgitação, refletindo uma sensibilidade diferente da espécie para C. serpentis.Cryptosporidium is a protozoan found in a wide variety of animal species which is responsible for gastritis and enteritis, but its epidemiology is poorly known in wild animals. The present investigation aimed to evaluate the prevalence of Cryptosporidium serpentis in gastric aspirate of captive snakes from the public serpentarium of the Butantan Institute (São Paulo, Brazil. Sampling was performed preferably one week after feeding, thereby preventing regurgitation due to manipulation. Smears were done from the gastric

  16. Crystallization and preliminary X-ray crystallographic studies of a Lys49-phospholipase A2 homologue from Bothrops pirajai venom complexed with rosmarinic acid

    International Nuclear Information System (INIS)

    Santos, Juliana I. dos; Santos-Filho, Norival A.; Soares, Andreimar M.; Fontes, Marcos R. M.

    2010-01-01

    PrTX-I, a noncatalytic and myotoxic Lys49-phospholipase A 2 from B. pirajai venom, was cocrystallized with the inhibitor rosmarinic acid from C. verbenacea. The crystals diffracted X-rays to 1.8 Å resolution and the structure was solved, indicating a remarkable electronic density for the ligand at the entrance to the hydrophobic channel. PrTX-I, a noncatalytic and myotoxic Lys49-phospholipase A 2 from Bothrops pirajai venom, was crystallized in the presence of the inhibitor rosmarinic acid (RA). This is the active compound in the methanolic extract of Cordia verbenacea, a plant that is largely used in Brazilian folk medicine. The crystals diffracted X-rays to 1.8 Å resolution and the structure was solved by molecular-replacement techniques, showing electron density that corresponds to RA molecules at the entrance to the hydrophobic channel. The crystals belong to space group P2 1 2 1 2 1 , indicating conformational changes in the structure after ligand binding: the crystals of all apo Lys49-phospholipase A 2 structures belong to space group P3 1 21, while the crystals of complexed structures belong to space groups P2 1 or P2 1 2 1 2 1

  17. Effect of the metalloproteinase inhibitor batimastat in the systemic toxicity induced by Bothrops asper snake venom: understanding the role of metalloproteinases in envenomation.

    Science.gov (United States)

    Rucavado, Alexandra; Escalante, Teresa; Gutiérrez, José María

    2004-03-15

    The peptidomimetic hydroxamate metalloproteinase inhibitor batimastat (BB-94) was assessed for its ability to neutralize the systemic effects (lethality, hemorrhage and coagulopathy) induced by the venom of Bothrops asper, the most important snake from a medical standpoint in Central America. Batimastat inhibited lethality when a venom challenge dose of two LD(50)s was used by intraperitoneal and intravenous routes, with ED(50)s of 250 and 22 microM, respectively. With a challenge dose of three LD(50)s, lethality was not abrogated, but a conspicuous and dose-dependent delay in the time of death was observed in mice injected with mixtures of venom plus batimastat. Upon incubation with 500 microM batimastat, venom LD(50) increased 2.86-fold (intraperitoneal route) and 2.37-fold (intravenous route), when compared with LD(50) of venom alone. Batimastat also inhibited the hemorrhagic effect induced by venom in the lungs after intravenous injection. Moreover, batimastat exerted a significant inhibition of in vitro coagulant and in vivo defibrinogenating effects of venom, evidencing that metalloproteinases play a key role in the coagulopathy characteristic of B. asper envenomation. The remaining uninhibited coagulant effect is due to serine proteinases, i.e. thrombin-like enzymes, since this effect was completely abrogated by the combination of batimastat and PMSF. Our results stress the view that metalloproteinases play a relevant role in the systemic pathophysiology of B. asper envenomation and that metalloproteinase inhibitors may become a therapeutic alternative in this pathology.

  18. Role of TNF-alpha, IL-1beta and IL-6 in the local tissue damage induced by Bothrops asper snake venom: an experimental assessment in mice.

    Science.gov (United States)

    Chaves, Fernando; Teixeira, Catarina F P; Gutiérrez, José María

    2005-02-01

    The role of the cytokines TNF-alpha, IL-1beta and IL-6 in the acute local pathological effects induced by Bothrops asper snake venom was assessed in mice. Intramuscular injection of this venom induced increments in IL-1beta and IL-6 in muscle, but no elevations of TNF-alpha were detected. Pentoxifylline (PTX), a methylxanthine derivative that inhibits the synthesis of TNF-alpha, and antibodies against these three cytokines were used to assess the role of these cytokines in venom-induced effects. As a control, PTX pretreatment was effective at abrogating lethality and serum TNF-alpha increments in mice subjected to endotoxemia induced by injection of Escherichia coli lipopolysaccharide, although it did not affect the increment in IL-1beta and IL-6 in such endotoxic model. PTX failed to reduce lethality, hemorrhage, myonecrosis, dermonecrosis and edema induced by B. asper venom. Moreover, pretreatment with anti-cytokine antibodies was also ineffective at reducing venom-induced myonecrosis and hemorrhage. It is concluded that TNF-alpha, IL-1beta and IL-6 do not have a significant role in the pathogenesis of the acute local pathological effects induced by B. asper venom in mice, although this does not exclude the possibility that these cytokines play a role in other aspects of venom-induced local pathology, as well as in the reparative and regenerative responses that take place after the onset of tissue damage.

  19. Effects of Bothrops asper snake venom on the expression of cyclooxygenases and production of prostaglandins by peritoneal leukocytes in vivo, and by isolated neutrophils and macrophages in vitro.

    Science.gov (United States)

    Moreira, Vanessa; Gutiérrez, José María; Amaral, Rafaela Bacci; Zamunér, Stella Regina; Teixeira, Catarina de Fátima Pereira

    2009-01-01

    In this study, the ability of Bothrops asper snake venom (BaV) to increase the production of prostaglandins PGE(2) and PGD(2) was assessed in a mouse model in vivo and in inflammatory cells in vitro. In addition, the expressions of COX-1 and COX-2 were assessed. BaV induced an increment in the in vivo synthesis of PGE(2) and PGD(2), together with an enhanced expression of COX-2, but not of COX-1. However, enzymatic activities of COX-1 and COX-2 were increased. Incubation of isolated macrophages and neutrophils with a sub-cytotoxic concentration of BaV in vitro resulted in increased release of PGE(2) and PGD(2) by macrophages and PGE(2) by neutrophils, concomitantly with an increment in the expression of COX-2, but not of COX-1 by both cell types. Our results demonstrate the ability of BaV to promote the expression of COX-2 and to induce the synthesis of proinflammatory prostaglandins. Macrophages and neutrophils may be important targets for this venom under in vivo situation.

  20. [Evaluation of the inhibitory effect of extracts from leaves of Renealmia alpinia Rottb. Maas (Zingiberaceae) on the venom of Bothrops asper (mapaná)].

    Science.gov (United States)

    Patiño, Arley Camilo; López, Jéssica; Aristizábal, Mónica; Quintana, Juan Carlos; Benjumea, Dora

    2012-09-01

    Traditional medicine is an invaluable source of research into new medicines as a supplement for the treatment of snakebite, considered as a serious public health problem worldwide. The extracts of the medicinal plant, Renealmia alpina, have been used traditionally by indigenous people of Chocó (Colombia) against Bothrops asper snakebite, a snake responsible for the majority of snakebite accidents in Colombia. The ability of extracts of R. alpinia leaves was tested for its ability to neutralize the hemorrhagic, coagulant and proteolytic effects of the snakebite venom of B. asper. The acute toxicity tests and analgesic activity of R. alpina were evaluated in vivo. In addition, tests were undertaken in in vitro conditions to demonstrate inhibition of coagulant, haemolytic and proteolytic activity of the B. asper venom. Results. Renealmia alpinia extracts had no toxic effects in experimental animals and also provided analgesic and antiophidian effects and protection against the lethal effects of the venom of B. asper. Renealmia. alpinia was an effective therapeutic alternative in association with antivenom treatment in the event of a B. asper snakebite accident. It was demonstrated to protect against the lethal effects and provided analgesic properties as well.

  1. A Lys49 phospholipase A(2) homologue from Bothrops asper snake venom induces proliferation, apoptosis and necrosis in a lymphoblastoid cell line.

    Science.gov (United States)

    Mora, Rodrigo; Valverde, Berta; Díaz, Cecilia; Lomonte, Bruno; Gutiérrez, José María

    2005-04-01

    Lys49 phospholipase A(2) homologues are abundant in viperid snake venoms. These proteins have substitutions at the calcium-binding loop and catalytic center which render them enzymatically inactive; however, they display a series of toxic activities, particularly cytotoxicity upon various cell lines in vitro. In this study we explored whether myotoxin II (MT-II), a Lys49 phospholipase A(2) homologue from the venom of the snake Bothrops asper, is capable of inducing various effects in a single cell type, using the lymphoblastoid B cell line CRL-8062 as a model. Cells were incubated with varying concentrations of MT-II for 24 and 48 h, time intervals that are more prolonged than the usual incubation times previously used in the characterization of this toxin. Results indicate that MT-II induces proliferation at low concentrations (0.5-5.0 microg/mL). Apoptosis was predominant at higher toxin levels (5-25 microg/mL), whereas necrosis, associated with overt plasma membrane disruption, occurred at concentrations > or =25 microg/mL, and was the predominant effect at higher MT-II concentrations (50 microg/mL). It is concluded that a single phospholipase A(2) homologue can induce markedly different effects on a single cell line, depending on the concentration used, an observation that may have implications for the action of this type of venom component in vivo.

  2. Inflammatory events induced by Lys-49 and Asp-49 phospholipases A2 isolated from Bothrops asper snake venom: role of catalytic activity.

    Science.gov (United States)

    Zuliani, Juliana P; Fernandes, Cristina M; Zamuner, Stella R; Gutiérrez, José M; Teixeira, Catarina F P

    2005-03-01

    The inflammatory events induced in the peritoneal cavity of mice by two PLA2s isolated from Bothrops asper snake venom were investigated. MT-III, an Asp-49 catalytically active enzyme and MT-II, a catalytically inactive Lys-49 variant induced increase in vascular permeability. Inhibition of enzymatic activity of MT-III with p-bromophenacyl bromide reduced this effect. MT-III induced a larger neutrophil infiltrate than MT-II. This activity was significantly reduced after inhibition of catalytic activity. Reduction in the number of neutrophils was observed when antibodies against L-selectin, CD18 or LFA-1 were used, suggesting the involvement of these adhesion molecules in the effects of both PLA2s. There was no effect with antibodies against ICAM-1 and PECAM-1. Increase in the levels of LTB4 and TXA2, as well as of IL-1, IL-6 and TNF-alpha, were observed in the peritoneal exudates induced by MT-III. MT-II did not enhance levels of eicosanoids but increased those of cytokines. It is concluded that both PLA2s induce inflammatory events in this model. Since MT-III exerts a stronger proinflammatory effect, the enzymatic phospholipid hydrolysis may be relevant for these phenomena. However, the fact that MT-II induced inflammation suggests that molecular regions distinct from the catalytic site elicit inflammatory events perhaps by interacting with specific cell membrane acceptors.

  3. Comparative study of the efficacy and safety of two polyvalent, caprylic acid fractionated [IgG and F(ab')2] antivenoms, in Bothrops asper bites in Colombia.

    Science.gov (United States)

    Otero-Patiño, Rafael; Segura, Alvaro; Herrera, María; Angulo, Yamileth; León, Guillermo; Gutiérrez, José María; Barona, Jacqueline; Estrada, Sebastián; Pereañez, Andrés; Quintana, Juan Carlos; Vargas, Leidy J; Gómez, Juan Pablo; Díaz, Abel; Suárez, Ana María; Fernández, Jorge; Ramírez, Patricia; Fabra, Patricia; Perea, Monica; Fernández, Diego; Arroyo, Yobana; Betancur, Dalila; Pupo, Lady; Córdoba, Elkin A; Ramírez, C Eugenio; Arrieta, Ana Berta; Rivero, Alcides; Mosquera, Diana Carolina; Conrado, Nectty Lorena; Ortiz, Rosina

    2012-02-01

    The efficacy and safety of two polyvalent horse-derived antivenoms in Bothrops asper envenomings were tested in a randomized, double-blind, clinical trial performed in Colombia. Both antivenoms were manufactured from the same pool of hyperimmune plasma. Antivenom A was made of F(ab')2 fragments, generated by pepsin digestion and caprylic acid precipitation, whereas antivenom B consisted of whole IgG molecules produced by caprylic acid precipitation followed by ion-exchange chromatography. Besides the different nature of the active substance, antivenom B had higher protein concentration, slightly higher turbidity and aggregate content. No significant differences were observed in the efficacy of antivenoms. Both halted local and systemic bleeding (P = 0.40) within 6-12 h of treatment in 100% of the cases, and restored blood coagulation (P = 0.87) within 6-24 h in 84.7% of patients, and within 48 h in all of them, in agreement with restoration of plasma fibrinogen concentration. Venom concentrations in serum dropped significantly (P asper envenomings in Colombia. Copyright © 2011 Elsevier Ltd. All rights reserved.

  4. Thrombocytopenia and platelet hypoaggregation induced by Bothrops asper snake venom. Toxins involved and their contribution to metalloproteinase-induced pulmonary hemorrhage.

    Science.gov (United States)

    Rucavado, Alexandra; Soto, Mónica; Escalante, Teresa; Loría, Gilbert D; Arni, Raghuvir; Gutiérrez, José María

    2005-07-01

    Thrombocytopenia and platelet dysfunction occur in patients bitten by Bothrops sp snakes in Latin America. An experimental model was developed in mice to study the effects of B. asper venom in platelet numbers and function. Intravenous administration of this venom induces rapid and prominent thrombocytopenia and ex vivo platelet hypoaggregation. The drop in platelet numbers was primarily due to aspercetin, a protein of the C-type lectin family which induces von Willebrand factor-mediated platelet aggregation/agglutination. In addition, the effect of class P-III hemorrhagic metalloproteinases on the microvessel wall also contributes to thrombocytopenia since jararhagin, a P-III metalloproteinase, reduced platelet counts. Hypoaggregation was associated with the action of procoagulant and defibrin(ogen)ating proteinases jararacussin-I (a thrombin-like serine proteinase) and basparin A (a prothrombin activating metalloproteinase). At the doses which induced hypoaggregation, these enzymes caused defibrin(ogen)ation, increments in fibrin(ogen) degradation products and D-dimer and prolongation of the bleeding time. Incubation of B. asper venom with batimastat and alpha2-macroglobulin abrogated the hypoaggregating activity, confirming the role of venom proteinases in this effect. Neither aspercetin nor the defibrin(ogen)ating and hypoaggregating components induced hemorrhage upon intravenous injection. However, aspercetin, but not the thrombin-like or the prothrombin-activating proteinases, potentiated the hemorrhagic activity of two hemorrhagic metalloproteinases in the lungs.

  5. Extracts of Renealmia alpinia (Rottb.) MAAS Protect against Lethality and Systemic Hemorrhage Induced by Bothrops asper Venom: Insights from a Model with Extract Administration before Venom Injection.

    Science.gov (United States)

    Patiño, Arley Camilo; Quintana, Juan Carlos; Gutiérrez, José María; Rucavado, Alexandra; Benjumea, Dora María; Pereañez, Jaime Andrés

    2015-04-30

    Renealmia alpinia (Rottb.) MAAS, obtained by micropropagation (in vitro) and wild forms have previously been shown to inhibit some toxic activities of Bothrops asper snake venom if preincubated before injection. In this study, assays were performed in a murine model in which extracts were administered for three days before venom injection. R. alpinia extracts inhibited lethal activity of B. asper venom injected by intraperitoneal route. Median Effective Dose (ED50) values were 36.6 ± 3.2 mg/kg and 31.7 ± 5.4 mg/kg (p > 0.05) for R. alpinia wild and in vitro extracts, respectively. At a dose of 75 mg/kg, both extracts totally inhibited the lethal activity of the venom. Moreover, this dose prolonged survival time of mice receiving a lethal dose of venom by the intravenous route. At 75 mg/kg, both extracts of R. alpinia reduced the extent of venom-induced pulmonary hemorrhage by 48.0% (in vitro extract) and 34.7% (wild extract), in agreement with histological observations of lung tissue. R. alpinia extracts also inhibited hemorrhage in heart and kidneys, as evidenced by a decrease in mg of hemoglobin/g of organ. These results suggest the possibility of using R. alpinia as a prophylactic agent in snakebite, a hypothesis that needs to be further explored.

  6. Neutralization of venom-induced hemorrhage by equine antibodies raised by immunization with a plasmid encoding a novel P-II metalloproteinase from the lancehead pitviper Bothrops asper.

    Science.gov (United States)

    Arce-Estrada, Viviana; Azofeifa-Cordero, Gabriela; Estrada, Ricardo; Alape-Girón, Alberto; Flores-Díaz, Marietta

    2009-01-14

    In this work, the cDNA encoding a novel P-II type metalloproteinase from Bothrops asper venom glands was cloned, sequenced and used for DNA immunization of animals with accelerated DNA-coated tungsten microparticles and the helius Gene Gun system. Specific antibodies against B. asper venom antigens were induced in mice co-immunized with the plasmid encoding the P-II metalloproteinase together with an expression plasmid encoding the murine IL-2. Similarly, specific antibodies against B. asper venom antigens were also induced in a horse co-immunized with the plasmid encoding the P-II metalloproteinase, together with a plasmid encoding the equine IL-6. The equine antibodies induced by immunization with the P-II metalloproteinase encoding plasmid cross react with several proteins of B. asper, Crotalus durissus durissus, and Lachesis stenophrys venoms in western blot, demonstrating antigenic similarity between the cloned metalloproteinase and other metalloproteinases present in these venoms. Furthermore, the equine antibodies induced by immunization with the P-II metalloproteinase encoding plasmid completely neutralized the hemorrhagic activity of the whole B. asper venom and partially the hemorrhagic activity of C. durissus durissus venom. The neutralizing ability of the produced antibodies raises, for the first time, the possibility of developing therapeutic antivenoms in horses by DNA immunization using tungsten microparticles.

  7. Geographical variability of the venoms of four populations of Bothrops asper from Panama: Toxicological analysis and neutralization by a polyvalent antivenom.

    Science.gov (United States)

    Vélez, Sara María; Salazar, Marcos; Acosta de Patiño, Hildaura; Gómez, Leandra; Rodriguez, Abdiel; Correa, David; Saldaña, Julio; Navarro, Deyvi; Lomonte, Bruno; Otero-Patiño, Rafael; Gutiérrez, José María

    2017-06-15

    Bothrops asper is the medically most important venomous snake in Central America. In Panama, the country having the highest incidence of snakebites in Latin America, B. asper is widely distributed throughout the country and is responsible for the vast majority of snakebites. This study was performed to analyze whether there are variations in the toxicological profile and in some biochemical parameters between the venoms of B. asper from four different regions in Panama. The venoms showed a similar profile of lethal, hemorrhagic, in vitro coagulant, defibrinogenating, edema-forming, myotoxic and indirect hemolytic activities, with subtle quantitative variations between samples of some regions. The venoms also had similar SDS-PAGE patterns and reverse phase HPLC profiles. A polyvalent antivenom manufactured in Costa Rica, and regularly used in Panama, was effective in the neutralization of lethal activity of the venoms of the four populations, with Mean Effective Doses (ED 50 ) ranging from 5.98 to 9.72 mg venom/mL antivenom. In agreement, a widespread pattern of cross-reactivity between this antivenom and the four venoms was observed by immunoblotting. Overall, results highlight the lack of marked differences between the venoms of the various populations of B. asper in Panama, and that the antivenom from Costa Rica is effective in neutralizing lethality. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Study of the design and analytical properties of the lethality neutralization assay used to estimate antivenom potency against Bothrops asper snake venom.

    Science.gov (United States)

    Solano, Gabriela; Segura, Alvaro; Herrera, María; Gómez, Aarón; Villalta, Mauren; Gutiérrez, José María; León, Guillermo

    2010-09-01

    The lethality neutralization assay performed in mice is the standard recommended by the World Health Organization to estimate antivenom potency. The interpretation of its results without considering its analytical capacity may lead to erroneous conclusions. Therefore, laboratories that manufacture or control antivenoms must demonstrate the appropriateness of their models. A study of the method used at Instituto Clodomiro Picado, Costa Rica, to estimate the potency of antivenoms against Bothrops asper snake venom was performed. Results show that venom doses ranging from 2 to 6 Median Lethal Doses (LD50) are appropriate to be used as challenge in this test. Variables such as the injection route, number of mice used per venom/antivenom level, and weight of the animals are critical in the estimation of the Median Effective Dose (ED50), whereas incubation time is not. The assay has an acceptable selectivity, linearity, and limits of detection and quantification. Accuracy of the lethality neutralization assay, expressed as percentage recovery, was between 71% and 127%. Intermediate precision, expressed as relative standard deviation, was < or = 17%. It is concluded that the analytical characteristics of this assay are adequate enough to prove product compliance and to have statistical control over an industrial line of antivenom serial production. Copyright 2010 The International Association for Biologicals. Published by Elsevier Ltd. All rights reserved.

  9. A Lys49-PLA2 myotoxin of Bothrops asper triggers a rapid death of macrophages that involves autocrine purinergic receptor signaling.

    Science.gov (United States)

    Tonello, F; Simonato, M; Aita, A; Pizzo, P; Fernández, J; Lomonte, B; Gutiérrez, J M; Montecucco, C

    2012-07-05

    Lys49-PLA(2) myotoxins, an important component of various viperid snake venoms, are a class of PLA(2)-homolog proteins deprived of catalytic activity. Similar to enzymatically active PLA(2) (Asp49) and to other classes of myotoxins, they cause severe myonecrosis. Moreover, these toxins are used as tools to study skeletal muscle repair and regeneration, a process that can be very limited after snakebites. In this work, the cytotoxic effect of different myotoxins, Bothrops asper Lys49 and Asp49-PLA(2), Notechis scutatus notexin and Naja mossambica cardiotoxin, was evaluated on macrophages, cells that have a key role in muscle regeneration. Only the Lys49-myotoxin was found to trigger a rapid asynchronous death of mouse peritoneal macrophages and macrophagic cell lines through a process that involves ATP release, ATP-induced ATP release and that is inhibited by various purinergic receptor antagonists. ATP leakage is induced also at sublytical doses of the Lys49-myotoxin, it involves Ca(2+) release from intracellular stores, and is reduced by inhibitors of VSOR and the maxi-anion channel. The toxin-induced cell death is different from that caused by high concentration of ATP and appears to be linked to localized purinergic signaling. Based on present findings, a mechanism of cell death is proposed that can be extended to other cytolytic proteins and peptides.

  10. In Vitro Antiplasmodial Activity of Phospholipases A2 and a Phospholipase Homologue Isolated from the Venom of the Snake Bothrops asper

    Directory of Open Access Journals (Sweden)

    Juan Carlos Alarcón Pérez

    2012-12-01

    Full Text Available The antimicrobial and antiparasite activity of phospholipase A2 (PLA2 from snakes and bees has been extensively explored. We studied the antiplasmodial effect of the whole venom of the snake Bothrops asper and of two fractions purified by ion-exchange chromatography: one containing catalytically-active phospholipases A2 (PLA2 (fraction V and another containing a PLA2 homologue devoid of enzymatic activity (fraction VI. The antiplasmodial effect was assessed on in vitro cultures of Plasmodium falciparum. The whole venom of B. asper, as well as its fractions V and VI, were active against the parasite at 0.13 ± 0.01 µg/mL, 1.42 ± 0.56 µg/mL and 22.89 ± 1.22 µg/mL, respectively. Differences in the cytotoxic activity on peripheral blood mononuclear cells between the whole venom and fractions V and VI were observed, fraction V showing higher toxicity than total venom and fraction VI. Regarding toxicity in mice, the whole venom showed the highest lethal effect in comparison to fractions V and VI. These results suggest that B. asper PLA2 and its homologue have antiplasmodial potential.

  11. In vitro antiplasmodial activity of phospholipases A2 and a phospholipase homologue isolated from the venom of the snake Bothrops asper.

    Science.gov (United States)

    Castillo, Juan Carlos Quintana; Vargas, Leidy Johana; Segura, Cesar; Gutiérrez, José María; Pérez, Juan Carlos Alarcón

    2012-12-14

    The antimicrobial and antiparasite activity of phospholipase A(2) (PLA(2)) from snakes and bees has been extensively explored. We studied the antiplasmodial effect of the whole venom of the snake Bothrops asper and of two fractions purified by ion-exchange chromatography: one containing catalytically-active phospholipases A(2) (PLA(2)) (fraction V) and another containing a PLA(2) homologue devoid of enzymatic activity (fraction VI). The antiplasmodial effect was assessed on in vitro cultures of Plasmodium falciparum. The whole venom of B. asper, as well as its fractions V and VI, were active against the parasite at 0.13 ± 0.01 µg/mL, 1.42 ± 0.56 µg/mL and 22.89 ± 1.22 µg/mL, respectively. Differences in the cytotoxic activity on peripheral blood mononuclear cells between the whole venom and fractions V and VI were observed, fraction V showing higher toxicity than total venom and fraction VI. Regarding toxicity in mice, the whole venom showed the highest lethal effect in comparison to fractions V and VI. These results suggest that B. asper PLA(2) and its homologue have antiplasmodial potential.

  12. Gamma radiation affects the anti-Leishmania activity of Bothrops moojeni venom and correlates with L-amino acid oxidase activity

    Energy Technology Data Exchange (ETDEWEB)

    Tempone, A.G.; Lourenco, C.O.; Spencer, P.J.; Rogero, J.R.; Nascimento, N. [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil). Div. de Radiobiologia; Andrade Junior, H.F. [Sao Paulo Univ., SP (Brazil). Faculdade de Medicina. Inst. de Medicina Tropical

    1999-11-01

    Leishmania causes human disfiguring skin disease in endemic areas of Amazon and North Eastern Brazil. Those parasites present a remarkable resistance to most treatments, except those using toxic antimonial salts. We detected a specific anti-Leishmania activity in snake venoms, using an in vitro promastigote assay. In this report, we analyzed the activity of Bothrops moojeni venom against L. Amazonensis, using whole venom or fractions of L-amino acid oxidase (L-AO). Crude venom of B.moojeni, was fractionated by molecular exclusion chromatography. Activity against promastigotes was detected by respiratory oxidative conversion of MTT in a colorimetric assay and L-AO activity was detected by a colorimetric assay with peroxidase and OPD as revealing reagents. Crude venom was irradiated with 500, 1000, and 2000 Gy in a {sup 60} Co gamma radiation source. The venom had an anti-Leishmania activity of 33 pg/promastigote and the active fraction migrates as 100-150 kDa, close to the size described for L-AOs, and also presented L-AO activity. The radiation reduces both the L-AO and anti-Leishmania activity in a dose dependent effect. Those data suggests the anti-Leishmania activity in this venom is closely related to the L-amino acid oxidase activity and also that radiation could be used as a tool to detect specific activities reduction in water solutions, similarly to observed in dry preparations. (author) 13 refs., 3 figs.

  13. Alkylation of histidine residues of Bothrops jararacussu venom proteins and isolated phospholipases A2: a biotechnological tool to improve the production of antibodies.

    Science.gov (United States)

    Guimarães, C L S; Andrião-Escarso, S H; Moreira-Dill, L S; Carvalho, B M A; Marchi-Salvador, D P; Santos-Filho, N A; Fernandes, C A H; Fontes, M R M; Giglio, J R; Barraviera, B; Zuliani, J P; Fernandes, C F C; Calderón, L A; Stábeli, R G; Albericio, F; da Silva, S L; Soares, A M

    2014-01-01

    Crude venom of Bothrops jararacussu and isolated phospholipases A2 (PLA2) of this toxin (BthTX-I and BthTX-II) were chemically modified (alkylation) by p-bromophenacyl bromide (BPB) in order to study antibody production capacity in function of the structure-function relationship of these substances (crude venom and PLA2 native and alkylated). BthTX-II showed enzymatic activity, while BthTX-I did not. Alkylation reduced BthTX-II activity by 50% while this process abolished the catalytic and myotoxic activities of BthTX-I, while reducing its edema-inducing activity by about 50%. Antibody production against the native and alkylated forms of BthTX-I and -II and the cross-reactivity of antibodies to native and alkylated toxins did not show any apparent differences and these observations were reinforced by surface plasmon resonance (SPR) data. Histopathological analysis of mouse gastrocnemius muscle sections after injection of PBS, BthTX-I, BthTX-II, or both myotoxins previously incubated with neutralizing antibody showed inhibition of the toxin-induced myotoxicity. These results reveal that the chemical modification of the phospholipases A2 (PLA2) diminished their toxicity but did not alter their antigenicity. This observation indicates that the modified PLA2 may provide a biotechnological tool to attenuate the toxicity of the crude venom, by improving the production of antibodies and decreasing the local toxic effects of this poisonous substance in animals used to produce antivenom.

  14. Neutralization of the neuromuscular activity of bothropstoxin-i, a myotoxin from Bothrops jararacussu snake venom, by a hydroalcoholic extract of Casearia sylvestris Sw. (guaçatonga

    Directory of Open Access Journals (Sweden)

    Y. Oshima-Franco

    2005-12-01

    Full Text Available Numerous plants are used as snakebite antidotes in Brazilian folk medicine, including Casearia sylvestris Swartz, popularly known as guaçatonga. In this study, we examined the action of a hydroalcoholic extract from C. sylvestris on the neuromuscular blockade caused by bothropstoxin-I (BthTX-I, a myotoxin from Bothrops jararacussu venom, in mouse isolated phrenic nerve-diaphragm (PND preparations. Aqueous (8 and 12 mg/ml, n=4 and 5, respectively and hydroalcoholic (12 mg/ml, n=12 extracts of the leaves of C. sylvestris caused facilitation in PND preparations followed by partial neuromuscular blockade. BthTX-I (20 µg/ml, n=4 caused 50% paralysis after 65±15 min (mean ± S.E.M. Preincubation (30 min at 37° C of BthTX-I (20 µg/ml, n=4 with a concentration of the hydroalcoholic extract (4 mg/ml that had no neuromuscular activity, such as the control (n=5, prevented the neuromuscular blockade caused by the toxin. This protection may be mediated by compounds such as flavonoids and phenols identified by thin-layer chromatography and colorimetric assays.

  15. Gamma radiation affects the anti-Leishmania activity of Bothrops moojeni venom and correlates with L-amino acid oxidase activity

    International Nuclear Information System (INIS)

    Tempone, A.G.; Lourenco, C.O.; Spencer, P.J.; Rogero, J.R.; Nascimento, N.; Andrade Junior, H.F.

    1999-01-01

    Leishmania causes human disfiguring skin disease in endemic areas of Amazon and North Eastern Brazil. Those parasites present a remarkable resistance to most treatments, except those using toxic antimonial salts. We detected a specific anti-Leishmania activity in snake venoms, using an in vitro promastigote assay. In this report, we analyzed the activity of Bothrops moojeni venom against L. Amazonensis, using whole venom or fractions of L-amino acid oxidase (L-AO). Crude venom of B.moojeni, was fractionated by molecular exclusion chromatography. Activity against promastigotes was detected by respiratory oxidative conversion of MTT in a colorimetric assay and L-AO activity was detected by a colorimetric assay with peroxidase and OPD as revealing reagents. Crude venom was irradiated with 500, 1000, and 2000 Gy in a 60 Co gamma radiation source. The venom had an anti-Leishmania activity of 33 pg/promastigote and the active fraction migrates as 100-150 kDa, close to the size described for L-AOs, and also presented L-AO activity. The radiation reduces both the L-AO and anti-Leishmania activity in a dose dependent effect. Those data suggests the anti-Leishmania activity in this venom is closely related to the L-amino acid oxidase activity and also that radiation could be used as a tool to detect specific activities reduction in water solutions, similarly to observed in dry preparations. (author)

  16. Purification and partial characterization of phospholipases A2 from Bothrops asper (barba amarilla snake venom from Chiriguaná (Cesar, Colombia

    Directory of Open Access Journals (Sweden)

    J. Ramírez-Avila

    2004-01-01

    Full Text Available Components with phospholipase A2 activity were isolated by gel filtration and cationic exchange chromatography from the venom of Bothrops asper snakes from Chiriguaná, Colombia (9°22´N; 73°37´W. Five fractions were obtained by the gel filtration, and PLA2 activity was found in fraction 3 (F3. In the cationic exchange chromatography, F3 showed eight components with PLA2 activity. Six of these components appeared as one band in polyacrylamide gel electrophoresis (SDS-PAGE. Fractions II and VII exhibited an optimal activity at pH 9 and 52ºC. The optimum calcium concentration for fraction II was 48 mM and for fraction VII, 384 mM. Both fractions showed thermal stability. Fraction II was stable at pH values between 2.5 and 9, and fraction VII, between 2.5 and 8. The Michaelis Menten constant (K M was 3.5x10-3 M for fraction II and 1.6x10-3 M for fraction VII. The molecular weight was 16,000 Dalton for fraction II and 17,000 Dalton for fraction VII. Both isoenzymes did not show any toxic activity (DL50 at 5.3 and 4 µg/g. The two fractions showed different kinetic constant (K M, calcium requirement, and substrate specificity for haemolytic activity.

  17. The lethality test used for estimating the potency of antivenoms against Bothrops asper snake venom: pathophysiological mechanisms, prophylactic analgesia, and a surrogate in vitro assay.

    Science.gov (United States)

    Chacón, Francisco; Oviedo, Andrea; Escalante, Teresa; Solano, Gabriela; Rucavado, Alexandra; Gutiérrez, José María

    2015-01-01

    The potency of antivenoms is assessed by analyzing the neutralization of venom-induced lethality, and is expressed as the Median Effective Dose (ED50). The present study was designed to investigate the pathophysiological mechanisms responsible for lethality induced by the venom of Bothrops asper, in the experimental conditions used for the evaluation of the neutralizing potency of antivenoms. Mice injected with 4 LD50s of venom by the intraperitoneal route died within ∼25 min with drastic alterations in the abdominal organs, characterized by hemorrhage, increment in plasma extravasation, and hemoconcentration, thus leading to hypovolemia and cardiovascular collapse. Snake venom metalloproteinases (SVMPs) play a predominat role in lethality, as judged by partial inhibition by the chelating agent CaNa2EDTA. When venom was mixed with antivenom, there was a venom/antivenom ratio at which hemorrhage was significantly reduced, but mice died at later time intervals with evident hemoconcentration, indicating that other components in addition to SVMPs also contribute to plasma extravasation and lethality. Pretreatment with the analgesic tramadol did not affect the outcome of the neutralization test, thus suggesting that prophylactic (precautionary) analgesia can be introduced in this assay. Neutralization of lethality in mice correlated with neutralization of in vitro coagulant activity in human plasma. Copyright © 2014 Elsevier Ltd. All rights reserved.

  18. Extracts of Renealmia alpinia (Rottb. MAAS Protect against Lethality and Systemic Hemorrhage Induced by Bothrops asper Venom: Insights from a Model with Extract Administration before Venom Injection

    Directory of Open Access Journals (Sweden)

    Arley Camilo Patiño

    2015-04-01

    Full Text Available Renealmia alpinia (Rottb. MAAS, obtained by micropropagation (in vitro and wild forms have previously been shown to inhibit some toxic activities of Bothrops asper snake venom if preincubated before injection. In this study, assays were performed in a murine model in which extracts were administered for three days before venom injection. R. alpinia extracts inhibited lethal activity of B. asper venom injected by intraperitoneal route. Median Effective Dose (ED50 values were 36.6 ± 3.2 mg/kg and 31.7 ± 5.4 mg/kg (p > 0.05 for R. alpinia wild and in vitro extracts, respectively. At a dose of 75 mg/kg, both extracts totally inhibited the lethal activity of the venom. Moreover, this dose prolonged survival time of mice receiving a lethal dose of venom by the intravenous route. At 75 mg/kg, both extracts of R. alpinia reduced the extent of venom-induced pulmonary hemorrhage by 48.0% (in vitro extract and 34.7% (wild extract, in agreement with histological observations of lung tissue. R. alpinia extracts also inhibited hemorrhage in heart and kidneys, as evidenced by a decrease in mg of hemoglobin/g of organ. These results suggest the possibility of using R. alpinia as a prophylactic agent in snakebite, a hypothesis that needs to be further explored.

  19. Morphometric and phylogenetic analyses of Serpentirhabdias viperidicus n. sp. (Nematoda: Rhabdiasidae) from the lancehead snake Bothrops moojeni Hoge, 1966 (Reptilia: Serpentes: Viperidae) in Brazil.

    Science.gov (United States)

    Morais, D H; Aguiar, A; Müller, M I; Narciso, R B; da Silva, L A F; da Silva, R J

    2017-05-01

    Serpentirhabdias viperidicus n. sp. (Nematoda: Rhabdiasidae) is described from the lungs of the 'Brazilian lancehead' Bothrops moojeni (Hoge, 1966) from the savannah in São Paulo State, Brazil. The new species is the eighth species of Serpentirhabdias described in the Neotropical region, and differs from other species mainly by a combination of characters: lips slightly notable, presence of fine striations at posterior ends, presence of two parallel lines with intercalated pores, a pore-shaped phasmid situated at the level of the anal aperture and another two in the posterior half of the tail. It is the first species of Serpentirhabdias reported in this snake host and the second species of this genus found parasitizing South American viperidian snakes. Molecular phylogenetic analysis using ribosomal (ITS and 28S partial) genes confirms Serpentirhabdias viperidicus n. sp. as a new species that clustered in the Serpentirhabdias clade, sister taxon to Serpentirhabdias fuscovenosa and Serpentirhabdias elaphe. This is the first description of Serpentirhabdias species from Brazil using molecular approaches and morphological characters to confirm the monophyly of this recent genus.

  20. Alkylation of Histidine Residues of Bothrops jararacussu Venom Proteins and Isolated Phospholipases A2: A Biotechnological Tool to Improve the Production of Antibodies

    Directory of Open Access Journals (Sweden)

    C. L. S. Guimarães

    2014-01-01

    Full Text Available Crude venom of Bothrops jararacussu and isolated phospholipases A2 (PLA2 of this toxin (BthTX-I and BthTX-II were chemically modified (alkylation by p-bromophenacyl bromide (BPB in order to study antibody production capacity in function of the structure-function relationship of these substances (crude venom and PLA2 native and alkylated. BthTX-II showed enzymatic activity, while BthTX-I did not. Alkylation reduced BthTX-II activity by 50% while this process abolished the catalytic and myotoxic activities of BthTX-I, while reducing its edema-inducing activity by about 50%. Antibody production against the native and alkylated forms of BthTX-I and -II and the cross-reactivity of antibodies to native and alkylated toxins did not show any apparent differences and these observations were reinforced by surface plasmon resonance (SPR data. Histopathological analysis of mouse gastrocnemius muscle sections after injection of PBS, BthTX-I, BthTX-II, or both myotoxins previously incubated with neutralizing antibody showed inhibition of the toxin-induced myotoxicity. These results reveal that the chemical modification of the phospholipases A2 (PLA2 diminished their toxicity but did not alter their antigenicity. This observation indicates that the modified PLA2 may provide a biotechnological tool to attenuate the toxicity of the crude venom, by improving the production of antibodies and decreasing the local toxic effects of this poisonous substance in animals used to produce antivenom.

  1. Sexual dimorphism in development and venom production of the insular threatened pit viper Bothrops insularism (Serpentes: Viperidae of Queimada Grande Island, Brazil

    Directory of Open Access Journals (Sweden)

    S.R. Travaglia-Cardoso

    2010-09-01

    Full Text Available Bothrops insularis is a threatened snake endemic to Queimada Grande Island, southern coast of São Paulo, Brazil, and the occurrence of sexual abnormalities in females (females with functional ovaries and rudimentary hemipenis has been reported in this population. To date there are few data regarding developmental features of this particular species. The aim of this study was to follow some developmental features in specimens maintained in captivity for seven years in the Herpetology Laboratory at Instituto Butantan, São Paulo, Brazil. We verified a pronounced sexual dimorphism in development and venom production in the specimens analyzed. In this regard, females showed greater length, mass and amount of venom in comparison to males. Our results suggest a possible niche partitioning between the sexes that reduces (or minimizes intraspecific disharmonic interactions (eg. competition on their small living area (Queimada Grande Island. Taken together, our data suggest that males and females probably are divergent in their diets, with females feeding preferentially on endothermic prey (such as migratory birds, while males maintain the juvenile diet (with the major items being ectothermic prey.

  2. Quadros clínico-patológicos do envenenamento ofídico por Crotalus durissus terrificus e Bothrops spp. em animais de produção

    Directory of Open Access Journals (Sweden)

    Carlos Hubinger Tokarnia

    2014-04-01

    Full Text Available Foi realizada uma revisão dos quadros clínico-patológicos causados pelos venenos de Crotalus durissus terrificus e Bothrops spp. em bovinos, búfalos, ovinos equinos e suínos. Foram compilados os dados obtidos pela experimentação em animais de produção encontrados na literatura e os obtidos através de experimentação realizada por nossa equipe. Também foram revisados os casos naturais de envenenamento ofídico comunicados. Em dois Quadros foram lançados os mais importantes dados dessas revisões, que revelou diversos aspectos interessantes: 1 em nossos experimentos, o veneno de Crotalus durissus terrificus, quando injetado por via subcutânea em cavalos, causou um edema acentuado no local da aplicação, ao contrário do que tem sido observado em todas as outras espécies animais, aspecto não relatado na literatura; 2 em nossos experimentos, o veneno de diversas espécies de Bothrops, quando injetado por via subcutânea em bovinos, ovinos e equinos, não causou edema como em geral é relatado na literatura, e sim hemorragias subcutâneas acentuadas no local da aplicação. Nos casos não fatais este sangue era reabsorvido em poucos dias sem deixar sequelas. Exceção foi a reação ao veneno de Bothrops jararacussu, que causou edema nos ovinos experimentais, e tumefação acentuada que resultou em fístula com eliminação de líquido seroso nos equinos experimentais. O objetivo do presente estudo visa contribuir para o aperfeiçoamento do diagnóstico de acidentes ofídicos em animais de produção.

  3. Cross-reactivity and cross-immunomodulation between venoms of the snakes Bothrops asper, Crotalus simus and Lachesis stenophrys, and its effect in the production of polyspecific antivenom for Central America.

    Science.gov (United States)

    Arroyo, Cynthia; Solano, Sergio; Segura, Álvaro; Herrera, María; Estrada, Ricardo; Villalta, Mauren; Vargas, Mariángela; Gutiérrez, José María; León, Guillermo

    2017-11-01

    A mixture of the venoms of Bothrops asper, Crotalus simus and Lachesis stenophrys is used as immunogen to produce the polyspecific Central American antivenom (PoliVal-ICP). In this work, we studied the ability of each of these venoms to modulate the antibody response induced by the other two venoms included in the immunization mixture. For that, equine monospecific, bispecific and polyspecific antivenoms were prepared and compared regarding their ability to neutralize the phospholipase A 2 , coagulant and lethal activities of each venom, and their anti-venom antibodies concentration. Results indicate that there is low cross-reactivity and cross-neutralization between venoms of B. asper, C. simus and L. stenophrys, hence justifying the use of all of them as immunogens for the production of the Central American antivenom. It was also found that the venom of B. asper reduces the anti-crotalic response while the venom of C. simus does not affect the anti-bothropic response. On the other hand, the venoms of B. asper and C. simus increase the anti-lachesic response, and L. stenoprhys venom reduced both the anti-bothropic and anti-crotalic responses. On the basis of these results, the immunization strategy can be adjusted by preventing or taking advantage of cross-immunomodulation between venoms, in order to maximize the antibody response towards all venoms. Immune responses can be improved by injecting horses with several immunogen mixtures, composed by one or two of the three venoms, and administering them at different times during the immunization, eventually generating a high titer against the three venoms. Our results suggest that addressing the issue of immunomodulation by venoms might improve antivenom manufacture worldwide. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. Comparison of the effect of Crotalus simus and Crotalus durissus ruruima venoms on the equine antibody response towards Bothrops asper venom: implications for the production of polyspecific snake antivenoms.

    Science.gov (United States)

    Dos-Santos, Maria Cristina; Arroyo, Cynthia; Solano, Sergio; Herrera, María; Villalta, Mauren; Segura, Alvaro; Estrada, Ricardo; Gutiérrez, José María; León, Guillermo

    2011-02-01

    Antivenoms are preparations of immunoglobulins purified from the plasma of animals immunized with snake venoms. Depending on the number of venoms used during the immunization, antivenoms can be monospecific (if venom from a single species is used) or polyspecific (if venoms from several species are used). In turn, polyspecific antivenoms can be prepared by purifying antibodies from the plasma of animals immunized with a mixture of venoms, or by mixing antibodies purified from the plasma of animals immunized separately with single venom. The suitability of these strategies to produce polyspecific antibothropic-crotalic antivenoms was assessed using as models the venoms of Bothrops asper, Crotalus simus and Crotalus durissus ruruima. It was demonstrated that, when used as co-immunogen, C. simus and C. durissus ruruima venoms exert a deleterious effect on the antibody response towards different components of B. asper venom and in the neutralization of hemorrhagic and coagulant effect of this venom when compared with a monospecific B. asper antivenom. Polyspecific antivenoms produced by purifying immunoglobulins from the plasma of animals immunized with venom mixtures showed higher antibody titers and neutralizing capacity than those produced by mixing antibodies purified from the plasma of animals immunized separately with single venom. Thus, despite the deleterious effect of Crotalus sp venoms on the immune response against B. asper venom, the use of venom mixtures is more effective than the immunization with separate venoms for the preparation of polyspecific bothropic-crotalic antivenoms. Copyright © 2010 Elsevier Ltd. All rights reserved.

  5. Snake venomics and antivenomics of Bothrops atrox venoms from Colombia and the Amazon regions of Brazil, Perú and Ecuador suggest the occurrence of geographic variation of venom phenotype by a trend towards paedomorphism.

    Science.gov (United States)

    Núñez, Vitelbina; Cid, Pedro; Sanz, Libia; De La Torre, Pilar; Angulo, Yamileth; Lomonte, Bruno; Gutiérrez, José María; Calvete, Juan J

    2009-11-02

    The venom proteomes of Bothrops atrox from Colombia, Brazil, Ecuador, and Perú were characterized using venomic and antivenomic strategies. Our results evidence the existence of two geographically differentiated venom phenotypes. The venom from Colombia comprises at least 26 different proteins belonging to 9 different groups of toxins. PI-metalloproteinases and K49-PLA(2) molecules represent the most abundant toxins. On the other hand, the venoms from Brazilian, Ecuadorian, and Peruvian B. atrox contain predominantly PIII-metalloproteinases. These toxin profiles correlate with the venom phenotypes of adult and juvenile B. asper from Costa Rica, respectively, suggesting that paedomorphism represented a selective trend during the trans-Amazonian southward expansion of B. atrox through the Andean Corridor. The high degree of crossreactivity of a Costa Rican polyvalent (Bothrops asper, Lachesis stenophrys, Crotalus simus) antivenom against B. atrox venoms further evidenced the close evolutionary kinship between B. asper and B. atrox. This antivenom was more efficient immunodepleting proteins from the venoms of B. atrox from Brazil, Ecuador, and Perú than from Colombia. Such behaviour may be rationalized taking into account the lower content of poorly immunogenic toxins, such as PLA(2) molecules and PI-SVMPs in the paedomorphic venoms. The immunological profile of the Costa Rican antivenom strongly suggests the possibility of using this antivenom for the management of snakebites by B. atrox in Colombia and the Amazon regions of Ecuador, Perú and Brazil.

  6. Amino acid sequence and crystal structure of BaP1, a metalloproteinase from Bothrops asper snake venom that exerts multiple tissue-damaging activities

    Science.gov (United States)

    Watanabe, Leandra; Shannon, John D.; Valente, Richard H.; Rucavado, Alexandra; Alape-Girón, Alberto; Kamiguti, Aura S.; Theakston, R. David G.; Fox, Jay W.; Gutiérrez, José María; Arni, Raghuvir K.

    2003-01-01

    BaP1 is a 22.7-kD P-I-type zinc-dependent metalloproteinase isolated from the venom of the snake Bothrops asper, a medically relevant species in Central America. This enzyme exerts multiple tissue-damaging activities, including hemorrhage, myonecrosis, dermonecrosis, blistering, and edema. BaP1 is a single chain of 202 amino acids that shows highest sequence identity with metalloproteinases isolated from the venoms of snakes of the subfamily Crotalinae. It has six Cys residues involved in three disulfide bridges (Cys 117–Cys 197, Cys 159–Cys 181, Cys 157–Cys 164). It has the consensus sequence H142E143XXH146XXGXXH152, as well as the sequence C164I165M166, which characterize the “metzincin” superfamily of metalloproteinases. The active-site cleft separates a major subdomain (residues 1–152), comprising four α-helices and a five-stranded β-sheet, from the minor subdomain, which is formed by a single α-helix and several loops. The catalytic zinc ion is coordinated by the Nɛ2 nitrogen atoms of His 142, His 146, and His 152, in addition to a solvent water molecule, which in turn is bound to Glu 143. Several conserved residues contribute to the formation of the hydrophobic pocket, and Met 166 serves as a hydrophobic base for the active-site groups. Sequence and structural comparisons of hemorrhagic and nonhemorrhagic P-I metalloproteinases from snake venoms revealed differences in several regions. In particular, the loop comprising residues 153 to 176 has marked structural differences between metalloproteinases with very different hemorrhagic activities. Because this region lies in close proximity to the active-site microenvironment, it may influence the interaction of these enzymes with physiologically relevant substrates in the extracellular matrix. PMID:14500885

  7. Inflammatory effects of BaP1 a metalloproteinase isolated from Bothrops asper snake venom: leukocyte recruitment and release of cytokines.

    Science.gov (United States)

    Fernandes, Cristina Maria; Zamuner, Stella Regina; Zuliani, Juliana Pavan; Rucavado, Alexandra; Gutiérrez, José Maria; Teixeira, Catarina de Fátima Pereira

    2006-04-01

    The inflammatory events induced by BaP1, a 22.7 kDa metalloproteinase isolated from Bothrops asper snake venom, were studied. BaP1 i.p. injection in mice induced a marked inflammatory cell infiltrate into peritoneal cavity of animals with predominance of neutrophils in the early phase followed by mononuclear cells in the late period. Inhibition of enzymatic activity of BaP1 by chelation with EDTA resulted in a drastic reduction of this effect. In addition, BaP1 induced a significant increase of blood neutrophil numbers before its accumulation in peritoneal cavity, thus suggesting a stimulatory action of BaP1 on mechanisms of cell mobilization from bone marrow reserve compartments. A reduction in the number of neutrophils was observed in the exudate when antibodies against LECAM-1, CD18 and LFA-1 were used, suggesting the involvement of these adhesion molecules in the effects of BaP1. In contrast, there was no effect with antibodies against ICAM-1 and PECAM-1. Moreover, a conspicuous increment in the levels of IL-1 and TNF-alpha, but not of LTB4, was observed in peritoneal washes collected from mice injected with BaP1. It is concluded that BaP1 induces in vivo a marked leukocyte influx, which parallels an increased number of these cells in the blood, and is associated to the expression of specific leukocyte adhesion molecules and release of chemotactic inflammatory cytokines. Since BaP1 is a P-I class metalloproteinase, these results indicate that the proteolytic domain of metalloproteinases per se can trigger specific inflammatory events.

  8. Hyperalgesia induced by Asp49 and Lys49 phospholipases A2 from Bothrops asper snake venom: pharmacological mediation and molecular determinants.

    Science.gov (United States)

    Chacur, M; Longo, I; Picolo, G; Gutiérrez, J M; Lomonte, B; Guerra, J L; Teixeira, C F P; Cury, Y

    2003-05-01

    The ability of Lys49 and Asp49 phospholipases A(2) (PLA(2)), from Bothrops asper snake venom, to cause hyperalgesia was investigated in rats, using the paw pressure test. Intraplantar injection of both toxins (5-20 micro g/paw) caused hyperalgesia, which peaked 1h after injections. Incubation of both proteins with heparin, prior to their injection, partially reduced this response. Chemical modification of Asp49 PLA(2) with p-bromophenacyl bromide (p-BPB), which abrogates its PLA(2) activity, also abolished hyperalgesia. Intraplantar injection of a synthetic peptide corresponding to the C-terminal sequence 115-129 of Lys49 PLA(2), caused hyperalgesia of similar time course, but varying magnitude, than that induced by the native protein. In contrast, a homologous peptide derived from the Asp49 PLA(2) did not show any nociceptive effect. Hyperalgesia induced by both PLA(2)s was blocked by the histamine and serotonin receptor antagonists promethazine and methysergide, respectively, by the bradykinin B(2) receptor antagonist HOE 140 and by antibodies to tumor necrosis factor alfa (TNFalpha) and interleukin 1 (IL-1). Pretreatment with guanethidine, atenolol, prazosin and yohimbine, inhibitors of sympathomimetic amines, or with indomethacin, inhibitor of the cyclo-oxygenase pathway, reduced Lys49 PLA(2)-induced hyperalgesia without interfering with the nociceptive activity of Asp49 PLA(2). The hyperalgesic response to both myotoxins was not modified by pretreatment with celecoxib, an inhibitor of the cyclo-oxygenase type II, by zileuton, an inhibitor of the lipoxygenase pathway or by N(g)-methyl-L-arginine (LNMMA), an inhibitor of nitric oxide synthase. These results suggest that Asp49 and Lys49 PLA(2)s are important hyperalgesic components of B. asper venom, and that Lys49 and Asp49 PLA(2)s exert their algogenic actions through different molecular mechanisms.

  9. Characterization of 'basparin A,' a prothrombin-activating metalloproteinase, from the venom of the snake Bothrops asper that inhibits platelet aggregation and induces defibrination and thrombosis.

    Science.gov (United States)

    Loría, Gilbert D; Rucavado, Alexandra; Kamiguti, Aura S; Theakston, R David G; Fox, Jay W; Alape, Alberto; Gutiérrez, José María

    2003-10-01

    A prothrombin activator, named 'basparin A,' was isolated from the venom of the crotaline snake Bothrops asper, the species responsible for the majority of snakebite cases in Central America. It is an acidic (pI 5.4), 70kDa, single chain P-III metalloproteinase comprising, in addition to the metalloproteinase domain, disintegrin-like, and high-cysteine domains. Basparin A is a glycoprotein displaying immunological cross-reactivity with BaH1, a P-III hemorrhagic metalloproteinase isolated from the same venom. It activates prothrombin through the formation of meizothrombin, without requiring additional cofactors; it is, therefore, a class A snake venom prothrombin activator. In contrast with most venom metalloproteinases, it does not degrade components of the extracellular matrix. Apart from its clotting activity, basparin A inhibits collagen-dependent platelet aggregation in vitro, an effect that does not depend on proteolytic activity. Clotting activity on human plasma is not abrogated by the plasma proteinase inhibitors alpha(2) macroglobulin and murinoglobulin, whereas activity is completely inhibited by Costa Rican polyvalent (Crotalinae) anti-venom. Basparin A does not induce local tissue alterations, such as hemorrhage, myonecrosis, and edema, in mice. Moreover, it does not induce systemic hemorrhage, thrombocytopenia nor prolongation of the bleeding time following intravenous administration. At low doses, the only observed effect induced by basparin A, when injected intravenously or intramuscularly into mice, is defibrin(ogen)ation. At higher doses, intravenous administration resulted in sudden death due to numerous occluding thrombi in pulmonary vessels. Basparin A is likely to play an important role in the coagulopathy associated with B. asper envenoming.

  10. Signaling pathways involved in zymosan phagocytosis induced by two secreted phospholipases A2isolated from Bothrops asper snake venom in macrophages.

    Science.gov (United States)

    Zuliani, Juliana Pavan; Gutiérrez, José María; Teixeira, Catarina

    2018-03-03

    Phagocytosis, a process involved in host defense, requires coordination of a variety of signaling reactions. MT-II, a catalytically-inactive Lys49-PLA 2 ¸ and MT-III, an active Asp49-PLA 2 isolated from Bothrops asper snake venom, activate phagocytosis in macrophages. In this study the signal pathways mediating zymosan phagocytosis, focusing in lipidic second messengers, were investigated. Macrophages collected from male Swiss mouse peritoneum were obtained 96h after i.p. injection of thioglycollate. Phagocytosis was evaluated with non-opsonized zymosan in the presence or absence of specific inhibitors. Data showed that both venom PLA 2 s increased phagocytosis. Zileuton, Etoricoxib, PACOCF 3 (5-LO, COX-2 and iPLA 2 inhibitors, respectively), as well as WEB2170 (PAF receptor antagonist) significantly reduced phagocytosis induced by both venom PLA 2 s. However, Indomethacin (COX-1/COX-2 inhibitor) and Montelukast (CysL receptor antagonist) did not affect the toxins-induced phagocytosis. Moreover, while PACOCF3 (iPLA 2 inhibitor), reduced the phagocytosis induced by MT-II and MT-III, AACOCF 3 (cPLA 2 inhibitor) significantly reduced the MT-II, but not MT-III-induced phagocytosis. These data suggest the effect of both sPLA 2 s depends on iPLA 2 and that the effect of MT-II depends on activation of cPLA 2 . COX-2 and 5-LO-derived metabolites as well as PAF are involved in the signaling events required for phagocytosis induced by both venom sPLA 2 s. Copyright © 2018 Elsevier B.V. All rights reserved.

  11. Neutralization of the edema-forming, defibrinating and coagulant effects of Bothrops asper venom by extracts of plants used by healers in Colombia.

    Science.gov (United States)

    Núñez, V; Otero, R; Barona, J; Saldarriaga, M; Osorio, R G; Fonnegra, R; Jiménez, S L; Díaz, A; Quintana, J C

    2004-07-01

    We determined the neutralizing activity of 12 ethanolic extracts of plants against the edema-forming, defibrinating and coagulant effects of Bothrops asper venom in Swiss Webster mice. The material used consisted of the leaves and branches of Bixa orellana (Bixaceae), Ficus nymphaeifolia (Moraceae), Struthanthus orbicularis (Loranthaceae) and Gonzalagunia panamensis (Rubiaceae); the stem barks of Brownea rosademonte (Caesalpiniaceae) and Tabebuia rosea (Bignoniaceae); the whole plant of Pleopeltis percussa (Polypodiaceae) and Trichomanes elegans (Hymenophyllaceae); rhizomes of Renealmia alpinia (Zingiberaceae), Heliconia curtispatha (Heliconiaceae) and Dracontium croatii (Araceae), and the ripe fruit of Citrus limon (Rutaceae). After preincubation of varying amounts of each extract with either 1.0 microg venom for the edema-forming effect or 2.0 microg venom for the defibrinating effect, the mixture was injected subcutaneously (sc) into the right foot pad or intravenously into the tail, respectively, to groups of four mice (18-20 g). All extracts (6.2-200 microg/mouse) partially neutralized the edema-forming activity of venom in a dose-dependent manner (58-76% inhibition), with B. orellana, S. orbicularis, G. panamensis, B. rosademonte, and D. croatii showing the highest effect. Ten extracts (3.9-2000 microg/mouse) also showed 100% neutralizing ability against the defibrinating effect of venom, and nine prolonged the coagulation time induced by the venom. When the extracts were administered either before or after venom injection, the neutralization of the edema-forming effect was lower than 40% for all extracts, and none of them neutralized the defibrinating effect of venom. When they were administered in situ (sc at the same site 5 min after venom injection), the neutralization of edema increased for six extracts, reaching levels up to 64% for C. limon.

  12. Amino acid sequence and crystal structure of BaP1, a metalloproteinase from Bothrops asper snake venom that exerts multiple tissue-damaging activities.

    Science.gov (United States)

    Watanabe, Leandra; Shannon, John D; Valente, Richard H; Rucavado, Alexandra; Alape-Girón, Alberto; Kamiguti, Aura S; Theakston, R David G; Fox, Jay W; Gutiérrez, José María; Arni, Raghuvir K

    2003-10-01

    BaP1 is a 22.7-kD P-I-type zinc-dependent metalloproteinase isolated from the venom of the snake Bothrops asper, a medically relevant species in Central America. This enzyme exerts multiple tissue-damaging activities, including hemorrhage, myonecrosis, dermonecrosis, blistering, and edema. BaP1 is a single chain of 202 amino acids that shows highest sequence identity with metalloproteinases isolated from the venoms of snakes of the subfamily Crotalinae. It has six Cys residues involved in three disulfide bridges (Cys 117-Cys 197, Cys 159-Cys 181, Cys 157-Cys 164). It has the consensus sequence H(142)E(143)XXH(146)XXGXXH(152), as well as the sequence C(164)I(165)M(166), which characterize the "metzincin" superfamily of metalloproteinases. The active-site cleft separates a major subdomain (residues 1-152), comprising four alpha-helices and a five-stranded beta-sheet, from the minor subdomain, which is formed by a single alpha-helix and several loops. The catalytic zinc ion is coordinated by the N(epsilon 2) nitrogen atoms of His 142, His 146, and His 152, in addition to a solvent water molecule, which in turn is bound to Glu 143. Several conserved residues contribute to the formation of the hydrophobic pocket, and Met 166 serves as a hydrophobic base for the active-site groups. Sequence and structural comparisons of hemorrhagic and nonhemorrhagic P-I metalloproteinases from snake venoms revealed differences in several regions. In particular, the loop comprising residues 153 to 176 has marked structural differences between metalloproteinases with very different hemorrhagic activities. Because this region lies in close proximity to the active-site microenvironment, it may influence the interaction of these enzymes with physiologically relevant substrates in the extracellular matrix.

  13. Contributions of the snake venoms of Bothrops asper, Crotalus simus and Lachesis stenophrys to the paraspecificity of the Central American polyspecific antivenom (PoliVal-ICP).

    Science.gov (United States)

    Solano, Gabriela; Gómez, Aarón; Corrales, Greivin; Chacón, Danilo; Estrada, Ricardo; León, Guillermo

    2018-03-15

    PoliVal-ICP antivenom is produced from plasma of horses immunized toward the venoms of Bothrops asper, Crotalus simus and Lachesis stenophrys. The antibody response induced by these venoms confers PoliVal-ICP the capacity to neutralize the venoms of the most important Central American viperids, including not only homologous venoms (i.e., venoms used as immunogen), but many heterologous venoms (i.e., venoms not used as immunogen). In this work, the individual contributions of homologous venoms to the paraspecificity of PoliVal-ICP were inferred from the capacity of experimental monospecific antivenoms toward venoms of B. asper (anti-Ba), C. simus (anti-Cs) and L. stenophrys (anti-Ls), and an experimental polyspecific antivenom (anti-Ba/Cs/Ls) to neutralize the lethality induced by different venoms in mice. It was found that all antivenoms neutralized their corresponding homologous venoms. Moreover, the anti-Ba antivenom cross-neutralized the venoms of Agkistrodon howardgloydi, Atropoides picadoi, Bothriechis lateralis, Bothriechis supraciliaris and Porthidium ophryomegas; the anti-Cs antivenom cross-neutralized the venoms of B. lateralis, B. supraciliaris, Cerrophidion sasai and Porthidium nasutum; and the anti-Ls antivenom cross-neutralized the venoms of B. lateralis, B. supraciliaris, C. sasai and Lachesis melanocephala. All venoms neutralized by any monospecific antivenom were also neutralized by the anti-Ba/Cs/Ls antivenom. Venoms of Atropoides mexicanus, Bothriechis nigroviridis and Bothriechis schlegelii were not neutralized by any experimental antivenom, thus explaining the limitations of PoliVal-ICP to neutralize these venoms. Consequently, an enlargement of the neutralization scope of PoliVal-ICP could be achieved by including these venoms in the group of those used as immunogens. Copyright © 2018 Elsevier Ltd. All rights reserved.

  14. Neutrophils do not contribute to local tissue damage, but play a key role in skeletal muscle regeneration, in mice injected with Bothrops asper snake venom.

    Science.gov (United States)

    Teixeira, C F P; Zamunér, S R; Zuliani, J P; Fernandes, C M; Cruz-Hofling, M A; Fernandes, I; Chaves, F; Gutiérrez, J M

    2003-10-01

    Local tissue damage induced by crotaline snake venoms includes edema, myonecrosis, hemorrhage, and an inflammatory response associated with a prominent cellular infiltrate. The role of neutrophils in the local tissue damage induced by Bothrops asper snake venom and by myotoxin I, a phospholipase A2 isolated from this venom, was investigated. Male Swiss mice were pretreated with either an antimouse granulocyte rat monoclonal immunoglobulin G (IgG) antibody or with isotype-matched control antibody. No significant differences in these local effects were observed between mice pretreated with antigranulocyte antibodies and those receiving control IgG. Moreover, myotoxicity induced by B. asper myotoxin I was similar in neutrophil-depleted and control mice. The role of neutrophils in the process of skeletal muscle regeneration was also assessed. Muscle regeneration was assessed by quantifying the muscle levels of creatine kinase and by morphometric histological analysis of the area comprised by regenerating cells in damaged regions of skeletal muscle. Mice depleted of neutrophils and then injected with B. asper venom showed a more deficient regenerative response than mice pretreated with control IgG. Moreover, a drastic difference in the regenerative response was observed in mice injected with myotoxin I, because animals pretreated with control IgG showed a successful regeneration, whereas those depleted of neutrophils had abundant areas of necrotic tissue that had not been removed 7 days after injection, associated with reduced contents of creatine kinase. It is concluded that (1) neutrophils do not play a significant role in the acute local pathological alterations induced by the venom of B. asper, and (2) neutrophils play a prominent role in the process of skeletal muscle regeneration after injection of B. asper venom and myotoxin I, probably related to the phagocytosis of necrotic material and the recruitment of other inflammatory cells, two events directly associated

  15. Membrane cholesterol modulates the cytolytic mechanism of myotoxin II, a Lys49 phospholipase A2 homologue from the venom of Bothrops asper.

    Science.gov (United States)

    Rangel, José; Quesada, Orestes; Gutiérrez, José María; Angulo, Yamileth; Lomonte, Bruno

    2011-07-01

    Lys49 phospholipase A2 (PLA2) homologues present in crotalid snake venoms lack enzymatic activity, yet they induce skeletal muscle necrosis by a membrane permeabilizing mechanism whose details are only partially understood. The present study evaluated the effect of altering the membrane cholesterol content on the cytolytic activity of myotoxin II, a Lys49 PLA2 isolated from the venom of Bothrops asper, using the myogenic cell line C2C12 as a model target. Cell membrane cholesterol depletion by methyl-β-cyclodextrin (MβCD) treatment enhanced the cytolytic action of myotoxin II, as well as of its bioactive C-terminal synthetic peptide p(115-129) . Conversely, cell membrane cholesterol enrichment by preformed cholesterol-MβCD complexes reduced the cytolytic effect of myotoxin II. The toxic actions of myotoxin I, a catalytically active PLA2 from the same venom, as well as of the cytolytic peptide melittin from bee venom, also increased in cholesterol-depleted cells. Although physical and functional changes resulting from variations in membrane cholesterol are complex, these findings suggest that membrane fluidity could be a relevant parameter to explain the observed modulation of the cytolytic mechanism of myotoxin II, possibly influencing bilayer penetration. In concordance, the cytolytic effect of myotoxin II decreased in direct proportion to lower temperature, a physical factor that affects membrane fluidity. In conclusion, physicochemical properties that depend on membrane cholesterol content significantly influence the cytolytic mechanism of myotoxin II, reinforcing the concept that the primary site of action of Lys49 PLA2 myotoxins is the plasma membrane. Copyright © 2011 John Wiley & Sons, Ltd.

  16. Food resources influence spatial ecology, habitat selection, and foraging behavior in an ambush-hunting snake (Viperidae: Bothrops asper): an experimental study.

    Science.gov (United States)

    Wasko, Dennis K; Sasa, Mahmood

    2012-06-01

    Prey availability affects many aspects of predators' life history and is considered a primary factor influencing individuals' decisions regarding spatial ecology and behavior, but few experimental data are currently available. Snakes may represent ideal model organisms relative to other animal groups for addressing such resource dependency, due to a presumably more direct link between food resources and many aspects of behavior and natural history. We experimentally investigated the relationship between food intake and spatial behavior in a population of the snake Bothrops asper in a Costa Rican lowland rainforest. Six adult snakes were allowed to forage naturally while six were offered supplemental food in the field, with both groups monitored using radiotelemetry. Mean home range size did not differ between groups presumably due to small sample size, but supplementally fed snakes demonstrated altered patterns of macro- and microhabitat selection, shorter and less frequent movements, and increased mass acquisition. Fed snakes also devoted less time to foraging efforts, instead more frequently remaining inactive and utilizing shelter. Because snakes were always fed in situ and not at designated feeding stations, observed shifts in habitat selection are not explained by animals simply moving to areas of higher food availability. Rather, B. asper may have moved to swamps in order to feed on amphibians when necessary, but remained in preferred forest habitat when food was otherwise abundant. The strong behavioral and spatiotemporal responses of snakes in this population may have been influenced by an overall scarcity of mammalian prey during the study period. Copyright © 2012 Elsevier GmbH. All rights reserved.

  17. Identification of linear B-cell epitopes on myotoxin II, a Lys49 phospholipase A₂ homologue from Bothrops asper snake venom.

    Science.gov (United States)

    Lomonte, Bruno

    2012-10-01

    Knowledge on toxin immunogenicity at the molecular level can provide valuable information for the improvement of antivenoms, as well as for understanding toxin structure-function relationships. The aims of this study are two-fold: first, to identify the linear B-cell epitopes of myotoxin II from Bothrops asper snake venom, a Lys49 phospholipase A₂ homologue; and second, to use antibodies specifically directed against an epitope having functional relevance in its toxicity, to probe the dimeric assembly mode of this protein in solution. Linear B-cell epitopes were identified using a library of overlapping synthetic peptides spanning its complete sequence. Epitopes recognized by a rabbit antiserum to purified myotoxin II, and by three batches of a polyvalent (Crotalidae) therapeutic antivenom (prepared in horses immunized with a mixture of B. asper, Crotalus simus, and Lachesis stenophrys venoms) were mapped using an enzyme-immunoassay based on the capture of biotinylated peptides by immobilized streptavidin. Some of the epitopes identified were shared between the two species, whereas others were unique. Differences in epitope recognition were observed not only between the two species, but also within the three batches of equine antivenom. Epitope V, located at the C-terminal region of this protein, is known to be relevant for toxicity and neutralization. Affinity-purified rabbit antibodies specific for this site were able to immunoprecipitate myotoxin II, suggesting that the two copies of epitope V are simultaneously available to antibody binding, which would be compatible with the mode of dimerization known as "conventional" dimer. Copyright © 2012 Elsevier Ltd. All rights reserved.

  18. Neutralization of the edema-forming, defibrinating and coagulant effects of Bothrops asper venom by extracts of plants used by healers in Colombia

    Directory of Open Access Journals (Sweden)

    V. Núñez

    2004-07-01

    Full Text Available We determined the neutralizing activity of 12 ethanolic extracts of plants against the edema-forming, defibrinating and coagulant effects of Bothrops asper venom in Swiss Webster mice. The material used consisted of the leaves and branches of Bixa orellana (Bixaceae, Ficus nymphaeifolia (Moraceae, Struthanthus orbicularis (Loranthaceae and Gonzalagunia panamensis (Rubiaceae; the stem barks of Brownea rosademonte (Caesalpiniaceae and Tabebuia rosea (Bignoniaceae; the whole plant of Pleopeltis percussa (Polypodiaceae and Trichomanes elegans (Hymenophyllaceae; rhizomes of Renealmia alpinia (Zingiberaceae, Heliconia curtispatha (Heliconiaceae and Dracontium croatii (Araceae, and the ripe fruit of Citrus limon (Rutaceae. After preincubation of varying amounts of each extract with either 1.0 µg venom for the edema-forming effect or 2.0 µg venom for the defibrinating effect, the mixture was injected subcutaneously (sc into the right foot pad or intravenously into the tail, respectively, to groups of four mice (18-20 g. All extracts (6.2-200 µg/mouse partially neutralized the edema-forming activity of venom in a dose-dependent manner (58-76% inhibition, with B. orellana, S. orbicularis, G. panamensis, B. rosademonte, and D. croatii showing the highest effect. Ten extracts (3.9-2000 µg/mouse also showed 100% neutralizing ability against the defibrinating effect of venom, and nine prolonged the coagulation time induced by the venom. When the extracts were administered either before or after venom injection, the neutralization of the edema-forming effect was lower than 40% for all extracts, and none of them neutralized the defibrinating effect of venom. When they were administered in situ (sc at the same site 5 min after venom injection, the neutralization of edema increased for six extracts, reaching levels up to 64% for C. limon.

  19. Action of Bothrops moojeni venom and its L-amino acid oxidase fraction, treated with 60Co gamma rays, in Leishmania spp

    International Nuclear Information System (INIS)

    Cardoso, Andre Gustavo Tempone

    1999-01-01

    Bothrops moojeni venom showed an anti leishmania activity in vitro, as determined by a cell viability assay using the reduction of MTT. After venom purification, by chromatography techniques, the fractions with anti leishmania and L-amino acid oxidase activities, eluted in the same positions. The molecular weight of the enzyme was estimated to be 140 kDa by molecular exclusion chromatography, and 69 kDa, by SDS-PAGE, migrating as a single band, with an isoelectric point of 4.8 as determined by isoelectric focusing. The purified LAO from B. moojeni venom, 135-fold more active than crude venom, showed homo dimeric constitution, and was active against Leishmania spp from the New World, with an effective concentration against L(L). amazonensis of 1.80 μg/ml (EC 50 ), L.(V.) panamensis (0.78 |μg/ml) and L.(L.) chagasi (0.63 (μg/ml). Ultrastructural studies of promastigotes affected by LAO demonstrated cell death, with edema in several organelles such as mitochondria and nuclear membrane, before cell disruption and necrosis. The action of LAO was demonstrated to be hydrogen peroxide-dependent. Studies with LLCMK-2 cells, treated with LAO, showed a toxic effect, with an EC 50 of 11|μg/ml. Irradiation of LAO with 6 0C o gamma rays, did not affect its whole oxidative activity, neither detoxified the enzyme. Amastigotes treated with LAO were not affected by its hydrogen peroxide, otherwise, the exogenous product, killed amastigotes with an EC 50 of 0.67mM. These data could be of help in the development of alternative therapeutic approaches to the treatment of leishmaniasis. (author)

  20. Increase of the cytotoxic effect of Bothrops jararacussu venom on mouse extensor digitorum longus and soleus by potassium channel blockers and by Na(+)/K(+)-ATPase inhibition.

    Science.gov (United States)

    Tomaz, Marcelo A; Fernandes, Fabrício F A; El-Kik, Camila Z; Moraes, Raphael A M; Calil-Elias, Sabrina; Saturnino-Oliveira, Jeison; Martinez, Ana Maria B; Ownby, Charlotte L; Melo, Paulo A

    2008-09-15

    We investigated the myotoxicity of Bothrops jararacussu crude venom and other cytolytic agents on mouse isolated extensor digitorum longus (EDL) and soleus (SOL) muscles, which present distinct properties: EDL is a fast-twitch, white muscle with predominantly glycolytic fibers, while SOL is slow-twitch, red muscle with predominantly oxidative fibers. Muscles were exposed to B. jararacussu crude venom (25 microg/ml) and other crotaline venoms (Agkistrodon contortrix laticinctus; Crotalus viridis viridis; Crotalus durissus terrificus) at the same concentration. Basal creatine kinase (CK) release to bathing solution was 0.43+/-0.06 for EDL and 0.29+/-0.06 for SOL (U g(-)(1) h(-)(1), n=36 for each muscle). Sixty minutes after exposure to B. jararacussu venom, EDL presented higher increase in the rate of CK release than SOL, respectively, 13.2+/-1.5 and 2.9+/-0.7 U g(-)(1)h(-)(1), n=10-12. Muscle denervation, despite decreasing CK content, did not affect sensitivities to B. jararacussu venom. Ouabain and potassium channel blockers (TEA; clotrimazole; glibenclamide) increased the rate of CK release by B. jararacussu in EDL and SOL muscles, decreasing and almost abolishing the different sensitivity. When we exposed EDL or SOL muscles to Naja naja, Apis mellifera venoms (25 microg/ml), or Triton X-100 (0.01%), they showed similar rate of CK release. Our present data suggest that a mechanism involving intracellular calcium regulation or potassium channels may participate in the different sensitivity of EDL and SOL to B. jararacussu venom.

  1. Actividad in vitro de los venenos de Lachesis muta y Bothrops atrox sobre la viabilidad y desarrollo embrionario de los huevos de Ascaris suum

    Directory of Open Access Journals (Sweden)

    Claudia Paredes

    2014-06-01

    Full Text Available Los venenos de serpientes son concentrados polienzimáticos cuya actividad biológica sobre algunas bacterias y protozoos ha sido comprobada. El objetivo principal del presente trabajo fue estudiar la actividad in vitro de los venenos totales de las serpientes Lachesis muta y Bothrops atrox sobre la viabilidad y el desarrollo embrionario de los huevos de Ascaris suum. Se emplearon los venenos totales en concentraciones de 2, 4, 8 y 16 mg/mL sobre huevos no embrionados y larvados in vitro. Se comparó la actividad de los venenos con la de otras sustancias como el hipoclorito de sodio al 5,25%, Albendazol (solución comercial y solución salina. Ambos venenos, en concentraciones de 4, 8 y 16 mg/mL, inhibieron la blastulación de estos huevos; hasta el sexto día de incubación; en cambio en concentración de 2 mg/mL la inhibición se dio hasta el cuarto día. Posteriormente iniciaron un proceso de embrionación aparentemente normal hasta la formación del estadio infectante. El veneno de B. atrox fue el que presentó el mayor efecto inhibitorio en concentración de 16 mg/mL. El hipoclorito de sodio destruyó el 100% de los huevos, mientras el albendazol ocasionó que los huevos iniciaran un proceso de segmentación anormal que originó su degeneración. Se concluye que los venenos de L. muta y B. atrox muestran actividad inhibitoria al inicio de la blastulación de los huevos de A. suum y no ejercen ningún efecto en los huevos larvados.

  2. A novel fibrinolytic metalloproteinase, barnettlysin-I from Bothrops barnetti (Barnett´s pitviper) snake venom with anti-platelet properties.

    Science.gov (United States)

    Sanchez, Eladio Flores; Richardson, Michael; Gremski, Luiza Helena; Veiga, Silvio Sanches; Yarleque, Armando; Niland, Stephan; Lima, Augusto Martins; Estevao-Costa, Maria Inácia; Eble, Johannes Andreas

    2016-03-01

    Viperid snake venoms contain active components that interfere with hemostasis. We report a new P-I class snake venom metalloproteinase (SVMP), barnettlysin-I (Bar-I), isolated from the venom of Bothrops barnetti and evaluated its fibrinolytic and antithrombotic potential. Bar-I was purified using a combination of molecular exclusion and cation-exchange chromatographies. We describe some biochemical features of Bar-I associated with its effects on hemostasis and platelet function. Bar-I is a 23.386 kDa single-chain polypeptide with pI of 6.7. Its sequence (202 residues) shows high homology to other members of the SVMPs. The enzymatic activity on dimethylcasein (DMC) is inhibited by metalloproteinase inhibitors e.g. EDTA, and by α2-macroglobulin. Bar-I degrades fibrin and fibrinogen dose- and time-dependently by cleaving their α-chains. Furthermore, it hydrolyses plasma fibronectin but not laminin nor collagen type I. In vitro Bar-I dissolves fibrin clots made either from purified fibrinogen or from whole blood. In contrast to many other P-I SVMPs, Bar-I is devoid of hemorrhagic activity. Also, Bar-I dose- and time-dependently inhibits aggregation of washed human platelets induced by vWF plus ristocetin and collagen (IC50=1.3 and 3.2 μM, respectively), presumably Bar-I cleaves both vWF and GPIb. Thus, it effectively inhibits vWF-induced platelet aggregation. Moreover, this proteinase cleaves the collagen-binding α2-A domain (160 kDa) of α2β1-integrin. This explains why it additionally inhibits collagen-induced platelet activation. A non-hemorrhagic but fibrinolytic metalloproteinase dissolves fibrin clots in vitro and impairs platelet function. This study provides new opportunities for drug development of a fibrinolytic agent with antithrombotic effect. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. BmajPLA2-II, a basic Lys49-phospholipase A2 homologue from Bothrops marajoensis snake venom with parasiticidal potential.

    Science.gov (United States)

    Grabner, Amy N; Alfonso, Jorge; Kayano, Anderson M; Moreira-Dill, Leandro S; Dos Santos, Ana Paula de A; Caldeira, Cleópatra A S; Sobrinho, Juliana C; Gómez, Ana; Grabner, Fernando P; Cardoso, Fabio F; Zuliani, Juliana Pavan; Fontes, Marcos R M; Pimenta, Daniel C; Gómez, Celeste Vega; Teles, Carolina B G; Soares, Andreimar M; Calderon, Leonardo A

    2017-09-01

    Snake venoms contain various proteins, especially phospholipases A 2 (PLA 2 s), which present potential applications in diverse areas of health and medicine. In this study, a new basic PLA 2 from Bothrops marajoensis with parasiticidal activity was purified and characterized biochemically and biologically. B. marajoensis venom was fractionated through cation exchange followed by reverse phase chromatographies. The isolated toxin, BmajPLA 2 -II, was structurally characterized with MALDI-TOF (Matrix-assisted laser desorption/ionization-time of flight) mass spectrometry, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), followed by two-dimensional electrophoresis, partial amino acid sequencing, an enzymatic activity assay, circular dichroism, and dynamic light scattering assays. These structural characterization tests presented BmajPLA 2 -II as a basic Lys49 PLA 2 homologue, compatible with other basic snake venom PLA 2 s (svPLA 2 ), with a tendency to form aggregations. The in vitro anti-parasitic potential of B. marajoensis venom and of BmajPLA 2 -II was evaluated against Leishmania infantum promastigotes and Trypanosoma cruzi epimastigotes, showing significant activity at a concentration of 100μg/mL. The venom and BmajPLA 2 -II presented IC 50 of 0.14±0.08 and 6.41±0.64μg/mL, respectively, against intraerythrocytic forms of Plasmodium falciparum with CC 50 cytotoxicity values against HepG2 cells of 43.64±7.94 and >150μg/mL, respectively. The biotechnological potential of these substances in relation to leishmaniasis, Chagas disease and malaria should be more deeply investigated. Copyright © 2017. Published by Elsevier B.V.

  4. Bothrops pirajai snake venom L-amino acid oxidase: in vitro effects on infection of Toxoplasma gondii in human foreskin fibroblasts

    Directory of Open Access Journals (Sweden)

    Luiz F. M. Izidoro

    2011-06-01

    Full Text Available The effect of an L-amino acid oxidase isolated from Bothrops pirajai snake venom (BpirLAAO-I was investigated on infection of Toxoplasma gondii in human foreskin fibroblasts (HFF. The cytotoxic activity of BpirLAAO-I on HFF cells showed a dose-dependent toxicity with median cytotoxic dose (TD50 of 11.8 µg/mL. BpirLAAO-I induced considerable dose-dependent decrease in the T. gondii infection index under two different conditions, treatment of tachyzoites before infection or treatment of HFF cells after infection. A maximal inhibition of infection (56% was found for treatment before infection, with a median inhibitory dose (ID50 at 1.83 µg/mL and selectivity index (SI at 6.45. For treatment after infection, it was observed a maximal inhibition of infection at 65%, ID50 of 1.20 µg/mL and SI of 9.83. The treatment before infection was also effective to reduce intracellular parasitism up to 62%, although presenting higher values of ID50 (3.14 µg/mL and lower values of SI (3.76. However, treatment after infection was not effective, suggesting that the enzyme seems to have no effect on the parasite intracellular replication for this condition. In conclusion, BpirLAAO-I was more effective to inhibit the infection of neighboring cells and consequently parasite dissemination than primary infection and parasite replication. Thus, the effect of BpirLAAO-I described herein could be taken into account for the development of new synthetic anti-parasite therapeutic agents.

  5. Comparison of venoms from wild and long-term captive Bothrops atrox snakes and characterization of Batroxrhagin, the predominant class PIII metalloproteinase from the venom of this species.

    Science.gov (United States)

    Freitas-de-Sousa, L A; Amazonas, D R; Sousa, L F; Sant'Anna, S S; Nishiyama, M Y; Serrano, S M T; Junqueira-de-Azevedo, I L M; Chalkidis, H M; Moura-da-Silva, A M; Mourão, R H V

    2015-11-01

    Comparisons between venoms from snakes kept under captivity or collected at the natural environment are of fundamental importance in order to obtain effective antivenoms to treat human victims of snakebites. In this study, we compared composition and biological activities of Bothrops atrox venom from snakes collected at Tapajós National Forest (Pará State, Brazil) or maintained for more than 10 years under captivity at Instituto Butantan herpetarium after have been collected mostly at Maranhão State, Brazil. Venoms from captive or wild snakes were similar except for small quantitative differences detected in peaks correspondent to phospholipases A2 (PLA2), snake venom metalloproteinases (SVMP) class PI and serine proteinases (SVSP), which did not correlate with fibrinolytic and coagulant activities (induced by PI-SVMPs and SVSPs). In both pools, the major toxic component corresponded to PIII-SVMPs, which were isolated and characterized. The characterization by mass spectrometry of both samples identified peptides that matched with a single PIII-SVMP cDNA characterized by transcriptomics, named Batroxrhagin. Sequence alignments show a strong similarity between Batroxrhagin and Jararhagin (96%). Batroxrhagin samples isolated from venoms of wild or captive snakes were not pro-coagulant, but inhibited collagen-induced platelet-aggregation, and induced hemorrhage and fibrin lysis with similar doses. Results suggest that in spite of environmental differences, venom variability was detected only among the less abundant components. In opposition, the most abundant toxin, which is a PIII-SVMP related to the key effects of the venom, is structurally conserved in the venoms. This observation is relevant for explaining the efficacy of antivenoms produced with venoms from captive snakes in human accidents inflicted at distinct natural environments. Copyright © 2015 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

  6. Neutralizing activities of ethanolic extracts of six plants traditionally used in Guatemala as antidotes for the envenomation caused by the snake Bothrops asper

    Directory of Open Access Journals (Sweden)

    Patricia Saravia-Otten

    2015-07-01

    Full Text Available Many plants are reported to be used in Guatemalan traditional medicine as antidotes against various effects of the snakebite; however, very few attempts have been made to evaluate their neutralizing capacity in controlled experiments. Six plants (Acacia hindsii, Cissampelos pareira; Hamelia patens, Piper peltatum, Sansevieria hyacinthoides and Aristolochia maxima were evaluated in vitro for their ability to neutralize phospholipase A2(PLA2 and proteolytic effects of the venom of Bothrops asper, the snake responsible for approximately half of the snakebite envenomations in Central America. These effects are indicatives of the ability of B. asper venom to produce myotoxicity, hemorrhage and inflammation. Plants were collected, dried and extracted by maceration with ethanol. After pre-incubation of several amounts of each extract with a challenge dose of venom, S. hyacinthoides demonstrated a low neutralizing capacity (< DE 50 of the PLA2 effect (13.90 ± 6.41%; C. pareira (32.98 ± 5.51% and P. peltatum (24.52 ± 7.45% neutralized less than 50% of the proteolytic effect. The results suggest that neither of the tested plants should be used individually to treat the main effects of B. asper envenomation. However, the three low-active extracts might be potentiated when used in mixtures composed of several plants, as prepared by traditional healers. Given the complexity of the venom components and the multiple pathologic effects produced by B. asper envenomation, more tests are required to fully investigate the ability of this plants to neutralize the coagulant, fibrin(ogenolytic, edematizing and myotoxic effects of the venom.

  7. Alternagin-C, a disintegrin-like protein from the venom of Bothrops alternatus, modulates a2ß1 integrin-mediated cell adhesion, migration and proliferation

    Directory of Open Access Journals (Sweden)

    Selistre-de-Araujo H.S.

    2005-01-01

    Full Text Available The alpha2ß1 integrin is a major collagen receptor that plays an essential role in the adhesion of normal and tumor cells to the extracellular matrix. Alternagin-C (ALT-C, a disintegrin-like protein purified from the venom of the Brazilian snake Bothrops alternatus, competitively interacts with the alpha2ß1 integrin, thereby inhibiting collagen binding. When immobilized in plate wells, ALT-C supports the adhesion of fibroblasts as well as of human vein endothelial cells (HUVEC and does not detach cells previously bound to collagen I. ALT-C is a strong inducer of HUVEC proliferation in vitro. Gene expression analysis was done using an Affimetrix HU-95A probe array with probe sets of ~10,000 human genes. In human fibroblasts growing on collagen-coated plates, ALT-C up-regulates the expression of several growth factors including vascular endothelial growth factor, as well as some cell cycle control genes. Up-regulation of the vascular endothelial growth factor gene and other growth factors could explain the positive effect on HUVEC proliferation. ALT-C also strongly activates protein kinase B phosphorylation, a signaling event involved in endothelial cell survival and angiogenesis. In human neutrophils, ALT-C has a potent chemotactic effect modulated by the intracellular signaling cascade characteristic of integrin-activated pathways. Thus, ALT-C acts as a survival factor, promoting adhesion, migration and endothelial cell proliferation after binding to alpha2ß1 integrin on the cell surface. The biological activities of ALT-C may be helpful as a therapeutic strategy in tissue regeneration as well as in the design of new therapeutic agents targeting alpha2ß1 integrin.

  8. Intoxicação por veneno de cobra: necrose symetrica da cortex renal: uremia

    Directory of Open Access Journals (Sweden)

    A. Penna de Azevedo

    1938-01-01

    Full Text Available Em um caso fatal de ophidismo, em individuo de 15 annos de edade, picado por uma cobra jararaca (Bothrops jararaca na face externa da perna direita e que veio a fallecer 26 dias apoz o accidente, os A.A, descrevem as lesões anatomo-pathologicas encontradas e as modificações do metabolismo, evidenciadas pelos exames chimicos do sangue. As principaes alterações existentes, acham-se localisadas nos rins os quaes apresentam lesões de glomerulonephrite diffusa e o aspecto typico da necrose cortical symmetrica. Como alterações de maior significação observam-se ainda lesões vasculares de grande intensidade e constituidas essencialmente por processo de endoarterite productiva. A necrose symmetrica da cortex renal, a vista das intensas alterações vasculares (endoarterite productiva que acarretaram a obliteração das arterías, é considerada como a consequencia immediata de taes lesões vasculares. Os vasos renaes, séde do processo inflammatorio, são as arterias interlobar, arciforme e interlobular, mas principalmente as arteriolares da camada cortical. O processo de endoarterite assume sempre o carater progressivo, de modo que a luz vascular vae sendo aos poucos, totalmente obstruida. Ao contrario do que se tem observado nos casos de necrose cortical symmetrica, citados na literatura, em que as alterações parenchymatosas são consequentes a thrombose dos vasos reanes, no caso presente esse aspecto não foi verificado mas tão sómente a existencia da endoarterite productiva obliterante. Consideram os A.A. as lesões renaes no caso que estudaram, como a resultante da actuação lenta e prolongada do veneno de cobra sobre as estructuras renaes, baseados nos seguintes factos já conhecidos e admittidos: eliminação do veneno de cobra pelos rins; capacidade do mesmo veneno, determinar a glomerulo-nephrite diffusa e acção do veneno de cobra sobre o endothelio vascular, facilitada essencialmente pela funcção especifica do orgão. As

  9. Caracterização individual do veneno de Bothrops alternatus Duméril, Bibron & Duméril em função da distribuição geográfica no Brasil (Serpentes,Viperidae Individual characterization of Bothrops alternatus Duméril, Bibron & Duméril venoms, according to their geographic distribution in Brazil (Serpentes, Viperidae

    Directory of Open Access Journals (Sweden)

    Marisa M. T. da Rocha

    2005-06-01

    Full Text Available Bothrops alternatus Duméril, Bibron & Duméril, 1854 é uma serpente de importância em saúde pública, com ampla distribuição geográfica, desde o Mato Grosso do Sul até o sudeste do Brasil, chegando até a Argentina e Uruguai, ocupando vários domínios morfoclimáticos. Neste trabalho investigou-se a variação do veneno de adultos de Bothrops alternatus, em função de sua distribuição geográfica no Brasil, comparativamente ao veneno elaborado sob a forma de "pool" desta espécie (veneno referência, que inclui serpentes, em sua maioria, da região do estado de São Paulo. Foram analisadas as atividades letal, coagulante sobre o plasma, proteolítica sobre a caseína e miotóxica, bem como os padrões eletroforéticos de 61 amostras individuais de veneno contrapostas ao "pool". Os resultados mostraram que o veneno de B. alternatus é pouco ativo, comparativamente ao de outros Bothrops Wagler, 1824. A variação individual prevaleceu, não apresentando correlação com as áreas de distribuição geográfica e domínios morfoclimáticos, porém a atividade coagulante das amostras de veneno provenientes do nordeste da distribuição geográfica apresentaram-se menos ativas comparativamente às da porção central da distribuição. Os venenos provenientes das bordas da distribuição apresentaram ações proteolíticas e miotóxicas mais intensas, que estatisticamente não foram significativamente diferentes. As variações individuais prevaleceram.Bothrops alternatus Duméril, Bibron & Duméril, 1854 snakebites are an important public health problem in Brazil. Such snakes are found from Mato Grosso do Sul (central Brazil to southeastern Brazil, reaching even Argentina and Uruguay and thereby occupying different morphoclimatic domains. This work investigated venom variation occurring in adult specimens of B. alternatus specimens, according to their geographic distribution in Brazil. The standard venom pool (reference venom produced by

  10. A neutralizing recombinant single chain antibody, scFv, against BaP1, A P-I hemorrhagic metalloproteinase from Bothrops asper snake venom.

    Science.gov (United States)

    Castro, J M A; Oliveira, T S; Silveira, C R F; Caporrino, M C; Rodriguez, D; Moura-da-Silva, A M; Ramos, O H P; Rucavado, A; Gutiérrez, J M; Magalhães, G S; Faquim-Mauro, E L; Fernandes, I

    2014-09-01

    BaP1 is a P-I class snake venom metalloproteinase (SVMP) relevant in the local tissue damage associated with envenomings by Bothrops asper, a medically important snake species in Central America and parts of South and North America. The main treatment for these accidents is the passive immunotherapy using antibodies raised in horses. In order to obtain more specific and batch-to-batch consistent antivenons, recombinant antibodies are considered a good option compared to animal immunization. We constructed a recombinant single chain variable fragment (scFv) from a monoclonal antibody against BaP1 (MABaP1) formerly secreted by a hybridoma clone. This recombinant antibody was cloned into pMST3 vector in fusion with SUMO protein and contains VH and VL domains linked by a flexible (G4S)3 polypeptide (scFvBaP1). The aim of this work was to produce scFvBaP1 and to evaluate its potential concerning the neutralization of biologically important activities of BaP1. The cytoplasmic expression of this construct was successfully achieved in C43 (DE3) bacteria. Our results showed that scFvBaP1-SUMO fusion protein presented an electrophoretic band of around 43 kDa from which SUMO alone corresponded to 13.6 kDa, and only the scFv was able to recognize BaP1 as well as the whole venom by ELISA. In contrast, neither an irrelevant scFv anti-LDL nor its MoAb partner recognized it. BaP1-induced fibrinolysis was significantly neutralized by scFvBaP1, but not by SUMO, in a concentration-dependent manner. In addition, scFvBaP1, as well as MaBaP1, completely neutralized in vivo hemorrhage, muscle necrosis, and inflammation induced by the toxin. Docking analyses revealed possible modes of interaction of the recombinant antibody with BaP1. Our data showed that scFv recognized BaP1 and whole B. asper venom, and neutralized biological effects of this SVMP. This scFv antibody can be used for understanding the molecular mechanisms of neutralization of SVMPs, and for exploring the potential of

  11. Identificación molecular y actividad sobre sustratos cromogénicos de la venombina A del veneno de la serpiente peruana Bothrops atrox

    Directory of Open Access Journals (Sweden)

    Gustavo A. Sandoval

    2011-05-01

    Full Text Available En el presente trabajo se ha realizado la identificación molecular de la enzima similar a trombina (EST del veneno de Bothrops atrox y se ha evaluado su actividad enzimática sobre diversos sustratos sintéticos. La enzima fue purificada utilizando tres pasos cromatrográficos, sobre Sephadex G-75, CM-Sephadex C-50 y Agarosa-PAB, determinándose su peso molecular por PAGE-SDS. La identificación molecular de la enzima aislada se realizó por la técnica de peptide mass fingerprinting basada en espectrometría de masas MALDI-TOF y posterior análisis in silico. Las actividades fibrinocoagulante y amidolítica fueron ensayadas sobre fibrinó- geno bovino y BApNA, respectivamente, así como la hidrólisis sobre los sustratos cromogénicos específicos S-2238, S-2251 y S-2266. Como resultado de los ensayos bioquímicos y estructurales, la EST del veneno de B. atrox, presentó un peso molecular de 29,6 kDa. El análisis mediante espectrometría de masas de los péptidos obtenidos, permitió identificar a esta enzima como una venombina A, presentando una identidad del 75%. Del análisis de actividad enzimática, se obtuvo que la EST de B. atrox produjo coagulación del fibrinógeno bovino y presentó actividad sobre BApNA, S-2238 y S-2266, siendo incapaz de hidrolizar el sustrato S-2251. El empleo de estas aproximaciones estructurales y funcionales ha permitido lograr la identificación molecular del principal componente del veneno de B. atrox relacionado con su acción coagulante, así como evaluar en detalle la naturaleza de su actividad enzimática sobre diversos sustratos.

  12. Increments in cytokines and matrix metalloproteinases in skeletal muscle after injection of tissue-damaging toxins from the venom of the snake Bothrops asper

    Directory of Open Access Journals (Sweden)

    Alexandra Rucavado

    2002-01-01

    Full Text Available Envenomations by the snake Bothrops asper are characterized by prominent local tissue damage (i.e. myonecrosis, blistering, hemorrhage and edema. Various phospholipases A2 and metalloproteinases that induce local pathological alterations have been purified from this venom. Since these toxins induce a conspicuous inflammatory response, it has been hypothesized that inflammatory mediators may contribute to the local pathological alterations described. This study evaluated the local production of cytokines and matrix metalloproteinases (MMPs as a consequence of intramuscular injections of an Asp-49 myotoxic phospholipase A2 (myotoxin III (MT-III and a P-I type hemorrhagic metalloproteinase (BaP1 isolated from B. asper venom. Both enzymes induced prominent tissue alterations and conspicuous increments in interleukin (IL-1β, IL-6 and a number of MMPs, especially gelatinase MMP-9, rapidly after injection. In contrast, no increments in tumor necrosis factor-α (TNF-α and interferon-γ were detected. In agreement, MT-III and BaP1 did not induce the synthesis of TNF-α by resident peritoneal macrophages in vitro. Despite the conspicuous expression of latent forms of MMPs in muscle, evidenced by zymography, there were no increments in activated MMP-2 and only a small increase in activated MMP-9, as detected by a functional enzymatic assay. This suggests that MMP activity was regulated by a highly controlled activation of latent forms and, probably, by a concomitant synthesis of MMP inhibitors. Since no hemorrhage nor dermonecrosis were observed after injection of MT-III, despite a prominent increase in MMP expression, and since inflammatory exudate did not enhance hemorrhage induced by BaP1, it is suggested that endogenous MMPs released in the tissue are not responsible for the dermonecrosis and hemorrhage characteristic of B. asper envenomation. Moreover, pretreatment of mice with the peptidomimetic MMP inhibitor batimastat did not reduce myotoxic nor

  13. Efeitos da MT-I, uma fosfolipase A2, isolada do veneno de Bothrops asper em mastócitos: ativação e sinalização intracelular envolvida na desgranulação.

    OpenAIRE

    Marlos Cortez Sampaio

    2015-01-01

    Os efeitos da MT-I, uma fosfolipase A2 isolada do veneno de Bothrops asper (VBa), foram avaliados em mastócitos (MC) em cultura, quanto à: i) desgranulação e liberação de prostaglandina E2 (PGE2); ii) papel da atividade catalítica na desgranulação; iii) papel da PLD, PLC, cPLA2, iPLA2, PI3K, MAPK, PKC, PTK, ERK1/2, Junk, Gαi, Gαq e do cálcio na desgranulação; iv) expressão gênica de citocinas Th1 e Th2, e v) alterações ultraestruturais em MC. Os resultados mostraram que a MT-I, e...

  14. Evaluación biológica preliminar de extractos vegetales utilizados en la medicina tradicional de la Sierra Nevada deSanta Marta contra el veneno de la Bothrops asper

    Directory of Open Access Journals (Sweden)

    Willinton Barranco Pérez

    2013-10-01

    Full Text Available Title: Preliminary biological evaluation of plants extracts used in the Sierra Nevada de Santa Marta against the snake Bothrops asper venom.ResumenLa mordedura de serpientes constituye un problema de salubridad importante en muchos países tropicales y subtropicales, con un estimado de 2,5 millones de personas envenenadas cada año. En Colombia las especies Bothropsasper y Bothropsatrox son las causantes del 70 al 90 % de los accidentes registrados. Se estima que el 60% de estos accidentes son inicialmente tratados por curanderos tradicionales utilizando plantas medicinales en diferentes preparaciones. Este estudio evaluó la capacidad inhibitoria de cinco especies contra el efecto proteolítico y hemolítico indirecto inducido por el veneno de B. asper en ensayos in vitro.Las especies, que fueron seleccionadas de acuerdo a su uso en la medicina tradicional por parte de las comunidades campesinas de la Sierra Nevada de Santa Marta, fueron, Aristolochia máxima, Cissampelospareira, Equisetumbogotense, Mucunacfpruriens y una especie de la familia Asteraceae. La planta E. bogotense mostró los mayores porcentaje de inhibición contra la actividad de las fosfolipasas A2(42,29 %, así como la mayor precipitación de las proteínas en un rango de masas moleculares de 28,2 y 94,43 KDa. Al fraccionar el extracto de E. bogotense se obtuvieron cinco fracciones, las cuales presentaron un porcentaje de inhibición de 36,6 ± 1,07 a 46,1 ± 13,6. Adicionalmente se detectaron por métodos cualitativos núcleos como, alcaloides, esteroides y/o triterpenos, taninos, cumarinas y leucoantocianidinas. En estudio se reporta la actividad antiofídica en ensayos in vitro de la especie E. bogotense contra el veneno de la especie B.asper. (DUAZARY 2012 No. 2, 140 - 150AbstractIn Colombia the species Bothrops asper and Bothrops atrox are responsible for 70 to 90% of the snakebite accidents. Around 60% of these injuries are initially treated by traditional healers; they

  15. Action of Bothrops moojeni venom and its L-amino acid oxidase fraction, treated with {sup 60}Co gamma rays, in Leishmania spp; Acao do veneno de Bothrops moojeni e sua fracao L-aminoacido oxidase, submetida ao tratamento com raios gama de {sup 60}Co, em Leishmania spp

    Energy Technology Data Exchange (ETDEWEB)

    Cardoso, Andre Gustavo Tempone

    1999-07-01

    Bothrops moojeni venom showed an anti leishmania activity in vitro, as determined by a cell viability assay using the reduction of MTT. After venom purification, by chromatography techniques, the fractions with anti leishmania and L-amino acid oxidase activities, eluted in the same positions. The molecular weight of the enzyme was estimated to be 140 kDa by molecular exclusion chromatography, and 69 kDa, by SDS-PAGE, migrating as a single band, with an isoelectric point of 4.8 as determined by isoelectric focusing. The purified LAO from B. moojeni venom, 135-fold more active than crude venom, showed homo dimeric constitution, and was active against Leishmania spp from the New World, with an effective concentration against L(L). amazonensis of 1.80 {mu}g/ml (EC{sub 50}), L.(V.) panamensis (0.78 |{mu}g/ml) and L.(L.) chagasi (0.63 ({mu}g/ml). Ultrastructural studies of promastigotes affected by LAO demonstrated cell death, with edema in several organelles such as mitochondria and nuclear membrane, before cell disruption and necrosis. The action of LAO was demonstrated to be hydrogen peroxide-dependent. Studies with LLCMK-2 cells, treated with LAO, showed a toxic effect, with an EC{sub 50} of 11|{mu}g/ml. Irradiation of LAO with 6{sup 0C}o gamma rays, did not affect its whole oxidative activity, neither detoxified the enzyme. Amastigotes treated with LAO were not affected by its hydrogen peroxide, otherwise, the exogenous product, killed amastigotes with an EC{sub 50} of 0.67mM. These data could be of help in the development of alternative therapeutic approaches to the treatment of leishmaniasis. (author)

  16. The potential of aqueous extracts of Bellucia dichotoma Cogn. (Melastomataceae) to inhibit the biological activities of Bothrops atrox venom: A comparison of specimens collected in the states of Pará and Amazonas, Brazil.

    Science.gov (United States)

    de Moura, Valéria Mourão; de Souza, Luana Yamille Andrade; da Costa Guimarães, Noranathan; Dos Santos, Ilia Gilmara Carvalho; de Almeida, Patrícia Danielle Oliveira; de Oliveira, Ricardo Bezerra; Mourão, Rosa Helena Veras; Dos-Santos, Maria Cristina

    2017-01-20

    The effectiveness of aqueous extract of Bellucia dichotoma Cogn. (Melastomataceae) specimems collected in Santarém, PA, against some biological activities of Bothrops atrox venom (BaV) has been scientifically proven. Here, we analyzed the components and assessed the anti-snakebite potential of aqueous extracts of bark of B. dichotoma collected in Manaus, AM, (AEBd-MAO) and Santarém, PA, (AEBd-STM), both in Brazil. The phytochemical profiles of the aqueous extracts were identified using thin layer chromatography (TLC), and the concentrations of phenolics were determined by colorimetric assay. The inhibitory potential of the extracts was tested against the phospholipase A 2 , coagulant and gelatinolytic activities of BaV in vitro and its defibrinating and edema-inducing activities in vivo. Interaction between BaV and the extracts was investigated using SDS-Page electrophoresis and Western blotting. Extract cytotoxicity and antioxidant potential were assessed using the human fibroblast cell line MRC-5 and the DPPH assay in cell culture, respectively. While there was no difference between the phytochemical profiles of the extracts, AEBd-MAO had higher concentrations of total phenolics, total tannins and hydrolysable tannins. The extracts inhibited 100% of the phospholipase and coagulant activity of BaV when pre-incubated. Without pre-incubation, however, there was no reduction in phospholipase activity, although significant inhibition of coagulant activity was observed. In the doses used in folk medicine, without pre-incubation, both extracts inhibited 100% of the coagulant activity of BaV. In vivo, the extracts were unable to inhibit the defibrinating activity of the venom but were effective in inhibiting its edema-inducing activity. In the profiles of the extracts pre-incubated with BaV, not all the protein bands revealed by SDS-PAGE and Western blot were observed. Both extracts had a high antioxidant potential and neither had a cytotoxic effect. Although the

  17. Efficacy and safety of two whole IgG polyvalent antivenoms, refined by caprylic acid fractionation with or without beta-propiolactone, in the treatment of Bothrops asper bites in Colombia.

    Science.gov (United States)

    Otero, Rafael; León, Guillermo; Gutiérrez, José María; Rojas, Gustavo; Toro, María Fabiola; Barona, Jacqueline; Rodríguez, Verónica; Díaz, Abel; Núñez, Vitelbina; Quintana, Juan Carlos; Ayala, Shirley; Mosquera, Diana; Conrado, Lesdy L; Fernández, Diego; Arroyo, Yobana; Paniagua, Carlos A; López, Mercedes; Ospina, Carlos E; Alzate, Claudia; Fernández, Jorge; Meza, Jazmín J; Silva, Juan F; Ramírez, Patricia; Fabra, Patricia E; Ramírez, Eugenio; Córdoba, Elkin; Arrieta, Ana B; Warrell, David A; Theakston, R David G

    2006-12-01

    The efficacy and safety of two whole IgG polyvalent antivenoms (A and B) were compared in a randomised, blinded clinical trial in 67 patients systemically envenomed by Bothrops asper in Colombia. Both antivenoms were fractionated by caprylic acid precipitation and had similar neutralising potencies, protein concentrations and aggregate contents. Antivenom B was additionally treated with beta-propiolactone to lower its anticomplementary activity. Analysing all treatment regimens together, there were no significant differences between the two antivenoms (A=34 patients; B=33 patients) in the time taken to reverse venom-induced bleeding and coagulopathy, to restore physiological fibrinogen concentrations and to clear serum venom antigenaemia. Blood coagulability was restored within 6-24 h in 97% of patients, all of whom had normal coagulation and plasma fibrinogen levels 48 h after the start of antivenom treatment. Two patients (3.0%) had recurrent coagulopathy and eight patients suffered recurrence of antigenaemia within 72 h of treatment. None of the dosage regimens of either antivenom used guaranteed resolution of venom-induced coagulopathy within 6 h, nor did they prevent recurrences. A further dose of antivenom at 6 h also did not guarantee resolution of coagulopathy within 12-24 h in all patients. The incidence of early adverse reactions (all mild) was similar for both antivenoms (15% and 24%; P>0.05).

  18. Estabilidad de la actividad neutralizante del antiveneno ofídico conservado a 4°C y 16°C contra el veneno de tayas (complejo Bothrops atrox-asper de Colombia

    Directory of Open Access Journals (Sweden)

    J. Ramírez

    1994-12-01

    Full Text Available En este trabajo se informa acerca de la determinación de la actividad neutralizante de la letalidad de varios lotes de antiveneno ofídico en forma liquida, producidos en el lnstituto Nacional de Salud y almacenados en completa oscuridad a 4°C y a 16°C. El veneno usado para retar el antiveneno fue una mezcla de venenos de tayas (complejo Bothrops atrox-asper, procedentes de diferentes regiones de Colombia. Se estableció el porcentaje de la potencia remanente de los lotes para cada año y para cada temperatura de almacenamiento, para así corroborar la fecha de expiración del producto en forma líquida. Los resultados de este estudio muestran que el antiveneno conservado en las condiciones de oscuridad a 4ºC, luego de 3 años de almacenamiento mantiene casi la totalidad de la capacidad neutralizante (95%, mientras que el antiveneno almacenado bajo condiciones de oscuridad a 16ºC, luego de 2 años mantiene el 87% de su capacidad neutralizante. Así, recomendamos desechar el producto mantenido bajo estas condiciones a los dos años de fabricación.

  19. Effect of heparin treatment on the expression and activity of different ion-motive P-type ATPase isoforms from mouse extensor digitorum longus muscle during degeneration and regeneration after Bothrops jararacussu venom injection.

    Science.gov (United States)

    Schaffazick, Naiara; Amaral, Luciana S; Fonseca, Tatiane F; Tomaz, Marcelo A; Gaban, Glauco A; Borges, Paula A; Calil-Elias, Sabrina; Noël, François; Melo, Paulo A; Quintas, Luis Eduardo M; Cunha, Valéria M N

    2010-01-01

    Ca(2+) ions are essential to myonecrosis, a serious complication of snake envenomation, and heparin seems to counteract this effect. We investigated the effect of local injection of Bothrops jararacussu venom in mouse fast-twitch extensor digitorum longus (EDL) muscle, without or with heparin, on functional/molecular alterations of two central proteins involved in intracellular Ca(2+) homeostasis, sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA) and Na(+)/K(+)-ATPase. EDL-specific SERCA1 isoform expression dropped significantly just after venom administration (up to 60% compared to control EDL values at days 1 and 3; p<0.05) while SERCA2 and Na(+)/K(+)-ATPase alpha(1) isoform expression increased at the same time (3-6- and 2-3-fold, respectively; p<0.05). Although not significant, Na(+)/K(+)-ATPase alpha(2) isoform followed the same trend. Except for SERCA2, all proteins reached basal levels at the 7th day. Intravenous heparin treatment did not affect these profiles. Ca(2+)-ATPase activity was also decreased during the first days after venom injection, but here heparin was effective to reinstate activity to control levels within 3 days. We also showed that B. jararacussu venom directly inhibited Ca(2+)-ATPase activity in a concentration-dependent manner. Our results indicate that EDL SERCA and Na(+)/K(+)-ATPase are importantly affected by B. jararacussu venom and heparin has protective effect on activity but not on protein expression.

  20. Purificación y caracterización de la l-amino ácido oxidasa del veneno de la serpiente Bothrops brazili "jergón shushupe"

    Directory of Open Access Journals (Sweden)

    Christian Solís

    2014-06-01

    Full Text Available Se ha purificado y caracterizado parcialmente la L-aminoácido oxidasa de la serpiente Bothrops brazili. El aislamiento se realizó usando técnicas cromatográficas en Sephadex G-100 y CM-Sephadex C-50, utilizando como eluyente buffer acetato de amonio 0,1M pH 6. La enzima fue purificada 29,3 veces con un rendimiento de 30,9%. Usando electroforesis en gel de poliacrilamida con dodecil sulfato de sodio (PAGE-SDS en condiciones reductoras y no reductoras, así como las técnicas de inmunodifusión e inmunoelectroforesis, se demostró la presencia de una sola banda proteica. La enzima fue caracterizada como una glicoproteína ácida con un peso molecular de 125,7 kd formada por dos subunidades de 59,9 kd unidas por enlaces débiles, con un pH óptimo entre 7,5 y 9, dependiendo del aminoácido usado como substrato; siendo termoestable hasta los 550 C y lábil a pH alcalino. Asimismo, los ensayos por el método del cilindro en placa de Grove demostraron el efecto antibacteriano de la proteína aislada en cepas estandarizadas de Staphylococcus aureus, Vibrio cholerae y Streptococcus faecalis.

  1. Secretory phospholipases A(2) isolated from Bothrops asper and from Crotalus durissus terrificus snake venoms induce distinct mechanisms for biosynthesis of prostaglandins E2 and D2 and expression of cyclooxygenases.

    Science.gov (United States)

    Moreira, Vanessa; Gutiérrez, José Maria; Soares, Andreimar Martins; Zamunér, Stella Regina; Purgatto, Eduardo; Teixeira, Catarina de Fátima Pereira

    2008-09-01

    The effects of myotoxin III (MT-III), a phospholipase A(2) (sPLA2) from Bothrops asper snake venom, and crotoxin B (CB), a neurotoxic and myotoxic sPLA2 from the venom of Crotalus durissus terrificus, on cyclooxygenases (COXs) expression and biosynthesis of prostaglandins (PGs) were evaluated, together with the mechanisms involved in these effects. Upon intraperitoneal injection in mice, both sPLA(2)s promoted the synthesis of PGD2 and PGE2, with a different time-course. MT-III, but not CB, induced COX-2 expression by peritoneal leukocytes without modification on COX-1 constitutive expression, whereas CB increased the constitutive activity of COX-1. MT-III increased the enzymatic activity of COX-1 and COX-2. Similar effects were observed when these sPLA(2)s were incubated with isolated macrophages, evidencing a direct effect on these inflammatory cells. Moreover, both toxins elicited the release of arachidonic acid from macrophages in vitro. Inhibition of cPLA2 by AACOCF3, but not of iPLA2 by PACOCF3 or BEL, significantly reduced PGD2, PGE2 and arachidonic acid (AA) release promoted by MT-III. These inhibitors did not affect MT-III-induced COX-2 expression. In contrast, cPLA2 inhibition did not modify the effects of CB, whereas iPLA2 inhibition reduced PGD2 and AA production induced by CB. These findings imply that distinct regulatory mechanisms leading to PGs' synthesis are triggered by these snake venom sPLA(2)s. Such differences are likely to explain the dissimilar patterns of inflammatory reaction elicited by these sPLA(2)s in vivo.

  2. Cytotoxicity induced in myotubes by a Lys49 phospholipase A2 homologue from the venom of the snake Bothrops asper: evidence of rapid plasma membrane damage and a dual role for extracellular calcium.

    Science.gov (United States)

    Villalobos, Juan Carlos; Mora, Rodrigo; Lomonte, Bruno; Gutiérrez, José María; Angulo, Yamileth

    2007-12-01

    Acute muscle tissue damage, myonecrosis, is a typical consequence of envenomations by snakes of the family Viperidae. Catalytically-inactive Lys49 phospholipase A(2) homologues are abundant myotoxic components in viperid venoms, causing plasma membrane damage by a mechanism independent of phospholipid hydrolysis. However, the precise mode of action of these myotoxins remains unsolved. In this work, a cell culture model of C2C12 myotubes was used to assess the action of Bothrops asper myotoxin II (Mt-II), a Lys49 phospholipase A(2) homologue. Mt-II induced a dose- and time-dependent cytotoxic effect associated with plasma membrane disruption, evidenced by the release of the cytosolic enzyme lactate dehydrogenase and the penetration of propidium iodide. A rapid increment in cytosolic Ca(2+) occurred after addition of Mt-II. Such elevation was associated with hypercontraction of myotubes and blebbing of plasma membrane. An increment in the Ca(2+) signal was observed in myotube nuclei. Elimination of extracellular Ca(2+) resulted in increased cytotoxicity upon incubation with Mt-II, suggesting a membrane-protective role for extracellular Ca(2+). Chelation of cytosolic Ca(2+) with BAPTA-AM did not modify the cytotoxic effect, probably due to the large increment induced by Mt-II in cytosolic Ca(2+) which overrides the chelating capacity of BAPTA-AM. It is concluded that Mt-II induces rapid and drastic plasma membrane lesion and a prominent Ca(2+) influx in myotubes. Extracellular Ca(2+) plays a dual role in this model: it protects the membrane from the cytolytic action of the toxin; at the same time, the Ca(2+) influx that occurs after membrane disruption is likely to play a key role in the intracellular degenerative events associated with Mt-II-induced myotube damage.

  3. Moojenactivase, a novel pro-coagulant PIIId metalloprotease isolated from Bothrops moojeni snake venom, activates coagulation factors II and X and induces tissue factor up-regulation in leukocytes.

    Science.gov (United States)

    Sartim, Marco A; Costa, Tassia R; Laure, Helen J; Espíndola, Milena S; Frantz, Fabiani G; Sorgi, Carlos A; Cintra, Adélia C O; Arantes, Eliane C; Faccioli, Lucia H; Rosa, José C; Sampaio, Suely V

    2016-05-01

    Coagulopathies following snakebite are triggered by pro-coagulant venom toxins, in which metalloproteases play a major role in envenomation-induced coagulation disorders by acting on coagulation cascade, platelet function and fibrinolysis. Considering this relevance, here we describe the isolation and biochemical characterization of moojenactivase (MooA), a metalloprotease from Bothrops moojeni snake venom, and investigate its involvement in hemostasis in vitro. MooA is a glycoprotein of 85,746.22 Da, member of the PIIId group of snake venom metalloproteases, composed of three linked disulfide-bonded chains: an N-glycosylated heavy chain, and two light chains. The venom protease induced human plasma clotting in vitro by activating on both blood coagulation factors II (prothrombin) and X, which in turn generated α-thrombin and factor Xa, respectively. Additionally, MooA induced expression of tissue factor (TF) on the membrane surface of peripheral blood mononuclear cells (PBMC), which led these cells to adopt pro-coagulant characteristics. MooA was also shown to be involved with production of the inflammatory mediators TNF-α, IL-8 and MCP-1, suggesting an association between MooA pro-inflammatory stimulation of PBMC and TF up-regulation. We also observed aggregation of washed platelets when in presence of MooA; however, the protease had no effect on fibrinolysis. Our findings show that MooA is a novel hemostatically active metalloprotease, which may lead to the development of coagulopathies during B. moojeni envenomation. Moreover, the metalloprotease may contribute to the development of new diagnostic tools and pharmacological approaches applied to hemostatic disorders.

  4. Variaciones ontogénicas y geográficas en los efectos tóxicos y en las características inmunoquímicas del veneno de serpientes del grupo Bothrops atrox del Meta y Antioquia

    Directory of Open Access Journals (Sweden)

    María Fabiola Toro

    2000-02-01

    Full Text Available

    El accidente ofídico es un problema de salud pública frecuente en algunos lugares del mundo. En Latinoamérica, el género Bothrops es el causante del 90% de los accidentes ofídicos y en Colombia el grupo Bothrops atrox es responsable del 50% o más de los casos que se reportan anualmente (2000 casos. Este grupo se encuentra distribuido en la mayor parte del territorio nacional colombiano, incluyendo el este y oeste de los Andes, vertiente Pacífica y Atlántica, además de los valles interandinos.

    El veneno del grupo Bothrops atrox induce un complejo cuadro de alteraciones fisiopatológicas locales y sistémicas, como mionecrosis, dermonecrosis, edema, hemorragia, dolor, coagulopatías, choque cardiovascular, nefrotoxicidad, actividad proteolítica y hemolítica indirecta.

    En diversos países se han realizado estudios sobre la variabilidad de los venenos de diferentes especies de serpientes. Se han encontrado diferencias en todos los niveles taxonómicos, siendo de mayor importancia la variabilidad intraespecífica. Con este estudio se pretende determinar la variabilidad ontogénica y geográfica del veneno de Bothrops atrox procedente de dos regiones colombianas, mediante pruebas biológicas e inmunoquímicas. Los resultados previos muestran que existe una variación ontogénica en el veneno de la especie Bothrops atrox del departamento del Meta con respecto a la letalidad, y a las actividades hemorrágica, coagulante y edematizante. Las serpientes recién nacidas tienen una mayor letalidad (DL50 = 51.8 ± 5.6 mg de veneno con respecto a los adultos (81.4±

  5. Ac2-26 Mimetic Peptide of Annexin A1 Inhibits Local and Systemic Inflammatory Processes Induced by Bothrops moojeni Venom and the Lys-49 Phospholipase A2 in a Rat Model.

    Directory of Open Access Journals (Sweden)

    Bruna Stuqui

    Full Text Available Annexin A1 (AnxA1 is an endogenous glucocorticoid regulated protein that modulates anti-inflammatory process and its therapeutic potential has recently been recognized in a range of systemic inflammatory disorders. The effect of the N-terminal peptide Ac2-26 of AnxA1 on the toxic activities of Bothrops moojeni crude venom (CV and its myotoxin II (MjTX-II were evaluated using a peritonitis rat model. Peritonitis was induced by the intraperitoneal injection of either CV or MjTX-II, a Lys-49 phospholipase A2. Fifteen minutes after the injection, the rats were treated with either Ac2-26 or PBS. Four hours later, the CV and MjTX-II-induced peritonitis were characterized by neutrophilia (in the peritoneal exudate, blood and mesentery and increased number of mesenteric degranulated mast cells and macrophages. At 24 hours post-injection, the local inflammatory response was attenuated in the CV-induced peritonitis while the MjTX-II group exhibited neutrophilia (peritoneal exudates and blood. Ac2-26 treatment prevented the influx of neutrophils in MjTX-II-induced peritonitis and diminished the proportion of mesenteric degranulated mast cells and macrophages in CV-induced peritonitis. Additionally, CV and MjTX-II promoted increased levels of IL-1β and IL-6 in the peritoneal exudates which were significantly reduced after Ac2-26 treatment. At 4 and 24 hours, the endogenous expression of AnxA1 was upregulated in the mesenteric neutrophils (CV and MjTX-II groups and mast cells (CV group. In the kidneys, CV and MjTX-II administrations were associated with an increased number of macrophages and morphological alterations in the juxtamedullary nephrons in proximal and distal tubules. Ac2-26 promoted significant recovery of the juxtamedullary structures, decreased the number of macrophages and diminished the AnxA1 in epithelial cells from distal tubules and renal capsules. Our results show that Ac2-26 treatment significantly attenuates local and systemic

  6. Glycolic acid inhibits enzymatic, hemorrhagic and edema-inducing activities of BaP1, a P-I metalloproteinase from Bothrops asper snake venom: insights from docking and molecular modeling.

    Science.gov (United States)

    Pereañez, Jaime Andrés; Patiño, Arley Camilo; Rey-Suarez, Paola; Núñez, Vitelbina; Henao Castañeda, Isabel Cristina; Rucavado, Alexandra

    2013-09-01

    Glycolic acid (GA) (2-Hydroxyethanoic acid) is widely used as chemical peeling agent in Dermatology and, more recently, as a therapeutic and cosmetic compound in the field of skin care and disease treatment. In this work we tested the inhibitory ability of glycolic acid on the enzymatic, hemorrhagic and edema-inducing activities of BaP1, a P-I metalloproteinase from Bothrops asper venom, which induces a variety of toxic actions. Glycolic acid inhibited the proteolytic activity of BaP1 on azocasein, with an IC₅₀ of 1.67 mM. The compound was also effective at inhibiting the hemorrhagic activity of BaP1 in skin and muscle in experiments involving preincubation of enzyme and inhibitor prior to injection. When BaP1 was injected i.m. and then, at the same site, different concentrations of glycolic acid were administered at either 0 or 5 min, 7 mM solutions of the inhibitor partially abrogated hemorrhagic activity when administered at 0 min. Moreover, glycolic acid inhibited, in a concentration-dependent manner, edema-forming activity of BaP1 in the footpad. In order to have insights on the mode of action of glycolic acid, UV-vis and intrinsic fluorescence studies were performed. Results of these assays suggest that glycolic acid interacts directly with BaP1 and chelates the Zn²⁺ ion at the active site. These findings were supported by molecular docking results, which suggested that glycolic acid forms hydrogen bonds with residues Glu143, Arg110 and Ala111 of the enzyme. Additionally, molecular modeling results suggest that the inhibitor chelates Zn²⁺, with a distance of 3.58 Å, and may occupy part of substrate binding cleft of BaP1. Our results suggest that glycolic acid is a candidate for the development of inhibitors to be used in snakebite envenomation. Copyright © 2013 Elsevier Ltd. All rights reserved.

  7. Características bioquímicas y capacidad neutralizante de cuatro antivenenos polivalentes frente a los efectos farmacológicos y enzimáticos del veneno de Bothrops asper y Porthidium nasutum de Antioquia y Chocó

    Directory of Open Access Journals (Sweden)

    Mónica Saldarriaga

    2002-01-01

    Full Text Available En Colombia, el 90-95% de las 3000 mordeduras de serpientes informadas cada año, son ocasionadas por Bothrops spp, con una elevada mortalidad y secuelas. Siguiendo recomendaciones de la OMS, se evaluó la capacidad neutralizante de los efectos farmacológicos y enzimáticos de los venenos de Bothrops asper y Porthidium nasutum de Antioquia y Chocó por cuatro antivenenos; 2 de ellos de IgG completa (polivalente antibothrópico, anticrotálico del Instituto Nacional de Salud INS -Colombia; polivalente antibothrópico, anticrotálico, antilachésico de Laboratorios Probiol -Colombia y 2 antivenenos de fragmentos F(ab’2 (polivalente antibothrópico, anticrotálico del Centro de Biotecnología de la Universidad Central de Venezuela; y el polivalente antibothrópico, anticrotálico Antivipmyn® del Instituto Bioclón -México. Se determinó la actividad letal, hemorrágica, desfibrinante, edematizante, mionecrosante y hemolítica indirecta de cada veneno, siguiendo métodos ya estandarizados. Las pruebas de neutralización in vitro e in vivo se realizaron por el método de preincubación a 370C de dosis fijas de veneno y dosis variables de antiveneno. Los antivenenos Antivipmyn® de México y polivalente INS de Colombia tuvieron la mayor potencia neutralizante de todos los efectos farmacológicos y enzimáticos del veneno de B. asper y P. nasutum. El antiveneno polivalente Probiol fue el de menor capacidad neutralizante y mayor concentración de proteínas. Los antivenenos de fragmentos F(ab’2 tuvieron más baja concentración de proteínas y solo cantidades menores de proteínas no inmunes por electroforesis. Ninety to 95% of the snakebites reported yearly in Colombia are inflicted by Bothrops spp with high mortality and sequelae. Following recommendations of the World Health Organization, the neutralizing ability of four polyvalent antivenoms against several pharmacological and enzymatic effects of Bothrops asper and Porthidium nasutum snake

  8. Bothrops bites in Colombia: a multicenter study on the efficacy and safety of Antivipmyn-Tri®, a polyvalent antivenom produced in Mexico Accidente bothrópico en Colombia: estudio multicéntrico de la eficacia y seguridad de Antivipmyn-Tri®, un antiveneno polivalente producido en México

    Directory of Open Access Journals (Sweden)

    María Eugenia Espinal Saldarriaga

    2007-08-01

    Full Text Available Introduction: Colombia is a country with two whole IgG antivenom producers, but the expectancy of all the market are not fulfilled by different technical reasons. Objectives: To evaluate the efficacy and safety of a F(ab´2 polyvalent antivenom produced in Mexico and of a new dosage regimen for Bothrops bites in Colombia. Methods: A clinical trial, including serum venom and antivenom measurements (ELISA, was performed during 9 months in 53 patients. Results: forty four patients were bitten by Bothrops asper in Antioquia and Chocó and 9 by B. atrox in Amazonas; on admission, all of them had nonclottable blood, 30 (56.6% presented local and 24 (45.3% systemic bleeding. The final envenoming grade was mild in 13 (24.5%, moderate in 30 (56.6% and severe in 10 patients (18.9%. At the antivenom doses used in this study (5 vials for mild / moderate and 10 for severe envenoming, Antivipmyn Triwas 100% efficient to decrease significantly serum venom concentrations within the first treatment hour, and to stop local and systemic bleeding within 6-12 hours, 96.2% efficient to restore blood coagulation within 24 hours and 100% within 48 hours. Two patients (3.8% had recurrence of coagulopathy without bleeding, and there were 12 recurrences of antigenaemia without clinical relevance. Ten (18.9% patients suffered early mild adverse reactions to fabotherapy. There were no deaths and four patients (7.5% presented sequelae. Conclusion: at the doses used in this study, Antivipmyn Tri® was efficient and safe for the treatment of Bothrops bites in Colombia. Introducción: Colombia es un país con dos productores de antivenenos de IgG, pero hay un mercado insatisfecho por diferentes razones técnicas. Objetivos: evaluar la eficacia y la seguridad de un antiveneno F(ab´2 polivalente (Antivipmyn-Tri® producido en México, y un nuevo esquema de dosis en accidente bothrópico en Colombia. Métodos: se realizaron durante 9 meses un ensayo clínico-terapéutico y

  9. VARIACIONES EN L COMPOSICIÓN PROTEICA, ACTIVIDADES ENZIMÁTICAS Y BIOLÓGICAS DEL VENENO DE LA SERPIENTE BOTHROPS ATROX (VIPEREIDAE EN RELACIÓN CON LA EDAD

    Directory of Open Access Journals (Sweden)

    O. Málaga

    2014-06-01

    Full Text Available La serpiente Bothrops atrox, que habita en la Selva Amazónica del Perú, es responsable del mayor número de accidentes ofídicos y constituye un problema de salud pública. Por esta razón, se ha realizado un estudio de las variaciones en la composición y actividad del veneno en tres grupos de especimenes en cautiverio, que corresponden a juveniles de 1 y 2 años, así como ejemplares adultos mayores de 5 años. Los venenos obtenidos fueron liofilizados para su conservación y diluidos en concentraciones iniciales de 1 mg/ml en solución salina o en el buffer apropiado. Con estas muestras se efectuaron los análisis de concentración de proteína por D. 0. 280 nm y por el método de Lowry, determinándose además el número de bandas proteicas por electroforesis en gel de poliacrilamida con dodecil sulfato de sodio (PAGE-SIDS. Con las mismas muestras se hicieron ensayos para medir las siguientes actividades enzimáticas: caseinolítica, amidolftica, coagulante, hialuronidasa, L-aminocácido oxidasa (LAO y fosfolipasa A, adicionalmente se determinó la actividad hemorrágica y el efecto edemático con ratones albinos. Los resultados mostraron que la mayor concentración proteica (0,938mg de proteína/mg de veneno así como el número máximo de bandas proteicas (8 bandas se obtuvieron con los venenos de juveniles de 2 años. Asimismo se encontró que algunas actividades enzimáticas como la amidolítica, fosfolipásica y L-aminoácido oxidasa fueron más altas en los juveniles de 2 años, además de los efectos hemorrágico y edemático. En cambio, las actividades coagulante y proteolítica sobre caseína se elevaron progresivamente con la edad de los ejemplares. Sin embargo, la enzima hialuronidasa tuvo su máximo valor en los juveniles de 1 año, decreciendo notablemente en los adultos. Es interesante señalar también que fosfolipasa A2 en los adultos sólo tuvo una actividad de 6,4% con respecto al valor más alto encontrado en el veneno

  10. Aspectos epidemiológicos e clínicos dos acidentes ofídicos ocorridos nos municípios do Estado do Amazonas Epidemiological and clinical aspects of snake accidentes in the municipalities of the State of Amazonas, Brazil

    Directory of Open Access Journals (Sweden)

    Célio Campos Borges

    1999-12-01

    Full Text Available No Amazonas, o acidente ofídico é um problema de saúde pública pouco conhecido. Por este motivo, foi realizado um estudo descritivo dos acidentes ofídicos atendidos nas Unidades de Saúde de 34 municípios, um distrito e dois pelotões de fronteira do Estado do Amazonas. As características mais comuns encontradas dentre os pacientes foram: agricultor (50,4%, do sexo masculino (81,3%, em idade produtiva (72,1%, picado no membro inferior (88,5%, por jararaca (48,6% ou surucucu (46,8%, na zona rural de seu município (70,2% e que só recebeu atendimento médico em tempo superior a seis horas, após acidente (57,3%. As manifestações locais mais freqüentes foram: edema (76,9%, dor (68,7%, eritema (10,2% e hemorragia (9,3%. Hemorragia (18,8% foi a manifestação sistêmica mais freqüente. O antiveneno foi administrado em apenas 65,9% dos pacientes. A via mais utilizada foi a endovenosa (52,3%, sendo relevante o uso de vias não mais recomendadas (47,7%. O antiveneno administrado, na maioria dos pacientes, foi o antibotrópico (66,7%. As complicações mais freqüentes foram abcesso (13,7%, necrose (12,3%, infecção secundária (8,3%, insuficiência renal (2,5% e gangrena (2,5%. Os procedimentos médicos mais usados para o tratamento das complicações foram: drenagem (52,6%, debridamento (28,9%, amputação (10,5%, limpeza cirúrgica (5,3% e diálise peritoneal (2,6%. A letalidade foi de 1%.In the State of Amazonas, accidents with snakes are a public health problem. For this reasons, the objective of this work was to carry out a descriptive study of the snake accidents attended in the health units of 34 municipalities, one district and two border platoons in the State of Amazonas. The characteristics most commonly observed among those involved in snake accidents were: farmers (50.4%, male (81.3%, belonging to the working age-group (72.1%, bitten on an upper limb (88.5% by a "jararaca" (48.6% or a "surucucu" (46.8% in the rural part of the

  11. Acções phylacticas não especificas em relação aos venenos ophidicos: tratamento auxiliar dos accidentes ophidicos

    Directory of Open Access Journals (Sweden)

    J. Vellard

    1929-01-01

    Full Text Available Dans l'intoxication expérimentale du pigeon avec les venins de Crotalus terrificus, de Lachesis jararaca et de Naya tripudians, nous avons étudié l'action de 12 médicaments, possédant des affinités neurotropes plus ou moins accusées, arrivant aux conclusions suivantes: 1 - Quelques-uns des ces médicaments exercent une action protectrice spéciale contre ces venins, permettant, injectés une heure avant le vénin, d'eviter tout symptôme d'intoxication chez le pigeon. Certains n'agissent qu'injectés em même temps que le vénin. D'autres sont complètement inactifs. Aucun d'êntre eux ne possède de propriétés curatives. 2 - En plus de ce pouvoir protecteur, ou phylactique, un certain nombre d'entre eux possède sur les venins, une action neutralisante "in vitro", s'accentuant après une heure de contact. 3 - Les conditions dans lesquelles s'exercent ces deux actions, protectrice et neutralisante, sont très variables suivant les venins et les médicaments. Sur leur mécanisme très complexe, on ne peut actuellement formuler que des hypothèses. 4 - Au point de vue pratique, certains de ces médicaments, surtout la spartéine et l'adrénaline, peuvent être utilisés avec avantage comme complément du traitement sérotérapique spécifique dans les accidents déterminés par les animaux venimeux.

  12. Cell death induced by Bothrops asper snake venom metalloproteinase on endothelial and other cell lines.

    Science.gov (United States)

    Brenes, Oscar; Muñóz, Eduardo; Roldán-Rodríguez, Raquel; Díaz, Cecilia

    2010-06-01

    Two adherent cell lines, BAEC and HeLa, and non-adherent Jurkat, were treated with snake venom metalloproteinase BaP1 to determine whether cytotoxicity, previously reported for this toxin, could be mediated by the process of anoikis. It was observed that there was no correlation between the ability of this toxin to induce loss of adherence, and the cytotoxic effect, since concentrations that do not induce loss of adherence (3-6 microg/mL), were able to trigger 50% of cytotoxicity in BAEC. In the case of HeLa, where toxicity was very low (less than 20% at maximun concentrations and times of exposure), significant detachment and no toxicity was observed at concentrations of 1.5 microg/mL, showing also no correlation between both events. We also observed differences between BAEC toxicity measured by XTT reduction and DNA fragmentation determined by flow cytometry (as an indicator of apoptosis), since concentrations that induce 100% of cytotoxicity barely showed any DNA fragmentation (12% at 24h), suggesting that if apoptosis was involved, DNA damage is still not present, although chromatin condensation, another indicator of apoptosis, is observed in 40% of the cells. Inhibition of BAEC cytotoxicity by caspase inhibitors indicate that apoptosis is playing a role in this process, but other mechanisms of cell death could be participating also. Another way to determine whether the mechanism of cell death was related to anoikis was using a non-adherent cell line, which should show substrate independence. We determined by TUNEL that at 50 microg/ml BaP1 triggered 50% of apoptosis at 96 h, an effect that was seen earlier, suggesting also that if this toxin was inducing apoptosis in a non-adherent cell line, the mechanism could not be related to loss of attachment. Cell cycle arrest in S phase was also observed in Jurkat cells, an effect that could be leading to apoptosis. In conclusion, since there was no correlation between cell detachment and cytotoxicity (and apoptosis) in adherent cell lines and due to the ability of BaP1 to induce apoptosis in a non-adherent cell line, we suggest that this enzyme is toxic by a mechanism not related to anoikis, and that in the case of Jurkat cells, it is likely to be related to its ability to induce cell cycle arrest. Processes other than apoptosis could be also involved in the cell death mechanism mediated by BaP1 on BAEC. Copyright 2010 Elsevier Inc. All rights reserved.

  13. ACIDENTES POR SERPENTE (BOTHROPS SPP. E CROTALLUS SPP.) EM CRIANÇAS: RELATO DE DOIS CASOS

    OpenAIRE

    Beatriz Ferreira Martins; Ana Paula dos Santos Campos; Maycon Rogério Seleghim; Tanimária da Silva Lira Ballani; Erika Okuda Tavares; Magda Lúcia Félix de Oliveira

    2012-01-01

    Los accidentes provocados por serpientes poseen importancia sanitaria debido a la frecuencia y gravedad. La estandarización de la atención es imprescindible para auxiliar a los equipos de salud en el diagnóstico precoz y en el tratamiento específico a las víctimas. Pero, la evolución clínica del caso presenta particularidades, principalmente en accidentes infantiles y en ancianos. El objetivo fue describir y analizar dos casos clínicos de accidentes infantiles con serpientes de los géneros Bo...

  14. Producción de Suero Antiveneno de Bothrops Alternatus en ovinos

    OpenAIRE

    Stamatti, Guillermo; Baravalle, Cristian Gerardo; Peñalva, Pablo; Gorchs, Carolina; Bini, Ricardo Bruno

    2012-01-01

    p. 101-106 La producción de suero antiofídico como único tratamiento comprobadamente efectivo contra el accidente producido por la mordedura de serpientes venenosas, se ha venido realizando en equinos desde finales del siglo pasado. En la actualidad, poco se conoce del porqué de la escasez de investigaciones en otras especies y además relativamente, se ha hecho poco en términos de conseguir una nueva manera de producir un antiveneno más efectivo. Los sueros equinos usados para el tratamien...

  15. Rapid Purification of a New P-I Class Metalloproteinase from Bothrops moojeni Venom with Antiplatelet Activity

    Directory of Open Access Journals (Sweden)

    Mayara R. de Queiroz

    2014-01-01

    Full Text Available The present study aimed to evaluate the proteolytic and biological activities of a new metalloproteinase from B. moojeni venom. The purification of BmooMPα-II was carried out through two chromatographic steps (ion-exchange and affinity. BmooMPα-II is a monomeric protein with an apparent molecular mass of 22.5 kDa on SDS-PAGE 14% under nonreducing conditions. The N-terminal sequence (FSPRYIELVVVADHGMFTKYKSNLN revealed homology with other snake venom metalloproteinases, mainly among P-I class. BmooMPα-II cleaves Aα-chain of fibrinogen followed by Bβ-chain, and does not show any effect on the γ-chain. Its optimum temperature and pH for the fibrinogenolytic activity were 30–50°C and pH 8, respectively. The inhibitory effects of EDTA and 1,10-phenantroline on the fibrinogenolytic activity suggest that BmooMPα-II is a metalloproteinase. This proteinase was devoid of haemorrhagic, coagulant, or anticoagulant activities. BmooMPα-II caused morphological alterations in liver, lung, kidney, and muscle of Swiss mice. The enzymatically active protein yet inhibited collagen, ADP, and ristocetin-induced platelet aggregation in a concentration-dependent manner. Our results suggest that BmooMPα-II contributes to the toxic effect of the envenomation and that more investigations to elucidate the mechanisms of inhibition of platelet aggregation may contribute to the studies of snake venom on thrombotic disorders.

  16. Clinical safety evaluation of F(ab')₂ antivenom (Crotalus durissus-Bothrops asper) administration in dogs.

    Science.gov (United States)

    Woods, Craig; Young, David

    2011-10-01

    Antivenoms consisting of selective antigen binding antibody fragments, or F(ab')(2), are becoming more popular in human and veterinary medicine, owing to their preferred kinetics, tissue distribution, and removal of the Fc binding portion of IgG. Consequences of antivenom administration can include acute and delayed reactions, dependent, in part, on the antivenom's donor source, purity, and composition. This study evaluated an equine-derived polyvalent F(ab')(2) pit viper antivenom in healthy dogs of various size, age, and breed under controlled conditions. Dogs were allocated into 6 treatment groups (n = 10 per group) based on weight (3 weight groups) and dose (2 dose groups) and administered F(ab')(2) antivenom over 1 hour by IV infusion. Dogs were observed for adverse events at 3, 6, 12, and 24 hours after administration and blood was collected for CBC and serum biochemistry before and at 24 hours postadministration. No abnormalities were noted in the 3-vial group. In the 6-vial group, 3 dogs developed mild self-limiting edema around the head or neck, suggestive of a type 1 hypersensitivity, while another dog vomited and developed mild hypocalcemia at 24 hours, reflecting a 13% reaction rate at high doses. However, no dogs exhibited clinical signs of hypocalcemia or had any severe adverse events. CBC remained normal in all groups. This study demonstrated that the polyvalent F(ab')(2) pit viper antivenom is well tolerated in dogs given large doses in a short period of time. © Veterinary Emergency and Critical Care Society 2011.

  17. Efecto inhibitorio de extractos de Renealmia alpinia Rottb. Maas (Zingiberaceae sobre el veneno de Bothrops asper (mapaná

    Directory of Open Access Journals (Sweden)

    Arley Camilo Patiño

    2012-03-01

    Full Text Available Introducción. La medicina tradicional es una invaluable fuente de investigación de nuevos remedioscomo complemento para el tratamiento del accidente ofídico, considerado como un grave problema desalud pública a nivel mundial. Objetivo. Este trabajo de investigación pretende comprobar la capacidad de neutralizar los efectoshemorrágicos, coagulantes y proteolíticos, de los extractos de hojas de Renealmia alpinia, usadatradicionalmente por los indígenas del Chocó (Colombia contra la mordedura de la serpiente Bothropsasper, causante de la gran mayoría de los accidentes ofídicos en nuestro país. Materiales y métodos. Se llevaron a cabo ensayos de toxicidad aguda y de actividad analgésica in vivo de R. alpinia. Además, se hicieron ensayos in vitro sobre inhibición de las actividades coagulante,hemolítica y proteolítica del veneno de Resultados. El presente estudio demuestra que R. alpinia no produce efectos tóxicos en animales deexperimentación; además, presenta efectos analgésicos in vivo y antiofídicos in vitro, y protege contralos efectos letales del veneno de B. asper, in vivo. Conclusión. Renealmia alpinia puede ser una buena alternativa terapéutica como complemento altratamiento con antiveneno en el accidente ofídico, por sus efectos analgésicos y antiofídicos. doi: http://dx.doi.org/10.7705/biomedica.v32i3.591

  18. Effects of a low-level semiconductor gallium arsenide laser on local pathological alterations induced by Bothrops moojeni snake venom.

    Science.gov (United States)

    Aranha de Sousa, Elziliam; Bittencourt, José Adolfo Homobono Machado; Seabra de Oliveira, Nayana Keyla; Correia Henriques, Shayanne Vanessa; dos Santos Picanço, Leide Caroline; Lobato, Camila Pena; Ribeiro, José Renato; Pereira, Washington Luiz Assunção; Carvalho, José Carlos Tavares; da Silva, Jocivânia Oliveira

    2013-10-01

    Antivenom therapy has been ineffective in neutralizing the tissue damage caused by snakebites. Among therapeutic strategies to minimize effects after envenoming, it was hypothesized that a low level laser would reduce complications and reduce the severity of local snake venom effects. In the current study, the effect of a low-level semiconductor gallium arsenide (GaAs) laser on the local pathological alterations induced by B. moojeni snake venom was investigated. The experimental groups consisted of five male mice, each administered either B. moojeni venom (VB), B. moojeni venom + antivenom (VAV), B. moojeni venom + laser (VL), B. moojeni venom + antivenom + laser (VAVL), or sterile saline solution (SSS) alone. Paw oedema was induced by intradermal administration of 0.05 mg kg(-1) of B. moojeni venom and was expressed in mm of directly induced oedema. Mice received by subcutaneous route 0.20 mg kg(-1) of venom for evaluating nociceptive activity and the time (in seconds) spent in licking and biting the injected paw was taken as an indicator of pain response. Inflammatory infiltration was determined by counting the number of leukocytes present in the gastrocnemius muscle after venom injection (0.10 mg kg(-1)). For histological examination of myonecrosis, venom (0.10 mg kg(-1)) was administered intramuscularly. The site of venom injection was irradiated by the GaAs laser and some animals received antivenom intraperitoneally. The results indicated that GaAs laser irradiation can help in reducing some local effects produced by the B. moojeni venom in mice, stimulating phagocytosis, proliferation of myoblasts and the regeneration of muscle fibers.

  19. Extratos de Curcuma longa L. e Kalanchoe brasiliensis Camb. no tratamento local do envenenamento por Bothrops alternatus

    Directory of Open Access Journals (Sweden)

    F. V. Fonseca

    Full Text Available Este trabalho teve como objetivo a utilização dos extratos aquosos das plantas Curcuma longa e Kalanchoe brasiliensis na terapêutica tópica complementar do envenenamento botrópico experimental em camundongos, visando a antagonização dos efeitos locais (edema, hemorragia e necrose provocados pelo veneno. O experimento mostrou que os melhores resultados foram obtidos com o extrato de Kalanchoe brasiliensis.

  20. Histological characterization of Sticholecitha serpentis Prudhoe, 1949 (digenea, bieriidae, sticholecithinae, parasite of Bothrops moojeni Hoge, 1966 (serpentes, viperidae

    Directory of Open Access Journals (Sweden)

    TB. Silva

    2005-12-01

    Full Text Available The aim of this study was to evaluate the morphology of the species Sticholecitha serpentis Prudhoe, 1949 by means of histological procedures. Helminths were previously fixed in Railliet & Henry solution, uncompressed and were included in 2-hydroxyethyl-methacrylate. Longitudinal and transverse serial sections with a 4-mum thickness were performed in a microtome (Leica RM 2165, stained with haematoxylin-eosin and then analyzed in a computerized image analysis system (Qwin Lite 2.5, Leica. Structures of systematic value, such as oral sucker, acetabulum, prepharynx, pharynx, esophagus, intestinal caeca, vitelline glands, ovary, uterus, cirrus pouch and testicles were described. Structures that were poorly visible in total preparations were also observed and described, such as efferent ducts, ejaculatory duct, prostate, seminal vesicle, seminal receptacle, Laurer’s channel, Mehlis’ gland, vitelline ducts, metraterm, genital atrium, digestive glands and excretory vesicle. We demonstrated that histological analysis can supply important data regarding the morphological characterization of S. serpentis and will be able to contribute to systematic studies of trematodes.

  1. IDENTIFICAÇÃO DA ATIVIDADE ANTIMICROBIANA NO VENENO DA SERPENTE Bothrops moojeni EM BACTÉRIAS GRAM NEGATIVAS.

    OpenAIRE

    Queiroz, Silvio José de

    2010-01-01

    As infecções causadas por microrganismos multirresistentes estão associadas com bactérias adquiridas no meio ambiente hospitalar. A pressão seletiva dos antimicrobianos é um importante fator de seleção e disseminação dos genes de resistência no meio ambiente. O uso de moléculas produzidas por diferentes seres vivos tem demonstrado atividade contra microrganismos. OBJETIVOS: Avaliar a atividade do veneno bruto da Bothrps moojeni sobre bactérias gram-negativas produtoras e não...

  2. Role of enzymatic activity in muscle damage and cytotoxicity induced by Bothrops asper Asp49 phospholipase A2 myotoxins: are there additional effector mechanisms involved?

    Science.gov (United States)

    Mora-Obando, Diana; Díaz, Cecilia; Angulo, Yamileth; Gutiérrez, José María; Lomonte, Bruno

    2014-01-01

    Viperid venoms often contain mixtures of Asp49 and Lys49 PLA2 myotoxin isoforms, relevant to development of myonecrosis. Given their difference in catalytic activity, mechanistic studies on each type require highly purified samples. Studies on Asp49 PLA2s have shown that enzyme inactivation using p-bromophenacyl bromide (p-BPB) drastically affects toxicity. However, based on the variable levels of residual toxicity observed in some studies, it has been suggested that effector mechanisms independent of catalysis may additionally be involved in the toxicity of these enzymes, possibly resembling those of the enzymatically inactive Lys49 myotoxins. A possibility that Lys49 isoforms could be present in Asp49 PLA2 preparations exists and, if undetected in previous studies, could explain the variable residual toxicity. This question is here addressed by using an enzyme preparation ascertained to be free of Lys49 myotoxins. In agreement with previous reports, inactivation of the catalytic activity of an Asp49 myotoxin preparation led to major inhibition of toxic effects in vitro and in vivo. The very low residual levels of myotoxicity (7%) and cytotoxicity (4%) observed can be attributed to the low, although detectable, enzyme remaining active after p-BPB treatment (2.7%), and would be difficult to reconcile with the proposed existence of additional catalytic-independent toxic mechanisms. These findings favor the concept that the effector mechanism of toxicity of Asp49 PLA2 myotoxins from viperids fundamentally relies on their ability to hydrolyze phospholipids, arguing against the proposal that membrane disruption may also be caused by additional mechanisms that are independent of catalysis.

  3. Role of enzymatic activity in muscle damage and cytotoxicity induced by Bothrops asper Asp49 phospholipase A2 myotoxins: are there additional effector mechanisms involved?

    Directory of Open Access Journals (Sweden)

    Diana Mora-Obando

    2014-09-01

    Full Text Available Viperid venoms often contain mixtures of Asp49 and Lys49 PLA2 myotoxin isoforms, relevant to development of myonecrosis. Given their difference in catalytic activity, mechanistic studies on each type require highly purified samples. Studies on Asp49 PLA2s have shown that enzyme inactivation using p-bromophenacyl bromide (p-BPB drastically affects toxicity. However, based on the variable levels of residual toxicity observed in some studies, it has been suggested that effector mechanisms independent of catalysis may additionally be involved in the toxicity of these enzymes, possibly resembling those of the enzymatically inactive Lys49 myotoxins. A possibility that Lys49 isoforms could be present in Asp49 PLA2 preparations exists and, if undetected in previous studies, could explain the variable residual toxicity. This question is here addressed by using an enzyme preparation ascertained to be free of Lys49 myotoxins. In agreement with previous reports, inactivation of the catalytic activity of an Asp49 myotoxin preparation led to major inhibition of toxic effects in vitro and in vivo. The very low residual levels of myotoxicity (7% and cytotoxicity (4% observed can be attributed to the low, although detectable, enzyme remaining active after p-BPB treatment (2.7%, and would be difficult to reconcile with the proposed existence of additional catalytic-independent toxic mechanisms. These findings favor the concept that the effector mechanism of toxicity of Asp49 PLA2 myotoxins from viperids fundamentally relies on their ability to hydrolyze phospholipids, arguing against the proposal that membrane disruption may also be caused by additional mechanisms that are independent of catalysis.

  4. Caracterização funcional e estrutural de uma fosfolipase A2 ácida isolada do veneno de Bothrops pirajai

    OpenAIRE

    Sabrina Schaaf Teixeira

    2009-01-01

    A caracterização proteômica de venenos de serpentes contribui significativamente para o avanço das Ciências na área Biomédica. Várias destas moléculas, utilizadas como instrumentos de investigação de mecanismos celulares e moleculares, estão envolvidas em diversos processos fisiopatológicos e de intoxicação. No entanto, os venenos de serpentes ainda requerem caracterização funcional e/ou biológica adicionais. As fosfolipases A2s (PLA2s) são enzimas que induzem vários efeitos farmacológicos, e...

  5. Efecto neutralizador del extracto acuoso de Dracontium loretense (JERGÓN SACHA sobre la actividad letal del veneno de Bothrops atrox

    Directory of Open Access Journals (Sweden)

    Amanda Lovera

    2006-07-01

    Full Text Available Objetivo: Determinar la capacidad del extracto acuoso D. loretense (jergon sacha para neutralizar la actividad letal del veneno de la serpiente B. atrox. Materiales y metodos: Se realizo el screening fitoquimico del extracto acuoso del bulbo desecado de D. loretense, se calculo la dosis letal 50(DL50 para el extracto y el veneno de B. atrox. Luego se realizaron enfrentamientos de diferentes dosis de extracto y veneno inyectadas intraperitonealmente en ratones, previa incubacion, para calcular la dosis eficaz 50% (DE50 del extracto para neutralizar el efecto letal del veneno por el metodo de Probits. Resultados: El analisis fitoquimico del extracto permitio identificar compuestos fenolicos, taninos, saponinas, proteinas, terpenoides y esteroides. No sobrevivio ningun raton que recibio el veneno sin D. loretense, y no fallecio ninguno al que se le dio solo el extracto acuoso de D. loretense. Se determino la DE50 en 91,15 µg/raton del extracto para neutralizar 2DL50 del veneno. Se encontro que a mayor dosis del veneno de B.atrox se necesito menores dosis del extracto acuoso de D. loretense. Conclusiones: el extracto acuoso de D. loretense neutraliza la actividad letal de veneno de B. atrox. Es necesario realizar estudios que permitan identificar los metabolitos del extracto que tienen esta accion.

  6. Presynaptic Neurotoxins: Biochemistry, Molecular Biology, Immunology and Other Exploratory Studies

    Science.gov (United States)

    1994-04-01

    analysis (Bieber). Primary sequences have also been determined for two myotoxins from the venom of Bothrops asper . One, a potent PLA2 with 67% sequence...fieldi, myotoxin II from Bothrops asper , PLA 2 from C. atrox) as opposed to monomeric PLA 2s. Once our MacQuadra 800 computer becomes available, we...on the isolation of Bothrops asper phospholipase A2s, we observed that these venoms also have metal-ion dependent proteases and high concentrations of

  7. A Lys49 phospholipase A2, isolated from Bothrops asper snake venom, induces lipid droplet formation in macrophages which depends on distinct signaling pathways and the C-terminal region.

    Science.gov (United States)

    Giannotti, Karina Cristina; Leiguez, Elbio; Moreira, Vanessa; Nascimento, Neide Galvão; Lomonte, Bruno; Gutiérrez, José Maria; Lopes de Melo, Robson; Teixeira, Catarina

    2013-01-01

    MT-II, a Lys49PLA2 homologue devoid of catalytic activity from B. asper venom, stimulates inflammatory events in macrophages. We investigated the ability of MT-II to induce formation of lipid droplets (LDs), key elements of inflammatory responses, in isolated macrophages and participation of protein kinases and intracellular PLA2s in this effect. Influence of MT-II on PLIN2 recruitment and expression was assessed, and the effects of some synthetic peptides on LD formation were further evaluated. At noncytotoxic concentrations, MT-II directly activated macrophages to form LDs. This effect was reproduced by a synthetic peptide corresponding to the C-terminal sequence 115-129 of MT-II, evidencing the critical role of C-terminus for MT-II-induced effect. Moreover, MT-II induced expression and recruitment of PLIN2. Pharmacological interventions with specific inhibitors showed that PKC, PI3K, ERK1/2, and iPLA2, but not P38(MAPK) or cPLA2, signaling pathways are involved in LD formation induced by MT-II. This sPLA2 homologue also induced synthesis of PGE2 that colocalized to LDs. In conclusion, MT-II is able to induce formation of LDs committed to PGE2 formation in a process dependent on C-terminal loop engagement and regulated by distinct protein kinases and iPLA2. LDs may constitute an important inflammatory mechanism triggered by MT-II in macrophages.

  8. A Lys49 Phospholipase A2, Isolated from Bothrops asper Snake Venom, Induces Lipid Droplet Formation in Macrophages Which Depends on Distinct Signaling Pathways and the C-Terminal Region

    Directory of Open Access Journals (Sweden)

    Karina Cristina Giannotti

    2013-01-01

    Full Text Available MT-II, a Lys49PLA2 homologue devoid of catalytic activity from B. asper venom, stimulates inflammatory events in macrophages. We investigated the ability of MT-II to induce formation of lipid droplets (LDs, key elements of inflammatory responses, in isolated macrophages and participation of protein kinases and intracellular PLA2s in this effect. Influence of MT-II on PLIN2 recruitment and expression was assessed, and the effects of some synthetic peptides on LD formation were further evaluated. At noncytotoxic concentrations, MT-II directly activated macrophages to form LDs. This effect was reproduced by a synthetic peptide corresponding to the C-terminal sequence 115–129 of MT-II, evidencing the critical role of C-terminus for MT-II-induced effect. Moreover, MT-II induced expression and recruitment of PLIN2. Pharmacological interventions with specific inhibitors showed that PKC, PI3K, ERK1/2, and iPLA2, but not P38MAPK or cPLA2, signaling pathways are involved in LD formation induced by MT-II. This sPLA2 homologue also induced synthesis of PGE2 that colocalized to LDs. In conclusion, MT-II is able to induce formation of LDs committed to PGE2 formation in a process dependent on C-terminal loop engagement and regulated by distinct protein kinases and iPLA2. LDs may constitute an important inflammatory mechanism triggered by MT-II in macrophages.

  9. Aspectos citoquímicos e morfológicos de elementos sanguíneos das serpentes dos gêneros Bothrops e Crotalus mantidas em cativeiro no serpentário do Instituto Vital Brasil

    Directory of Open Access Journals (Sweden)

    L.M. Kindlovits

    Full Text Available RESUMO A criação de serpentes peçonhentas em cativeiro vem se tornando prática cada vez mais difundida no país. Dessa forma, o conhecimento do manejo e da clínica de serpentes se torna prioritário, a fim de permitir maior sobrevida dos animais. No que concerne a serpentes peçonhentas, dados hematológicos já foram descritos na literatura, no entanto, apesar dos recursos utilizados, os dados analisados ainda são insipientes. Com o objetivo de caracterizar, morfologicamente, as células sanguíneas e de esclarecer diferenças morfológicas e funcionais, foram coletadas amostras de sangue de 50 serpentes pertencentes ao plantel do Instituto Vital Brasil. Foram confeccionados e analisados por microscopia óptica e citoquimicamente os esfregaços sanguíneos corados por métodos de Romanowsky e citoquímicos. Foi possível diferenciar as células, caracterizar e confirmar a existência de eosinófilo em B. atrox e C. durissus. Concluiu-se que a caracterização celular pode fornecer evidências indispensáveis ao entendimento da fisiologia de serpentes.

  10. Características bioquímicas y evaluación preclínica de un antiveneno botrópico liofilizado contra el veneno de la serpiente Bothrops atrox

    Directory of Open Access Journals (Sweden)

    Patricia J. García

    2008-10-01

    Full Text Available Se han estudiado las características bioquímicas y la capacidad neutralizante del antiveneno botrópico liofilizado producido por el Instituto Nacional de Salud (Lima, Perú, se encontró que posee 51,4 mg/mL de proteínas, las preparaciones liofilizadas se reconstituyen en un periodo de 10 min alcanzando valores de Abs600nm y pH de 0,091 y 7,0, respectivamente. Para el caso de las actividades tóxicas del veneno en estudio se obtuvieron valores de toxicidad DL50: 3,33 μg/g ratσn, dosis hemorrαgica mínima: 4,10 ± 0,64 μg, dosis miotóxica mínima 30,2 ± 2,5 μg, dosis coagulante mνnima: 4,50 ± 0,6 μg y dosis defibrinante mνnima: 8 μg, y valores de dosis efectiva del antiveneno evaluado de 140,48 (120,09-164,33, 230,67 ±11,78, 316,56 ± 40,31, 105,5 ± 4,2 y 500 μL antiveneno/mg veneno, respectivamente, lo cual indica que posee capacidad para neutralizar tales parαmetros. Por estas razones se concluye que el producto biológico investigado cumple con los requerimientos de la Organización Mundial de la Salud (OMS para ser considerado un antiveneno neutralizante de las principales actividades biológicas antes señaladas.

  11. Development of a sensitive enzyme immunoassay (ELISA for specific identification of Lachesis acrochorda venom

    Directory of Open Access Journals (Sweden)

    V Núñez Rangel

    2012-01-01

    Full Text Available The snake genus Lachesis provokes 2 to 3% of snakebites in Colombia every year. Two Lachesis species, L. acrochorda and L. muta, share habitats with snakes from another genus, namely Bothrops asper and B. atrox. Lachesis venom causes systemic and local effects such as swelling, hemorrhaging, myonecrosis, hemostatic disorders and nephrotoxic symptoms similar to those induced by Bothrops, Portidium and Bothriechis bites. Bothrops antivenoms neutralize a variety of Lachesis venom toxins. However, these products are unable to avoid coagulation problems provoked by Lachesis snakebites. Thus, it is important to ascertain whether the envenomation was caused by a Bothrops or Lachesis snake. The present study found enzyme linked immunosorbent assay (ELISA efficient for detecting Lachesis acrochorda venom in a concentration range of 3.9 to 1000 ng/mL, which did not show a cross-reaction with Bothrops, Portidium, Botriechis and Crotalus venoms. Furthermore, one fraction of L. acrochorda venom that did not show crossreactivity with B. asper venom was isolated using the same ELISA antibodies; some of its proteins were identified including one Gal-specific lectin and one metalloproteinase. This test may be useful to physicians, since it could be applicable for tracking the kinetic distribution of antigens in patients or experimentally envenomed animals.

  12. Management of venomous snakebites in dogs and cats in Brazil

    Directory of Open Access Journals (Sweden)

    R.S. Ferreira Júnior

    2004-01-01

    Full Text Available Snake envenoming is a major problem both to veterinary and human medicine in tropical countries due to high incidence, severity, and sequelae. In Brazil, most envenomings involving animals are caused by Bothrops and Crotalus snakes; these are the highest risk to animals. This study reports on Bothrops and Crotalus envenomings in dogs, the main species responsible for epidemiology, pathogenesis, venom action, clinical signs, sequelae and complications, clinical pathology, necropsy findings, diagnosis, and treatment. Veterinarians must be capable of identifying the snake not only by observing its characteristics but also symptom evolution.

  13. Canopy Venom: Proteomic Comparison among New World Arboreal Pit-Viper Venoms.

    Science.gov (United States)

    Debono, Jordan; Cochran, Chip; Kuruppu, Sanjaya; Nouwens, Amanda; Rajapakse, Niwanthi W; Kawasaki, Minami; Wood, Kelly; Dobson, James; Baumann, Kate; Jouiaei, Mahdokht; Jackson, Timothy N W; Koludarov, Ivan; Low, Dolyce; Ali, Syed A; Smith, A Ian; Barnes, Andrew; Fry, Bryan G

    2016-07-08

    Central and South American pitvipers, belonging to the genera Bothrops and Bothriechis, have independently evolved arboreal tendencies. Little is known regarding the composition and activity of their venoms. In order to close this knowledge gap, venom proteomics and toxin activity of species of Bothriechis, and Bothrops (including Bothriopsis) were investigated through established analytical methods. A combination of proteomics and bioactivity techniques was used to demonstrate a similar diversification of venom composition between large and small species within Bothriechis and Bothriopsis. Increasing our understanding of the evolution of complex venom cocktails may facilitate future biodiscoveries.

  14. Canopy Venom: Proteomic Comparison among New World Arboreal Pit-Viper Venoms

    Directory of Open Access Journals (Sweden)

    Jordan Debono

    2016-07-01

    Full Text Available Central and South American pitvipers, belonging to the genera Bothrops and Bothriechis, have independently evolved arboreal tendencies. Little is known regarding the composition and activity of their venoms. In order to close this knowledge gap, venom proteomics and toxin activity of species of Bothriechis, and Bothrops (including Bothriopsis were investigated through established analytical methods. A combination of proteomics and bioactivity techniques was used to demonstrate a similar diversification of venom composition between large and small species within Bothriechis and Bothriopsis. Increasing our understanding of the evolution of complex venom cocktails may facilitate future biodiscoveries.

  15. Estudos cristalográficos em macromoléculas biológicas: Aplicações em calgranulina C de granulócitos porcinos, tripanotiona redutase deTrypanosoma cruzi e fosfolipase A2 extraída do veneno da serpente Bothrops moojeni.

    OpenAIRE

    Maria Cristina Nonato Costa

    1997-01-01

    A cristalografia de raios-X e um método de singular importância para a determinação da estrutura de macromoléculas. A importância em resolver estruturas de proteínas continua a crescer em campos variando desde a bioquímica e biofísica básicas ate o desenvolvimento farmacêutico e biotecnologia. o presente trabalho esta relacionado com os estudos cristalográficos de três diferentes moléculas biológicas. Calgranulina C de granulócitos porcinos, uma proteína que liga cálcio, supostamente envolvid...

  16. Screening for Proteolytic Activities in Snake Venom by Means of a Multiplexing ESI-MS Assay Scheme

    NARCIS (Netherlands)

    Liesener, A.; Perchuc, Anna-Maria; Schöni, Reto; Wilmer, Marianne; Karst, U.

    2005-01-01

    A multiplexed mass spectrometry based assay scheme for the simultaneous determination of five different substrate/product pairs was developed as a tool for screening of proteolytic activities in snake venom fractions from Bothrops moojeni. The assay scheme was employed in the functional

  17. Selection of ligand peptides with the ability to detect antibodies in ...

    African Journals Online (AJOL)

    enoh

    2012-04-05

    Apr 5, 2012 ... infection include restrictions on the trading of the animal or its products such ... immunize animals. Some of them are capable of inducing new antibodies that present cross-reaction with the natural epitopes which are considered as mimetic immunogenic ...... Bothrops neuwiedi snake venom. Toxicon, 53(2): ...

  18. Blood Evaluation Of Cl and Na Concentration In Crioulo Breed Horses Using NAA: Comparison With Humans Levels

    International Nuclear Information System (INIS)

    Baptista, Tatyana S.; Zamboni, Cibele B.; Medeiros, Jose Agostinho G. de; Marcelino, Jose R.; Higashi, Hisako G.; Freitas, Monica G.

    2009-01-01

    Neutron Activation Analysis was utilized for determining the concentration of chlorine and sodium in blood of Crioulo breed horses used for hyperimmune sera production (Bothrops, Diphtheria and Tetanus) at Butantan Institute (Sao Paulo city, Brasil). These data are an important support for a toxicological control of adverse reactions in patients who will receive the hyperimmune serum.

  19. Cases of human envenoming caused by Philodryas olfersii and Philodryas patagoniensis (Serpentes: Colubridae).

    Science.gov (United States)

    de Araújo, M E; dos Santos, A C

    1997-01-01

    The present paper reports two cases of human envenoming by colubrid snakes of Philodryas, considered as not poisonous, showing evidence of the clinical aspects and the evolution of the symptoms of envenoming. The similarity of these cases with those caused by Bothrops suggests a more careful evaluation on the victims considering the medical treatment to be adopted.

  20. A novel synthetic quinolinone inhibitor presents proteolytic and hemorrhagic inhibitory activities against snake venom metalloproteases.

    Science.gov (United States)

    Baraldi, Patrícia T; Magro, Angelo J; Matioli, Fábio F; Marcussi, Silvana; Lemke, Ney; Calderon, Leonardo A; Stábeli, Rodrigo G; Soares, Andreimar M; Correa, Arlene G; Fontes, Marcos R M

    2016-02-01

    Metalloproteases play a fundamental role in snake venom envenomation inducing hemorrhagic, fibrigen(ogen)olytic and myotoxic effects in their victims. Several snake venoms, such as those from the Bothrops genus, present important local effects which are not efficiently neutralized by conventional serum therapy. Consequently, these accidents may result in permanent sequelae and disability, creating economic and social problems, especially in developing countries, leading the attention of the World Health Organization that considered ophidic envenomations a neglected tropical disease. Aiming to produce an efficient inhibitor against bothropic venoms, we synthesized different molecules classified as quinolinones - a group of low-toxic chemical compounds widely used as antibacterial and antimycobacterial drugs - and tested their inhibitory properties against hemorrhage caused by bothropic venoms. The results from this initial screening indicated the molecule 2-hydroxymethyl-6-methoxy-1,4-dihydro-4-quinolinone (Q8) was the most effective antihemorrhagic compound among all of the assayed synthetic quinolinones. Other in vitro and in vivo experiments showed this novel compound was able to inhibit significantly the hemorrhagic and/or proteolytic activities of bothropic crude venoms and isolated snake venom metalloproteases (SVMPs) even at lower concentrations. Docking and molecular dynamic simulations were also performed to get insights into the structural basis of Q8 inhibitory mechanism against proteolytic and hemorrhagic SVMPs. These structural studies demonstrated that Q8 may form a stable complex with SVMPs, impairing the access of substrates to the active sites of these toxins. Therefore, both experimental and structural data indicate that Q8 compound is an interesting candidate for antiophidic therapy, particularly for the treatment of the hemorrhagic and necrotic effects induced by bothropic venoms. Copyright © 2015 Elsevier B.V. and Société Française de

  1. Serpentes da área urbana de Cuiabá, Mato Grosso: aspectos ecológicos e acidentes ofídicos associados Snakes from the urban area of Cuiabá, Mato Grosso: ecological aspects and associated snakebites

    Directory of Open Access Journals (Sweden)

    Marcos André de Carvalho

    1998-10-01

    Full Text Available São apresentadas informações sobre as serpentes da área urbana do Município de Cuiabá, Mato Grosso, potencialmente causadoras de acidentes ofídicos. As informações estão baseadas em exemplares doados por populares ao Núcleo de Ofiologia Regional de Mato Grosso (Normat entre 1986 e 1993 e em registros efetuados pelo Centro de Informações Anti-Veneno (Ciave entre 1988 e 1993, que não fazem referência ao animal causador. Foram catalogadas 37 espécies de serpentes, em 25 gêneros e 3 famílias, com hábitos predominantemente diurnos, terrestres e com dietas baseadas em anfíbios e/ou lagartos. Dentre os 307 acidentes ofídicos registrados, 56% foram causados por serpentes sem interesse médico e 44% constituíram acidentes de importância médica. Cerca de 99% dos acidentes de importância médica foram atribuídos ao gênero Bothrops (Bothrops moojeni e Bothrops neuwiedi seriam as principais causadoras. Dentre as espécies sem interesse médico, Philodryas olfersii e Waglerophis merremii provavelmente foram as principais responsáveis pela maior parte dos acidentes.This study presents data on snakes recorded in the urban area of Cuiabá, Mato Grosso, Brazil. Sources of information included specimens captured by local residents (1986-1993 and turned over to the Mato Grosso Regional Ophiological Center (Normat, and data from the Anti-Venom Information Center (Ciave, regarding urban snakebites (1988-1993. Thirty-seven species of snakes from 25 genera and three families were recorded. Diurnal and terrestrial habits predominated, as well as a diet based on amphibians and/or lizards. From a total of 307 snakebites recorded, some 56% were of no clinical importance, caused by non-venomous snakes, whereas 44% were clinically relevant. Approximately 99% of the latter were attributed to vipers of the genus Bothrops, and especially the Bothrops moojeni and Bothrops neuwiedi species The colubrids Philodryas olfersii and Waglerophis merremii were

  2. Cases of human envenoming caused by Philodryas olfersii and Philodryas patagoniensis (serpentes: Colubridae

    Directory of Open Access Journals (Sweden)

    Maria Elisabeth de Araújo

    1997-12-01

    Full Text Available The present paper reports two cases of human envenoming by colubrid snakes of Philodryas, considered as not poisonous, showing evidence of the clinical aspects and the evolution of the symptoms of envenoming. The similarity of these cases with those caused by Bothrops suggests a more careful evaluation on the victims considering the medical treatment to be adopted.O presente trabalho é um relato de dois casos de acidentes com colubrídeos (Philodryas olfersii e P. patagoniensis considerados não peçonhentos, que destaca as manifestações clínicas e as suas evoluções. A semelhança de tais acidentes com aqueles causados por serpentes Bothrops indica a necessidade de uma melhor avaliação dos pacientes quanto à terapêutica a ser adotada.

  3. P-I snake venom metalloproteinase is able to activate the complement system by direct cleavage of central components of the cascade.

    Directory of Open Access Journals (Sweden)

    Giselle Pidde-Queiroz

    Full Text Available BACKGROUND: Snake Venom Metalloproteinases (SVMPs are amongst the key enzymes that contribute to the high toxicity of snake venom. We have recently shown that snake venoms from the Bothrops genus activate the Complement system (C by promoting direct cleavage of C-components and generating anaphylatoxins, thereby contributing to the pathology and spread of the venom. The aim of the present study was to isolate and characterize the C-activating protease from Bothrops pirajai venom. RESULTS: Using two gel-filtration chromatography steps, a metalloproteinase of 23 kDa that activates Complement was isolated from Bothrops pirajai venom. The mass spectrometric identification of this protein, named here as C-SVMP, revealed peptides that matched sequences from the P-I class of SVMPs. C-SVMP activated the alternative, classical and lectin C-pathways by cleaving the α-chain of C3, C4 and C5, thereby generating anaphylatoxins C3a, C4a and C5a. In vivo, C-SVMP induced consumption of murine complement components, most likely by activation of the pathways and/or by direct cleavage of C3, leading to a reduction of serum lytic activity. CONCLUSION: We show here that a P-I metalloproteinase from Bothrops pirajai snake venom activated the Complement system by direct cleavage of the central C-components, i.e., C3, C4 and C5, thereby generating biologically active fragments, such as anaphylatoxins, and by cleaving the C1-Inhibitor, which may affect Complement activation control. These results suggest that direct complement activation by SVMPs may play a role in the progression of symptoms that follow envenomation.

  4. Envenomation by the neotropical colubrid Boiruna maculata (Boulenger, 1896): a case report.

    Science.gov (United States)

    dos Santos-Costa, M C; Outeiral, A B; D'Agostini, F M; Cappellari, L H

    2000-01-01

    This is a case report of a Boiruna maculata snake bite in a child admitted to the Hospital Municipal de Pronto Socorro de Porto Alegre, Porto Alegre, RS, Brazil. The patient was bitten on the lower left limb, and exhibited pronounced local manifestations of envenomation. She was treated with Bothrops antivenom and was discharged from the hospital five days later with marked improvement of envenomation.

  5. P-I Snake Venom Metalloproteinase Is Able to Activate the Complement System by Direct Cleavage of Central Components of the Cascade

    Science.gov (United States)

    Pidde-Queiroz, Giselle; Magnoli, Fábio Carlos; Portaro, Fernanda C. V.; Serrano, Solange M. T.; Lopes, Aline Soriano; Paes Leme, Adriana Franco; van den Berg, Carmen W.; Tambourgi, Denise V.

    2013-01-01

    Background Snake Venom Metalloproteinases (SVMPs) are amongst the key enzymes that contribute to the high toxicity of snake venom. We have recently shown that snake venoms from the Bothrops genus activate the Complement system (C) by promoting direct cleavage of C-components and generating anaphylatoxins, thereby contributing to the pathology and spread of the venom. The aim of the present study was to isolate and characterize the C-activating protease from Bothrops pirajai venom. Results Using two gel-filtration chromatography steps, a metalloproteinase of 23 kDa that activates Complement was isolated from Bothrops pirajai venom. The mass spectrometric identification of this protein, named here as C-SVMP, revealed peptides that matched sequences from the P-I class of SVMPs. C-SVMP activated the alternative, classical and lectin C-pathways by cleaving the α-chain of C3, C4 and C5, thereby generating anaphylatoxins C3a, C4a and C5a. In vivo, C-SVMP induced consumption of murine complement components, most likely by activation of the pathways and/or by direct cleavage of C3, leading to a reduction of serum lytic activity. Conclusion We show here that a P-I metalloproteinase from Bothrops pirajai snake venom activated the Complement system by direct cleavage of the central C-components, i.e., C3, C4 and C5, thereby generating biologically active fragments, such as anaphylatoxins, and by cleaving the C1-Inhibitor, which may affect Complement activation control. These results suggest that direct complement activation by SVMPs may play a role in the progression of symptoms that follow envenomation. PMID:24205428

  6. Snake envenomations in northwest counties of the Rio de Janeiro State, Brazil

    Directory of Open Access Journals (Sweden)

    H. B. Tinoco

    2005-03-01

    Full Text Available The frequency of envenoming in Northwest counties of the Rio de Janeiro State, Brazil, from 1995 to 2000 was evaluated. Reports from the Municipal Secretariat of Health of these counties were used. The results demonstrated that, from 1997 to 1999, there was a shortage of notification, and 40 cases of envenomations caused by Bothrops snakes were registered. These cases were more common from February to October, and the lower limbs of male peasants were the mainly affected areas.

  7. Acción del antiveneno botrópico polivalente sobre las actividades proteolíticas presentes en los venenos de serpientes peruanas

    Directory of Open Access Journals (Sweden)

    Armando Yarlequé

    2008-04-01

    Full Text Available Los venenos de las serpientes peruanas causantes de la mayoría de accidentes ofídicos, contienen enzimas proteolíticas que pueden degradar proteínas tisulares y plasmáticas, así como causar hipotensión y coagulación sanguínea. Objetivos. Evaluar la capacidad inhibitoria del antiveneno botrópico polivalente al estado líquido producido por el Instituto Nacional de Salud del Perú (INS sobre las actividades caseinolítica, coagulante y amidolítica de los venenos de Bothrops atrox, Bothrops brazili, Bothrops pictus y Bothrops barnetti. Materiales y métodos. Se usaron en cada caso sustratos como caseína, fibrinógeno bovino y el cromógeno benzoil-arginil-p-nitroanilida (BApNA respectivamente, y se midieron los cambios en los valores de la actividad enzimática a ½, 1 y 2 dosis del antiveneno tanto al estado natural como calentado a 37 °C durante cinco días. Resultados. La actividad caseinolítica es la más resistente a la inhibición especialmente por el suero no calentado en tanto que, la actividad amidolítica fue severamente inhibida principalmente en los venenos de B. pictus y B. atrox. Así mismo la actividad coagulante fue totalmente inhibida en el veneno de B. pictus, mostrándose a su vez una elevada inhibición sobre los venenos de B. brazili y B. atrox. Para las actividades coagulante y amidolítica, los sueros calentados fueron menos efectivos que aquellos al estado natural. Conclusiones. El suero antibotrópico polivalente producido por el INS es efectivo para inhibir las actividades proteolíticas de los venenos de las serpientes peruanas ensayadas.

  8. Philodryas patagoniensis bite and local envenoming

    OpenAIRE

    Nishioka,Sérgio de Andrade; Silveira,Paulo Vitor Portella

    1994-01-01

    A 5-year-old boy bitten by a specimen of Philodryas patagoniensis, a colubrid snake currently classified as nonvenomous, developed signs of local envenoming characterized by swelling and warmth on the bitten limb. This is the first time that local envenoming following Philodryas patagoniensis bite is recognized. Based on the clinical findings and misidentification of the snake, the patient was treated as a victim of Bothrops bite, having received unnecessarily the specific antivenom. Educatio...

  9. Cases of human envenoming caused by Philodryas olfersii and Philodryas patagoniensis (serpentes: Colubridae)

    OpenAIRE

    Araújo,Maria Elisabeth de; Santos,Ana Cristina M.C.A. dos

    1997-01-01

    The present paper reports two cases of human envenoming by colubrid snakes of Philodryas, considered as not poisonous, showing evidence of the clinical aspects and the evolution of the symptoms of envenoming. The similarity of these cases with those caused by Bothrops suggests a more careful evaluation on the victims considering the medical treatment to be adopted.O presente trabalho é um relato de dois casos de acidentes com colubrídeos (Philodryas olfersii e P. patagoniensis) considerados n...

  10. Picada por Philodryas patagoniensis e envenenamento local

    OpenAIRE

    Nishioka, Sérgio de Andrade; Silveira, Paulo Vitor Portella

    1994-01-01

    A 5-year-old boy bitten by a specimen of Philodryas patagoniensis, a colubrid snake currently classified as nonvenomous, developed signs of local envenoming characterized by swelling and warmth on the bitten limb. This is the first time that local envenoming following Philodryas patagoniensis bite is recognized. Based on the clinical findings and misidentification of the snake, the patient was treated as a victim of Bothrops bite, having received unnecessarily the specific antivenom. Educatio...

  11. Crystallization and preliminary X-ray diffraction analysis of a Lys49-phospholipase A2 complexed with caffeic acid, a molecule with inhibitory properties against snake venoms

    International Nuclear Information System (INIS)

    Shimabuku, Patrícia S.; Fernandes, Carlos A. H.; Magro, Angelo J.; Costa, Tássia R.; Soares, Andreimar M.; Fontes, Marcos R. M.

    2011-01-01

    Piratoxin I, a noncatalytic and myotoxic Lys49-phospholipase A 2 from B. pirajai venom, was cocrystallized with the inhibitor caffeic acid and a data set was collected to a resolution of 1.65 Å. The electron-density map unambiguously indicated that three inhibitor molecules interact with the C-terminus of the protein. Phospholipases A 2 (PLA 2 s) are one of the main components of bothropic venoms; in addition to their phospholipid hydrolysis action, they are involved in a wide spectrum of pharmacological activities, including neurotoxicity, myotoxicity and cardiotoxicity. Caffeic acid is an inhibitor that is present in several plants and is employed for the treatment of ophidian envenomations in the folk medicine of many developing countries; as bothropic snake bites are not efficiently neutralized by conventional serum therapy, it may be useful as an antivenom. In this work, the cocrystallization and preliminary X-ray diffraction analysis of the Lys49-PLA 2 piratoxin I from Bothrops pirajai venom in the presence of the inhibitor caffeic acid (CA) are reported. The crystals diffracted X-rays to 1.65 Å resolution and the structure was solved by molecular-replacement techniques. The electron-density map unambiguously indicated the presence of three CA molecules that interact with the C-terminus of the protein. This is the first time a ligand has been observed bound to this region and is in agreement with various experiments previously reported in the literature

  12. Serpentes da área urbana de Cuiabá, Mato Grosso: aspectos ecológicos e acidentes ofídicos associados

    Directory of Open Access Journals (Sweden)

    Carvalho Marcos André de

    1998-01-01

    Full Text Available São apresentadas informações sobre as serpentes da área urbana do Município de Cuiabá, Mato Grosso, potencialmente causadoras de acidentes ofídicos. As informações estão baseadas em exemplares doados por populares ao Núcleo de Ofiologia Regional de Mato Grosso (Normat entre 1986 e 1993 e em registros efetuados pelo Centro de Informações Anti-Veneno (Ciave entre 1988 e 1993, que não fazem referência ao animal causador. Foram catalogadas 37 espécies de serpentes, em 25 gêneros e 3 famílias, com hábitos predominantemente diurnos, terrestres e com dietas baseadas em anfíbios e/ou lagartos. Dentre os 307 acidentes ofídicos registrados, 56% foram causados por serpentes sem interesse médico e 44% constituíram acidentes de importância médica. Cerca de 99% dos acidentes de importância médica foram atribuídos ao gênero Bothrops (Bothrops moojeni e Bothrops neuwiedi seriam as principais causadoras. Dentre as espécies sem interesse médico, Philodryas olfersii e Waglerophis merremii provavelmente foram as principais responsáveis pela maior parte dos acidentes.

  13. Isolation and characterization of ellagic acid derivatives isolated from Casearia sylvestris SW aqueous extract with anti-PLA(2) activity.

    Science.gov (United States)

    Da Silva, Saulo L; Calgarotto, Andrana K; Chaar, Jamal S; Marangoni, Sérgio

    2008-11-01

    The Casearia sylvestris SW (Flacourtiaceae) is utilized in folk medicine (Brazil and all Latin American) to treat several pathologic processes as inflammation, cancer, microbial infection and snake bites. Studies showed that C. sylvestris aqueous extract can inhibit many toxic effects caused by snake venoms (or caused by phospholipase A(2) isolated) from different species, mainly of Bothrops genus. Inhibition of enzymatic and myotoxic activities, decrease of edema formation and increase of the survival rate of rats injected with lethal doses of bothropic venoms are some toxic effects inhibited by C. sylvestris. In this study, four ellagic acid derivatives from aqueous extracts of C. sylvestris were isolated, characterized, and tested against effects from both total venom and PLA(2) (Asp 49 BthTX-II) from the venom of Bothrops jararacussu. The isolated compounds were as follows: ellagic acid (A), 3'-O-methyl ellagic acid (B), 3,3'-di-O-methyl ellagic acid (C), 3-O-methyl-3',4'-methylenedioxy ellagic acid (D). The inhibition constant values (Ki) for enzymatic activity, as well the IC(50) values found in the edematogenic and myotoxic activities, indicate that the ellagic acid is the best inhibitor of these activities, while compounds C and D are the substances with lowest capacity on inhibiting these same effects. Our results show that the presence of hydroxyls at position 3 or 3' (compounds A and B) increases the capacity of these derivatives on inhibiting these toxic effects. However, the presence of methoxyl groups at position 3 or 3' reduced, but did not completely inhibit the capacity of compounds C and D on inhibiting all the toxic effects studied.

  14. A snakebite caused by a bushmaster (Lachesis muta): report of a confirmed case in State of Pernambuco, Brazil.

    Science.gov (United States)

    Lima, Pedro Henrique Santana de; Haddad Junior, Vidal

    2015-01-01

    We report a case of envenomation caused by a bushmaster ( Lachesis muta) in a male child in State of Pernambuco, Brazil. The victim showed discrete local manifestations, but presented altered blood coagulation 2 hours after the bite. Ten ampoules of bothropic-lachetic antivenom therapy were administered, and 48 hours later, the patient showed discrete edema, pain, and ecchymosis around the bite and normal blood coagulation. The patient was discharged 5 days after the envenomation. The prompt administration of specific treatment was important for the favorable outcomes observed.

  15. Contribuição à biologia de serpentes da Bahia, Brasil: I. vivíparas Contribution to reproductive biology of snakes in Bahia, Brazil: I. viviparous

    Directory of Open Access Journals (Sweden)

    Rejane Maria Lira-da-Silva

    1994-01-01

    Full Text Available Great part of lhe avaiable data about snakes reprodution refers to species coming from subtropical and temperate regions. In Brazil, the data is rather rare and can be found in various works where information is restricted. Results from studies developed with five viviparous snakes - Crotalus durissus cascavella (Wagler, 1824. Bothrops erythromelas(Amaral, 1923, B. leucurus (Wagler, 1824, Helicops leopardinus (Schlegel, 1873 and Thamnodynastes strigilis (Thiinberg, 1787 - which come from the Northeast of Brazil (Bahia are described. Data about pregnancy and birth, number, sex ratio, length and weight of neonates is given and discussed.

  16. Inorganic elements in blood of mice immunized with snake venom using NAA and XRF techniques

    International Nuclear Information System (INIS)

    Metairon, S.; Zamboni, C.B.; Suzuki, M.F.; Lopes da Silva, L.F.F.; Rizzutto, M.A.

    2016-01-01

    Brazil has the greatest diversity of snakes in the world and a large portion of them are venomous. Nowadays, Instituto Butantan (research center, at Brazil) produces various types of antivenom to meet the large number of incidences. In this investigation, mice were immunized with different species of Bothrops snake venom to evaluate the inorganic elements concentration in their blood by using NAA and XRF techniques. The results were compared with the control group (mice not immunized) and with human estimative. The data allows to evaluate the toxicity of these elements, important for clinical screening of patients submitted to immunological therapy. (author)

  17. Efficacy of IgG and F(ab′)2 Antivenoms to Neutralize Snake Venom-induced Local Tissue Damage as Assessed by the Proteomic Analysis of Wound Exudate

    OpenAIRE

    Rucavado, Alexandra; Escalante Muñoz, Teresa; Shannon, John D.; Ayala Castro, Carla N.; Villalta, Mauren; Gutiérrez, José María; Fox, Jay W.

    2012-01-01

    2082-01 Embargo por política editorial Proteomic analysis of wound exudates represents a valuable tool to investigate tissue pathology and to assess the therapeutic success of various interventions. In this study, the ability of horse-derived IgG and F(ab0)2 antivenoms to neutralize local pathological effects induced by the venom of the snake Bothrops asper in mouse muscle was investigated by the proteomic analysis of exudates collected in the vicinity of affected tissue. In experiments...

  18. A snakebite caused by a bushmaster (Lachesis muta: report of a confirmed case in State of Pernambuco, Brazil

    Directory of Open Access Journals (Sweden)

    Pedro Henrique Santana de Lima

    2015-10-01

    Full Text Available ABSTRACTWe report a case of envenomation caused by a bushmaster ( Lachesis muta in a male child in State of Pernambuco, Brazil. The victim showed discrete local manifestations, but presented altered blood coagulation 2 hours after the bite. Ten ampoules of bothropic-lachetic antivenom therapy were administered, and 48 hours later, the patient showed discrete edema, pain, and ecchymosis around the bite and normal blood coagulation. The patient was discharged 5 days after the envenomation. The prompt administration of specific treatment was important for the favorable outcomes observed.

  19. Analysis of Cl and Na in Hyperimmune Sera by NAA

    International Nuclear Information System (INIS)

    Baptista, T. S.; Zamboni, C. B.; Marcelino, J. R.

    2011-01-01

    The Cl and Na concentration values in four types of hyperimmune sera (anti-Bothrops, anti-Diphtheria, anti-Rabies and anti-Tetanus) used for immunological therapy were determined using Neutron Activation Analysis (NAA). These data were compatible with the specifications established by the Word Health Organization (WHO-OMS) and with the Brazilian Official Pharmacopea (Pharmaceutical Code Official of the Country). These data are an important support for quality control of hyperimmune sera production at Butantan Institute (Sao Paulo city, Brazil), responsible for supplying the Brazilian market.

  20. Analysis of biological activities from Philodryas olfersii (Lichtenstein) and P. patagoniensis (Girard) venoms (Serpents, Colubridae)

    OpenAIRE

    Rocha, Marisa M. T. da; Furtado, Maria de F. D.

    2007-01-01

    Philodryas olfersii (Lichtenstein, 1823) e P.patagonienis (Girard, 1857) são serpentes colubrídeas da série opistóglifa, restritas à América do Sul. Vários acidentes ocasionados por estas serpentes têm sido relatados, caracterizando-se por ação local importante: dor, edema e hemorragia. É um acidente muito semelhante àquele causado por serpentes do gênero Bothrops Wagler, 1824 e muitas vezes os pacientes são tratados com soro antibotrópico. Poucos estudos tratam da caracterização destes venen...

  1. Aspects of the immune response against proteic antigens submitted to the effects of 60Co gamma radiation

    International Nuclear Information System (INIS)

    Alves, Janaina Baptista.

    2004-01-01

    Considering the effects of gamma radiation on proteins and the capacity of the immune system to recognize modified macromolecules, we decided to evaluate some immunological aspects of B10.PL mice exposed to native or irradiated ovalbumin and bothrops toxin-1 (BthTx-1). In order to evaluate possible structural modifications of the molecules after being irradiated ( 60 Co gamma rays), bothrops toxin-1 was analysed by electrophoresis, while ovalbumin was submitted to analytical size exclusion chromatography. The toxin was also analysed by ESI-mass spectrometry. Our results indicate that radiation promoted modifications on both the molecules. Aiming to compare the toxicity of the native and irradiated forms of the toxin, an in vitro cytotoxicity assay, using CHO cells, was performed. According to our results, the modified toxin was 5 folds less toxic than its native counterpart. Sera of animals immunized with the native and irradiated proteins were analyzed in order to evaluate levels of IgG, as well as to quantify specific isotopes. While the native proteins induced a predominant Th2 response, the irradiated molecules apparently promoted a switch towards a Th1 pattern. We also performed a cell proliferation assay with splenocytes from mice immunized with either the native or the irradiated proteins, cultured in the presence of the antigens. Our results indicate that both the forms of the proteins induced a similar proliferative response. These data indicate a potential use of detoxified proteins as antigens for immunization. (author)

  2. Occurrence of Microcerella halli (Engel (Diptera, Sarcophagidae in snake carrion in southeastern Brazil Ocorrência de Microcerella halli (Engel (Diptera, Sarcophagidae em uma Carcaça de Cobra no Sudeste Brasileiro

    Directory of Open Access Journals (Sweden)

    Thiago de C. Moretti

    2009-06-01

    Full Text Available The occurrence of 27 second-instar larvae of the flesh fly Microcerella halli (Engel, 1931 (Diptera, Sarcophagidae in a carcass of a snake usually called as Urutu, Bothrops alternatus (Duméril, Bibron & Duméril, 1854 (Serpentes, Viperidae, Crotalinae is reported. The snake was kept in captivity in a snake farm in Morungaba, São Paulo state, Brazil. Descriptions of reptile carcass colonization by insects and general biological data of this flesh fly are scarce and this necrophagic behavior is described for the first time in literature.A ocorrência de 27 larvas de segundo estádio do sarcofagídeo Microcerella halli (Engel, 1931 (Diptera, Sarcophagidae em uma carcaça de urutu Bothrops alternatus (Duméril, Bibron & Duméril, 1854 (Serpentes, Viperidae, Crotalinae é relatada. A cobra era mantida em cativeiro em um serpentário no município de Morungaba, estado de São Paulo, Brasil. Descrições de colonização de carcaças de répteis por insetos e dados gerais da biologia deste sarcofagídeo são escassos, e este comportamento necrófago é descrito pela primeira vez na literatura.

  3. Social Perception and Encounters with Snakes in Costa Rica: An Analysis through the Social Network Facebook

    Directory of Open Access Journals (Sweden)

    Katherine Sánchez-Paniagua

    2017-12-01

    Full Text Available Snakes are among the animals most affected by the ignorance of humans towards wildlife In Costa Rica, little effort has been made to know how snakes are affected by human actions. In the present study, we used information supplied to the Serpientes de Costa Rica page of the social network Facebook to find out: which species are reported most frequently? Which of these suffer the greatest number of deaths? and what ability to recognize them have those who send the reports? A total of 484 reports were obtained, 389 of which were non —venomous snakes and 95 were venomous, with 66 species being identified. The most frequent species were Ninia maculata, Senticolis triaspis, Leptodeira rhombifera, Boa imperator, Bothrops asper, Bothriechis schlegelii and Mastigodryas melanolomus. The genera with higher mortality were Mastigodryas, Bothrops, Senticolis, Geophis and Leptodeira. People killed one in four snakes, regardless of whether it was venomous or not. Species of the genera Mastigodryas, Geophis, Senticolis, Ninia and Leptodeira were among the least identified. The best identified species were B. imperator, B. asper and B. schlegelii. Only one in five people could correctly recognize whether a snake was poisonous or not. The reports were given with a small group of species, which seem to be the ones that are in closest contact with the studied population; education for the conservation of these animals can start with those species that seem to be more frequent and so people will gradually recognize and protect the wild life that surrounds them.

  4. Antibothropic action of Casearia sylvestris Sw. (Flacourtiaceae) extracts.

    Science.gov (United States)

    Cintra-Francischinelli, M; Silva, M G; Andréo-Filho, N; Gerenutti, M; Cintra, A C O; Giglio, J R; Leite, G B; Cruz-Höfling, M A; Rodrigues-Simioni, L; Oshima-Franco, Y

    2008-06-01

    Casearia sylvestris Sw., popularly known in Brazil as 'guaçatonga', has been used as antitumor, antiseptic, antiulcer, local anaesthetic and healer in folk medicine. Snakebite envenomation by Bothrops jararacussu (Bjssu) constitutes a relevant public health hazard capable of inducing serious local damage in victims. This study examined the pharmacological action of apolar and polar C. sylvestris leaf extracts in reverting the neuromuscular blockade and myonecrosis, which is induced by Bjssu venom and its major toxin bothropstoxin-I on the mouse phrenic nerve-diaphragm preparations. The polar methanol extract (ME) was by far the most efficacious. ME not only prevented myonecrosis and abolished the blockade, but also increased ACh release. Such facilitation in neuromuscular transmission was observed with ME alone, but was accentuated in preparations incubated with ME plus venom or toxin. This established synergy opens an interesting point of investigation because the venom or toxin in contact with ME changes from a blocking to a facilitating effect. It is suggested that rutin, known to have potent antioxidant properties, and one of the components present in the ME, could have a role in the observed effects. Since commercial rutin did not reproduce the ME effects, it is likely that a rutin-containing phytocomplex is neutralizing the bothropic envenoming effects. Copyright (c) 2008 John Wiley & Sons, Ltd.

  5. Evaluación comparativa de dos métodos para determinar la actividad de fosfolipasa A en venenos de serpientes

    Directory of Open Access Journals (Sweden)

    Fanny Lazo

    2014-06-01

    Full Text Available Se ha evaluado la actividad de fosfolipasa A en seis venenos de serpientes comparando las ventajas de un método espectrofotométrico, previamente adaptado a nuestras condiciones de laboratorio, frente al método macroscópico de retardo en la coagulación de una emulsión lipoproteica. En ambos casos, la actividad de fosfolipasa A estuvo presente en los venenos de Micrurus spixii, Crotalus durissus, Bothrops brazili, Lachesis muta y Bothrops atrox y la actividad decreció en orden. En cambio, la actividad de la enzima en el veneno de Micrurus surinamensis sólo fue detectada por el método espectrofotométrico. Así mismo, los resultados basados en la cantidad de veneno utilizado para medir la actividad, mostraron una mayor sensibilidad con el método espectrofotométrico en comparación con el método macroscópico ya que se requirieron cantidades menores en el orden de 2 a 10 veces.

  6. Disseminated intravascular coagulation caused by moojenactivase, a procoagulant snake venom metalloprotease.

    Science.gov (United States)

    Sartim, Marco A; Cezarette, Gabriel N; Jacob-Ferreira, Anna L; Frantz, Fabiani G; Faccioli, Lucia H; Sampaio, Suely V

    2017-10-01

    Snake venom toxins that activate coagulation factors are key players in the process of venom-induced coagulopathy, and account for severe clinical manifestations. The present study applies a variety of biochemical, hematological, and histopathological approaches to broadly investigate the intravascular and systemic effects of moojenactivase (MooA), the first described PIIId subclass metalloprotease isolated from Bothrops sp. venom that activates coagulation factors. MooA induced consumption coagulopathy with high toxic potency, characterized by prolongation of prothrombin and activated partial thromboplastin time, consumption of fibrinogen and the plasma coagulation factors X and II, and thrombocytopenia. MooA promoted leukocytosis and expression of the proinflammatory cytokines interleukin-6 and tumor necrosis factor-α, accompanied by tissue factor-dependent procoagulant activity in peripheral blood mononuclear cells. This metalloprotease also caused intravascular hemolysis, elevated plasma levels of creatine kinase-MB, aspartate transaminase, and urea/creatinine, and induced morphopathological alterations in erythrocytes, heart, kidney, and lungs associated with thrombosis and hemorrhage. Diagnosis of MooA-induced disseminated intravascular coagulation represents an important approach to better understand the pathophysiology of Bothrops envenomation and develop novel therapeutic strategies targeting hemostatic disturbances. Copyright © 2017. Published by Elsevier B.V.

  7. Unraveling the distinctive features of hemorrhagic and non-hemorrhagic snake venom metalloproteinases using molecular simulations

    Science.gov (United States)

    de Souza, Raoni Almeida; Díaz, Natalia; Nagem, Ronaldo Alves Pinto; Ferreira, Rafaela Salgado; Suárez, Dimas

    2016-01-01

    Snake venom metalloproteinases are important toxins that play fundamental roles during envenomation. They share a structurally similar catalytic domain, but with diverse hemorrhagic capabilities. To understand the structural basis for this difference, we build and compare two dynamical models, one for the hemorrhagic atroxlysin-I from Bothrops atrox and the other for the non-hemorraghic leucurolysin-a from Bothrops leucurus. The analysis of the extended molecular dynamics simulations shows some changes in the local structure, flexibility and surface determinants that can contribute to explain the different hemorrhagic activity of the two enzymes. In agreement with previous results, the long Ω-loop (from residue 149 to 177) has a larger mobility in the hemorrhagic protein. In addition, we find some potentially-relevant differences at the base of the S1' pocket, what may be interesting for the structure-based design of new anti-venom agents. However, the sharpest differences in the computational models of atroxlysin-I and leucurolysin-a are observed in the surface electrostatic potential around the active site region, suggesting thus that the hemorrhagic versus non-hemorrhagic activity is probably determined by protein surface determinants.

  8. Comparative study of anticoagulant and procoagulant properties of 28 snake venoms from families Elapidae, Viperidae, and purified Russell's viper venom-factor X activator (RVV-X).

    Science.gov (United States)

    Suntravat, Montamas; Nuchprayoon, Issarang; Pérez, John C

    2010-09-15

    Snake venoms consist of numerous molecules with diverse biological functions used for capturing prey. Each component of venom has a specific target, and alters the biological function of its target. Once these molecules are identified, characterized, and cloned; they could have medical applications. The activated clotting time (ACT) and clot rate were used for screening procoagulant and anticoagulant properties of 28 snake venoms. Crude venoms from Daboia russellii siamensis, Bothrops asper, Bothrops moojeni, and one Crotalus oreganus helleri from Wrightwood, CA, had procoagulant activity. These venoms induced a significant shortening of the ACT and showed a significant increase in the clot rate when compared to the negative control. Factor X activator activity was also measured in 28 venoms, and D. r. siamensis venom was 5-6 times higher than those of B. asper, B. moojeni, and C. o. helleri from Wrightwood County. Russell's viper venom-factor X activator (RVV-X) was purified from D. r. siamensis venom, and then procoagulant activity was evaluated by the ACT and clot rate. Other venoms, Crotalus atrox and two Naja pallida, had anticoagulant activity. A significant increase in the ACT and a significant decrease in the clot rate were observed after the addition of these venoms; therefore, the venoms were considered to have anticoagulant activity. Venoms from the same species did not always have the same ACT and clot rate profiles, but the profiles were an excellent way to identify procoagulant and anticoagulant activities in snake venoms.

  9. Ethnobotanic study of Randia aculeata (Rubiaceae in Jamapa, Veracruz, Mexico, and its anti-snake venom effects on mouse tissue

    Directory of Open Access Journals (Sweden)

    CA Gallardo-Casas

    2012-01-01

    Full Text Available In Mexico, medicinal plants are widely used. The use of Randia aculeata by healers against snakebites has never been scientifically tested in relation to possible effects on blood parameters and muscle tissue damage. Interviews were carried out in Jamapa, Veracuz, Mexico, with local residents to collect information about the traditional use of Randia aculeata. In this locality, seven pieces of fruit from the plant are mixed in a liter of alcohol, and then administered orally against snakebites. By using histological techniques and a murine model, we explored its cytoprotective properties against the effects of Crotalus simus and Bothrops asper venoms. Possible protections provided by the plant against tissue damage to skeletal and cardiac muscles and against the typical loss of red blood cells were analyzed. Randia aculeata caused an increase in microhematocrit and total hemoglobin, parameters that are often decremented in association with the loss of red blood cells, which is a characteristic effect of animal venom. Randia aculeata was also shown to protect against the lowering of platelet levels caused by Bothrops asper venom. Finally, Randia aculeata produced a partial inhibition of necrosis following administration of snake venom in skeletal and myocardial muscles. The present results provide solid evidence for the traditional use of Randia aculeata against snakebites, as demonstrated by protection against muscular tissue damage and the diminution of red blood cells.

  10. A importância dos acidentes ofídicos como causa de mortes em bovinos no Brasil The importance of snake bites as cause of cattle death in Brazil

    Directory of Open Access Journals (Sweden)

    Carlos Hubinger Tokarnia

    2006-06-01

    Full Text Available A revisão da literatura pertinente indica que as opiniões sobre a importância dos acidentes ofídicos, como causa de mortes em bovinos no Brasil, são divergentes no meio veterinário. Enquanto alguns acreditam que são pouco importantes, ou que têm menor significado do que lhes é atribuído, outros são da opinião que esses acidentes são freqüentes. Verificou-se que só foi relatado diagnóstico fundamentado de dois casos fatais de envenenamento por Bothrops spp em bovinos, e de nenhum por Crotalus spp. Um questionário por nós submetido a patologistas e clínicos veterinários que atuam em diversos Estados do país, revelou apenas raros casos suspeitos de envenenamento ofídico fatal em bovinos no Brasil. Em nossas viagens de estudo e nos trabalhos de diagnóstico nunca estabelecemos o diagnóstico de morte por acidente ofídico em bovinos. Os casos tidos como envenenamento ofídico, na sua grande maioria, são apenas suposições, sem embasamento. Esses "diagnósticos", em geral, são feitos à distância dos animais que morreram, à revelia de exame clínico, necropsia e estudo histopatológico. Importante foi a constatação de que, no Brasil, embora algumas serpentes do gênero Bothrops possam, teoricamente, produzir quantidades suficientes de veneno para matar um bovino adulto, em experimentos realizados, apenas Bothrops alternatus foi capaz de levar a morte um dos três bovinos experimentalmente por ela picados; esse animal tinha apenas 279 kg. Já as serpentes do gênero Crotalus poderiam inocular quantidades letais de veneno para bovinos adultos. Mesmo assim, tanto para Bothrops spp, como para Crotalus spp, há que se considerar que as serpentes, em geral, só inoculam parte do veneno disponível. Esse estudo indica que é necessário melhor investigar as mortes suspeitas de terem sido causadas por acidente ofídico em bovinos no Brasil. O estabelecimento do diagnóstico de morte por envenenamento ofídico, porém, só é poss

  11. Epidemiologia dos acidentes ofídicos notificados pelo Centro de Assistência e Informação Toxicológica de Campina Grande (Ceatox-CG, Paraíba Epidemiologic profile of snakebites reported by the Poison Information Center of Campina Grande, Paraíba

    Directory of Open Access Journals (Sweden)

    Josiverton de Carvalho Lemos

    2009-03-01

    Full Text Available Os acidentes causados por animais peçonhentos ainda constituem problema de Saúde Pública no Brasil. Embora a produção e a distribuição dos soros no país encontrem-se estabilizadas, há problemas relacionados à notificação de acidentes deste tipo em várias regiões. Considerando esta realidade, foi realizado um estudo transversal entre janeiro e dezembro de 2005, utilizando documentação indireta sobre os acidentes ofídicos ocorridos em Campina Grande e 80 municípios adjacentes à região, com o objetivo de conhecer o perfil epidemiológico e clínico deste tipo de caso. Todos os pacientes atendidos tiveram diagnóstico médico de acidente por serpentes realizado pelo Centro de Atendimento Toxicológico de Campina Grande (Ceatox-CG. Os dados foram coletados através da ficha de notificação do Sinan (Sistema Nacional de Notificação de Agravo do Ministério da Saúde. Para análise dos dados, foi utilizada estatística descritiva e os dados foram tabulados no programa Microsoft Excel 2003. Dos 1.443 atendimentos no Centro, 737 foram causados por animais peçonhentos e, destes, 277 foram provocados por serpentes peçonhentas e não peçonhentas. Os acidentes predominaram no sexo masculino, na faixa etária de 10 a 29 anos, principalmente em agricultores na zona rural, entre maio e novembro de 2005. O gênero Bothrops foi responsável pelo maior número de casos (71,5%, e as extremidades superior e inferior do corpo foram os locais de maior predominância de ataques. Na extremidade inferior, foi atingido principalmente o pé. Em relação à gravidade, foram mais frequentes os acidentes classificados como leves causados pelo gênero Bothrops e apenas um paciente evoluiu para óbito. A média do tempo de atendimento, em horas, foi superior a 12 horas nos acidentes considerados graves, causados pelos gêneros Bothrops e Crotalus. Nossos resultados concordam com o perfil epidemiológico nacional dos acidentes ofídicos, atingindo com

  12. Serpentes peçonhentas e ofidismo em Cruzeiro do Sul, Alto Juruá, Estado do Acre, Brasil Venomous snakes and ophidism in Cruzeiro do Sul, Alto Juruá, State of Acre, Brazil

    Directory of Open Access Journals (Sweden)

    Paulo Sérgio Bernarde

    2012-03-01

    Full Text Available Apesar da importância dos acidentes ofídicos na Saúde Pública, são relativamente poucas as pesquisas realizadas sobre esse tema no Brasil. Devido aos poucos estudos sobre ofidismo na Amazônia e especialmente no estado do Acre, trabalhos epidemiológicos são de grande relevância. Esse estudo apresenta a lista de serpentes peçonhentas e aspectos epidemiológicos dos acidentes ofídicos em Cruzeiro do Sul, região do Alto Juruá (Acre, verificando quais gêneros de serpentes são responsáveis pelos envenenamentos e aspectos que envolvem o acidente e o atendimento hospitalar. Os dados epidemiológicos foram coligidos a partir do SINAN (Sistema de Informações de Agravos de Notificação, no setor de vigilância epidemiológica do Hospital Regional do Juruá, no município de Cruzeiro do Sul. Sete espécies de serpentes peçonhentas foram registradas nesse município: três viperídeos (Bothrops atrox, Bothriopsis bilineata e Lachesis muta e quatro elapídeos (Micrurus hemprichii, M. lemniscatus, M. remotus and M. surinamensis. Durante o período de dois anos (agosto de 2007 a julho de 2009 foram registrados 195 casos de acidentes ofídicos. Cinquenta e um por cento dos acidentes foi classificado como laquético (Lachesis, seguido pelo botrópico (Bothrops e Bothriopsis com 38% e crotálico (Crotalus com 2%. Em 9% dos casos o gênero da serpente envolvida não foi informado. A maioria dos acidentes envolveu indivíduos adultos do gênero masculino em área rural, afetados principalmente nos membros inferiores. Os casos ocorreram mais frequentemente nos meses de novembro a abril, coincidindo com os maiores níveis pluviométricos. A maioria dos acidentes foi atribuída equivocadamente à serpente L. muta, tendo sido provavelmente causados por B. atrox.Despite the importance of accidents involving snakebites, research on this theme in Brazil is relatively rare, especially in the Amazon region. Due to the poor number of studies about ophidism

  13. Photobiomodulation Protects and Promotes Differentiation of C2C12 Myoblast Cells Exposed to Snake Venom.

    Directory of Open Access Journals (Sweden)

    Luciana Miato Gonçalves Silva

    Full Text Available Snakebites is a neglected disease and in Brazil is considered a serious health problem, with the majority of the snakebites caused by the genus Bothrops. Antivenom therapy and other first-aid treatments do not reverse local myonecrose which is the main sequel caused by the envenomation. Several studies have shown the effectiveness of low level laser (LLL therapy in reducing local myonecrosis induced by Bothropic venoms, however the mechanism involved in this effect is unknown. In this in vitro study, we aimed to analyze the effect of LLL irradiation against cytotoxicity induced by Bothrops jararacussu venom on myoblast C2C12 cells.C2C12 were utilized as a model target and were incubated with B. jararacussu venom (12.5 μg/mL and immediately irradiated with LLL at wavelength of red 685 nm or infrared 830 nm with energy density of 2.0, 4.6 and 7.0 J/cm2. Effects of LLL on cellular responses of venom-induced cytotoxicity were examined, including cell viability, measurement of cell damage and intra and extracellular ATP levels, expression of myogenic regulatory factors, as well as cellular differentiation.In non-irradiated cells, the venom caused a decrease in cell viability and a massive release of LDH and CK levels indicating myonecrosis. Infrared and red laser at all energy densities were able to considerably decrease venom-induced cytotoxicity. Laser irradiation induced myoblasts to differentiate into myotubes and this effect was accompanied by up regulation of MyoD and specially myogenin. Moreover, LLL was able to reduce the extracellular while increased the intracellular ATP content after venom exposure. In addition, no difference in the intensity of cytotoxicity was shown by non-irradiated and irradiated venom.LLL irradiation caused a protective effect on C2C12 cells against the cytotoxicity caused by B. jararacussu venom and promotes differentiation of these cells by up regulation of myogenic factors. A modulatory effect of ATP synthesis may

  14. Influence of gamma-radiation on the biological activity of snake venoms in Peru

    International Nuclear Information System (INIS)

    Yarleque Ch, A.

    1986-03-01

    Effects of Co-60 gamma radiation on enzymatic, haemorragic and necrotic activities of Lachesis muta and Bothrops atrox venoms was studied at several ranges of irradiation lower than 1.0 Mrad. The radiation produced changes on its enzymatic activities. Irradiation at 0.1 Mrad resulted in the partial or complete inactivation of the following enzymes that are listed in order of increasing sensitivity: exonuclease, phospholipase A, caseinolytic enzyme, thrombinolytic enzyme, fibrinolytic enzyme, 5'-nucleotidase and endonuclease. The enzymatic inactivation was increased with 0.5 and 1.0 Mrad although not in a linear manner. Exonuclease was found to be the most radioresistant. The haemorragic activity was decreased to a greater extent than the necrotic activity. The probable mechanism for the changes in the enzymatic, haemorragic and necrotic activities are discussed

  15. Ochetosoma heterocoelium (Digenea: Plagiorchiidae in snakes from Colombia Ochetosoma heterocoelium (Digenea: Plagiorchiidae en ofidios de Colombia

    Directory of Open Access Journals (Sweden)

    Carolina Lenis

    2009-12-01

    Full Text Available Ochetosoma heterocoelium Travassos, 1921 is a parasite that has been identified in snakes from different locations in Colombia and is considered of veterinary importance. Here, we present a redescription based on morphological traits of juveniles and adults. Parasite burden of O. heterocoelium in the host varied between 3-207 individuals which were localized in the oral cavity of snakes, causing damage that range from small infections to mechanical obstruction of the esophagus and Jacobson's organ. The low levels of abundance and prevalence in the snakes Leptodeira septentrionalis, Bothriechis schlegelli, Bothrops asper and Porthidium nasutum coming from the localities of Maceo, Vegachí and Acandí regions, suggest that the parasite is not a threat to the snakes of Colombia. We propose a treatment program that consists of manual removal of Digenea as well as treating snakes with antihelminthics, the applaying of soft antiseptics and monitoring recovery. The prevalence of O. heterocoelium was established in new regions, namely Middle Magdalena, Valle de Aburrá, Urabá Chocoano and northern Colombia. This extends the known geographical distribution from Brazil and Venezuela. New hosts were also identified: Atractus lasallei, Bothriechis schlegelli, Bothrops asper, Chironius carinatus, Leptodeira septentrionalis, Leptophis ahaetulla and Porthidium nasutum.Se identifica Ochetosoma heterocoelium Travassos, 1921 como un parásito de importancia veterinaria en ofidios de diferentes localidades de Colombia y se redescribe con base en las características morfológicas de individuos juveniles y adultos. La carga parasitaria de O. heterocoelium varió de 3-207 individuos alojados en la cavidad bucal de los ofidios, causando desde infecciones leves hasta obstrucción mecánica en el esófago y órgano de Jacobson. Los bajos valores de abundancia y prevalencia en las localidades de Maceo, Vegachí y Acandí y en los ofidios Leptodeira septentrionalis

  16. Envenomation by the neotropical colubrid Boiruna maculata (Boulenger, 1896: a case report Envenenamento por Colubrídeo Neotropical Boiruna maculata (Boulenger, 1896: registro de um caso

    Directory of Open Access Journals (Sweden)

    Maria Cristina dos SANTOS-COSTA

    2000-10-01

    Full Text Available This is a case report of a Boiruna maculata snake bite in a child admitted to the Hospital Municipal de Pronto Socorro de Porto Alegre, Porto Alegre, RS, Brazil. The patient was bitten on the lower left limb, and exhibited pronounced local manifestations of envenomation. She was treated with Bothrops antivenom and was discharged from the hospital five days later with marked improvement of envenomation.Este trabalho relata o envenenamento por serpente do gênero Boiruna maculata em criança admitida e posteriormente hospitalizada no Hospital Municipal de Pronto Socorro de Porto Alegre, RS, Brasil. A paciente foi mordida no membro inferior esquerdo e apresentou sinais de envenenamento local pronunciado, foi tratada como acidente botrópico e permaneceu no hospital por cinco dias, recebendo alta após melhora.

  17. Chemical constituents and antiedematogenic activity of Peltodon radicans (Lamiaceae); Constituintes quimicos e atividade antiedematogenica de Peltodon radicans (Lamiaceae)

    Energy Technology Data Exchange (ETDEWEB)

    Costa, Habdel Nasser Rocha da [Universidade Federal de Roraima, Boa Vista, RR (Brazil). Centro de Ciencias e Tecnologia. Dept. de Quimica; Santos, Maria Cristina dos [Universidade Federal do Amazonas, Manaus, AM (Brazil). Inst. de Ciencias Biologicas. Dept. de Parasitologia; Alcantara, Antonio Flavio de Carvalho; Silva, Marilda Conceicao; Franca, Roberta Cabral; Pilo-Veloso, Dorila [Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, MG (Brazil). Inst. de Ciencias Exatas. Dept. de Quimica]. E-mail: aalcantara@zeus.qui.ufmg.br

    2008-07-01

    Most of the snakebite incidents in the Amazon region involve Bothrops atrox, whose venom presents the most potent edematogenic and necrotic activities in the genus. This work describes the studies of isolation of the chemical constituents and antiedematogenic activity of the species Peltodon radicans (Lamiaceae), which is used in the treatment of snakebites and scorpion stings in the region. The extracts presented aliphatic hydrocarbons, 3{beta}-OH,{beta}-amirin (1), 3{beta}-OH,a-amirin (2), {beta}-sitosterol (3), stigmasterol (4), ursolic acid (5), 2{alpha},3{beta},19{alpha}- trihydroxy-urs-12-en-28-oic acid (tormentic acid, 6), methyl 3{beta}-hydroxy,28-methyl-ursolate (7), sitosterol-3-O-{beta}-D-glucopyranoside (8), and stigmasterol-3-O-{beta}-D-glucopyranoside (9). The flower extracts presented the higher antiedematogenic activity. This is the first report on the study of the flowers, stem, and roots of this plant. (author)

  18. Noteworthy records of reptiles from natural open vegetation areas in plateau and coastal areas of the state of Santa Catarina, southern Brazil

    Directory of Open Access Journals (Sweden)

    Ivo Rohling Ghizoni-Jr.

    2009-09-01

    Full Text Available The composition and geographic distribution of reptiles in Santa Catarina are little known, particularly in the open areas of its plateau (covered with ombrophylous steppe and Atlantic littoral (coastal dunes with thin, low, and predominant herbaceous vegetation known as restinga. Aiming to contribute to the knowledge of reptiles that inhabit these areas, this paper presents sixteen records of previously unknown or uncommon species in Santa Catarina: Acanthochelys spixii, Anops kingii, Cnemidophorus lacertoides, C. vacariensis, Leptotyphlops munoai, Mastigodryas b. bifossatus, Tantilla aff. melanocephala, Atractus reticulatus, Gomesophis brasiliensis, Lygophis flavifrenatus, Oxyrhopus r. rhombifer, Phalotris reticulatus, Philodryas agassizii, Xenodon dorbignyi, Micrurus altirostris and Bothrops pubescens. Aspects of the distribution and conservation of these species are discussed briefly.

  19. Faculdade de Saúde e Desenvolvimento Humano Santo Agostinho

    Directory of Open Access Journals (Sweden)

    Beatriz Ferreira Martins

    2012-09-01

    Full Text Available Accidents caused by snakes should be dealt with attention by public health organs, for these accidents are very frequent and serious. The treatment standardization is indispensable for helping health workers to diagnose and treat the victims. However, sometimes the case clinical evolution presents peculiarities, mainly in accidents with children and elderly people. This study aims at describing and analyzing two clinical cases of accidents with children and snakes of the genera Bothrops spp. and Crotallus spp., which were notified at an information and toxicological assistance center in northwestern Parana (Brazil. We could find some positive aspects in the assistance, such as the precocious access to the health unit, identification of symptoms and antivenom treatment and the lack of antivenom adverse reactions. The negative aspects noticed are the lack of recommended biochemical examinations, for a better diagnosis, as well as an early hospital discharge, disconsidering the possibility of adverse reactions.

  20. Professor José Roberto Giglio and toxinnology in Brazil: 48 years in research

    Directory of Open Access Journals (Sweden)

    A. M. Soares

    2007-01-01

    Full Text Available This work succinctly describes the professional and scientific life of Dr. José R. Giglio, one of the most outstanding Brazilian researchers in the field of Toxinology. During his long and successful career, he has made major contributions, especially in elucidating the function, structure, and mechanisms of action of animal venom proteins (from snakes, scorpions and spiders as well as the characterization of antibodies and several inhibitors of venoms and toxins. We present here a brief history of Dr. Giglio’s personal and professional life, also reporting some of his numerous published scientific articles on venoms from snakes (Bothrops, Crotalus, and other genera, scorpions (Tityus sp, spiders (Phoneutria sp, their isolated toxins and natural inhibitors. Thus, this work is a tribute to Dr. Giglio in his 73rd birthday, having devoted 48 years of his life studying animal venoms, an effort that has continued even after his formal retirement from university duties.

  1. Hemorrhagic activity of the Duvernoy's gland secretion of the xenodontine colubrid Philodryas patagoniensis from the north-east region of Argentina.

    Science.gov (United States)

    Acosta, O; Leiva, L C; Peichoto, M E; Maruñak, S; Teibler, P; Rey, L

    2003-06-01

    Colubrid snakes belonging to Philodryas genus, widespread all over South America, bring about lesions (swelling, ecchymosis, transient bleeding from the bite site punctures), that are similar to those produced by Bothrops species (yarará). In the present work we began the characterization of Philodryas patagoniensis venom. We examined if this venom produces hemorrhagic lesions as those observed in victims bitten by Philodryas olfersii. Hemorrhagic, proteolytic and fibrinogenolytic activities were evaluated, and histological observations in samples of gastrocnemius muscle were carried out. Inhibition studies were carried out in metal chelator (ethylenediaminetetraacetic acid) presence. Our results show a small Minimum Hemorrhagic Dose (MHD=0.035 microg) and a high proteolytic activity (143 U/mg), and prove the capacity of this venom to degrade fibrinogen in vitro rendering it unclottable by thrombin, supporting the presence of proteases, principally metalloproteases, in P. patagoniensis venom that are able to alterate the vascular wall and degrade fibrinogen, being both activities responsible of a high hemorrhagic activity.

  2. Philodryas patagoniensis bite and local envenoming.

    Science.gov (United States)

    Nishioka, S A; Silveira, P V

    1994-01-01

    A 5-year-old boy bitten by a specimen of Philodryas patagoniensis, a colubrid snake currently classified as nonvenomous, developed signs of local envenoming characterized by swelling and warmth on the bitten limb. This is the first time that local envenoming following Philodryas patagoniensis bite is recognized. Based on the clinical findings and misidentification of the snake, the patient was treated as a victim of Bothrops bite, having received unnecessarily the specific antivenom. Educational efforts to make doctors and health workers capable to identify correctly venomous snakes are necessary, to avoid inappropriate indication of antivenom and decrease the risk of its potentially harmful untoward effects. Examination of the bite site can be useful to the differential diagnosis between pit viper and colubrid bites.

  3. Análise das atividades biológicas dos venenos de Philodryas olfersii (Lichtenstein e P. patagoniensis (Girard (Serpentes, Colubridae Analysis of biological activities from Philodryas olfersii (Lichtenstein and P. patagoniensis (Girard venoms (Serpents, Colubridae

    Directory of Open Access Journals (Sweden)

    Marisa M. T. da Rocha

    2007-06-01

    Full Text Available Philodryas olfersii (Lichtenstein, 1823 e P.patagonienis (Girard, 1857 são serpentes colubrídeas da série opistóglifa, restritas à América do Sul. Vários acidentes ocasionados por estas serpentes têm sido relatados, caracterizando-se por ação local importante: dor, edema e hemorragia. É um acidente muito semelhante àquele causado por serpentes do gênero Bothrops Wagler, 1824 e muitas vezes os pacientes são tratados com soro antibotrópico. Poucos estudos tratam da caracterização destes venenos, assim tivemos como objetivo de trabalho o estudo dos venenos de P.olfersii e P.patagonienis. Os venenos apresentaram teor de proteínas entre 75 e 90%. A atividade desfibrinante não foi detectada quando testada em camundongos. O quadro de dor causado pelo envenenamento experimental, em camundongos, mostrou que os venenos de P.olfersii e P.patagoniensis causaram intensa reatividade, sendo que o veneno de P.patagoniensis foi o mais ativo. Ambos os venenos apresentaram dose mínima edematogênica em torno de 1 µg/camundongo com ação máxima em 30 minutos.A ação hemorrágica se instalou rapidamente, com doses mínimas semelhantes. As atividades tóxicas foram semelhantes, com valores em torno de 60,0 µg/camundongo, comparáveis aos venenos botrópicos.Philodryas olfersii (Lichtenstein, 1823 and P. patagoniesis (Girard, 1857 are species of colubrid snakes of the opisthoglyphous series restricted to South America. Several accidents caused by these snakes have been reported and they are mainly characterized by marked effects at the site of bite, such as pain, swelling and hemorrhage. Such accidents are similar to those caused by snakes of the genus Bothrops, and thereby patients bitten by Philodryas sp. are frequently treated with Bothrops antivenom. Since few studies have dealt with the characterization of these venoms, our aim was to study the venoms of P.olfersii and P.patagoniensis. They presented protein contents between 75 and 90

  4. Distribution of the herpetofauna community associated to four areas with different interference degree in Gorgona Island, Colombian pacific

    International Nuclear Information System (INIS)

    Urbina C, Jose Nicolas; Londono M, Maria Cecilia

    2003-01-01

    A total of 1840 individuals from 28 species (19 reptiles and 9 amphibians) were found in Gorgona Island, during June and July 2001. Based on 32 transects placed in four areas with different antropic perturbation degree (Prison, palm plantations, secondary forest and primary forest) it was found that the species richness was higher at the secondary forest. The species registered at primary and secondary forest where very similar as well as the species present at the prison and the palm plantations. A Canonical Correspondence Analysis (CCA) showed that Boa constricto1; Basiliscus galeritus. Ameiva bridgesii and Epipedobates boulengeri were found to be associated to open areas and their distribution was hardly affected by the environmental temperature. From the following species associated with forested areas, the canopy cover over the micro habitat influenced the distribution of Eleutherodactylus gularis. Eleutherodactylus achatinus and Bothrops atrox. While the understory cover influenced the distribution of Atelopus elegans. Bufo typhonius. Micrurus mipartitus y Enyalioides heterolepis

  5. Heterologous expression of the antimyotoxic protein DM64 in Pichia pastoris

    Science.gov (United States)

    Vieira, Saulo Martins; da Rocha, Surza Lucia Gonçalves; Neves-Ferreira, Ana Gisele da Costa; Almeida, Rodrigo Volcan; Perales, Jonas

    2017-01-01

    Snakebite envenomation is a neglected condition that constitutes a public health problem in tropical and subtropical countries, including Brazil. Interestingly, some animals are resistant to snake envenomation due to the presence of inhibitory glycoproteins in their serum that target toxic venom components. DM64 is an acidic glycoprotein isolated from Didelphis aurita (opossum) serum that has been characterized as an inhibitor of the myotoxicity induced by bothropic toxins bearing phospholipase A2 (PLA2) structures. This antitoxic protein can serve as an excellent starting template for the design of novel therapeutics against snakebite envenomation, particularly venom-induced local tissue damage. Therefore, the aim of this work was to produce a recombinant DM64 (rDM64) in the methylotrophic yeast Pichia pastoris and to compare its biological properties with those of native DM64. Yeast fermentation in the presence of Pefabloc, a serine protease inhibitor, stimulated cell growth (~1.5-fold), increased the rDM64 production yield approximately 10-fold and significantly reduced the susceptibility of rDM64 to proteolytic degradation. P. pastoris fermentation products were identified by mass spectrometry and Western blotting. The heterologous protein was efficiently purified from the culture medium by affinity chromatography (with immobilized PLA2 myotoxin) and/or an ion exchange column. Although both native and recombinant DM64 exhibit different glycosylation patterns, they show very similar electrophoretic mobilities after PNGase F treatment. rDM64 formed a noncovalent complex with myotoxin II (Lys49-PLA2) from Bothrops asper and displayed biological activity that was similar to that of native DM64, inhibiting the cytotoxicity of myotoxin II by 92% at a 1:1 molar ratio. PMID:28759578

  6. Heterologous expression of the antimyotoxic protein DM64 in Pichia pastoris.

    Directory of Open Access Journals (Sweden)

    Saulo Martins Vieira

    2017-07-01

    Full Text Available Snakebite envenomation is a neglected condition that constitutes a public health problem in tropical and subtropical countries, including Brazil. Interestingly, some animals are resistant to snake envenomation due to the presence of inhibitory glycoproteins in their serum that target toxic venom components. DM64 is an acidic glycoprotein isolated from Didelphis aurita (opossum serum that has been characterized as an inhibitor of the myotoxicity induced by bothropic toxins bearing phospholipase A2 (PLA2 structures. This antitoxic protein can serve as an excellent starting template for the design of novel therapeutics against snakebite envenomation, particularly venom-induced local tissue damage. Therefore, the aim of this work was to produce a recombinant DM64 (rDM64 in the methylotrophic yeast Pichia pastoris and to compare its biological properties with those of native DM64. Yeast fermentation in the presence of Pefabloc, a serine protease inhibitor, stimulated cell growth (~1.5-fold, increased the rDM64 production yield approximately 10-fold and significantly reduced the susceptibility of rDM64 to proteolytic degradation. P. pastoris fermentation products were identified by mass spectrometry and Western blotting. The heterologous protein was efficiently purified from the culture medium by affinity chromatography (with immobilized PLA2 myotoxin and/or an ion exchange column. Although both native and recombinant DM64 exhibit different glycosylation patterns, they show very similar electrophoretic mobilities after PNGase F treatment. rDM64 formed a noncovalent complex with myotoxin II (Lys49-PLA2 from Bothrops asper and displayed biological activity that was similar to that of native DM64, inhibiting the cytotoxicity of myotoxin II by 92% at a 1:1 molar ratio.

  7. Atividade antiinflamatória do extrato aquoso de Arrabidaea chica (Humb. & Bonpl. B. Verl. sobre o edema induzido por venenos de serpentes amazônicas

    Directory of Open Access Journals (Sweden)

    Denys Paixão Costa de Oliveira

    Full Text Available O estudo investigou o efeito antiinflamatório do extrato aquoso da Arrabidaea chica (Humb. & Bonpl. B. Verl., Bignoniaceae, popularmente conhecida como "crajiru", sobre o edema induzido por venenos de serpentes amazônicas dos gêneros Brothrops e Crotalus, em camundongos albinos, por via oral, intraperitoneal e subcutânea. O efeito anti-edematogênico foi avaliado pela medição do diâmetro dos coxins das patas posteriores, sendo medidos as 1, 3, 6, 12 e 24 horas, para B. atrox e 1, 3 e 6 horas para Crotalus durissus ruruima, e também avaliado por histopatologia. O estudo mostrou que o efeito inibitório do extrato aquoso para o gênero Bothrops, pelas vias subcutânea e intraperitoneal (12 horas foi de 55,87% e 65,70%, respectivamente. Para o gênero Crotalus o efeito inibitório do extrato pela via subcutânea após 3 horas foi de 33,55% e após 6 horas de 79,81%. Pela via intraperitoneal após 3 horas foi de 48,02% e após 6 horas de 92,52%. Na análise histopatológica, o infiltrado de granulócitos e a miocitólise foram os efeitos inflamatórios inibidos mais significativamente. Os resultados sugerem a presença no extrato aquoso de A. chica de substâncias com atividade inibitória sobre os efeitos inflamatórios dos venenos das serpentes Bothrops atrox e Crotalus durissus ruruima.

  8. Liver dysfunction in patients bitten by Crotalus durissus terrificus (Laurenti, 1768 snakes in Botucatu (State of São Paulo, Brazil Alterações hepáticas em doentes picados por serpentes Crotalus durissus terrificus (Laurenti, 1768 na região de Botucatu, Estado de São Paulo, Brasil

    Directory of Open Access Journals (Sweden)

    Benedito Barra Viera

    1995-02-01

    Full Text Available Thirty-two patients bitten by venomous snakes sixteen by Bothrops spp. and sixteen by Crotalus durissus terrificus were studied. The group comprised thirty males and two females, aged eight to sixty-three years (mean 33±15. Bromsulphalein tests were increased in the majority of patients bitten by Crotalus durissus terrificus. The correlation coefficient of Spearman was positive between bromsulphalein tests and alanine aminotransferase levels, and between alanine aminotransferase and aspartate aminotransferase levels only in the Crotalus group. The only patient who died was bitten by Crotalus durissus terrificus and showed hydropic degeneration and mitochondrial injury in the liver. It was concluded that the hepatic damage might have been caused by at least two possible mechanisms: venom effect on liver mitochondria and cytokine effects on hepatocyte, specially interleukin-6.Os autores estudaram 32 doentes picados por serpentes venenosas, sendo 16 picados por Bothrops spp. e 16 por Crotalus durissus terrificus. Trinta doentes eram do sexo masculino e dois do feminino com idades variando entre 8 e 63 anos (méda 33±15. A prova da retenção da bromosulfaleína apresentou-se aumentada na maioria dos doentes picados por serpentes Crotalus durissus terrificus. Houve correlação positiva entre a retenção da bromosulfaleína e os níveis séricos de alanina aminotransferase e entre alanina e aspartato aminotransferase apenas nos doentes do grupo Crotalus. Um dos doentes evoluiu para o óbito e apresentou no exame anatomopatológico do fígado degeneração hidrópica e lesões mitocondriais. Os autores concluem que as alterações hepáticas são causadas por pelo menos dois mecanismos a saber: lesão mitocondrial por efeito do veneno crotálico; efeito das citoquinas, especialmente a interleucina-6.

  9. Envenenamento ofídico em crianças: frequência de reações precoces ao antiveneno em pacientes que receberam pré-tratamento com antagonistas H1 e H2 da histamina e hidrocortisona Snake envenomation in children: early reactions frequency at antivenom in patients pretreated with histamine antagonists HI and H2 and hydrocortisone

    Directory of Open Access Journals (Sweden)

    Fábio Bucarethi

    1994-10-01

    Full Text Available Foram estudadas 24 crianças, com idade entre 2 a 14 anos, de 1989 a 1993, vítimas de acidentes ofídicos, submetidas a pré-tratamento com antagonistas H1 (dextroclorfeniramina e H2 (cimetidine ou ranitidina da histamina e hidrocortisona, com objetivo de avaliar a frequência e o tipo de reações precoces (RP ao antiveneno (AV. Em nenhum paciente havia antecedente de atopia ou uso prévio de algum tipo de antiveneno ou antitoxina heteróloga. Das 24 crianças 15 receberam AV botrópico (RP em 5, 7 AV crotálico (RP em 5, 1 AV crotálico e AV botrópico-crotálico e 1 AV elapídico (RP. Foram observadas RP graves em 3 crianças, as 3 classificadas como acidente crotálico grave. A análise dos resultados sugere que o pré-tratamento realizado não ofereceu uma proteção segura quanto ao aparecimento de RP.Type and frequency of early reactions (ER were studied in 24 children aging 2-14 years victims of snake bites who received pretreatment with histamine antagonists H1 (dextrochlorfenirarnine and H2 (cimetidine or ranitidine and hydrocortisone from 1989 to 1993. None of them had atopy nor received any type of anti-venoms(AV and antitoxins before. Of 24 children, 15 received bothropic AV (ER in 5, 7 crotalic AV (ER in 5, 1 crotalic plus crotalic-bothropic AV, and 1 elapidic AV (ER in 1. In 3 children severe early reactions were observed and they were classified as severe crotalic accident. Results suggest that pre-treatment did not offer safety protection at the appearance of early reactions.

  10. Epidemiology of snakebite accidents in the municipalities of the state of Paraíba, Brazil Epidemiologia dos acidentes ofídicos ocorridos nos municípios do Estado da Paraíba, Brasil

    Directory of Open Access Journals (Sweden)

    Rafaella Moreno Barros

    2013-05-01

    Full Text Available Accidents involving venomous animals represent an important, albeit neglected, public health issue worldwide. A descriptive study was made of snakebite cases attended and recorded between 2007 and 2010 in the health units of the municipalities of Cariri, State of Paraíba, northeastern Brazil. Data was collected from the Injury Notification Information System data banks of the Health Ministry and a total of 351 records of snakebite victims were reviewed. Victims were predominantly male farm workers over 50. The highest incidence of snakebites occurred in rural areas, between April and June of 2007 and 2010. Snakes of the genus Bothrops were responsible for most cases, and victims were mostly bitten on the feet. The majority of the victims received medical assistance within 1 to 3 hours after being bitten. The most common clinical manifestations were pain, edema and ecchymosis, which were mainly classified as mild or moderate. Two deaths were reported. It was concluded that there is a significant impact of seasonality in snakebites, the prevalence of attacks caused by Bothrops, affecting the lower limbs of adult male farmers in rural areas. The findings of this study may contribute to identify the conditions that increase the risk of snake attacks in the northeastern region.Acidentes por animais peçonhentos representam um importante, embora negligenciado, problema de saúde pública mundial. Neste sentido, foi realizado um estudo descritivo dos acidentes ofídicos atendidos e registrados, entre 2007 e 2010, nas unidades de saúde dos municípios do Cariri, Estado da Paraíba, Nordeste do Brasil. As informações foram coletadas do banco de dados do Sistema de Informação de Agravos de Notificação do Ministério da Saúde. Um total de 351 registros de vítimas de ataques por serpentes peçonhentas foram analisados. As vítimas foram predominantemente trabalhadores rurais do sexo masculino com mais de 50 anos. As maiores incidências de

  11. Influenza em animais heterotérmicos Influenza in heterothermics

    Directory of Open Access Journals (Sweden)

    Dalva Assunção Portari Mancini

    2004-06-01

    Full Text Available O objetivo foi pesquisar Ortomyxovirus em animais heterotérmicos. Coletou-se sangue de serpentes dos gêneros Bothrops e Crotalus e de sapo e rãs dos gêneros Bufo e Rana, para a detecção dos receptores de hemácias e anticorpos específicos, ao vírus influenza, pelos testes de hemaglutinação e inibição da hemaglutinação, respectivamente. Pelo teste de hemaglutinação, verificou-se que serpentes e sapos em cativeiro apresentaram receptores em suas hemácias para o vírus influenza, humano e eqüino do tipo A e tipo B. O mesmo ocorreu com serpentes recém chegadas. Quanto ao teste de inibição da hemaglutinação dos soros dos répteis observou-se títulos protetores de anticorpos aos vírus influenza tipo A (origens humana e eqüina e tipo B. Com soro de sapo não se observou reação de inibição da hemaglutinação porém, 83,3% das rãs obtiveram médias de 40UIH para algumas cepas. Conclui-se que animais heterotérmicos podem oferecer condições de hospedeiros aos vírus influenza, assim como susceptibilidade à infecção.The objective was to study Orthomyxovirus in heterothermic animals. Blood samples from snakes (genus Bothrops and Crotalus and from toads and frogs (genus Bufo and Rana were collected to evaluate the red cell receptors and antibodies specific to influenza virus by the hemagglutination and hemagglutination inhibition tests, respectively. Both snakes and toads kept in captivity presented receptors in their red cells and antibodies specific to either influenza virus type A (human and equine origin or influenza type B. The same was observed with recently captured snakes. Concerning the influenza hemagglutination inhibition antibodies protective levels were observed in the reptiles' serum, against influenza type A and type B. Unlike the toads, 83.3% of the frogs presented mean levels of Ab 40HIU for some influenza strains. It was concluded that heterothermic animals could offer host conditions to the influenza

  12. Involvement of Nitric Oxide on Bothropoides insularis Venom Biological Effects on Murine Macrophages In Vitro.

    Directory of Open Access Journals (Sweden)

    Ramon R P P B de Menezes

    Full Text Available Viperidae venom has several local and systemic effects, such as pain, edema, inflammation, kidney failure and coagulopathy. Additionally, bothropic venom and its isolated components directly interfere on cellular metabolism, causing alterations such as cell death and proliferation. Inflammatory cells are particularly involved in pathological envenomation mechanisms due to their capacity of releasing many mediators, such as nitric oxide (NO. NO has many effects on cell viability and it is associated to the development of inflammation and tissue damage caused by Bothrops and Bothropoides venom. Bothropoides insularis is a snake found only in Queimada Grande Island, which has markedly toxic venom. Thus, the aim of this work was to evaluate the biological effects of Bothropoides insularis venom (BiV on RAW 264.7 cells and assess NO involvement. The venom was submitted to colorimetric assays to identify the presence of some enzymatic components. We observed that BiV induced H2O2 production and showed proteolytic and phospholipasic activities. RAW 264.7 murine macrophages were incubated with different concentrations of BiV and then cell viability was assessed by MTT reduction assay after 2, 6, 12 and 24 hours of incubation. A time- and concentration-dependent effect was observed, with a tendency to cell proliferation at lower BiV concentrations and cell death at higher concentrations. The cytotoxic effect was confirmed after lactate dehydrogenase (LDH measurement in the supernatant from the experimental groups. Flow cytometry analyses revealed that necrosis is the main cell death pathway caused by BiV. Also, BiV induced NO release. The inhibition of both proliferative and cytotoxic effects with L-NAME were demonstrated, indicating that NO is important for these effects. Finally, BiV induced an increase in iNOS expression. Altogether, these results demonstrate that B. insularis venom have proliferative and cytotoxic effects on macrophages, with

  13. Ofidismo: Estudio de 30 casos en Brasil Ophidism: Study of 30 cases in Brazil

    Directory of Open Access Journals (Sweden)

    Moisés Morejón García

    2006-06-01

    Full Text Available Se realizó un estudio de 30 pacientes ingresados por mordedura de cobras, en el Hospital Municipal de Paraiso de Tocantins, Estado de Tocantins, Brasil, en el período comprendido entre 1998-2001. Los resultados arrojaron un predominio del accidente ofídico en pacientes masculinos (76,6 %, con edades entre los 15 y 50 años (80 %. Hubo una participación mayoritaria de reptiles del género Bothrops (63,3 %, que provocó la mayor parte de las mordeduras en los miembros inferiores (80 %. Las manifestaciones clínicas más evidentes fueron el edema y el dolor (93,3 %, de moderado a severo en 63,3 % de ellos, por lo que la dosis media de suero antiofídico utilizada fue de 10 ámpulas en dosis única. El coagulograma reflejó alteración de la coagulación en el 70 % de los pacientes, pero se recuperó totalmente en 48 h. La complicación que más se presentó fue el fallo renal en 9 pacientes, de los cuales 5 se recuperaron en nuestro hospital y 4 fueron remitidos a unidad de terapia intensiva. Fallecieron 2 pacientes (6,6 %, ambos por fallo renal y shock . Del total de pacientes, 25 (83,3 % tuvieron evolución satisfactoria.Thirty patients admitted in the Municipal Hospital of Paraiso de Tocantins, State of Tocantins , Brazil , due to cobra bite, were studied between 1998 and 2001 . According to the results, there was a predominance of ophidic accidents in male patients (76.6 % aged 15-50 (80 %. Most of the reptiles corresponded to Bothrops genus (63.3 %, and the greatest number of bites occurred in the lower limbs (80 %. The most evident clinical manifestations were edema and pain (93.3 %, which were from moderate to severe in 63.3 % of them. Therefore, the mean dose of anti-ophidic serum used was 10 ampules in a single dose. The coagulogram showed coagulation alteration in 70 % of the patients, but it was totally recovered in 48 hrs. The most frequent complication observed was a renal failure in 9 patients, of whom 5 had a complete recovery in

  14. Avaliação das pressões venosa e arterial em cães submetidos a diferentes tipos de hipotensão Evaluation of venous and arterial blood pressures in dogs submitted to hypotension

    Directory of Open Access Journals (Sweden)

    R.C. Rabelo

    2005-12-01

    Full Text Available Estabeleceram-se a pressão venosa periférica (PVP, a pressão venosa central (PVC, a pressão arterial invasiva (PAI e a pressão arterial não invasiva (PANI em cães após diferentes eventos de hipotensão. Foram utilizados 15 cães adultos, distribuídos aleatoriamente em três grupos (G com cinco animais cada, submetidos aos seguintes eventos hipotensores: GI - cloridrato de xilazina a 2%, GII - choque hipovolêmico agudo e GIII - veneno da serpente Bothrops moojeni. Os animais, avaliados durante 30 minutos após o início do evento hipotensor, foram tratados com cloridrato de ioimbina (GI, amido hidroxietílico a 6% (GII e cetoprofeno (GIII e reavaliados por mais 30 minutos. Somente os animais do GII apresentaram redução da PVP após o evento hipotensor e aumento, 25 minutos após tratamento. Os cães dos grupos II e III mostraram redução da PVC após o evento hipotensor, e somente os animais do GII exibiram discreto aumento cinco minutos imediatamente após o tratamento. Houve diminuição da PAI e PANI nos dos grupos II e III após o evento hipotensor, com recuperação gradativa imediata, após o tratamento, somente da PAI.The peripheral venous pressure (PVP, the central venous pressure (CVP, the invasive (IAP and non-invasive blood pressure (NIAP in dogs submitted to different hypotensive events were studied. Fifteen adult mongrel dogs were randomly divided in three groups with five animals each, and submitted to hypotensive event as follow: GI - xylazine chloride 2%, GII - acute hypovolemic shock and GIII - snake venom (Bothrops moojeni. All animals were evaluated for 30 minutes after starting hypotensive event, treated with yoimbine chloride (GI, colloid hetastarch 6% (GII and ketoprofen (GIII and reevaluated for more 30 minutes. Only the group II dogs showed PVP decrease after hypotensive event, and increase 25 minutes after treatment. In animals of groups II and III, the CVP decreased after hypotensive event and only in GII

  15. Low-Level Laser Therapy (904 nm Counteracts Motor Deficit of Mice Hind Limb following Skeletal Muscle Injury Caused by Snakebite-Mimicking Intramuscular Venom Injection.

    Directory of Open Access Journals (Sweden)

    Willians Fernando Vieira

    Full Text Available Myotoxins present in Bothrops venom disrupt the sarcolemma of muscle fibers leading to the release of sarcoplasmic proteins and loss of muscle homeostasis. Myonecrosis and tissue anoxia induced by vascularization impairment can lead to amputation or motor functional deficit. The objective of this study was to investigate the dynamic behavior of motor function in mice subjected to injection of Bothrops jararacussu venom (Bjssu and exposed to low-level laser therapy (LLLT. Male Swiss mice received Bjssu injection (830 μg/kg into the medial portion of the right gastrocnemius muscle. Three hours later the injected region was irradiated with diode semiconductor Gallium Arsenide (GaAs- 904 nm, 4 J/cm² laser following by irradiation at 24, 48 and 72 hours. Saline injection (0.9% NaCl was used as control. Gait analysis was performed 24 hours before Bjssu injection and at every period post-Bjssu using CatWalk method. Data from spatiotemporal parameters Stand, Maximum Intensity, Swing, Swing Speed, Stride Length and Step Cycle were considered. The period of 3 hours post venom-induced injury was considered critical for all parameters evaluated in the right hindlimb. Differences (p<0.05 were concentrated in venom and venom + placebo laser groups during the 3 hours post-injury period, in which the values of stand of most animals were null. After this period, the gait characteristics were re-established for all parameters. The venom + laser group kept the values at 3 hours post-Bjssu equal to that at 24 hours before Bjssu injection indicating that the GaAs laser therapy improved spatially and temporally gait parameters at the critical injury period caused by Bjssu. This is the first study to analyze with cutting edge technology the gait functional deficits caused by snake envenoming and gait gains produced by GaAs laser irradiation. In this sense, the study fills a gap on the field of motor function after laser treatment following snake envenoming.

  16. Perfil dos acidentes ofídicos no norte do Estado de Minas Gerais, Brasil Profile of snakebite accidents in the north of the State of Minas Gerais, Brazil

    Directory of Open Access Journals (Sweden)

    Juliano Santos Lima

    2009-10-01

    Full Text Available O objetivo deste estudo foi descrever o perfil epidemiológico dos acidentes ofídicos da macrorregião de saúde do Norte do Estado de Minas Gerais, Brasil. Foram analisadas informações sobre os acidentes ofídicos relativos ao período compreendido entre janeiro de 2002 a dezembro de 2006, por meio de bancos de dados. Os resultados demonstraram 10.553 casos notificados, com ênfase para a maior casuística em meses de tempo quente e chuvoso, em áreas urbanas (54,1%, faixa etária menor de 20 anos (39,7%, acometendo mais homens e estudantes (53,1% e 29,1% respectivamente. Os membros inferiores (pé, dedo do pé, perna e coxa foram os locais mais afetados (35,9%, as serpentes prevalentes foram do gênero Bothrops (82,9% e a gravidade da maioria dos acidentes foi leve (66,2%. Observou-se nesse estudo um importante impacto da sazonalidade, urbanização, subnotificação das espécies envolvidas nesses acidentes e busca rápida pelo pronto atendimento. Espera-se que os dados inéditos da casuística obtida possam servir de substrato para o planejamento e execução de medidas voltadas para vigilância em saúde e atendimento.The aim of this study was to describe the epidemiological profile of snakebite accidents in the healthcare macroregion of the north of the State of Minas Gerais, Brazil. Database information on snakebite accidents covering the period from January 2002 to December 2006 was analyzed. It was found that 10,553 cases were notified, and that the samples were noticeably larger in the months of hot and rainy weather, in urban areas (54.1%, at ages less then 20 years (39.7% and among men and students (53.1% and 29.1% respectively. The lower limbs (feet, toes, legs and thighs were the locations most affected (35.9%. The most prevalent snakes were in the genus Bothrops (82.9% and most of the accidents were mild (66.2%. In this study, it was seen that the seasonality, urbanization and undernotification of the species involved in these

  17. Role of collagens and perlecan in microvascular stability: exploring the mechanism of capillary vessel damage by snake venom metalloproteinases.

    Directory of Open Access Journals (Sweden)

    Teresa Escalante

    Full Text Available Hemorrhage is a clinically important manifestation of viperid snakebite envenomings, and is induced by snake venom metalloproteinases (SVMPs. Hemorrhagic and non-hemorrhagic SVMPs hydrolyze some basement membrane (BM and associated extracellular matrix (ECM proteins. Nevertheless, only hemorrhagic SVMPs are able to disrupt microvessels; the mechanisms behind this functional difference remain largely unknown. We compared the proteolytic activity of the hemorrhagic P-I SVMP BaP1, from the venom of Bothrops asper, and the non-hemorrhagic P-I SVMP leucurolysin-a (leuc-a, from the venom of Bothrops leucurus, on several substrates in vitro and in vivo, focusing on BM proteins. When incubated with Matrigel, a soluble extract of BM, both enzymes hydrolyzed laminin, nidogen and perlecan, albeit BaP1 did it at a faster rate. Type IV collagen was readily digested by BaP1 while leuc-a only induced a slight hydrolysis. Degradation of BM proteins in vivo was studied in mouse gastrocnemius muscle. Western blot analysis of muscle tissue homogenates showed a similar degradation of laminin chains by both enzymes, whereas nidogen was cleaved to a higher extent by BaP1, and perlecan and type IV collagen were readily digested by BaP1 but not by leuc-a. Immunohistochemistry of muscle tissue samples showed a decrease in the immunostaining of type IV collagen after injection of BaP1, but not by leuc-a. Proteomic analysis by LC/MS/MS of exudates collected from injected muscle revealed higher amounts of perlecan, and types VI and XV collagens, in exudates from BaP1-injected tissue. The differences in the hemorrhagic activity of these SVMPs could be explained by their variable ability to degrade key BM and associated ECM substrates in vivo, particularly perlecan and several non-fibrillar collagens, which play a mechanical stabilizing role in microvessel structure. These results underscore the key role played by these ECM components in the mechanical stability of