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Sample records for bothrops neuwiedii diporus

  1. 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

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    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

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

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

    2016-01-01

    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

  3. Cross-neutralization of the coagulant activity of Crotalus durissus terrificus venom from the northeast of Argentina by bivalent bothropic antivenom

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    JP Rodríguez

    2012-01-01

    Full Text Available Cross-neutralization of Crotalus durissus terrificus venom coagulant activity was tested using bivalent horse antivenom against Bothrops alternatus and Bothrops diporus venoms. Our in vitro and in vivo experiments showed that bothropic antivenom neutralizes the thrombin-like activity of crotalic snake venom and this cross-reaction was demonstrated by immunoassays either with whole venom or a purified thrombin-like enzyme. These results suggest common antigenic properties and, consequently, similar molecular structure among venom thrombin-like enzymes. Besides, they provide information that could be further used in the development of new antivenom formulations.

  4. Two related thrombin-like enzymes present in Bothrops atrox venom

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    J.H. Petretski

    2000-11-01

    Full Text Available This article describes the presence of two new forms of a thrombin-like enzyme, both with apparent molecular masses of 38 kDa, in Bothrops atrox venom. Both share the ability to cleave fibrinogen into fibrin and to digest casein. Both present identical Km on the substrate BApNA. Their N-terminal amino acid sequences are identical for 26 residues, sharing 80% homology with batroxobin and flavoxobin. Two groups of monoclonal antibodies (mAbs raised against the purified enzyme forms recognized different epitopes of the putative corresponding enzymes present in B. atrox crude venom. On Western blotting analysis of B. atrox crude venom, mAbs 5DB2C8, 5AA10 and 5CF11, but not mAbs 6CC5 and 6AD2-G5, revealed two or more protein bands ranging from 25 to 38 kDa. By immunoprecipitation assays, the 6AD2-G5 mAb was able to precipitate protein bands of 36-38 kDa from B. atrox, B. leucurus, B. pradoi, B. moojeni, B. jararaca and B. neuwiedii crude venoms. Fibrinogen-clotting activity was inhibited when the same venom specimens were pre-incubated with mAb 6AD2-G5, except for B. jararaca and B. neuwiedii.

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

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    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.

  6. 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. PMID:20621114

  7. 21 CFR 864.8100 - Bothrops atrox reagent.

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    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Bothrops atrox reagent. 864.8100 Section 864.8100...) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Reagents § 864.8100 Bothrops atrox reagent. (a) Identification. A Bothrops atrox reagent is a device made from snake venom and used to determine blood...

  8. Heparin-antivenom association: differential neutralization effectiveness in Bothrops atrox and Bothrops erythromelas envenoming

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    Antônio Luiz R. BOECHAT; Paiva, Cristiano S.; FRANÇA Francisco Oscar; DOS-SANTOS Maria Cristina

    2001-01-01

    Heparin, in some regions of Brazil has been used in the treatment of bothropic accidents, but the data found in the literature are inconclusive about its effectiveness. The venoms of Bothrops atrox and of B. erythromelas were characterized according to their biological activities. The capacity of heparin in neutralizing these activities was tested with doses of 3 and 6 IU in isolated form and associated to Antibothropic Serum (ABS). It was verified that heparin, in doses of 3 and 6 IU, was no...

  9. Trichomoniasis in Bothrops jararaca (serpentes, viperidae)

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    F.C. Vilela; M.G. da Silva; T. H. Barrella; R. J. DA SILVA

    2003-01-01

    We describe a case of trichomoniasis in a Bothrops jararaca (Serpentes, Viperidae) donated to the Center for the Study of Venoms and Venomous Animals - CEVAP/UNESP. The animal had diarrhea with great quantity of flagellated protozoa in the feces. Microscopic examination of fecal smears stained with Giemsa revealed the presence of trichomonads, morphologically similar to Trichomonas acosta. Trichomonads were not detected in fecal exams after treatment with a single dose of 40 mg/kg metronidazo...

  10. Trichomoniasis in Bothrops jararaca (serpentes, viperidae

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    F. C. Vilela

    2003-01-01

    Full Text Available We describe a case of trichomoniasis in a Bothrops jararaca (Serpentes, Viperidae donated to the Center for the Study of Venoms and Venomous Animals - CEVAP/UNESP. The animal had diarrhea with great quantity of flagellated protozoa in the feces. Microscopic examination of fecal smears stained with Giemsa revealed the presence of trichomonads, morphologically similar to Trichomonas acosta. Trichomonads were not detected in fecal exams after treatment with a single dose of 40 mg/kg metronidazole (Flagyl®.

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

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    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

  12. 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

  13. Accidente ofídico causado por Bothrops Asper

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    Galofre-Ruiz Mario David

    2013-01-01

    Introduction: The snake bite of the genus Bothrops is an important cause of ophidic accident in Colombia, with high morbidity and mortality. Clinical case: A case of bite by the snake Bothrops Asper, which was classified as mild grade of poisoning at the entrance of the hospital center is presented. It was managed with antiophidic serum in doses lower than the recommended one, with progression of the symptoms of poisoning. Adjustments in the doses were done and total recovery w...

  14. [Effect of reduction in the Bothrops antivenin dose administrated in patients bitten by the Bothrops snake].

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    Jorge, M T; Ribeiro, L A

    1994-01-01

    Seven hundred and thirty accidents by Bothrops snake bite attended at Hospital Vital Brazil-Instituto Butantan (HVB-IB) were studied: 411 of in 1983/84 (group A) and 319 in 1986 (group B). All the patients were seen in the Hospital 6 hours after the bite. As an average, all the 411 patients of the group A and the 319 patients of the group B received antivenom capable of neutralizing 215 mg and 117 mg of Bothrops jararaca, respectively. Both groups were similar concerning to: sex and age of the patients; anatomic region bitten; frequency of incision and tourniquet use at the site bite; time in between of the accident and the admission at HVB-IB; classification of B. jararaca either young or adult; frequency of the coagulation impairment. There was no difference in the evolution of the local envenoming manifestations and there were no cases of death. PMID:8061698

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

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    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.

  16. First report of hepatic hematoma after presumed Bothrops envenomation

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    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.

  17. Heparin-antivenom association: differential neutralization effectiveness in Bothrops atrox and Bothrops erythromelas envenoming.

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    Boechat, A L; Paiva, C S; França, F O; Dos-Santos, M C

    2001-01-01

    Heparin, in some regions of Brazil has been used in the treatment of bothropic accidents, but the data found in the literature are inconclusive about its effectiveness. The venoms of Bothrops atrox and of B. erythromelas were characterized according to their biological activities. The capacity of heparin in neutralizing these activities was tested with doses of 3 and 6 IU in isolated form and associated to Antibothropic Serum (ABS). It was verified that heparin, in doses of 3 and 6 IU, was not effective in neutralizing the desfibrinating and edema-forming activities of B. atrox venom and the hemorrhagic and coagulant actions of both venoms. Heparin diminished the effectiveness of the ABS in the neutralization of the hemorrhagic and edema-forming activities of the B. atrox venom. However, heparin in the 6 IU dose was capable of neutralize the edema-forming of the B. erythromelas and increase the effectiveness of the ABS. Heparin also neutralized the phospholipasic A2 activity of B. atrox (14.3%) and B. erythromelas (28.0%) venoms. For B. erythromelas venom, the associated treatment, heparin and ABS, was more effective in the neutralization of its lethal activity. PMID:11246276

  18. Heparin-antivenom association: differential neutralization effectiveness in Bothrops atrox and Bothrops erythromelas envenoming

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    BOECHAT Antônio Luiz R.

    2001-01-01

    Full Text Available Heparin, in some regions of Brazil has been used in the treatment of bothropic accidents, but the data found in the literature are inconclusive about its effectiveness. The venoms of Bothrops atrox and of B. erythromelas were characterized according to their biological activities. The capacity of heparin in neutralizing these activities was tested with doses of 3 and 6 IU in isolated form and associated to Antibothropic Serum (ABS. It was verified that heparin, in doses of 3 and 6 IU, was not effective in neutralizing the desfibrinating and edema-forming activities of B. atrox venom and the hemorrhagic and coagulant actions of both venoms. Heparin diminished the effectiveness of the ABS in the neutralization of the hemorrhagic and edema-forming activities of the B. atrox venom. However, heparin in the 6 IU dose was capable of neutralize the edema-forming of the B. erythromelas and increase the effectiveness of the ABS. Heparin also neutralized the phospholipasic A2 activity of B. atrox (14.3% and B. erythromelas (28.0% venoms. For B. erythromelas venom, the associated treatment, heparin and ABS, was more effective in the neutralization of its lethal activity.

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

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    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 <1 year of age, with a change towards bands having lower mol. mass as snakes aged. Gel filtration chromatography showed five peaks in the venoms of B. 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. PMID:14505941

  20. Postprandial thermogenesis in Bothrops moojeni (Serpentes: Viperidae

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    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.

  1. Envenomation by Bothrops atrox in a traveler to Manaus, Brazil.

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    Larréché, Sébastien; Imbert, Patrick; Mornand, Pierre; Andriamanantena, Dina; Mion, Georges; Rapp, Christophe

    2013-01-01

    Snakebites are an infrequent but real risk for travelers. We report a case of envenomation by Bothrops atrox in a traveler to Manaus, Brazil. Rapid administration of specific antivenom prevented the expected systemic disorders. This case gives opportunity to review prevention and first aid measures of snakebites in travelers. PMID:23570867

  2. Brainstem ischemic stroke after to Bothrops atrox snakebite.

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    Cañas, Carlos A

    2016-09-15

    We report case of a 48 years old woman bitten on her right foot by a Bothrops atrox viper. As a result, she developed a severe coagulopathy which improved with application of polyvalent antivenom. Four days after bite she suffered a devastating brainstem ischemic stroke. Possible pathogenetic mechanisms are discussed. PMID:27527269

  3. Neuromuscular activity of Bothrops alcatraz snake venom in chick biventer cervicis preparations.

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    de Moraes, Delkia Seabra; Aparecido de Abreu, Valdemir; Rostelato-Ferreira, Sandro; Leite, Gildo B; Alice da Cruz-Höfling, Maria; Travaglia-Cardoso, Silvia R; Hyslop, Stephen; Rodrigues-Simioni, Léa

    2012-02-01

    Venom (10-100 μg/ml) from Bothrops alcatraz, a pitviper from the Alcatrazes Archipelago off the coast of southeastern Brazil, caused progressive, irreversible neuromuscular blockade in chick isolated biventer cervicis preparations. The venom also inhibited contractures to exogenous ACh (110 μM) and KCl (20 mM), caused myofiber damage and increased creatine kinase release. Commercial bothropic antivenom raised against mainland Bothrops species neutralized the neuromuscular activity, depending on the venom concentration. PMID:22155137

  4. Accidente ofídico causado por Bothrops Asper

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    Galofre-Ruiz Mario David

    2013-12-01

    Full Text Available Introduction: The snake bite of the genus Bothrops is an important cause of ophidic accident in Colombia, with high morbidity and mortality. Clinical case: A case of bite by the snake Bothrops Asper, which was classified as mild grade of poisoning at the entrance of the hospital center is presented. It was managed with antiophidic serum in doses lower than the recommended one, with progression of the symptoms of poisoning. Adjustments in the doses were done and total recovery was reached in five days. Conclusions: Antiophidic serums, named also antivenins, are the cornerstone of the treatment to minimize the local tissue damage and the systemic complications. Rev. cienc.biomed. 2013;4(2:353-357

  5. Bothrops lanceolatus Bites: Guidelines for Severity Assessment and Emergent Management

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    Resiere, Dabor; Mégarbane, Bruno; Valentino, Ruddy; Mehdaoui, Hossein; Thomas, Laurent

    2010-01-01

    Approximately 20-30 declared snakebite cases occurin Martinique each year. Bothrops lanceolatus, a member of the Crotalidae family, is considered to be the only involved snake. B. lanceolatus, commonly named “Fer-de-Lance”, is endemic and only found on this Caribbean island. Envenomation local features include the presence of fang marks, swelling, pain, bleeding from punctures, and ecchymosis. Severe envenomation is associated with multiple systemic thromboses appearing within 48 h of the bit...

  6. Capacidade reprodutiva de Bothrops jararaca (Serpentes, Viperidae Reproductive capacity of Bothrops jararaca (Serpentes, Viperidae

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    Thélia Rosana Forte Janeiro-Cinquini

    Full Text Available The relationship between female body size and egg number was examined in the snake Bothrops jararaca (Wied, 1924. Some authors have demonstrated that the estimated clutch size may be affected by the inclusion or exclusion of dead neonates or non-developed eggs. The reproductive system of 254 mature females newly arrived from nature was examined and 29.55% (n=75 had eggs in the oviducts in different stages of embryonic development. The presence of atresic eggs was observed in 34.5% of those females. A significant difference was observed in the number of eggs in the right and left oviducts (t=9.48, P<0.001, but not in the number of atresic eggs (t=1.110, P=0.278. The correlation of female body size and egg number was greater when all, the normal and atresic eggs were considered (r=0.63, P<0.001. The exclusion of atresic eggs from the clutch size decreased slightly this correlation (r=0.54, P<0.001.

  7. Therapeutic possibilities of Bothrops jararaca in high dilution

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    Eduardo Costa Gaia Nazareth

    2011-09-01

    Full Text Available Introduction: The knowledge and use of the venom of Bothrops jararaca in high dilutions is still quite limited. One of the important properties is the use of one of its components, bradykinin, for the development of antihypertensive medication known as captopril. Other situations, such as clinical, local and systemic should receive more depth to the composition of Materia Medica related to various medical actions on the man and mammals in general. The systemic action of the bite of this snake, includes hemostasis disorders, culminating as bleeding gums, in addition to sweating, hypertension, and hypothermia. The action includes local pain and swelling with bruising, bleeding and often blistering and tissue necrosis. The action on the immune system, through action on the complement C3 and other complement components may show its possible use in cases of bacterial infections, including mycobacteria, as presented in the study of 1970 Vanessa Birdsey, "Interactions of poisons toxic with the addition, "the journal of Immunology 1971. Today, this poison has a toxicology published by Anibal Melgarejo, "Venomous Animals of Brazil", 2003, which subsidizes the development of study for its use in high dilutions, and a comprehensive study of the biology of the animal itself. Published studies on biomolecular analysis add more details about the relations of the poison and mammals. All these characteristics suggest the use of poison as a homeopathic remedy. Objective: To investigate the therapeutic possibilities in high dilutions of the venom of the snake Bothrops jararaca, expanding its clinical use. Methodology: Methodological description of this poison in contemporary bases including: Origin, physical description chemistry, toxicology, pharmacology and medicine in preparation of high dilution, general action, specific actions on systems or organs, sensations, modalities, concomitants, etiological indications relations main clinics. Results: Defining

  8. Assessment of efficacy of bothropic antivenom therapy on microcirculatory effects induced by Bothrops jararaca snake venom.

    Science.gov (United States)

    Battellino, Carolina; Piazza, Roxane; da Silva, Ana M M; Cury, Yara; Farsky, Sandra H P

    2003-04-01

    Intravenous administration of antibothropic antivenom (BAv) neutralises the systemic effects, but does not efficiently reverse the local symptoms elicited by the Bothrops jararaca venom (BjV). The mechanisms involved in this poor protection have not been clarified. In this work, intravital microscopy studies were carried out to determine the efficacy of different schedules of BAv treatment on local effects evoked by topical application of BjV in the microcirculatory network of the internal spermatic fascia of Wistar rats. Results demonstrated that BAv administration 15 min before, simultaneously with, or 15 min after BjV application did not totally reverse the local symptoms, represented by disturbances of coagulation, development of haemorrhage lesions, vascular permeability increase and increment on leukocyte-endothelium interactions. This lack of effectiveness neither reflects an inadequate amount of specific antibodies in the antivenom against toxins responsible for local effects nor an insufficient dose of circulating BAv during the assays. Administration of fluorescein isothiocyanate (FITC) labelled-BAv showed the dynamics of distribution of the antivenom in the microcirculatory network. Images obtained from prior and simultaneously treated animals showed that the antivenom remains at luminal side of vessels before venom application, and the latency time to antivenom leakage is coincidental to that for local effects evoked by the venom. In addition, images from posterior treatment demonstrated that the intense alterations in the microcirculatory network impair antivenom distribution at the site of injection. Together, our data show that the lack of effectiveness of antivenom therapy is due to impaired and delayed venom and antivenom interaction at the site of injury.

  9. Isotopic analysis of Bothrops atrox in Amazonian forest

    Science.gov (United States)

    Martinez, M. G.; Silva, A. M.; Chalkidis, H.; de Oliveira Júnior, R. C.; Camargo, P. B.

    2012-12-01

    The poisoning of snakes is considered a public health problem, especially in populations from rural areas of tropical and subtropical countries. In Brazil, the 26,000 snakebites, 90% are of the genus Bothrops, and Bothrops atrox species predominant in the Amazon region including all the Brazilian Amazon. Research shows that using stable isotopes, we can verify the isotopic composition of tissues of animals that depend mainly on food, water ingested and inhaled gases. For this study, samples taken from Bothrops atrox (B. atrox), in forest using pitfall traps and fall ("Pitt-fall traps with drift fence"). The analyzes were performed by mass spectrometry, where the analytical error is 0.3‰ for carbon and 0.5‰ to nitrogen. The results of the forest animals are significantly different from results of animal vivarium. The average values of the tissues and venoms of snakes of the forest for carbon-13 and nitrogen-15 are: δ13C = -24.68‰ and δ15N = 14.22‰ and mean values of tissue and poisons snakes vivarium (Instituto Butantan) to carbon-13 and nitrogen-15 are δ13C = -20.47‰ and δ15N = 8.36‰, with a significantly different due to different sources of food animals. Based on all results isotopic δ13C and δ15N, we can suggest that changes as the power of the serpent, (nature and captivity), changes occur in relation to diet and environment as the means of the isotopic data are quite distinct, showing that these changes can also cause metabolic changes in the body of the animal itself and the different periods of turnover of each tissue analyzed.

  10. Action of anti-bothropic factor isolated from Didelphis marsupialis on renal effects of Bothrops erythromelas venom.

    Science.gov (United States)

    Martins, Alice M C; Sousa, Fabiola C M; Barbosa, Paulo S F; Toyama, Marcos H; Toyama, Daniela O; Aprígio, Cleidiana C; Queiroz, Maria G R; Guarnieri, Mirian C; Havt, Alexandre; de Menezes, Dalgimar B; Fonteles, Manassés C; Monteiro, Helena S A

    2005-11-01

    Acute renal failure is the most common complication in the lethal cases caused by snakebites in Brazil. Among the Brazilian venom snakes, Bothrops erythromelas is responsible for the majority of accidents in Northeastern Brazil. Didelphis marsupialis serum could inhibit myonecrotic, hemorrhagic, edematogenic hyperalgesic and lethal effects of envenomation determined by ophidian bites. In the present study, we evaluated the action of the anti-bothropic factor isolated from D. marsupialis on the renal effects promoted by B. erythromelas venom without systemic interference. Isolated kidneys from Wistar rats were perfused with Krebs-Henseleit solution containing 6% bovine serum albumin. We analyzed renal perfusion pressure (PP), renal vascular resistance (RVR), glomerular filtration rate (GFR), urinary flow (UF), and the percentages of sodium and potassium tubular transport (%TNa+, %TK+). The B. erythromelas venom (10 microg mL(-1)) decreased the PP (ct = 108.71+/-5.09 mmHg; BE = 65.21+/-5.6 mmHg*) and RVR (ct = 5.76+/-0.65 mmHg mL(-1) g(-1) min(-1); BE = 3.10+/-0.45 mmHg mL(-1) g(-1) min(-1)*). On the other hand, the GFR decreased at 60 min (ct60 = 0.76+/-0.07 mL g(-1) min(-1); BE60 = 0.42+/-0.12 mL g(-1) min(-1)*) and increased at 120 min (ct120 = 0.72+/-0.01 mL g(-1) min(-1); BE120 = 1.24+/-0.26 mL g(-1) min(-1)*). The UF increased significantly when compared with the control group (ct = 0.14+/-0.01 mL g(-1) min(-1); BE = 0.47+/-0.08 mL g(-1) min(-1)*). The venom reduced the %TNa(+) (ct90 = 79.18+/-0.88%; BE90 = 58.35+/-4.86%*) and %TK+ (ct90 = 67.20+/-4.04%; BE90 = 57.32+/-5.26%*) The anti-bothropic factor from D. marsupialis (10 microg mL(-1)) incubated with B. erythromelas venom (10 microg mL(-1)) blocked the effects on PP, RVR, %TNa+, and %TK+, but was not able to reverse the effects in UF and GFR promoted by venom alone. However, the highest concentration of D. marsupialis serum (30 microg mL(-1)) reversed all the renal effects induced by the venom. In

  11. Bothropic antivenom based on monoclonal antibodies, is it possible?

    Science.gov (United States)

    Frauches, Thiago S; Petretski, Jorge H; Arnholdt, Andrea C V; Lasunskaia, Elena B; de Carvalho, Eulógio C Q; Kipnis, Thereza L; da Silva, Wilmar D; Kanashiro, Milton M

    2013-09-01

    Neutralizing monoclonal antibodies against three major toxic components of Bothrops atrox venom were produced and tested. The mAbs against phospholipase A2, hemorrhagic metalloprotease, and thrombin-like enzymes were produced in large amounts and purified with caprylic acid followed by ammonium sulfate precipitation. Purified mAbs were analyzed by SDS-PAGE and their ability to neutralize the respective toxins was tested. Five Swiss mice were injected i.p. with 13.5 mg of pooled mAbs and challenged via s.c. route with venom. Survival rate was recorded for the next 48 h. All mice treated and challenged with venom survived, whereas only one mouse in the control group survived. Bleeding time in mice treated with mAbs was similar to that observed in control mice. Our results show that monoclonal antibodies neutralized the lethal toxicity of Bothrops venom and indicate that there is a reasonable possibility of developing antivenoms based on humanized mAbs to treat victims of venomous animals in the future. PMID:23732123

  12. Comparison of phylogeny, venom composition and neutralization by antivenom in diverse species of bothrops complex.

    Science.gov (United States)

    Sousa, Leijiane F; Nicolau, Carolina A; 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

  13. Comparison of phylogeny, venom composition and neutralization by antivenom in diverse species of bothrops complex.

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    Leijiane F Sousa

    Full Text Available 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

  14. Heparin-antivenom association: differential neutralization effectiveness in Bothrops atrox and Bothrops erythromelas envenoming Associação de heparina e antiveneno: eficácia da neutralização dos venenos de Bothrops atrox e Bothrops erythromelas

    OpenAIRE

    Antônio Luiz R. BOECHAT; Paiva, Cristiano S.; FRANÇA Francisco Oscar; DOS-SANTOS Maria Cristina

    2001-01-01

    Heparin, in some regions of Brazil has been used in the treatment of bothropic accidents, but the data found in the literature are inconclusive about its effectiveness. The venoms of Bothrops atrox and of B. erythromelas were characterized according to their biological activities. The capacity of heparin in neutralizing these activities was tested with doses of 3 and 6 IU in isolated form and associated to Antibothropic Serum (ABS). It was verified that heparin, in doses of 3 and 6 IU, was no...

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

    OpenAIRE

    LJ Martins; PMF de Araújo; Bon, C.; S. HYSLOP; AL de Araújo

    2009-01-01

    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...

  16. ELISA assays for the detection of Bothrops lanceolatus venom in envenomed patient plasmas.

    Science.gov (United States)

    Rodriguez-Acosta, A; Uzcategui, W; Azuaje, R; Giron, M E; Aguilar, I

    1998-01-01

    A double antibody sandwich enzyme linked immunosorbant assay (ELISA) was carried out to detect Bothrops Ianceolatus venom in plasma from envenomed patients at various time intervals (0, 6, 12, 18 and 24 hrs). The test could detect Bothrops lanceolatus levels up to 12 ng/mL of envenomed patient plasmas. Elaboration of an easy, fast and species-diagnostic based on this ELISA technique useful to physicians is discussed. PMID:11845439

  17. Pharmacological characterization and neutralization of the venoms used in the production of Bothropic antivenom in Brazil.

    Science.gov (United States)

    Camey, Kyoko U; Velarde, David T; Sanchez, Eladio F

    2002-05-01

    Some pharmacological effects of the venoms of five different Brazilian Bothrops species and a pool of these venoms (AgB) were quantified. The ability of polyspecific Bothropic antivenom produced at Fundação Ezequiel Dias (FUNED, Brazil) to neutralize the principal toxic and enzymatic activities was studied using in vivo and in vitro assays. The lethality, hemorrhagic, necrotizing, proteolytic, phospholipase, coagulant and fibrinolytic activities were measured for each of these venoms. Comparison of protein electrophoretic patterns showed significant differences such as the presence of common and also unique components. Furthermore, experimental studies revealed differences in their biological properties among individual samples. It was found that the Bothrops antivenom was highly effective in the neutralization of the toxic activities of all venoms tested. In addition, indirect ELISA was used to compare the antigenic cross-reactivity for each of the five Bothrops venoms as well as the venoms of B. atrox, B. leucurus and B. erythromelas which were not included in the antigenic pool (AgB). Therefore, the characterization of several toxic activities of snake venoms is necessary, if toxicity is to be properly evaluated. Results indicate that the Brazilian antivenom prepared at FUNED against Bothrops snakes is effective in neutralizing the main toxic effects of Bothrops venoms. PMID:11821121

  18. Transcriptome analysis of the Amazonian viper Bothrops atrox venom gland using expressed sequence tags (ESTs).

    Science.gov (United States)

    Neiva, Márcia; Arraes, Fabricio B M; de Souza, Jonso Vieira; Rádis-Baptista, Gandhi; Prieto da Silva, Alvaro R B; Walter, Maria Emilia M T; Brigido, Marcelo de Macedo; Yamane, Tetsuo; López-Lozano, Jorge Luiz; Astolfi-Filho, Spartaco

    2009-03-15

    Bothrops atrox is a highly dangerous pit viper in the Brazilian Amazon region. We produced a global catalogue of gene transcripts to identify the main toxin and other protein families present in the B. atrox venom gland. We prepared a directional cDNA library, from which a set of 610 high quality expressed sequence tags (ESTs) were generated by bioinformatics processing. Our data indicated a predominance of transcripts encoding mainly metalloproteinases (59% of the toxins). The expression pattern of the B. atrox venom was similar to Bothrops insularis, Bothrops jararaca and Bothrops jararacussu in terms of toxin type, although some differences were observed. B. atrox showed a higher amount of the PIII class of metalloproteinases which correlates well with the observed intense hemorrhagic action of its toxin. Also, the PLA2 content was the second highest in this sample compared to the other three Bothrops transcriptomes. To our knowledge, this work is the first transcriptome analysis of an Amazonian rain forest pit viper and it will contribute to the body of knowledge regarding the gene diversity of the venom gland of members of the Bothrops genus. Moreover, our results can be used for future studies with other snake species from the Amazon region to investigate differences in gene patterns or phylogenetic relationships. PMID:19708221

  19. Lance-headed viper (Bothrops moojeni bite wounding the eye Acidente botrópico (Bothrops moojeni com ferimento ocular

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    Érico Otaviano Brandão

    1993-08-01

    Full Text Available A 5-year-old girl was bitten in her left eye by a lance-headed viper identified as Bothrops moojeni, measuring 115 cm of length. There was severe facial swelling and left exophthalmus, and enucleation of the eye was necessary. The patient apparently had mild systemic envenoming, but local inflammatory signs and histological evidence of necrosis suggest that both the mechanical trauma and the local action of the venom had a role in the genesis of the eye lesion. It is arguable if the loss of the eye could be prevented even if the antivenom was administered earlier.Uma menina de 5 anos de idade foi picada no olho esquerdo por uma serpente da espécie Bothrops moojeni, que media 115 cm de comprimento. Houve intenso edema facial e ex-oftalmia; enucleação do olho foi necessária. A paciente aparentemente apresentou um envenenamento sistêmico leve, mas os sinais inflamatórios locais e a evidência histológica de necrose sugerem que tanto o traumatismo mecânico como a ação local do veneno tiveram um papel na gênese da lesão ocular. É questionável se o prognóstico da lesão ocular teria sido diferente se o antiveneno tivesse sido administrado em dose adequada mais precocemente.

  20. Clinical and immunological aspects of envenomations by Bothrops snakes

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    KPO Luna

    2011-01-01

    Full Text Available Accidents caused by snakes, especially in tropical and subtropical countries, still constitute a serious public health problem due to the lack of knowledge of health professionals and the precariousness of health systems in the regions where most accidents occur. Snake venoms contain a range of molecules that may provoke local swelling, pain, renal and respiratory insufficiencies. The study of the effects of each molecule on humans can help the development of complementary therapy. Similarly, the knowledge of clinical aspects of envenomations provides a better identification and implementation of appropriate treatment. In addition, to understand Bothrops envenomations and improve the therapeutic strategy, it is necessary to understand and study the role of important inflammatory mediators, particularly nitric oxide (NO, cytokines and the complement system.

  1. Occipital infarction revealed by quadranopsia following snakebite by Bothrops lanceolatus.

    Science.gov (United States)

    Merle, Harold; Donnio, Angélique; Ayeboua, Lucas; Plumelle, Yves; Smadja, Didier; Thomas, Laurent

    2005-09-01

    We report a case of snakebite in which envenomation was manifested through impairment of the visual field. The patient, a 46-year-old man, was bitten on the right thumb by Bothrops lanceolatus. Treatment with a specific equine antivenom (Bothrofav) was administered one hour after the bite. With the exception of fang marks, the results of a clinical examination, particularly the neurologic component, were normal. The day after the bite, the patient developed an inferior left lateral homonymous quadranopsia with macular epargne. T2 magnetic resonance imaging showed a right occipital infarction. His condition improved clinically and biologically. This observation of snakebite is the first in which envenomation was accompanied exclusively by an impairment of the visual field. Envenomation by B. lanceolatus is distinct in its incidence of significant thrombotic complications at a distance from the site of the bite. PMID:16172485

  2. Acidente ofídico em ovinos causado por Bothrops jararaca Snake bite accident in sheep by Bothrops jararaca

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    Carlos Hubinger Tokarnia

    2008-12-01

    Full Text Available Relata-se a ocorrência de um acidente ofídico em um rebanho de ovinos, mestiços da raça Bergamascia, no município de Miguel Pereira, Estado do Rio de Janeiro. Quatro ovelhas adultas, com cerca 35 kg de peso, foram picados na cabeça por um exemplar de Bothrops jararaca dentro de curto espaço de tempo. Os animais apresentaram a cabeça muito tumefeita, mas o estado geral era bom. Um ovino foi eutanasiado. Verificou-se que a tumefação da cabeça era causada por hemorragia embaixo da pele, que se estendia ao pescoço. Histologicamente foi verificada na face, hemorragia maciça, principalmente no tecido subcutâneo, com infiltração de hemácias no tecido muscular adjacente, que em áreas onde era mais intensa, provocou afastamento entre as fibras musculares, algumas com aspecto hialinizado. Discute-se a causa das diferenças sobre a ocorrência de edema/hemorragia e necrose no envenenamento causado pelas diversas serpentes do gênero Bothrops nas diferentes espécies animais.The occurrence of a snake bite accident in a herd of mixed Bergamascia sheep in the county of Miguel Pereira, State of Rio de Janeiro, is reported. Four adult sheep were bitten in the head by Bothrops jararaca within a short period of time. Their head was severely swollen, but the general state of them was good. One of the sheep was euthanasied. Masssive hemorrhages were found beneath the skin, which extended to the neck. Histologically, the subcutaneous hemorrhages infiltrated the adjacent muscles. In areas where the hemorrhages were more intense they caused separation of muscle fibers, some of which had a hyaline aspect. Differences in the occurrence of edema/hemorraghes and necrosis in snake bite accidents, caused by various snake species in different animal species, are discussed.

  3. Genotoxic effect of Bothrops snake venoms and isolated toxins on human lymphocyte DNA.

    Science.gov (United States)

    Marcussi, Silvana; Stábeli, Rodrigo G; Santos-Filho, Norival A; Menaldo, Danilo L; Silva Pereira, Luciana L; Zuliani, Juliana P; Calderon, Leonardo A; da Silva, Saulo L; Antunes, Lusânia M Greggi; Soares, Andreimar M

    2013-04-01

    In the present study, micronucleus with cytokinesis blocking and comet assays were used to evaluate the genotoxic potential of Bothrops jararacussu, Bothrops atrox, Bothrops moojeni, Bothrops alternatus (Rhinocerophis alternatus) and Bothrops brazili snake venoms, and also of some isolated toxins (MjTX-I, BthTX-I and II myotoxins, BjussuMP-II metalloprotease, and BatxLAAO l-amino acid oxidase) on human lymphocytes. Significant DNA damages were observed, indicating genotoxic potential after exposure of the lymphocytes to the toxins BthTX-I, II and BatxLAAO compared to untreated and Cisplatin-treated controls, which were able to induce greater formation of micronuclei. B. brazili, B. jararacussu and B. atrox crude venoms also presented genotoxic potential, and the latter two induced DNA breakage 5 times more often than in normal environmental conditions (control without treatment). B. jararacussu venom and its isolated toxins, as well as an LAAO from B. atrox, were able to cause lymphocyte DNA breakage in the comet test with more than 85% damage levels. The DNA damage evaluation allows a widening of the toxic-pharmacological characterization of snake venoms and their toxins and also contributes to the understanding of the mechanisms of action of these molecules in several human pathologies. PMID:23333649

  4. Cloning, Characterization and Anti-Inflammatory Properties of Bothrops jararaca Snake Antithrombin.

    Science.gov (United States)

    Morais-Zani, Karen de; Grego, Kathleen F; Torquato, Ricardo J S; Silva, Caroline S; Tanaka, Aparecida S; Tanaka-Azevedo, Anita M

    2015-01-01

    Antithrombin inhibits blood coagulation through the interaction with serine proteases in both intrinsic and extrinsic pathways. In addition, antithrombin also shows anti-inflammatory properties, which are independent of its effects on coagulation. This work shows for the first time the cloning and sequencing of antithrombin from a snake species. This predicted protein is composed by 430 amino acids and presents about 64.5% sequence identity to human antithrombin. Biacore experiments revealed that the binding affinity of Bothrops jararaca snake antithrombin to heparin was ~30 times higher than that of human antithrombin. Furthermore, Bothrops jararaca antithrombin is more effective in preventing acute inflammation induced by carrageenan when compared to human antithrombin. Hence, the results showed herein suggest that Bothrops jararaca antithrombin can play a key role in the control of acute inflammation and that this molecule might be used as a pharmacological tool and as a prototype for drug development. PMID:25687119

  5. Hemorrhagic stroke in children caused by Bothrops marajoensis envenoming: a case report.

    Science.gov (United States)

    de Oliveira Pardal, Pedro Pereira; Pinheiro, Augusto Cezar Jennings da Silva; Silva, Cristiane Tarcis Cunha; Santos, Paulo Roberto Silva Garcez; Gadelha, Maria Apolônia da Costa

    2015-01-01

    According to the World Health Organization, snakebites are considered neglected diseases. Bothrops, the genus most frequently implicated in envenomations in Brazil, includes the species B. marajoensis Hoge, 1966, part of the complex B. atrox, which is found in the savannas of Marajó Island, Pará state, Brazil, a region that presents scarce epidemiological data. This work reports the first case of hemorrhagic stroke in a child, attributed to delayed medical care after snakebite envenoming by Bothrops marajoensis in Anajás city, Marajó Island, Pará, Brazil, which led to permanent hemiplegia as a sequela. PMID:26672486

  6. Reproductive Biology of Bothrops erythromelas from the Brazilian Caatinga

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    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.

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

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    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.

  8. Bothrops lanceolatus bites: guidelines for severity assessment and emergent management.

    Science.gov (United States)

    Resiere, Dabor; Mégarbane, Bruno; Valentino, Ruddy; Mehdaoui, Hossein; Thomas, Laurent

    2010-01-01

    Approximately 20-30 declared snakebite cases occurin Martinique each year. Bothrops lanceolatus, a member of the Crotalidae family, is considered to be the only involved snake. B. lanceolatus, commonly named "Fer-de-Lance", is endemic and only found on this Caribbean island. Envenomation local features include the presence of fang marks, swelling, pain, bleeding from punctures, and ecchymosis. Severe envenomation is associated with multiple systemic thromboses appearing within 48 h of the bite and resulting in cerebral, myocardial or pulmonary infarctions. Diagnosis requires first of all identification of the snake. Coagulation tests are helpful to identify thrombocytopenia or disseminated intravascular coagulation. A clinical score based on 4 grades is helpful to assess envonimation severity. A specific monovalent equine anti-venom (Bothrofav(®), Sanofi-Pasteur, France) to neutralize B. lanceolatus venom is available. Its early administration within 6h from the biting in case of progressive local injures, general signs or coagulation disturbances is effective to prevent severe thrombosis and coagulopathy. Its tolerance is considered to be good. Despite an increasing incidence of bites, no deaths have been recently attributed to B. lanceolatus in Martinique, probably due to the currently recommended strategy of early antivenom administration when required. PMID:22069552

  9. 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.

  10. NAA TECHNIQUE FOR CLINICAL INVESTIGATION OF MICE IMMUNIZED WITH BOTHROP VENOM

    International Nuclear Information System (INIS)

    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.

  11. Antigenic cross-reactivity and immunogenicity of Bothrops venoms from snakes of the Amazon region.

    Science.gov (United States)

    Furtado, Maria de Fátima D; Cardoso, Silvia Travaglia; Soares, Oscar Espellet; Pereira, Aparecida Pietro; Fernandes, Daniel Silva; Tambourgi, Denise Vilarinho; Sant'Anna, Osvaldo Augusto

    2010-04-01

    Snakebites are still a critical public health problem in developing countries or isolated areas. In Brazil, the North Region has a high distribution coefficient worsened by the significant number of eventually unreported cases, due to difficulties in access to health services, to the natural geographic barriers and the vast territory. In the Rio Negro area, the species Bothrops atrox, Bothrops brazili, Lachesis muta muta and Bothriopsis taeniata are thought to be the major species responsible for snakebites. The aim of this study was to qualitatively and quantitatively determine the antigenic cross-reactivity and expression of toxins and the immunogenicity of Bothrops venom species of the Amazon and to evaluate the general efficacy of the therapeutic sera. The in vivo assays demonstrated that the defibrinating activity of B. taeniata venom was absent but that the lethal and hemorrhagic properties were more intense than in the B. atrox venom. The results evidence venom variability among the two B. atrox populations from two distinct Amazonian regions, which may reveal a subjacent speciation process. The results point to new aspects that may guide the improvement of anti-Bothropic therapeutic serum. PMID:20036275

  12. Natural resistance of opossum (Didelphis marsupialis) to the mapanare (Bothrops lanceolatus) snake venom.

    Science.gov (United States)

    Pifano, F; Aguilar, I; Giron, M E; Gamboa, N; Rodriguez-Acosta, A

    1993-01-01

    The inactivation of local and general effects of the Mapanare (Bothrops lanceolatus) venom by Opossum's (Didelphis marsupialis) serum fractions was tested using an in vivo assay and an in vitro preincubation experiment. A serum fraction of the Opossum serum has been obtained by immunochemical purification. It is only present in opossum's protective opossum serum fraction (F-0.1). PMID:8186456

  13. Purification and characterization of a hemorrhagic metalloproteinase from Bothrops lanceolatus (Fer-de-lance) snake venom.

    Science.gov (United States)

    Stroka, Alessandra; Donato, José L; Bon, Cassian; Hyslop, Stephen; de Araújo, Albetiza Lôbo

    2005-03-15

    Bothrops snake venoms contain metalloproteinases that contribute to the local effects seen after envenoming. In this work, a hemorrhagic metalloproteinase (BlaH1) was purified from the venom of the snake Bothrops lanceolatus by a combination of gel filtration, affinity (metal chelating) and hydrophobic interaction chromatographies. The hemorrhagin was homogeneous by SDS-PAGE and had a molecular mass of 28 kDa that was unaltered by treatment with beta-mercaptoethanol. BlaH1 gave a single band in immunoelectrophoresis and immunoblotting using commercial bothropic antivenom. BlaH1 had hemorrhagic, caseinolytic, fibrinogenolytic, collagenolytic and elastinolytic activities, but no phospholipase A(2) activity. The hemorrhagic and caseinolytic activities were inhibited by EDTA, indicating that they were metal ion-dependent. In contrast, aprotinin, benzamidine and PMSF did not affect these activities. The caseinolytic activity of BlaH1 had a pH optimum of 8.0 and was stable in solution at up to 40 degrees C; activity was completely lost at > or =70 degrees C. The hemorrhagic activity was neutralized by commercial bothropic antivenom. These properties suggest that this new hemorrhagin belongs to class P-I snake venom metalloproteinases. PMID:15733562

  14. 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. PMID:24950051

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

    International Nuclear Information System (INIS)

    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)

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

    International Nuclear Information System (INIS)

    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)

  17. 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)

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

    International Nuclear Information System (INIS)

    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 105 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)

  19. Tissue damage caused by Bothrops sp envenoming evaluated by magnetic resonance imaging (MRI)

    Energy Technology Data Exchange (ETDEWEB)

    Fonseca, M.G. [Faculdade do Norte Paulista, Bebedouro, SP (Brazil); Matias, M.R.C. [UNESP, Botucatu, SP (Brazil). Hospital Universitario. Unidade de Ressonancia Magnetica; Yamashita, S.; Morceli, J. [UNESP, Botucatu, SP (Brazil). Faculdade de Medicina. Dept. de Doencas Tropicais e Diagnostico por Imagem; Barraviera, B. [Universidade Estadual Paulista (UNESP), Botucatu, SP (Brazil). Centro de Estudos de Venenos e Animais Venenosos (CEVAP)]. E-mail: marilucefonseca@hotmail.com

    2002-07-01

    The objective of this clinical study was to evaluate local tissue damage caused by Bothrops sp envenoming in relation to lesion type and damaged tissues using magnetic resonance imaging (MRI). Fifteen patients bitten by Bothrops snakes were treated at the Emergency Unit of the Tropical Diseases Unit at the University Hospital, Botucatu School of Medicine, UNESP, Sao Paulo State, Brazil. After receiving specific sero therapy, the patients were submitted to MR of the bite site. T 1 spin-echo MRI were obtained revealing the following lesions: edema (n=9), edema associated with hemorrhage (n=5), and hemorrhage (n=1). Peri muscular areas (n=6) and subcutaneous tissues (n=5) were the most affected, followed by muscular tissues (n=4). It is important to mention that MRI did not show myonecrosis of the bite site, a widely reported finding in anatomical and histopathological experimental studies. (author)

  20. Detection of an antibothropic fraction in opossum (Didelphis marsupialis) milk that neutralizes Bothrops jararaca venom.

    Science.gov (United States)

    Jurgilas, P B; Neves-Ferreira, A G; Domont, G B; Moussatché, H; Perales, J

    1999-01-01

    An antibothropic fraction (ABF) from Didelphis marsupialis (opossum) serum, which is responsible for the neutralization of Bothrops jararaca venom was isolated by Perales et al. [Perales, J., Moussatché, H., Marangoni, S., Oliveira, B. and Domont, G. B. (1994). Isolation and partial characterization of an antibothropic complex from the serum of South American Didelphidae. Toxicon 32, 1237-1249]. The aim of this work was to verify the presence of this factor in opossum's milk, which could represent an additional protection for the neonatal opossum against bothropic venoms. An active milk fraction was isolated and showed similar physicochemical, structural, antigenic and biological properties when compared to ABF, indicating that they are probably the same protein. PMID:9920488

  1. The defensive strike of five species of lanceheads of the genus Bothrops (Viperidae).

    Science.gov (United States)

    Araújo, M S; Martins, M

    2007-05-01

    We studied the defensive strike of one species of each of five recognized lineages within the genus Bothrops, namely, B. alternatus, B. jararaca, B. jararacussu, B. moojeni and B. pauloensis. The defensive strike of the studied species was in general similar to that of Crotalus viridis and C. atrox, but some important differences were observed. Bothrops alternatus and B. pauloensis struck preferentially from a tight body posture, whereas B. jararaca and B. moojeni from a loose body posture. Defensive strikes were either true or false (during the latter, the mouth remains closed or partially open). Almost all strikes were successful; only on a few occasions snakes missed their target (flawed strikes). Strike variables were very conservative among the five species, especially strike distance and height, and one possible explanation may be related to constraints imposed on strike variables as a way of increasing strike accuracy. PMID:17876444

  2. 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

    2009-03-01

    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 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

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

    OpenAIRE

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

    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 chro...

  4. Cell adhesion molecules involved in the leukocyte recruitment induced by venom of the snake Bothrops jararaca

    OpenAIRE

    Catarina F. P. Teixeira; Stella R. Zamuner

    2002-01-01

    It has been shown that Bothrops jararaca venom (BjV) induces a significant leukocyte accumulation, mainly neutrophils, at the local of tissue damage. Therefore, the role of the adhesion molecules intercellular adhesion molecule-1 (ICAM-1), LECAM-1, CD18, leukocyte function-associated antigen-1 (LFA-1) and platelet endothelial cell adhesion molecule-1 (PECAM-1) on the BjV-induced neutrophil accumulation and the correlation with release of LTB4, TXA2, tumor necrosis factor-alpha, interleukin (I...

  5. Genetic relationships among species of the genus Bothrops based on RAPD markers

    Directory of Open Access Journals (Sweden)

    Felipe Grazziotin

    2005-05-01

    Full Text Available The genetic relationships among 11 species of Bothrops found in Brazil were determined using random amplified fragment (RAPD information. A total of 239 amplified bands were scored using 20 aleatory primers. Most of the bands (88.7% were polymorphic. Phylogenetic analysis of the data determined three similarity groups within Bothrops: Group I- B. alternatus, B. neuwiedi, B. cotiara, and B. jararacusu; Group II- B. insularis, B. jararaca, and B. erythromelas; and Group III- B. moojeni, B. leucurus, and B. atrox. These groups coincided with those obtained by other methods, indicating that RAPD's could be a useful tool for the evaluation of genetic relationships at the interspecific level.As relações genéticas entre 11 espécies de Bothrops (sensus stricto de ocorrência no Brasil foram determinadas através da comparação dos perfis de fragmentos amplificados aleatoriamente (RAPD. Um total de 239 bandas foram obtidas utilizando 20 sequencias iniciadoras (primers. A maior parte das bandas (88,7% foram polimórficas. A análise filogenética dos dados permitiu a identificação de três grupos dentro do gênero Bothrops: Grupo 1- B. alternatus, B. neuwiedi, B. cotiara, e B. jararacusu; Grupo II- B. insularis, B. jararaca, e B. erythromelas; e Grupo III- B. moojeni, B. leucurus, e B. atrox. Estes grupos coincidem com aqueles obtidos por outros métodos indicando que os marcadores de RAPD podem ser considerados ferramentas úteis para a avaliação de relações a nível interespecífico.

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

    OpenAIRE

    Sousa Marcelo V; Paba Jaime; López-Lozano Jorge L; Shevchenko Andrej; Shevchenko Anna; Guércio Rafael AP; Ricart Carlos AO

    2006-01-01

    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 (...

  7. Neutralizing capacity of a new monovalent anti-Bothrops atrox antivenom: comparison with two commercial antivenoms

    OpenAIRE

    Otero, R; V. Núñez; Gutiérrez, J M; Robles, A; R. Estrada; R.G. Osorio; G. Del-Valle; R. Valderrama; Giraldo, C. A.

    1997-01-01

    Three horse-derived antivenoms were tested for their ability to neutralize lethal, hemorrhagic, edema-forming, defibrinating and myotoxic activities induced by the venom of Bothrops atrox from Antioquia and Chocó (Colombia). The following antivenoms were used: a) polyvalent (crotaline) antivenom produced by Instituto Clodomiro Picado (Costa Rica), b) monovalent antibothropic antivenom produced by Instituto Nacional de Salud-INS (Bogotá), and c) a new monovalent anti-B. atrox antivenom produce...

  8. 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)

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

    Directory of Open Access Journals (Sweden)

    P. C. Lucas de Oliveira

    2007-01-01

    Full Text Available 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 60Co 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 60Co source. Twelve goats were divided into two groups of six animals: GINV, inoculated with 0.5mg/kg of natural venom; and GIIV, inoculated with 0.5mg/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 (CK, aspartate amino-transferase (AST and alanine amino-transferase (ALT; 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.

  10. 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.

  11. Intradermal injection of Bothrops cotiara venom in mice in an experimental wound model

    Directory of Open Access Journals (Sweden)

    JA Lopes

    2012-01-01

    Full Text Available Bothropic envenomation induces hemorrhage, coagulant disturbances and necrosis. Regarding therapies against the local damage caused by the venom, there is little information on tissue changes until the complete healing. In the current study, local damage was evaluated by examination of morphological inflammatory alterations, mast cell count, and analysis of collagen deposition. Bleeding was evident four hours after inoculation. After 24 hours, a large area of injury appeared presenting disorganized tissue, significant hemorrhage and acute inflammation. After three days, the damaged area was extensive, with a large amount of inflammatory cells and the presence of scab. In seven days, healing and reepithelization process started. And, 21 days later, the epithelium showed less infiltration and no skin appendages. The number of mast cells was similar to control after four hours, with a drop of 50% at 24 hours, followed by an increase until the 21st day. No differences of collagen deposition were observed among experimental groups. Taken together, wound healing after intradermal injection of Bothrops cotiara venom in mice follows similar parameters to wounds caused by other bothropic venoms. The present work reveals the importance of experimental wound models to the study of neutralizing agents against venom toxins.

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

    Energy Technology Data Exchange (ETDEWEB)

    Ferreira Junior, R.S.; Meira, D.A. [UNESP, Botucatu, SP (Brazil). Dept. de Doencas Tropicais; Barraviera, B. [UNESP, Botucatu, SP (Brazil). Center for the Study of Venoms and Venomous Animals - CEVAP; Nascimento, N.; Alves, J.B. [Instituto de Pesquisas Energeticas (IPEN/CNEN-SP), Sao Paulo, SP (Brazil); Martinez, J.C. [UNESP, Botucatu, SP (Brazil). Escola de Engenharia]. E-mail: rseabra@cevap.org.br

    2005-10-15

    ELISA was used to evaluate, accompany, and compare the humoral immune response of Swiss mice during hyperimmunization with native and Cobalt-60-irradiated ({sup 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 {sup 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)

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

    International Nuclear Information System (INIS)

    ELISA was used to evaluate, accompany, and compare the humoral immune response of Swiss mice during hyperimmunization with native and Cobalt-60-irradiated (60Co) 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)

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

    International Nuclear Information System (INIS)

    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 60Co 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 60Co 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)

  15. Nomenclatural instability in the venomous snakes of the Bothrops complex: Implications in toxinology and public health.

    Science.gov (United States)

    Carrasco, Paola Andrea; Venegas, Pablo Javier; Chaparro, Juan Carlos; Scrocchi, Gustavo José

    2016-09-01

    Since nomenclature is intended to reflect the evolutionary history of organisms, advances in our understanding of historical relationships may lead to changes in classification, and thus potentially in taxonomic instability. An unstable nomenclature for medically important animals like venomous snakes is of concern, and its implications in venom/antivenom research and snakebite treatment have been extensively discussed since the 90´s. The taxonomy of the pitvipers of the Bothrops complex has been historically problematic and different genus-level rearrangements were proposed to rectify the long-standing paraphyly of the group. Here we review the toxinological literature on the Bothrops complex to estimate the impact of recent proposals of classification in non-systematic research. This assessment revealed moderate levels of nomenclatural instability in the last five years, and the recurrence of some practices discussed in previous studies regarding the use of classifications and the information provided about the origin of venom samples. We briefly comment on a few examples and the implications of different proposals of classifications for the Bothrops complex. The aim of this review is to contribute to the reduction of adverse effects of current taxonomic instability in a group of medical importance in the Americas. PMID:27242040

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

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    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

  17. 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. PMID:16360723

  18. Acidente vascular cerebral hemorrágico associado à acidente ofídico por serpente do gênero bothrops: relato de caso Hemorrhagic stroke related to snakebite by bothrops genus: a case report

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    Amanda Silva Machado

    2010-10-01

    Full Text Available Este trabalho tem como objetivo relatar um caso de acidente vascular cerebral hemorrágico, associado à acidente ofídico por serpente do gênero bothrops e hipertensão arterial sistêmica grave. Apesar do ofidismo botrópico ser frequente no Estado do Pará, tais associações são incomuns, necessitando de uma abordagem especializada e precoce, visando menores complicações.This research reports a clinical case of hemorrhagic stroke due to envenomation by bothrops snakebite associated with severe hypertension. Although bothrops snakebites are frequent in the State of Pará, such associations are uncommon, requiring specialized and early management to avoid severe complications.

  19. [Pulmonary embolism and disseminated intravascular coagulation after being bitten by a Bothrops lanceolatus snake. Apropos of a case].

    Science.gov (United States)

    Estrade, G; Garnier, D; Bernasconi, F; Donatien, Y

    1989-11-01

    The authors report the case of a Bothrops lanceolatus snake bite complicated by severe pulmonary embolism a few hours after admission. This thromboembolic complication developed despite heparin therapy and was followed by disseminated intravascular coagulation (DIC). Vascular thrombosis and pulmonary embolism are rare after Bothrops lanceolatus snake bite as patients are usually hypocoagulable due to DIC. In this case, the thromboembolism was probably caused by the procoagulant effect of the thrombin-like enzymes of the snake venom which may have been injected directly into the vein of a young woman taking a contraceptive pill. A specific antivenin which has recently become available fort treatment may decrease the complications of Bothrops lanceolatus snake bite. PMID:2514645

  20. The role of TLR2 in the acute inflammatory response induced by Bothrops atrox snake venom.

    Science.gov (United States)

    Moreira, Vanessa; Teixeira, Catarina; Borges da Silva, Henrique; D'Império Lima, Maria Regina; Dos-Santos, Maria Cristina

    2016-08-01

    Envenomation by snakes of the species Bothrops atrox induces local and systemic effects. Local effects include drastic tissue damage and a marked inflammatory response as a result of the synthesis and release of a variety of protein and lipid mediators. Toll-like receptor (TLR) signaling pathways can play an important role in this response, leading to synthesis of these inflammatory mediators. This study investigated the influence of TLR2 on the acute inflammatory response induced by Bothrops atrox venom. Wild-type C57BL/6 mice (WT) and TLR2 gene knockout mice (TLR2(-/-)) were injected with Bothrops atrox venom (BaV), and the following responses to the venom were assessed in peritoneal exudate: leukocyte accumulation; release of mediators, including CCL-2, IL-10, IL-1β, IL-6 and LTB4; protein expression of COX-1 and COX-2; and quantification of their products PGE2 and TXA2. After injection with BaV, the TLR2(-/-) mice (TLR2(-/-)BaV) had higher levels of IL-6 and CCL-2 than WT animals kept under the same conditions (WTBaV), together with an accumulation of polymorphonuclear leukocytes (PMNs), inhibition of IL-1β and LTB4 and reduced mononuclear leukocyte influx. However, no significant differences in COX-2 protein expression or PGE2, TXA2 and IL-10 production between the TLR2(-/-)BaV and WTBav animals were observed. Together, these results indicate that the signaling pathway activated by TLR2 acts by modulating the induced inflammatory response to BaV through the direct action of venom-associated molecular patterns (VAMPs) or indirectly by forming damage-associated molecular patterns (DAMPs) and that this may have important therapeutic implications. PMID:27109323

  1. Pharmacological characterization of the rat paw edema induced by Bothrops lanceolatus (Fer de lance) venom.

    Science.gov (United States)

    de Faria L; Antunes, E; Bon, C; de Araújo, A L

    2001-06-01

    The inflammatory response induced by Bothrops lanceolatus venom (BLV) in the rat hind-paw was studied measuring paw edema. Non-heated BLV (75microg/paw) caused a marked paw edema accompanied by intense haemorrhage whereas heated venom (97 degrees C, 30s; 12.5-100microg/paw) produced a dose- and time-dependent non-haemorrhagic edema. The response with heated BLV was maximal within 15min disappearing over 24h. Heated venom was then routinely used at the dose of 75microg/paw. The prostacyclin analogue iloprost (0.1microg/paw) potentiated by 125% the venom-induced edema. The histamine H(1) receptor antagonist mepyramine (6mg/kg) or the serotonin/histamine receptor antagonist cyproheptadine (6mg/kg) partially inhibited BLV-induced edema whereas the combination of both compounds virtually abolished the edema. The lipoxygenase inhibitor BWA4C (10mg/kg), but not the cyclooxygenase inhibitor indomethacin (10mg/kg), significantly inhibited the edema (35% reduction; P<0.05). Dexamethasone (1mg/kg) also markedly (P<0.001) reduced venom-induced edema. The bradykinin B(2) receptor antagonist Hoe 140 (0.6mg/kg) reduced by 30% (P<0.05) the venom induced edema, whereas the angiotensin-converting enzyme inhibitor captopril (300microg/paw) potentiated by 42% (P<0.05) the edema. Bothrops lanceolatus antivenon (anti-BLV) reduced by 28% (P<0.05) the venom-induced edema while intravenous administration of antivenom failed to affect the edema. In conclusion, BLV-induced rat paw edema involves mast cell degranulation causing local release of histamine and serotonin, a phenomenon mediated mainly by kinins and lipoxygenase metabolites. Additionally, the use of a specific Bothrops lanceolatus antivenom, given subplantarily or intravenously, revealed to be little effective to prevent BLV-induced edema. PMID:11137542

  2. 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.

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

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    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.

  4. Two related thrombin-like enzymes present in Bothrops atrox venom

    OpenAIRE

    J.H. Petretski; Kanashiro, M; Silva, C. P.; E.W. Alves; Kipnis, T L

    2000-01-01

    This article describes the presence of two new forms of a thrombin-like enzyme, both with apparent molecular masses of 38 kDa, in Bothrops atrox venom. Both share the ability to cleave fibrinogen into fibrin and to digest casein. Both present identical Km on the substrate BApNA. Their N-terminal amino acid sequences are identical for 26 residues, sharing 80% homology with batroxobin and flavoxobin. Two groups of monoclonal antibodies (mAbs) raised against the purified enzyme forms recognized ...

  5. Estudo bacteriológico de abscessos causados por picada de serpentes do genero bothrops Bacterial flora of abscesses following Bothrops snakebite

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    João guimarães De Andrade

    1989-12-01

    Full Text Available Foi analisada a flora bacteriana de 99 abscessos causados por picadas de serpentes do gênero Bothrops, correspondendo a 61,1% dos casos que ocorreram em 1030 acidentes ofídicos atendidos no Hospital de Doenças Tropiciais (HDT de Goiânia, no período de janeiro de 1984 a abril de 1988. O exsudato dos abscessos foi estudado através de bacterioscopia, cultura e testes de sensibilidade para aeróbios. Os bacilos Gram negativos foram isolados em maior frequência, destacando-se a Morganella morganii, Escherichia coli e Providencia sp presentes respectivamente em 44,4%, 20,2% e 13,1% das amostras. Esta flora aeróbica foi semelhante à encontrada na cavidade oral e no veneno das serpentes em outros estudos, nos quais predominaram Morganella morganii. Foi sugerido o uso do cloranfenicol no tratamento dos abscessos que não respondam à simples drenagem, face à grande sensibilidade destes microorganismos demonstrada nos testes "in vitro".The bacterial flora of 99 cases of abscesses following Bothrops snakebite were analysed. They corresponded to 61.1% of all snakebite abs cesses observed in 1030 patients attending the Hospital de Doenças Tropicais de Goiânia in Goiás, Brazil, from January 1984 to April 1988. An exsudate sample of each abscess was examined by Gram stain, culture and susceptibility tests. The Gram negative bacillis, Morganella morganii, Escherichia coli and Providencia sp were the most frequent bacterias isolated. They were identified in 44.4%, 20.2% and 13.1% of the samples respectively. This flora was similar to those described in snake mouth and venom by other researchers. Based on the results of the susceptibility tests the authors suggested the use of chloramphenicol for the treatment of those abscesses which do not respond to simple drainage.

  6. Fatores associados à incoagulabilidade sangüínea no envenenamento por serpentes do gênero Bothrops Risk factors associated with coagulation abnormalities in Bothrops envenoming

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    Ricardo Borges de Oliveira

    2003-12-01

    Full Text Available Com o objetivo de conhecer fatores associados à incoagulabilidade sangüínea no envenenamento botrópico, foram obtidas informações de 2.991 prontuários médicos de pacientes atendidos no Instituto Butantan de 1981 a 1990. Associaram-se positivamente à incoagulabilidade sangüínea (p0,05: horário do acidente; presença de presa recém-deglutida no tubo digestivo da serpente; sexo e idade do paciente; ocorrência de bolha, necrose, abscesso e incisão local, amputação, insuficiência renal e óbito. Pode-se concluir que, embora a incoagulabilidade sangüínea apresente associação com manifestações precoces do envenenamento, não tem boa associação com a evolução clínica do paciente.This study aimed at assessing, in the envenoming by Bothrops, factors that are associated with blood incoagulability. Information was obtained from the charts of 2,991 patients admitted to Instituto Butantan, from 1981 to 1990. Factors positively associated with blood incoagulability (p0.05 were: time of the bite; presence of recently swallowed prey in the snake gut; gender and age of the patient; blister, necrosis, and abscess at the bite site; occurrence of amputation, renal failure and death; presence of an incision at the bite site. We conclude that although blood incoagulability is associated with early manifestations of Bothrops envenoming, it is not associated with the clinical outcome.

  7. Snake venomics of the Lesser Antillean pit vipers Bothrops caribbaeus and Bothrops lanceolatus: correlation with toxicological activities and immunoreactivity of a heterologous antivenom.

    Science.gov (United States)

    Gutiérrez, José María; Sanz, Libia; Escolano, José; Fernández, Julián; Lomonte, Bruno; Angulo, Yamileth; Rucavado, Alexandra; Warrell, David A; Calvete, Juan J

    2008-10-01

    The venom proteomes of the snakes Bothrops caribbaeus and Bothrops lanceolatus, endemic to the Lesser Antillean islands of Saint Lucia and Martinique, respectively, were 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 venoms contain proteins belonging to seven ( B. caribbaeus) and five ( B. lanceolatus) types of toxins. B. caribbaeus and B. lanceolatus venoms contain phospholipases A 2, serine proteinases, l-amino acid oxidases and zinc-dependent metalloproteinases, whereas a long disintegrin, DC-fragments and a CRISP molecule were present only in the venom of B. caribbaeus, and a C-type lectin-like molecule was characterized in the venom of B. lanceolatus. Compositional differences between venoms among closely related species from different geographic regions may be due to evolutionary environmental pressure acting on isolated populations. The venoms of these two species differed in the composition and the relative abundance of their component toxins, but they exhibited similar toxicological and enzymatic profiles in mice, characterized by lethal, hemorrhagic, edema-forming, phospholipase A 2 and proteolytic activities. The venoms of B. caribbaeus and B. lanceolatus are devoid of coagulant and defibrinogenating effects and induce only mild local myotoxicity in mice. The characteristic thrombotic effect described in human envenomings by these species was not reproduced in the mouse model. The toxicological profile observed is consistent with the abundance of metalloproteinases, PLA 2s and serine proteinases in the venoms. A polyvalent (Crotalinae) antivenom produced in Costa Rica was able to immunodeplete approximately 80% of the proteins from both B. caribbaeus and B. lanceolatus venoms, and was effective in neutralizing the lethal, hemorrhagic, phospholipase

  8. Biological and immunological properties of the venom of Bothrops alcatraz, an endemic species of pitviper from Brazil.

    Science.gov (United States)

    Furtado, M F D

    2005-06-01

    Bothrops alcatraz is a new pitviper species derived from the Bothrops jararaca group, whose natural habitat is situated in Alcatrazes Archipelago, a group of marine islands near São Paulo State coast in Brazil. Herein, the biological and biochemical properties of venoms of four adult specimens of B. alcatraz were examined comparatively to a reference pool of Bothrops jararaca venom. Both venoms showed similar activities and electrophoretic patterns, but B. alcatraz venom showed three protein bands of molecular masses of 97, 80 and 38 kDa that were not present in B. jararaca reference venom. The i.p. median lethal dose of B. alcatraz venom ranged from 5.1 to 6.6 mg/kg, while it was 1.5 mg/kg for B. jararaca venom. The minimum hemorrhagic dose of B. jararaca venom was 0.63, whereas 2.28 mug/mouse for B. alcatraz venom. In contrast, B. alcatraz venom was more potent in regard to procoagulant and proteolytic activities. These differences were supported by western blotting and neutralization tests, employing commercial bothropic antivenom, which showed that hemorrhagic and lethal activities of B. alcatraz venom were less effectively inhibited than B. jararaca venom. Such results evidence that B. alcatraz shows quantitative and qualitative differences in venom composition in comparison with its B. jararaca relatives, which might represent an optimization of venom towards a specialized diet. PMID:16002343

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

    International Nuclear Information System (INIS)

    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

  10. Purification and biological effects of a C-type lectin isolated from Bothrops moojeni

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    PSF Barbosa

    2010-01-01

    Full Text Available Snake venom proteins from the C-type lectin family have very distinct biological activities despite their highly conserved primary structure, which is homologous to the carbohydrate recognition region of true C-type lectins. We purified a lectin-like protein (BmLec from Bothrops moojeni venom and investigated its effect on platelet aggregation, insulin secretion, antibacterial activity, and isolated kidney cells. The BmLec was purified using two chromatographic steps: affinity chromatography and reverse phase high performance liquid chromatography (HPLC. BmLec showed a dose-dependent platelet aggregation and significantly decreased the bacterial growth rate in approximately 15%. During scanning electron microscopy, the profile of Xanthomonas axonopodis pv. passiflorae treated with lectin disclosed a high vesiculation and membrane rupture. BmLec induced a strong and significant increase in insulin secretion at 2.8 and 16.7 mM glucose concentrations, and this effect was seen in the presence of EGTA in both experiments. BmLec (10 µg/mL increased the perfusion pressure, renal vascular resistance and urinary flow. The glomerular filtration rate and percentages of sodium, potassium and chloride tubular transport were reduced at 60 minutes of perfusion. Renal alterations caused by BmLec were completely inhibited by indomethacin in all evaluated parameters. In conclusion, the C-type lectin isolated from Bothrops moojeni affected platelet aggregation, insulin secretion, antibacterial activity and isolated kidney function.

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

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    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.

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

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    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.

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

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    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.

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

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    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.

  15. 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; San Feliciano, Arturo; Oshima-Franco, Yoko

    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

  16. Purification and functional characterization of two fibrinogenolytic enzymes from Bothrops alternatus venom

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    J. O. Costa

    2007-01-01

    Full Text Available Two fibrinogenolytic enzymes, Bothrops alternatus metalloprotease isoform (BaltMP-I and II, were purified from Bothrops alternatus venom using Diethylaminoethyl (DEAE Sephacel, Sephadex G-75 and Heparin-Agarose column chromatography. Purified BaltMP-I and II ran as single protein bands on analytical polyacrylamide gel electrophoresis and showed molecular weights of 29000 and 36000, respectively, under reducing conditions in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE. BaltMP-II, but not BaltMP-I, displayed blood-clotting activity in bovine plasma, which was about 10-fold higher than that of the crude venom. Both enzymes were proteolytically active against bovine fibrinogen as substrate. When fibrinogen and each enzyme were incubated at 37°C, at a ratio of 1:100 (w/w, BaltMP-II cleaved preferentially the Aalpha -chain and more slowly the Bbeta -chain. The action of BaltMP-I was similar, but lower. None of the proteases degraded the gamma-chain of fibrinogen. The fibrinogenolytic activity of the enzymes was inhibited by 1,10-phenanthroline, suggesting they are metalloproteases. Since both enzymes were found to cause defibrinogenation when intraperitoneally (i.p. administered to mice, they can be of medical interest as a therapeutic agent in the treatment and prevention of arterial thrombosis.

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

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    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.

  18. 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

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    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.

  19. Purificación de fosfolipas A2 a partir de veneno de Bothrops atrox

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    Quevedo B.

    1999-06-01

    Full Text Available La fosfolipasa A2 (PLA2 fue purificada a partir de veneno de serpiente de Bothrops atrox (Sensu lato de Chiriguaná (Colombia por cromatografía de exclusión sobre Sephadex G-75, de donde se obtuvieron cinco fracciones, una de ellas con actividad fosfolipasa A2. Después de pasarla por una columna de intercambio catiónico Mono S, donde se separaron ocho fracciones con activi-dad PLA2, ésta se midió usando el método hemolítico. La electroforesis en gel de poliacrilamida en condiciones desnaturalizantes, mostró pesos moleculares entre 16.000 y 17.000 para las ocho fracciones, seis de las cuales pre-sentaron una sola banda. 

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

    International Nuclear Information System (INIS)

    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. A2, 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 A2 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

  1. Neutralizing capacity of a new monovalent anti-Bothrops atrox antivenom: comparison with two commercial antivenoms

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    R. Otero

    1997-03-01

    Full Text Available Three horse-derived antivenoms were tested for their ability to neutralize lethal, hemorrhagic, edema-forming, defibrinating and myotoxic activities induced by the venom of Bothrops atrox from Antioquia and Chocó (Colombia. The following antivenoms were used: a polyvalent (crotaline antivenom produced by Instituto Clodomiro Picado (Costa Rica, b monovalent antibothropic antivenom produced by Instituto Nacional de Salud-INS (Bogotá, and c a new monovalent anti-B. atrox antivenom produced with the venom of B. atrox from Antioquia and Chocó. The three antivenoms neutralized all toxic activities tested albeit with different potencies. The new monovalent anti-B. atrox antivenom showed the highest neutralizing ability against edema-forming and defibrinating effects of B. atrox venom (41 ± 2 and 100 ± 32 µl antivenom/mg venom, respectively, suggesting that it should be useful in the treatment of B. atrox envenomation in Antioquia and Chocó

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

    OpenAIRE

    Barros, S F; I. Friedlanskaia; V. L. Petricevich; Kipnis, T L

    1998-01-01

    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); (b)BALB/c mice (LD50=100.0 microg) were injected i.p. with 50 microg of venom produced IL-6, IL-10, INF-gamma, TNF-alpha 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-gamma and less amounts of IL-6; (c) e...

  3. Thrombocytin, a serine protease from Bothrops atrox venom. 1. Purification and characterization of the enzyme

    Energy Technology Data Exchange (ETDEWEB)

    Kirby, E.P. (Temple Univ. Health Sciences Center, Philadelphia, PA); Niewiarowski, S.; Stocker, K.; Kettner, C.; Shaw, E.; Brudzynsi, T.M.

    1979-08-07

    Thrombocytin, a platelet-activating enzyme from Bothrops atrox venom, has been purified to homogeneity by precipitation with sodium salicylate and chromatography on heparin-agarose. Thrombocytin is a single-chain glycoprotein with a molecular weight of 36,000 which contains 5.6% carbohydrate. It causes platelet aggregation, release of platelet serotonin, and activation of factor XIII. The most sensitive substrate for the amidolytic activity of thrombocytin was Tos-Gly-Pro-Arg-p-nitroanilide hydrochloride. The activity of thrombocytin on this substrate and on platelets was inhibited by diisopropyl fluorophosphate (DFP), soybean trypsin inhibitor, and several arginine chloromethyl ketones. Active site titration with nitrophenyl guanidinobenzoate demonstrated that approximately 86% of the preparation was in the active form. These experiments demonstrate the presence of serine and histidine in the active site of thrombocytin and suggest that thrombocytin is a classical serine protease with a platelet-activating activity similar to thrombin.

  4. Local and systemic biochemical alterations induced by Bothrops atrox snake venom in mice.

    Science.gov (United States)

    de Souza, Carlos At; Kayano, Anderson M; Setúbal, Sulamita S; Pontes, Adriana S; Furtado, Juliana L; Kwasniewski, Fábio H; Zaqueo, Kayena D; Soares, Andreimar M; Stábeli, Rodrigo G; Zuliani, Juliana P

    2012-01-01

    The local and systemic alterations induced by Bothrops atrox snake venom (BaV) injection in mice were studied. BaV induced superoxide production by migrated neutrophils, mast cell degranulation and phagocytosis by macrophages. Moreover, BaV caused hemorrhage in dorsum of mice after 2hr post- injection. Three hours post-injection in gastrocnemius muscle, we also observed myonecrosis, which was assessed by the determination of serum and tissue CK besides the release of urea, but not creatinine and uric acid, indicating kidney alterations. BaV also induced the release of LDH and transaminases (ALT and AST) indicating tissue and liver abnormalities. In conclusion, the data indicate that BaV induces events of local and systemic importance. PMID:23487552

  5. Glial cells of the central nervous system of Bothrops jararaca (Reptilia, Ofidae: an ultrastructural study

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    Eduardo F. Bondan

    2015-07-01

    Full Text Available Abstract Although ultrastructural characteristics of mature neuroglia in the central nervous system (CNS are very well described in mammals, much less is known in reptiles, especially serpents. In this context, two specimens of Bothrops jararaca were euthanized for morphological analysis of CNS glial cells. Samples from telencephalon, mesencephalon and spinal cord were collected and processed for light and transmission electron microscopy investigation. Astrocytes, oligodendrocytes, microglial cells and ependymal cells, as well as myelin sheaths, presented similar ultrastructural features to those already observed in mammals and tended to maintain their general aspect all over the distinct CNS regions observed. Morphological similarities between reptilian and mammalian glia are probably linked to their evolutionary conservation throughout vertebrate phylogeny.

  6. Phylogenetic analysis of β-defensin-like genes of Bothrops, Crotalus and Lachesis snakes.

    Science.gov (United States)

    Correa, Poliana G; Oguiura, Nancy

    2013-07-01

    Defensins are components of the vertebrate innate immune system; they comprise a diverse group of small cationic antimicrobial peptides. Among them, β-defensins have a characteristic β-sheet-rich fold plus six conserved cysteines with particular spacing and intramolecular bonds. They have been fully studied in mammals, but there is little information about them in snakes. Using a PCR approach, we described 13 β-defensin-like sequences in Bothrops and Lachesis snakes. The genes are organized in three exons and two introns, with exception of B.atrox_defensinB_01 which has only two exons. They show high similarities in exon 1, intron 1 and intron 2, but exons 2 and 3 have undergone accelerated evolution. The theoretical translated sequences encode a pre-β-defensin-like molecule with a conserved signal peptide and a mature peptide. The signal peptides are leucine-rich and the mature β-defensin-like molecules have a size around 4.5 kDa, a net charge from +2 to +11, and the conserved cysteine motif. Phylogenetic analysis was done using maximum parsimony, maximum likelihood and Bayesian analyses, and all resulted in similar topologies with slight differences. The genus Bothrops displayed two separate lineages. The reconciliation of gene trees and species tree indicated eight to nine duplications and 23 to 29 extinctions depending on the gene tree used. Our results together with previously published data indicate that the ancestral β-defensin-like gene may have three exons in vertebrates and that their evolution occurred according to a birth-and-death model. PMID:23500066

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

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    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.

  8. Comparative evaluation of adverse effects in the use of powder trivalent antivenom and liquid antivenoms in Bothrops snake bites

    OpenAIRE

    Iran Mendonça da Silva; Antônio Magela Tavares

    2012-01-01

    INTRODUCTION: Snake bite, a problem in public health, generally occurs where there is no electric power. METHODS: A comparative clinical study was conducted with 102 victims of Bothrops snake bite, from the state of Amazonas, Brazil; 58 victims were treated with liofilizated trivalent antivenom serum (SATL) and 44 victims treated with liquid bivalent and monovalent antivenom serum (SAMBL). RESULTS: 17% (10/58) of patients presented adverse effects with the SATL and 25% (11/44) with the SAMBL....

  9. Phylogeography of the Bothrops jararaca complex (Serpentes: Viperidae): past fragmentation and island colonization in the Brazilian Atlantic Forest.

    Science.gov (United States)

    Grazziotin, Felipe G; Monzel, Markus; Echeverrigaray, Sergio; Bonatto, Sandro L

    2006-11-01

    The Brazilian Atlantic Forest is one of the world's major biodiversity hotspots and is threatened by a severe habitat loss. Yet little is known about the processes that originated its remarkable richness of endemic species. Here we present results of a large-scale survey of the genetic variation at the mitochondrial cytochrome b gene of the pitviper, jararaca lancehead (Bothrops jararaca), and two closely related insular species (Bothrops insularis and Bothrops alcatraz), endemic of this region. Phylogenetic and network analyses revealed the existence of two well-supported clades, exhibiting a southern and a northern distribution. The divergence time of these two phylogroups was estimated at 3.8 million years ago, in the Pliocene, a period of intense climatic changes and frequent fragmentation of the tropical rainforest. Our data also suggest that the two groups underwent a large size expansion between 50,000 and 100,000 years ago. However, the southern group showed a more marked signal of population size fluctuation than the northern group, corroborating evidences that southern forests may have suffered a more pronounced reduction in area in the late Pleistocene. The insular species B. alcatraz and B. insularis presented very low diversity, each one sharing haplotypes with mainland individuals placed in different subclades. Despite their marked morphological and behavioural uniqueness, these two insular species seem to have originated very recently and most likely from distinct costal B. jararaca populations, possibly associated with late Pleistocene or Holocene sea level fluctuations. PMID:17054497

  10. Low-level laser therapy decreases local effects induced by myotoxins isolated from Bothrops jararacussu snake venom

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    AM Barbosa

    2010-01-01

    Full Text Available The prominent myotoxic effects induced by Bothrops jararacussu crude venom are due, in part, to its polycationic myotoxins, BthTX-I and BthTX-II. Both myotoxins have a phospholipase A2 structure: BthTX-II is an active enzyme Asp-49 PLA2, while BthTX-I is a Lys-49 PLA2 devoid of enzymatic activity. In this study, the effect of low-level laser therapy (LLLT, 685 nm laser at a dose of 4.2 J/cm2 on edema formation, leukocyte influx and myonecrosis caused by BthTX-I and BthTX-II, isolated from Bothrops jararacussu snake venom, was analyzed. BthTX-I and BthTX-II caused a significant edema formation, a prominent leukocyte infiltrate composed predominantly by neutrophils and myonecrosis in envenomed gastrocnemius muscle. LLLT significantly reduced the edema formation, neutrophil accumulation and myonecrosis induced by both myotoxins 24 hours after the injection. LLLT reduced the myonecrosis caused by BthTX-I and BthTX-II, respectively, by 60 and 43%; the edema formation, by 41 and 60.7%; and the leukocyte influx, by 57.5 and 51.6%. In conclusion, LLLT significantly reduced the effect of these snake toxins on the inflammatory response and myonecrosis. These results suggest that LLLT should be considered a potential therapeutic approach for treatment of local effects of Bothrops species venom.

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

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    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.

  12. Purification of an acidic phospholipase A2 from Bothrops lanceolatus (fer de lance) venom: molecular and enzymatic properties.

    Science.gov (United States)

    de Araújo, A L; Radvanyi, F; Bon, C

    1994-09-01

    The acidic phospholipase A2 from Bothrops lanceolatus venom has been purified by gel filtration on Sephadex G-50 and ion exchange chromatography on CM-cellulose. Analysis by FPLC on Mono-Q column of the purified phospholipase A2 indicated that it is a mixture of several isoenzymes. The two major isoforms consist of a single polypeptide chain with mol. wts of 14,500 and 15,000, which slightly differ in their isoelectric point (4.9 and 5.3) and amino acid composition. However, enzymatic and pharmacological properties of the various isoenzymes are identical. The phospholipase from B. lanceolatus venom is characterized by a progressive increase in the rate of hydrolysis when enzymatic activity is determined with crude egg yolk as substrate in the absence of detergent. This phenomenon, which is not observed with mixed micelles of lecithin-detergent, is not due to the presence of a phospholipase A2 inhibitor in the venom, as previously suggested by several investigators in the case of other Bothrops and Cobra venoms. It is rather a catalytic characteristics of B. lanceolatus venom phospholipase, the enzymatic activity of which depends on the physical state of phospholipids. Bothrops lanceolatus acidic phospholipase A2 is non-toxic. PMID:7801343

  13. Hemiparesia esquerda consecutiva a empeçonhamento: Por Bothrops Jararacussu

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    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.

  14. IgG antibodies against phospholipase A2 from Crotalus durissus terrificus: cross-reaction with venoms from Bothrops species from Argentina

    OpenAIRE

    JP Rodríguez; MC De Marzi; S Maruñak; P Teibler; O Acosta; EL Malchiodi; LC Leiva

    2009-01-01

    We examined the ability of IgG anti-crotalic PLA2 to cross-react with Bothrops spp. venoms, from snakes found in the northeast of Argentina. Immunoblotting and ELISA tests showed that IgG anti-crotalic PLA2 recognize antigens of bothropic venoms. Indirect hemolytic activity tests showed that the quantity of antibodies that neutralized 50% of Crotalus durissus terrificus venom (ED50: 2.1 mg IgG anti-crotalic PLA2/100 µg of venom) were also able to neutralize venom from other snakes in the foll...

  15. 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

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    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

  16. 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

    OpenAIRE

    Julio C. Mendoza; Dan Vivas; Edith Rodríguez; Rosio Inga; Gustavo Sandoval; Fanny Lazo; Armando Yarlequé

    2012-01-01

    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 ...

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

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    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.

  18. 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.

  19. [Bites by snakes in the genus Bothrops: a series of 3,139 cases].

    Science.gov (United States)

    Ribeiro, L A; Jorge, M T

    1997-01-01

    Medical records of 3,139 patients bitten by Bothrops snakes and attended at Vital Brazil Hospital (HVB) from 1981 to 1990 were reviewed. 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%), ecchymosis (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). PMID:9463193

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

    International Nuclear Information System (INIS)

    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. (author)

  1. Bothrops jararacussu venom-induced neuromuscular blockade inhibited by Casearia gossypiosperma Briquet hydroalcoholic extract

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    TM Camargo

    2010-01-01

    Full Text Available The hydroalcoholic extract of Casearia gossypiosperma Briquet (Flacourtiaceae was standardized for the first time through quality control procedures including pharmacognostic methods, fingerprint chromatograms, defined amounts of marker substances and physicochemical characteristics. The pharmacological activity of C. gossypiosperma (Cg hydroalcoholic extract was assayed by a traditional in vitro test, which involved irreversible neuromuscular blockade induced by Bothrops jararacussu (Bjssu venom (60 µg/mL in mouse phrenic nerve-diaphragm preparations. Bjssu venom blocked muscle activity for 26 (± 2.0 minutes (n = 6. Cg extract (0.1 mg/mL induced changes on the baseline muscle activity without impairing the muscle function and inhibited 87.6% (± 1.8 (n = 6 of the Bjssu venom-induced blockade. Both flavonoids (0.624 g% and polyphenols (4.63 g% from the extract were spectrophotometrically quantified. Therefore, the present study confirms the antibothropic activity of Cg extract, supporting the ethnomedical use of Casearia sp. in the treatment of snakebite victims.

  2. Purification and functional characterization of a new metalloproteinase (BleucMP) from Bothrops leucurus snake venom.

    Science.gov (United States)

    Gomes, Mário Sérgio R; de Queiroz, Mayara R; Mamede, Carla C N; Mendes, Mirian M; Hamaguchi, Amélia; Homsi-Brandeburgo, Maria I; Sousa, Marcelo V; Aquino, Elaine Nascimento; Castro, Mariana S; de Oliveira, Fábio; Rodrigues, Veridiana M

    2011-04-01

    A fibrino(geno)lytic nonhemorrhagic metalloproteinase (BleucMP) was purified from Bothrops leucurus snake venom by two chromatographic steps procedure on DEAE-Sephadex A-25 followed by CM-Sepharose Fast Flow column. BleucMP represented 1.75% (w/w) of the crude venom and was homogeneous on SDS-PAGE. BleucMP analyzed by MALDI TOF/TOF, showed a molecular mass of 23,057.54Da and when alkylated and reduced, the mass is 23,830.40Da. Their peptides analyzed in MS (MALDI TOF\\TOF) showed significant score when compared with those of other proteins by NCBI-BLAST2 alignment display. As regards their proteolytic activities, BleucMP efficiently acted on fibrinogen, fibrin, and was inhibited by EDTA and 1.10-phenanthroline. This enzyme was also able to decrease significantly the plasma fibrinogen level provoking blood incoagulability, however was devoid of hemorrhagic activity when tested in the mice skin and did not induce relevant biochemical, hematological and histopathological alterations in mice. The aspects addressed in this paper provide data on the effect of BleucMP in envenomation from B. leucurus snakes in order to better understand the effects caused by snake venom metalloproteinase. PMID:21130897

  3. 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)

  4. Effects of photobiostimulation on edema and hemorrhage induced by Bothrops moojeni venom.

    Science.gov (United States)

    Nadur-Andrade, Nikele; Barbosa, Ana Maria; Carlos, Fernando Pereira; Lima, Carlos José; Cogo, José Carlos; Zamuner, Stella Regina

    2012-01-01

    Antivenom (AV) treatment has been ineffective in neutralizing the severe local fast-developing tissue damage following snake-bite envenoming. We studied the effectiveness of low-level laser (LLL) and light-emitting diode (LED) irradiation alone or in combination with AV in reducing local edema formation and hemorrhage induced by Bothrops moojeni venom (BmV) in mice. Edema formation was induced by injection of 1 μg per paw of BmV into the right paw and was evaluated before and at several intervals after BmV intraplantar injection. Hemorrhagic activity was evaluated after intradermal injection of 20 μg of BmV by measuring the diameter of the hemorrhagic area on the inner side of the skin. The site of BmV injection was irradiated by LLL or LED 30 min after BmV inoculation. AV was also administered intravenously 30 min after BmV injection. Irradiation with LLL at a wavelength of 685 nm and a dose of 2.2 J/cm(2) and with a red LED and an infrared LED at wavelengths of 635 nm and 945 nm, respectively, and a dose of 4 J/cm(2) reduced edema formation and hemorrhage induced by BmV (p BmV may be clinically relevant.

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

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

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    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.

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

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    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.

  8. Coagulopathy after snake bite by Bothrops neuwiedi: case report and results of in vitro experiments.

    Science.gov (United States)

    Dempfle, C E; Kohl, R; Harenberg, J; Kirschstein, W; Schlauch, D; Heene, D L

    1990-12-01

    Coagulation studies were performed in a patient who had been bitten by a snake of the species Bothrops neuwiedi. The patient presented with hemorrhagic necrosis at the envenomization site and considerable bleeding from venous puncture sites. He developed a severe defibrination syndrome with a clottable fibrinogen level of approximately 0.1 g/l. Fibrinogen was not measurable by clotting time assay. Fibrin degradation products were greatly elevated. Treatment with antivenom caused an anaphylactic reaction within ten minutes and serum sickness after three days. In vitro experiments revealed that B. neuwiedi venom directly activates Factors II and X, but does not activate Factor XIII. In vivo consumption of Factor XIII after B. neuwiedi envenomization is ascribed to the action of Factor IIa. At low venom concentrations clotting is initiated by activation of prothrombin by the venom either directly or via Factor X activation. Treatment with heparin might be beneficial in coagulopathy secondary to snake bite by reducing circulating active thrombin. The venom contains thrombin-like proteases which cause slow clotting of fibrinogen, and plasmin-like components causing further proteolysis of fibrinogen and fibrin. Antivenom has no effect on the proteolytic action of the snake venom. The in vivo effects of antivenom are presumably caused by acceleration of the elimination of venom components from the circulation. Intravenous administration of antivenom caused normalization of blood coagulation parameters within 48 h. PMID:2291986

  9. Isolation, functional, and partial biochemical characterization of galatrox, an acidic lectin from Bothrops atrox snake venom.

    Science.gov (United States)

    Mendonça-Franqueiro, Elaine de Paula; Alves-Paiva, Raquel de Melo; Sartim, Marco Aurélio; Callejon, Daniel Roberto; Paiva, Helder Henrique; Antonucci, Gilmara Ausech; Rosa, José César; Cintra, Adélia Cristina Oliveira; Franco, João José; Arantes, Eliane Candiani; Dias-Baruffi, Marcelo; Sampaio, Suely Vilela

    2011-03-01

    Snake venom lectins have been studied in regard to their chemical structure and biological functions. However, little is known about lectins isolated from Bothrops atrox snake venom. We report here the isolation and partial functional and biochemical characterization of an acidic glycan-binding protein called galatrox from this venom. This lectin was purified by affinity chromatography using a lactosyl-sepharose column, and its homogeneity and molecular mass were evaluated by high-performance liquid chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. The purified galatrox was homogeneous and characterized as an acidic protein (pI 5.2) with a monomeric and dimeric molecular mass of 16.2 and 32.5 kDa, respectively. Alignment of N-terminal and internal amino acid sequences of galatrox indicated that this protein exhibits high homology to other C-type snake venom lectins. Galatrox showed optimal hemagglutinating activity at a concentration of 100 μg/ml and this effect was drastically inhibited by lactose, ethylenediaminetetraacetic acid, and heating, which confirmed galatrox's lectin activity. While galatrox failed to induce the same level of paw edema or mast cell degranulation as B. atrox crude venom, galatrox did alter cellular viability, which suggested that galatrox might contribute to venom toxicity by directly inducing cell death. PMID:21297119

  10. A rational protocol for the successful crystallization of L-amino-acid oxidase from Bothrops atrox.

    Science.gov (United States)

    Alves, Raquel Melo; Feliciano, Patricia Rosa; Sampaio, Suely Vilela; Nonato, Maria Cristina

    2011-04-01

    Despite the valuable contributions of robotics and high-throughput approaches to protein crystallization, the role of an experienced crystallographer in the evaluation and rationalization of a crystallization process is still crucial to obtaining crystals suitable for X-ray diffraction measurements. In this work, the difficult task of crystallizing the flavoenzyme L-amino-acid oxidase purified from Bothrops atrox snake venom was overcome by the development of a protocol that first required the identification of a non-amorphous precipitate as a promising crystallization condition followed by the implementation of a methodology that combined crystallization in the presence of oil and seeding techniques. Crystals were obtained and a complete data set was collected to 2.3 Å resolution. The crystals belonged to space group P2(1), with unit-cell parameters a = 73.64, b = 123.92, c = 105.08 Å, β = 96.03°. There were four protein subunits in the asymmetric unit, which gave a Matthews coefficient V(M) of 2.12 Å(3) Da(-1), corresponding to 42% solvent content. The structure has been solved by molecular-replacement techniques. PMID:21505245

  11. Isolation, functional, and partial biochemical characterization of galatrox, an acidic lectin from Bothrops atrox snake venom

    Institute of Scientific and Technical Information of China (English)

    Elaine de Paula Mendonca-Franqueiro; Eliane Candiani Arantes; Marcelo Dias-Baruffi; Suely Vilela Sampaio; Raquel de Melo Alves-Paiva; Marco Aurélio Sartim; Daniel Roberto Callejon; Helder Henrique Paiva; Gilmara Ausech Antonucci; José César Rosa; Adélia Cristina Oliveira Cintra; Jo(a)o José Franco

    2011-01-01

    Snake venom lectins have been studied in regard to their chemical structure and biological functions. However, little is known about lectins isolated from Bothrops atrox snake venom. We report here the isolation and partial functional and biochemical characterization of an acidic glycanbinding protein called galatrox from this venom. This lectin was purified by affinity chromatography using a lactosyl-sepharose column, and its homogeneity and molecular mass were evaluated by high-performance liquid chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. The purified galatrox was homogeneous and characterized as an acidic protein (pI 5.2) with a monomeric and dimeric molecular mass of 16.2 and 32.5 kDa, respectively. Alignment of N-terminal and internal amino acid sequences of galatrox indicated that this protein exhibits high homology to other C-type snake venom lectins. Galatrox showed optimal hemagglutinating activity at a concentration of 100 μg/ml and this effect was drastically inhibited by lactose, ethylenediaminetetraacetic acid, and heating, which confirmed galatrox's lectin activity. While galatrox failed to induce the same level of paw edema or mast cell degranulation as B. atrox crude venom, galatrox did alter cellular viability,which suggested that galatrox might contribute to venom toxicity by directly inducing cell death.

  12. Neuromuscular action of Bothrops lanceolatus (Fer de lance) venom and a caseinolytic fraction.

    Science.gov (United States)

    Lôbo de Araújo, Albetiza; Donato, José Luiz; Leite, Gildo Bernardo; Prado-Franceschi, Júlia; Fontana, Marcos Dias; Bon, Cassian; Rodrigues Simioni, Léa

    2002-09-01

    A protein capable of inducing neuromuscular blockade in avian preparations and of depolarizing mouse diaphragm muscle was isolated from Bothrops lanceolatus venom using gel filtration and ion-exchange chromatography. The purified protein was a single chain polypeptide with an estimated molecular mass of 27.5 kDa by SDS-PAGE and had caseinolytic activity (13.3 units/mg), but no phospholipase A(2). B.lanceolatus venom (50 micro g/ml) and the caseinolytic protein (20 micro g/ml) produced contracture and progressive irreversible blockade (50% in 25+/-5 min (SEM) and 45+/-15 min, respectively), in indirectly stimulated chick biventer cervicis preparations. The contractile responses to acetylcholine (ACh; 37 and 74 micro M, n=6) were inhibited by venom and the caseinolytic protein, whereas those to potassium (13.4mM, n=6) were not. Membrane resting potential measurements in mouse hemidiaphragm preparations showed that B.lanceolatus venom and the purified protein caused depolarization which was prevented by D-tubocurarine (14.6mM). The venom produced a slight increase in the amplitude and frequency of miniature end-plate potentials, but this effect was not seen with the purified fraction. These results suggest that the purified protein acts exclusively post-synaptically. PMID:12220713

  13. Hyperbaric oxygen therapy after Bothrops lanceolatus snake bites in Martinique: a brief report.

    Science.gov (United States)

    Hochedez, P; Thomas, L; Mehdaoui, H

    2010-01-01

    Every year 10 to 20 cases of snake bites are reported on the Caribbean island of Martinique. The only snake involved, Bothrops lanceolatus, is endemic on the island, and its bite may lead to systemic multifocal thrombotic complications in the'absence of the monospecific antivenom. Between January 1988 and January 2009, more than 250 snake bites have been reported, and five patients were treated with hyperbaric oxygen (HBO2) therapy for local complications. The patients were male, bitten on the leg or the hand, and presented with severe complications such as necrotizing soft tissue infections, compartment syndrome or abscesses despite prompt wound care and administration of antivenomous serum. Outcomes were favorable for these five patients, except for one who was left with a functional defect of the hand. Although snake bites are not part of the currently recommended indications for HBO2 therapy, local complications, namely compartment syndrome, necrotizing soft tissue infections and enhancement of healing in selected problem wounds, are approved uses of HBO2 therapy as defined by the Hyperbaric Oxygen Therapy Committee and would benefit from prospective studies. PMID:21226390

  14. Coagulant and anticoagulant activities of Bothrops lanceolatus (Fer de lance) venom.

    Science.gov (United States)

    Lôbo de Araújo, A; Kamiguti, A; Bon, C

    2001-01-01

    Bothrops lanceolatus venom contains caseinolytic, phospholipase, esterase and haemorrhagic activities. We have investigated the coagulant and anticoagulant actions of B. lanceolatus venom on human citrated plasma and on purified plasma components. Although B. lanceolatus venom up to 50 microg/ml was unable to clot citrated plasma, at concentrations > or = 5 microg/ml the venom dose-dependently clotted purified human fibrinogen, indicating the presence of a thrombin-like enzyme. Human plasma (final concentration > or = 12.5%) dose-dependently inhibited the venom-induced fibrinogen clotting. This finding suggested that endogenous plasma protease inhibitors can affect the venom's action on fibrinogen. To investigate this possibility, B. lanceolatus venom was incubated with different plasma protease inhibitors and the activity on fibrinogen tested. alpha(2)-Macroglobulin and alpha(1)-antitrypsin did not interfere with the coagulant activity of the venom whereas the antithrombin-III/heparin complex partially inhibited this activity. A non-toxic, acidic phospholipase A(2) purified from B. lanceolatus venom prolonged the activated partial thromboplastin time in human plasma from 39.7+/-0.5 s (control with saline) to 60.2+/-0.9 s with 50 microg of PLA(2) (p<0.001), suggesting an anticoagulant activity associated with this enzyme. This anticoagulant activity may account for some of the effects of the venom on blood coagulation. PMID:10978756

  15. Low-level laser therapy decreases local effects induced by myotoxins isolated from Bothrops jararacussu snake venom

    OpenAIRE

    AM Barbosa; AB Villaverde; L Guimarães-Sousa; AM Soares; SF Zamuner; JC Cogo; SR Zamuner

    2010-01-01

    The prominent myotoxic effects induced by Bothrops jararacussu crude venom are due, in part, to its polycationic myotoxins, BthTX-I and BthTX-II. Both myotoxins have a phospholipase A2 structure: BthTX-II is an active enzyme Asp-49 PLA2, while BthTX-I is a Lys-49 PLA2 devoid of enzymatic activity. In this study, the effect of low-level laser therapy (LLLT), 685 nm laser at a dose of 4.2 J/cm2 on edema formation, leukocyte influx and myonecrosis caused by BthTX-I and BthTX-II, isolated from Bo...

  16. Kinetics of venom and antivenom serum and clinical parameters and treatment efficacy in Bothrops alternatus envenomed dogs.

    Science.gov (United States)

    Jacome, Do; Melo, M M; Santos, M M B; Heneine, L G D

    2002-12-01

    Dogs envenomed with non-lethal doses of Bothrops alternatus venom received standard antivenom therapy, im injections of flunixin meglumine, or topical treatmentwith aqueous Curcuma longa plant extract. Biodistribution of the venom and antivenom were determined by ELISA. There was no significant difference in the efficacy of antivenom and plant extract on local effects; flunixin treatment had lower efficacy. Distribution of the venom was similar with all 3 treatments. Serum levels of the antivenom reached maximum 2-4 h after administration and were not detected after the 5th d.

  17. Inhibition of the hyperalgesic activity of Bothrops jararaca venom by an antibothropic fraction isolated from opossum (Didelphis marsupialis) serum.

    Science.gov (United States)

    Rocha, S L; Frutuoso, V S; Domont, G B; Martins, M A; Moussatché, H; Perales, J

    2000-06-01

    The antibothropic fraction (ABF) already isolated from Didelphis marsupialis serum, inhibits the haemorrhagic, oedematogenic, myonecrotic and lethal activities of Bothrops jararaca venom (Bjv). The aim of this work was to verify the capability of ABF to inhibit the hyperalgesic activity of Bjv. Intraplantar injection of Bjv induced hyperalgesia in a time- and dose-dependent manner and ABF administered in situ concomitantly with Bjv or i.v. 30 min before venom injection reduced the induced hyperalgesia. This same effect was observed when ABF was intravenously injected at 5 and 15 min after Bjv. Our results show that ABF inhibits also the hyperalgesia induced by Bjv. PMID:10695972

  18. Comparative evaluation of adverse effects in the use of powder trivalent antivenom and liquid antivenoms in Bothrops snake bites

    Directory of Open Access Journals (Sweden)

    Iran Mendonça da Silva

    2012-08-01

    Full Text Available INTRODUCTION: Snake bite, a problem in public health, generally occurs where there is no electric power. METHODS: A comparative clinical study was conducted with 102 victims of Bothrops snake bite, from the state of Amazonas, Brazil; 58 victims were treated with liofilizated trivalent antivenom serum (SATL and 44 victims treated with liquid bivalent and monovalent antivenom serum (SAMBL. RESULTS: 17% (10/58 of patients presented adverse effects with the SATL and 25% (11/44 with the SAMBL. CONCLUSIONS: There was no statistic difference in number of adverse effects between the two types of snake bite antivenom.

  19. N-terminal amino acid sequences and some characteristics of fibrinolytic/hemorrhagic metalloproteinases purified from Bothrops jararaca venom.

    Science.gov (United States)

    Maruyama, Masugi; Sugiki, Masahiko; Anai, Keita; Yoshida, Etsuo

    2002-08-01

    We determined the N-terminal amino acid sequences of the fibrinolytic/hemorrhagic metalloproteinases (jararafibrases I, III and IV) purified from Bothrops jararaca venom. The N-terminal amino acid sequences of jararafibrase I and its degradation products were identical to those of jararhagin, another hemorrhagic metalloproteinase purified from the same snake venom. Together with enzymatic and immunological properties, we concluded that those two enzymes are identical. The N-terminal amino acid sequence of jararafibrase III was quite similar to C-type lectin isolated from Crotalus atrox, and the protein had a hemagglutinating activity on intact rat red blood cells. PMID:12165326

  20. Neutralization of four Peruvian Bothrops sp. snake venoms by polyvalent antivenoms produced in Perú and Costa Rica: preclinical assessment.

    Science.gov (United States)

    Rojas, Ermila; Quesada, Lil; Arce, Viviana; Lomonte, Bruno; Rojas, Gustavo; Gutiérrez, José María

    2005-01-01

    Envenomations after bites inflicted by snakes of the genus Bothrops constitute a public health hazard in Perú, and the intravenous administration of equine-derived antivenoms represents the only scientifically validated treatment. This study presents a preclinical assessment of the efficacy of two whole IgG antivenoms, prepared in Perú and Costa Rica, to neutralize the most relevant toxic effects induced by the venoms of Bothrops atrox, B. brazili, B. barnetti and B. pictus from Perú. Peruvian antivenom is produced by immunizing horses with Bothrops sp. venoms from this country, whereas the production of Costa Rican antivenom involves immunization with venoms from Central American snakes. The neutralization of lethal, hemorrhagic, edema-forming, myotoxic, coagulant and defibrinating activities was evaluated in assays involving incubation of venom and antivenom prior to testing. Both antivenoms were effective in the neutralization of these effects, with quantitative variations in the values of effective dose 50% depending on the effects being studied. Peruvian antivenom was more effective in the neutralization of lethality induced by B. atrox and B. barnetti venoms. However, Peruvian antivenom failed to neutralize coagulant activity of B. barnetti venom and edema-forming activity of B. brazili venom, whereas neutralization was achieved by Costa Rican antivenom. It is concluded that an extensive immunological cross-reactivity exists between Bothrops sp. venoms from Perú and Costa Rica, and that both antivenoms are effective in the neutralization of these four venoms in a rodent model of envenoming. PMID:15589801

  1. Avaliação das lesões locais de cães envenenados experimentalmente com Bothrops alternatus após diferentes tratamentos Evaluation of local lesions in dogs experimentally envenomed by Bothrops alternatus after different treatments

    Directory of Open Access Journals (Sweden)

    M.M.B. Santos

    2003-10-01

    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.

  2. 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

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    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

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

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    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.

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

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    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.

  5. 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. PMID:16198389

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

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    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.

  7. Exploring the proteomes of the venoms of the Peruvian pit vipers Bothrops atrox, B. barnetti and B. pictus.

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    Kohlhoff, Markus; Borges, Marcia H; Yarleque, Armando; Cabezas, Cesar; Richardson, Michael; Sanchez, Eladio F

    2012-04-01

    We report the comparative proteomic characterization of the venoms of Bothrops atrox, B. barnetti and B. pictus. The venoms were subjected to RP-HPLC and the resulting fractions analyzed by SDS-PAGE. The proteins were cut from the gels, digested with trypsin and identified via peptide mass fingerprint and manual sequencing of selected peptides by MALDI-TOF/TOF mass spectrometry. Around 20-25 proteins were identified belonging to only 6-7 protein families. Metalloproteinases of the classes P-I and P-III were the most abundant proteins in all venoms (58-74% based on peak area A214 nm), followed by phospholipases-A(2) (6.4-14%), disintegrins (3.2-9%) and serine proteinases (7-11%), and some of these proteins occurred in several isoforms. In contrast cysteine-rich secretory proteins and L-amino acid oxidases appeared only as single isoforms and were found only in B. atrox and B. barnetti. C-type lectins were also detected in all venoms but at low levels (~ 5%). Furthermore, the venoms contain variable numbers of peptides (Bothrops species is in agreement with their pharmacological and pathological effects. PMID:22300577

  8. Isolation and biological characterization of Batx-I, a weak hemorrhagic and fibrinogenolytic PI metalloproteinase from Colombian Bothrops atrox venom.

    Science.gov (United States)

    Patiño, Arley C; Pereañez, Jaime A; Núñez, Vitelbina; Benjumea, Dora M; Fernandez, Maritza; Rucavado, Alexandra; Sanz, Libia; Calvete, Juan J

    2010-11-01

    A hemorrhagic metalloproteinase, named Batx-I, was isolated from the venom of Bothrops atrox specimens (from Southeastern Colombian region) by a combination of CM-Sephadex C25 ion-exchange and Affi-gel Blue affinity chromatographies. This enzyme accounts for about 45% of venom proteins, and it has an ESI-MS isotope-averaged molecular mass of 23296.2 Da and a blocked N-terminus. Two internal fragments sequenced by mass spectrometric analysis showed similarity to other SVMPs from Bothrops venoms. To investigate the possible participation of Batx-I in the envenomation pathophysiology, proteolytic, fibrinogenolytic, hemorrhagic, and other biological activities were evaluated. The minimal hemorrhagic dose obtained was 17 microg/20 g body weight. The enzyme showed proteolytic activity on azocasein, comparable with activity of BaP1. This activity was inhibited by EDTA and 1, 10 o-phenanthroline but not by aprotinin, pepstatin A or PMSF. Fibrinogenolytic activity was analyzed by SDS-PAGE, revealing a preference for degrading the A alpha- and B beta-chains, although partial degradation of the gamma-chain was also detected. The protein lacks coagulant and defibrinating activity. The CK levels obtained, clearly reflects a myotoxic activity induced by Batx-I. The hemorrhagic and fibrinogenolytic activities exhibited by the isolated PI-SVMP may play a role in the hemorrhagic and blood-clotting disorders observed in patients bitten by B. atrox in Colombia. PMID:20600221

  9. Spectroscopic and thermal characterization of alternative model biomembranes from shed skins of Bothrops jararaca and Spilotis pullatus

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    André Rolim Baby

    2009-09-01

    Full Text Available Recently, there has been an interest in the use of shed snake skin as alternative model biomembrane for human stratum corneum. This research work presented as objective the qualitative characterization of alternative model biomembranes from Bothrops jararaca and Spilotis pullatus by FT-Raman, PAS-FTIR and DSC. The employed biophysical techniques permitted the characterization of the biomembranes from shed snake skin of B. jararaca and S. pullatus by the identification of vibrational frequencies and endothermic transitions that are similar to those of the human stratum corneum.Existe atualmente interesse no uso da muda de pele de cobra como modelos alternativos de biomembranas da pele humana. O presente trabalho apresentou como objetivo a caracterização qualitativa de modelos alternativos de biomembranas provenientes de mudas de pele de cobra da Bothrops jararaca e Spilotis pullatus por espectroscopia Raman (FT-Raman, espectroscopia fotoacústica no infravermelho (PAS-FTIR e calorimetria exploratória diferencial (DSC. As técnicas biofísicas FT-Raman, PAS-FTIR e DSC permitiram caracterizar qualitativamente os modelos alternativos de biomembranas provenientes das mudas de pele de cobra da B. jararaca e S. pullatus e identificar freqüências vibracionais e transições endotérmicas similares ao estrato córneo humano.

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

    International Nuclear Information System (INIS)

    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 60Co 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

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

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    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

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

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    Javier Mora

    Full Text Available BACKGROUND: 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. METHODOLOGY/PRINCIPAL FINDINGS: 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. CONCLUSIONS/SIGNIFICANCE: 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.

  13. A transcriptomic view of the proteome variability of newborn and adult Bothrops jararaca snake venoms.

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    André Zelanis

    Full Text Available BACKGROUND: Snake bite is a neglected public health problem in communities in rural areas of several countries. Bothrops jararaca causes many snake bites in Brazil and previous studies have demonstrated that the pharmacological activities displayed by its venom undergo a significant ontogenetic shift. Similarly, the venom proteome of B. jararaca exhibits a considerable variation upon neonate to adult transition, which is associated with changes in diet from ectothermic prey in early life to endothermic prey in adulthood. Moreover, it has been shown that the Brazilian commercial antibothropic antivenom, which is produced by immunization with adult venom, is less effective in neutralizing newborn venom effects. On the other hand, venom gland transcripts of newborn snakes are poorly known since all transcriptomic studies have been carried out using mRNA from adult specimens. METHODS/PRINCIPAL FINDINGS: Here we analyzed venom gland cDNA libraries of newborn and adult B. jararaca in order to evaluate whether the variability demonstrated for its venom proteome and pharmacological activities was correlated with differences in the structure of toxin transcripts. The analysis revealed that the variability in B. jararaca venom gland transcriptomes is quantitative, as illustrated by the very high content of metalloproteinases in the newborn venom glands. Moreover, the variability is also characterized by the structural diversity of SVMP precursors found in newborn and adult transcriptomes. In the adult transcriptome, however, the content of metalloproteinase precursors considerably diminishes and the number of transcripts of serine proteinases, C-type lectins and bradykinin-potentiating peptides increase. Moreover, the comparison of the content of ESTs encoding toxins in adult male and female venom glands showed some gender-related differences. CONCLUSIONS/SIGNIFICANCE: We demonstrate a substantial shift in toxin transcripts upon snake development and a

  14. Snake venomics of Bothrops punctatus, a semiarboreal pitviper species from Antioquia, Colombia

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    Maritza Fernández Culma

    2014-01-01

    Full Text Available Bothrops punctatus is an endangered, semi-arboreal pitviper species distributed in Panamá, Colombia, and Ecuador, whose venom is poorly characterized. In the present work, the protein composition of this venom was profiled using the ‘snake venomics’ analytical strategy. Decomplexation of the crude venom by RP-HPLC and SDS-PAGE, followed by tandem mass spectrometry of tryptic digests, showed that it consists of proteins assigned to at least nine snake toxin families. Metalloproteinases are predominant in this secretion (41.5% of the total proteins, followed by C-type lectin/lectin-like proteins (16.7%, bradykinin-potentiating peptides (10.7%, phospholipases A2 (93%, serine proteinases (5.4%, disintegrins (38%, L-amino acid oxidases (3.1%, vascular endothelial growth factors (17%, and cysteine-rich secretory proteins (1.2%. Altogether, 6.6% of the proteins were not identified. In vitro, the venom exhibited proteolytic, phospholipase A2, and L-amino acid oxidase activities, as well as angiotensin-converting enzyme (ACE-inhibitory activity, in agreement with the obtained proteomic profile. Cytotoxic activity on murine C2C12 myoblasts was negative, suggesting that the majority of venom phospholipases A2 likely belong to the acidic type, which often lack major toxic effects. The protein composition of B. punctatus venom shows a good correlation with toxic activities here and previously reported, and adds further data in support of the wide diversity of strategies that have evolved in snake venoms to subdue prey, as increasingly being revealed by proteomic analyses.

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

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    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.

  16. Cell adhesion molecules involved in the leukocyte recruitment induced by venom of the snake Bothrops jararaca

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    Stella R. Zamuner

    2002-01-01

    Full Text Available It has been shown that Bothrops jararaca venom (BjV induces a significant leukocyte accumulation, mainly neutrophils, at the local of tissue damage. Therefore, the role of the adhesion molecules intercellular adhesion molecule-1 (ICAM-1, LECAM-1, CD18, leukocyte function-associated antigen-1 (LFA-1 and platelet endothelial cell adhesion molecule-1 (PECAM-1 on the BjV-induced neutrophil accumulation and the correlation with release of LTB4, TXA2, tumor necrosis factor-α, interleukin (IL-1 and IL-6 have been investigated. Anti-mouse LECAM-1, LFA-1, ICAM-1 and PECAM-1 monoclonal antibody injection resulted in a reduction of 42%, 80%, 66% and 67%, respectively, of neutrophil accumulation induced by BjV (250 μg/kg, intraperitoneal injection in male mice compared with isotype-matched control injected animals. The anti-mouse CD18 monoclonal antibody had no significant effect on venom-induced neutrophil accumulation. Concentrations of LTB4, TXA2, IL-6 and TNF-α were significant increased in the peritoneal exudates of animals injected with venom, whereas no increment in IL-1 was detected. This results suggest that ICAM-1, LECAM-1, LFA-1 and PECAM-1, but not CD18, adhesion molecules are involved in the recruitment of neutrophils into the inflammatory site induced by BjV. This is the first in vivo evidence that snake venom is able to up-regulate the expression of adhesion molecules by both leukocytes and endothelial cells. This venom effect may be indirect, probably through the release of the inflammatory mediators evidenced in the present study.

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

    Energy Technology Data Exchange (ETDEWEB)

    Gabriel, L.M.; Soares, M.A.; Bicalho, M.S.; Santos, R.G. [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil)], e-mail: marcellaaraugio@yahoo.com.br, e-mail: mbs@cdtn.br, e-mail: santosr@cdtn.br; Sanchez, E.O.F.; Silva, S.G. [Ezequiel Dias Foundation, Belo Horizonte, MG (Brazil)], e-mail: silea@funed.mg.gov.br, e-mail: eladio@funed.mg.gov.br

    2009-07-01

    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 {sup 125}I and evaluate the interaction of {sup 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 {sup 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 {sup 125}I-Leuc-B were 1.77 (tumor/normal paw) and 8.44 tumor/skeletal muscle. The results suggest that {sup 125}I-Leuc- B may constitute a good template for development of a tool for detection of solid tumors. (author)

  18. Biochemical and biological properties of phospholipases A(2) from Bothrops atrox snake venom.

    Science.gov (United States)

    Kanashiro, Milton M; de Cássia M Escocard, Rita; Petretski, Jorge H; Prates, Maura V; Alves, Elias W; Machado, Olga L T; da Silva, Wilmar Dias; Kipnis, Thereza L

    2002-10-01

    Phospholipases A(2) (PLA(2)s), of molecular mass 13-15kDa, are commonly isolated from snake venom. Two myotoxins with PLA(2) activity, BaPLA(2)I and BaPLA(2)III, with estimated molecular masses of 15kDa were isolated from the venom of Bothrops atrox using Sephacryl S-100-HR and reverse-phase chromatography. BaPLA(2)I was basic, with a pI of 9.1, while BaPLA(2)III was neutral with a pI of 6.9. On a molecular basis, BaPLA(2)III exhibited higher catalytic activity on synthetic substrates than BaPLA(2)I. Comparison of the N-terminal residues of BaPLA(2)I with other PLA(2) proteins from snake venoms showed that it has the highest homology (94%) with B. asper myotoxin II and homology with a PLA(2) Lys(49) from B. atrox (89%). In contrast, BaPLA(2)III demonstrated 75, 72, and 71% homology with PLA(2) from Vipera ammodytes meridionalis, B. jararacussu, and B. jararaca, respectively. BaPLA(2)I and BaPLA(2)III were capable, in vitro, of inducing mast cell degranulation and, in vivo, of causing creatine kinase release, edema, and myonecrosis typical of PLA(2)s from snake venoms, characterized by rapid disruption of the plasma membrane as indicated by clumping of myofilaments and necrosis of affected skeletal muscle cells. BaPLA(2)I- and BaPLA(2)III-specific monoclonal and polyclonal antibodies, although incapable of neutralizing PLA(2) edematogenic activity, blocked myonecrosis efficiently in an in vivo neutralization assay. The results presented herein suggest that the biological active site responsible for edema induction by these two PLA(2) enzymes is distinct from the myonecrosis active site and is not dependent upon the catalytic activity of the PLA(2) enzyme. PMID:12234622

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

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    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

  20. Isolation and characterization of a serine protease, Ba III-4, from Peruvian Bothrops atrox venom.

    Science.gov (United States)

    Ponce-Soto, L A; Bonfim, V L; Novello, J C; Navarro Oviedo, R; Yarlequé Chocas, A; Marangoni, S

    2007-09-01

    A serine protease from Bothrops atrox (Peruvian specimen's venom) was isolated in two chromatographic steps in LC molecular exclusion and reverse phase-HPLC. This protein was denominated Ba III-4 (33,080.265 Da determinated by MALDI-TOF mass spectrometry) and showed pI of 5.06, Km 0.2 x 10(-1 ) M and the V (máx) 4.1 x 10(-1 )nmoles p-NA/lt/min on the synthetic substrate BapNA. Ba III-4 also showed ability to coagulate bovine fibrinogen. The serine protease was inhibited by soyben trypsin inhibitor and DA2II, which is an anti-hemorrhagic factor isolated from the opossum specie Didelphis albiventris. The primary structure of Ba III-4 showed the presence of His(44), Asp(94) and Ser(193) residues in the corresponding positions to the catalytic triad established in the serine proteases and Ser(193) are inhibited by phenylmethylsulfonylfluoride (PMSF). Amino acid analysis showed a high content of Asp, Glu, Gly, Ser, Ala and Pro, as well as 12 half-cysteine residues. Ba III-4 contained 293 amino acid residues and the primary structure of VIGGDECDIN EHPFLAFMYY SPRYFCGMTL INQEWVLTAA HCRYFCGMTL IHLGVHRESE KANYDEVRRF PKEKYFIFCD NNFTDDEVDK DIMLIRLDKP VSNSEHIAPL SLPSNPPSVG SVCRIMGWGQ TTTSPIDVLS PDEPHCANIN LFDNTVCHTA HPQVANTRTS TDTLCAGDLQ GGRDTCNGDS GGPLICNEQL HGILSWGGDP CAQPNKPAFY TKVYYFDHPW IKSIIAGNKK TVNFTCPPLR SDAKDDSTTY INQEWDWVLT AEHCDRTHMR NSFYDYSSIN SDS. Titration experiments did not show the presence of free sulfhydryl groups after 4 h incubation, nor were differences found in relation to titration kinetics in the presence of nondenaturating buffer. The isolation of this protein, Ba III-4, is of potential interest for the understanding of the pathomechanism of the snake venom action and for the identification of new blood coagulation enzymes of natural sources. PMID:17522968

  1. Bothrops lanceolatus (Fer de lance) venom stimulates leukocyte migration into the peritoneal cavity of mice.

    Science.gov (United States)

    Arruda, Vanessa Alves; de Queiroz Guimarães, Alessandra; Hyslop, Stephen; de Araújo, Paulo Maria Ferreira; Bon, Cassian; de Araújo, Albetiza Lôbo

    2003-01-01

    The ability of Bothrops lanceolatus venom to induce neutrophil migration into the peritoneal cavity of mice was investigated. Intraperitoneal injection of venom caused dose- and time-dependent neutrophil migration, which peaked with 750 ng of venom/cavity 4h after venom injection. The neutrophil migration was significantly reduced by pretreatment with dexamethasone (0.5 mg/kg, s.c.), an indirect inhibitor of phospholipase A(2) (PLA(2)), and AA861 (0.01 mg/kg, s.c.), a 5-lipoxygenase inhibitor, but in contrast, was not modified by pretreatment with indomethacin (2 mg/kg, s.c.), an inhibitor of the cyclooxygenase pathway, meloxicam (5 mg/kg, s.c.), an inhibitor of the cyclooxygenase-2 pathway, or the PAF inhibitor WEB2086 (40 mg/kg, s.c.). Dexamethasone and AA861 also inhibited the neutrophil migration by 60% when administered immediately after venom injection, and the coadministration of these two drugs caused a 75% reduction in migration. BLV-induced neutrophil migration was not due to contamination by endotoxin since polymyxin B-treated venom retained its activity. Heating the venom (97 degrees C, 2 min) reduced the PLA(2) activity by 64% and this was accompanied by a corresponding reduction (68%) in neutrophil migration. These results suggest that arachidonate-derived lipoxygenase metabolites (possibly leukotriene B(4)) are involved in the chemotaxis observed. Macrophages may be an important source of these metabolites since the migratory response to venom was potentiated in mice pretreated with thioglycollate, but reduced when the peritoneal cavity was washed with sterile saline. PMID:12467667

  2. Annual changes in seminal variables of golden lanchead pitvipers (Bothrops insularis) maintained in captivity.

    Science.gov (United States)

    Silva, K B; Zogno, M A; Camillo, A B; Pereira, R J G; Almeida-Santos, S M

    2015-12-01

    Bothrops insularis is an endemic and critically endangered snake with an estimated population of 2000 individuals restricted to Queimada Grande Island, in southeastern Brazil. Brazilian researchers established a captive breeding program for the species that includes the application of assisted reproductive technologies. The present study, therefore, aimed to evaluate semen samples from captive B. insularis throughout the year to ascertain seasonal differences in semen traits as well as correlations with body size and weight. Eighteen males with snout-vent length (SVL) ranging from 43.5 to 73.7 cm were collected at quarterly basis between August 2012 and May 2013. Macroscopic analysis revealed semen volumes ranging from 0.5 to 6.0 μL with samples featuring whitish to yellowish color and creamy and thick consistency. Viable sperm was obtained from all males indicating that individuals with SVL equal to or greater than 43.5 cm are sexually developed. However, adult and immature males (estimated by SVL) exhibited different seasonal profiles for motility and progressive motility. Adult males had a decrease in sperm motility and progressive motility during summer and spring, respectively, whereas the same variables did not vary throughout the year in immature snakes. Sperm concentration in all individuals was less (0.5 × 10(9) μL) during the winter, but no seasonal fluctuations were detected in semen volume. These findings are of particular importance to the development of reproductive tools such as male selection, artificial insemination and sperm freezing for the genetic management of this critically endangered snake. PMID:26559333

  3. 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

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    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

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

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    C.M.L. Vasconcelos

    1998-03-01

    Full Text Available 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 131I 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 105 cpm/mouse of 131I-venom (N = 42, 131I-albumin or 131I (controls, N = 28 each. Thirty minutes and 1, 3, 6, 12, 18 and 24 h after inoculation, the animals were perfused with 0.85% NaCl and skin and various organs were collected in order to determine radioactivity content. There was a high rate of venom absorption in the 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 (3rd h, small intestine (3rd 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 3rd h postinjection (1.98%. There was a low binding of labeled venom in the liver (<0.54%, thyroid (<0.11% and lungs (<0.08%, and no iodinated venom was detected in brain, heart, diaphragm, spleen or bladder. The low venom binding observed in most internal organs, comparable to that of albumin, suggests that B. erythromelas venom does not specifically target most internal organs

  5. 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.

  6. Flora bacteriana da cavidade oral, presas e veneno de Bothrops jararaca: possível fonte de infecção no local da picada Bacterial flora of the oral cavity, fangs and venom of Bothrops jararaca: possible source of infection at the local bite

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    Miguel Tanús Jorge

    1990-02-01

    Full Text Available Materiais colhidos das presas, das bainhas das presas e do veneno de 15 Bothrops jararaca recém-capturadas, aparentemente saudáveis, foram submetidos a exame bacterioscópico e cultura aeróbia a anaeróbia. As bactérias mais freqüentemente isoladas foram os estreptococos do grupo D (1.2 serpentes, Enterobacter sp. (6, Providencia rettgeri (6, Providencia sp. (4, Escherichia coli (4, Morganella morganii (3 e Clostridium sp. (5. Como estas bactérias são semelhantes às encontradas nos abscessos de pacientes picados por serpentes do gênero Bothrops, é válido considerar a possibilidade de que bactérias da boca da serpente sejam inoculadas no momento da picada e, encontrando condições favoráveis de multiplicação, causem infecção.Culture of fang, fang sheath and venom of fifteen healthy freshly captured Bothrops jararaca were analised. The bacteria most frequently encountered were group D streptococci (12 snakes, Enterobacter sp. (6, Providencia rettgeri (6, Providencia sp. (4, Escherichia coli (4, Morganella morganii (3 and Clostridium sp. (5. The bacteria observed are similar to those found in the abscesses from Bothrops bitten patients. Since these snake mouth bacteria may be inoculated during the snake bite, bacterial multiplication and infection may occur under favorable conditions.

  7. Another new and threatened species of lancehead genus Bothrops (Serpentes, Viperidae) from Ilha dos Franceses, Southeastern Brazil.

    Science.gov (United States)

    Barbo, Fausto E; Gasparini, João Luiz; Almeida, Antonio P; Zaher, Hussam; Grazziotin, Felipe G; Gusmão, Rodrigo B; Ferrarini, José Mário G; Sawaya, Ricardo J

    2016-01-01

    A new insular species of the genus Bothrops is described from Ilha dos Franceses, a small island off the coast of Espírito Santo State, in southeastern Brazil. The new species differs from mainland populations of B. jararaca mainly by its small size, relative longer tail, relative smaller head length, and relative larger eyes. The new species is distinguished from B. alcatraz, B. insularis and B. otavioi by the higher number of ventral and subcaudal scales, relative longer tail and smaller head. The new species is highly abundant on the island, being nocturnal, semiarboreal, and feeding on small lizards and centipeds. Due its unique and restricted area of occurrence, declining quality of habitat, and constant use of the island for tourism, the new species may be considered as critically endangered. PMID:27394563

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

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    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.

  9. 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.

  10. Purification and Characterization of Jararassin-I,A Thrombin-like Enzyme from Bothrops jararaca Snake Venom

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    Débora F. VIEIRA; Leandra WATANABE; Carolina D. SANT'ANA; Silvana MARCUSSI; Suely V. SAMPAIO; Andreimar M. SOARES; Raghuvir K. ARNI

    2004-01-01

    A thrombin-like serine protease, jararassin-I, was isolated from the venom of Bothrops jararaca. The protein was obtained in high yield and purity by a single chromatographic step using the affinity resin Benzamidine-Sepharose CL-6B. SDS-PAGE and dynamic light scattering analyses indicated that the molecular mass of the enzyme was about 30 kD. The enzyme possessed fibrinogenolytic and coagulant activities. The jararassin-I degraded the Bβ chain of fibrinogen while the Aα chain and γ chain were unchanged.Proteases inhibitors, PMSF and benzamidine inhibited the coagulant activity. These results showed jararassinI is a serine protease similar to coagulating thrombin-like snake venom proteases, but it specifically cleaves Bβ chain of bovine fibrinogen. Single crystals of enzyme were obtained (0.2 mm×0.2 mm×0.2 mm) and used for X-ray diffraction experiments.

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

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    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.

  12. 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. PMID:26739285

  13. Crystallization and preliminary crystallographic studies of a phospholipase A2 from the venom of the Brazilian snake Bothrops moojeni.

    Science.gov (United States)

    Nonato, M C; Garratt, R C; Mascarenhas, Y P; Jesus, W D; Assakura, M T; Serrano, S M; Oliva, G

    2001-04-01

    A phospholipase A(2) purified from the venom of the snake Bothrops moojeni has been crystallized by vapour-diffusion techniques in hanging drops at 291 K. The crystals, which were grown in the absence of Ca(2+), belong to the cubic system, space group P432, with unit-cell parameters a = b = c = 91.86 A, and contain one molecule in the asymmetric unit (V(M) = 2.71 A(3) Da(-1)). X-ray diffraction experiments provide data to 2.35 A resolution collected on a rotating-anode home source at cryogenic temperatures. The structure has been solved via molecular-replacement techniques using a single monomer of the crystallographic structure of the phospholipase from the Western diamondback rattlesnake (Crotalus atrox) as a search model. PMID:11264594

  14. Preliminary assessment of Hedychium coronarium essential oil on fibrinogenolytic and coagulant activity induced by Bothrops and Lachesis snake venoms

    Science.gov (United States)

    2014-01-01

    Background The search for new inhibitors of snake venom toxins is essential to complement or even replace traditional antivenom therapy, especially in relation to compounds that neutralize the local effects of envenomations. Besides their possible use as alternative to traditional antivenom therapy, some plant species possess bioactive secondary metabolites including essential oils, which can be extracted from weeds that are considered substantial problems for agriculture, such as Hedychium coronarium. Methods The essential oils of leaves and rhizomes from H. coronarium were extracted by hydrodistillation, and their potential inhibitory effects on the coagulant and fibrinogenolytic activities induced by the venoms of Lachesis muta, Bothrops atrox and Bothrops moojeni were analyzed. Citrated human plasma was used to evaluate the clotting time whereas changes in fibrinogen molecules were visualized by electrophoresis in polyacrylamide gel. The experimental design used for testing coagulation inhibition was randomized in a 3 × 2 factorial arrangement (concentration × essential oils), with three replications. The essential oils were compared since they were extracted from different organs of the same botanical species, H. coronarium. Results The results suggest that the oils interact with venom proteases and plasma constituents, since all oils evaluated, when previously incubated with venoms, were able to inhibit the clotting effect, with less inhibition when oils and plasma were preincubated prior to the addition of venoms. Conclusions Thus, after extensive characterization of their pharmacological and toxicological effects, the essential oils can be used as an alternative to complement serum therapy, especially considering that these plant metabolites generally do not require specific formulations and may be used topically immediately after extraction. PMID:26413083

  15. Aqueous Leaf Extract of Jatropha gossypiifolia L. (Euphorbiaceae) Inhibits Enzymatic and Biological Actions of Bothrops jararaca Snake Venom

    Science.gov (United States)

    Félix-Silva, Juliana; Souza, Thiago; Menezes, Yamara A. S.; Cabral, Bárbara; Câmara, Rafael B. G.; Silva-Junior, Arnóbio A.; Rocha, Hugo A. O.; Rebecchi, Ivanise M. M.; Zucolotto, Silvana M.; Fernandes-Pedrosa, Matheus F.

    2014-01-01

    Snakebites are a serious public health problem due their high morbi-mortality. The main available specific treatment is the antivenom serum therapy, which has some disadvantages, such as poor neutralization of local effects, risk of immunological reactions, high cost and difficult access in some regions. In this context, the search for alternative therapies is relevant. Therefore, the aim of this study was to evaluate the antiophidic properties of Jatropha gossypiifolia, a medicinal plant used in folk medicine to treat snakebites. The aqueous leaf extract of the plant was prepared by decoction and phytochemical analysis revealed the presence of sugars, alkaloids, flavonoids, tannins, terpenes and/or steroids and proteins. The extract was able to inhibit enzymatic and biologic activities induced by Bothrops jararaca snake venom in vitro and in vivo. The blood incoagulability was efficiently inhibited by the extract by oral route. The hemorrhagic and edematogenic local effects were also inhibited, the former by up to 56% and the latter by 100%, in animals treated with extract by oral and intraperitoneal routes, respectively. The inhibition of myotoxic action of B. jararaca reached almost 100%. According to enzymatic tests performed, it is possible to suggest that the antiophidic activity may be due an inhibitory action upon snake venom metalloproteinases (SVMPs) and/or serine proteinases (SVSPs), including fibrinogenolytic enzymes, clotting factors activators and thrombin like enzymes (SVTLEs), as well upon catalytically inactive phospholipases A2 (Lys49 PLA2). Anti-inflammatory activity, at least partially, could also be related to the inhibition of local effects. Additionally, protein precipitating and antioxidant activities may also be important features contributing to the activity presented. In conclusion, the results demonstrate the potential antiophidic activity of J. gossypiifolia extract, including its significant action upon local effects, suggesting that

  16. Aqueous leaf extract of Jatropha gossypiifolia L. (Euphorbiaceae inhibits enzymatic and biological actions of Bothrops jararaca snake venom.

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    Juliana Félix-Silva

    Full Text Available Snakebites are a serious public health problem due their high morbi-mortality. The main available specific treatment is the antivenom serum therapy, which has some disadvantages, such as poor neutralization of local effects, risk of immunological reactions, high cost and difficult access in some regions. In this context, the search for alternative therapies is relevant. Therefore, the aim of this study was to evaluate the antiophidic properties of Jatropha gossypiifolia, a medicinal plant used in folk medicine to treat snakebites. The aqueous leaf extract of the plant was prepared by decoction and phytochemical analysis revealed the presence of sugars, alkaloids, flavonoids, tannins, terpenes and/or steroids and proteins. The extract was able to inhibit enzymatic and biologic activities induced by Bothrops jararaca snake venom in vitro and in vivo. The blood incoagulability was efficiently inhibited by the extract by oral route. The hemorrhagic and edematogenic local effects were also inhibited, the former by up to 56% and the latter by 100%, in animals treated with extract by oral and intraperitoneal routes, respectively. The inhibition of myotoxic action of B. jararaca reached almost 100%. According to enzymatic tests performed, it is possible to suggest that the antiophidic activity may be due an inhibitory action upon snake venom metalloproteinases (SVMPs and/or serine proteinases (SVSPs, including fibrinogenolytic enzymes, clotting factors activators and thrombin like enzymes (SVTLEs, as well upon catalytically inactive phospholipases A2 (Lys49 PLA2. Anti-inflammatory activity, at least partially, could also be related to the inhibition of local effects. Additionally, protein precipitating and antioxidant activities may also be important features contributing to the activity presented. In conclusion, the results demonstrate the potential antiophidic activity of J. gossypiifolia extract, including its significant action upon local effects

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

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    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

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

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    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.

  19. 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

    2013-02-01

    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

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

    Directory of Open Access Journals (Sweden)

    Leonardo A. Calderon

    2013-02-01

    Full Text Available 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.

  1. Analgesic Effect of Photobiomodulation on Bothrops Moojeni Venom-Induced Hyperalgesia: A Mechanism Dependent on Neuronal Inhibition, Cytokines and Kinin Receptors Modulation

    Science.gov (United States)

    Oliveira, Victoria Regina da Silva; Toniolo, Elaine Flamia; Feliciano, Regiane dos Santos; da Silva Jr., José Antonio; Zamuner, Stella Regina

    2016-01-01

    Background Envenoming induced by Bothrops snakebites is characterized by drastic local tissue damage that involves an intense inflammatory reaction and local hyperalgesia which are not neutralized by conventional antivenom treatment. Herein, the effectiveness of photobiomodulation to reduce inflammatory hyperalgesia induced by Bothrops moojeni venom (Bmv), as well as the mechanisms involved was investigated. Methodology/Principal Findings Bmv (1 μg) was injected through the intraplantar route in the right hind paw of mice. Mechanical hyperalgesia and allodynia were evaluated by von Frey filaments at different time points after venom injection. Low level laser therapy (LLLT) was applied at the site of Bmv injection at wavelength of red 685 nm with energy density of 2.2 J/cm2 at 30 min and 3 h after venom inoculation. Neuronal activation in the dorsal horn spinal cord was determined by immunohistochemistry of Fos protein and the mRNA expression of IL-6, TNF-α, IL-10, B1 and B2 kinin receptors were evaluated by Real time-PCR 6 h after venom injection. Photobiomodulation reversed Bmv-induced mechanical hyperalgesia and allodynia and decreased Fos expression, induced by Bmv as well as the mRNA levels of IL-6, TNF-α and B1 and B2 kinin receptors. Finally, an increase on IL-10, was observed following LLLT. Conclusion/Significance These data demonstrate that LLLT interferes with mechanisms involved in nociception and hyperalgesia and modulates Bmv-induced nociceptive signal. The use of photobiomodulation in reducing local pain induced by Bothropic venoms should be considered as a novel therapeutic tool for the treatment of local symptoms induced after bothropic snakebites. PMID:27749899

  2. An Isoflavone from Dipteryx alata Vogel is Active against the in Vitro Neuromuscular Paralysis of Bothrops jararacussu Snake Venom and Bothropstoxin I, and Prevents Venom-Induced Myonecrosis

    OpenAIRE

    Miriéle C. Ferraz; Edson H. Yoshida; Renata V.S. Tavares; Cogo, José C; Adélia C.O. Cintra; Dal Belo, Cháriston A.; Franco, Luiz M.; dos Santos, Márcio G; Flávia A Resende; Eliana A. Varanda; Stephen Hyslop; Pilar Puebla; Arturo San Feliciano; Yoko Oshima-Franco

    2014-01-01

    Snakebite is a neglected disease and serious health problem in Brazil, with most bites being caused by snakes of the genus Bothrops. Although serum therapy is the primary treatment for systemic envenomation, it is generally ineffective in neutralizing the local effects of these venoms. In this work, we examined the ability of 7,8,3'-trihydroxy-4'-methoxyisoflavone (TM), an isoflavone from Dipteryx alata, to neutralize the neurotoxicity (in mouse phrenic nerve-diaphragm preparations) and myoto...

  3. Neutralization of the edema-forming, defibrinating and coagulant effects of Bothrops asper venom by extracts of plants used by healers in Colombia

    OpenAIRE

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

    2004-01-01

    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 Pleop...

  4. Sexual dimorphism in development and venom production of the insular threatened pit viper Bothrops insularism (Serpentes: Viperidae) of Queimada Grande Island, Brazil

    OpenAIRE

    S.R. Travaglia-Cardoso; A. Zelanis; M. F. D. Furtado

    2010-01-01

    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 Labora...

  5. 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

  6. Description of a new ascaridoid parasite of Bothrops jararaca Wied (Reptilia, Ophidia in Brazil Descrição de um novo ascarídeo parasito de Bothrops jararaca Wied (Reptilia, Ophidia no Brasil

    Directory of Open Access Journals (Sweden)

    Liege R. Siqueira

    2005-09-01

    Full Text Available A new species of Ophidascaris Baylis, 1920 is described on the basis of specimens recovered from the pit viper Bothrops jararaca Wied, 1824 in Brazil. The new species referred here was compared to other of the genus that are the closest. The differentiation was based on reliable morphological characters, mainly on what refers to the spicular length, number and distribution of caudal papillae, location of the vulvar opening, eggs size and aspect of the labial indentations. Moreover, female specimens are the only to possess a most prominent and characteristic post-anal tubercle-like structure and when analyzed together with the males, that show a great number of pre-cloacal papillae, the presence of an unpaired papilla and also of a less conspicuous post-cloacal tubercle-like structure, permit the prompt identification of the new species.Uma nova espécie de Ophidascaris Baylis, 1920 é descrita, com base em exemplares parasitos de jararaca, Bothrops jararaca Wied, 1824 no Brasil. A nova espécie agora descrita foi comparada a outras do gênero que lhe são as mais próximas. A diferenciação foi baseada em caracteres morfológicos consistentes, principalmente nos que dizem respeito ao tamanho dos espículos, número e distribuição de papilas caudais, localização da abertura vulvar, tamanho dos ovos e aspecto das indentações labiais. Além disso, os exemplares fêmeas são os únicos no gênero a apresentarem uma estrutura em forma de tubérculo pós-anal bastante característica e proeminente, quando analisados junto aos exemplares machos, nos quais existe um grande número de papilas pré-cloacais, presença de papila caudal ímpar e, também, da estrutura pós-cloacal em forma de tubérculo, embora menos evidente que nas fêmeas, permitem a pronta identificação da espécie nova.

  7. Linear B-cell epitopes in BthTX-1, BthTX-II and BthA-1, phospholipase A₂'s from Bothrops jararacussu snake venom, recognized by therapeutically neutralizing commercial horse antivenom.

    Science.gov (United States)

    De-Simone, Salvatore G; Napoleão-Pego, Paloma; Teixeira-Pinto, Luiz A L; Santos, Jonathas D L; De-Simone, Thatiane S; Melgarejo, Anibal R; Aguiar, Aniesse S; Marchi-Salvador, Daniela P

    2013-09-01

    The benefits from treatment with antivenom sera are indubitable. However, the mechanism for toxin neutralization has not been completely elucidated. A mixture of anti-bothropic and anti-crotalic horse antivenom has been reported to be more effective in neutralizing the effects of Bothrops jararacussu snake venom than anti-bothropic antivenom alone. This study determined which regions in the three PLA₂s from B. jararacussu snake venom are bound by antibodies in tetravalent anti-bothropic and monovalent anti-crotalic commercial horse antivenom. Mapping experiments of BthTX-I, BthTX-II and BthA-I using two small libraries of 69 peptides each revealed six major IgG-binding epitopes that were recognized by both anti-bothropic and anti-crotalic horse antivenom. Two epitopes in BthTX-I were only recognized by the anti-bothropic horse antivenom, while anti-crotalic horse antivenom recognized four unique epitopes across the three PLA₂s. Our studies suggest that the harmful activities of the PLA₂s present in the venom of B. jararacussu are neutralized by the combinatorial treatment with both antivenom sera through their complementary binding sites, which provides a wide coverage on the PLA₂s. This is the first peptide microarray of PLA₂s from B. jararacussu snake venom to survey the performance of commercial horse antiophidic antivenom. Regions recognized by the protective antivenom sera are prime candidates for improved venom cocktails or a chimeric protein encoding the multiple epitopes to immunize animals as well as for designing future synthetic vaccines. PMID:23792452

  8. 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.

  9. 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.

  10. The crucial role of the MyD88 adaptor protein in the inflammatory response induced by Bothrops atrox venom.

    Science.gov (United States)

    Moreira, Vanessa; Teixeira, Catarina; Borges da Silva, Henrique; D'Império Lima, Maria Regina; Dos-Santos, Maria Cristina

    2013-06-01

    Most snake accidents in North Brazil are attributed to Bothrops atrox, a snake species of the Viperidae family whose venom simultaneously induces local and systemic effects in the victims. The former are clinically more important than the latter, as they cause severe tissue lesions associated with strong inflammatory responses. Although several studies have shown that inflammatory mediators are produced in response to B. atrox venom (BaV), there is little information concerning the molecular pathways involved in innate immune system signaling. Myeloid differentiation factor 88 (MyD88) is an adaptor molecule responsible for transmitting intracellular signals from most toll-like receptors (TLRs) after they interact with pathogen-associated molecular patterns (PAMPs) or other stimuli such as endogenous damage-associated molecular patterns (DAMPs). The MyD88-dependent pathway leads to activation of transcription factors, which in turn induce synthesis of inflammatory mediators such as eicosanoids, cytokines and chemokines. The aim of this study was to investigate the involvement of MyD88 on the acute inflammatory response induced by BaV. Wild-type (WT) C57BL/6 mice and MyD88 knockout (MyD88(-/-)) mice were intraperitoneally injected with BaV. Compared to WT mice, MyD88(-/-) animals showed an impaired inflammatory response to BaV, with lower influx of polymorphonuclear and mononuclear cells to the peritoneal cavity. Furthermore, peritoneal leukocytes from BaV-injected MyD88(-/-) mice did not induce COX-2 or LTB4 protein expression and released low concentrations of PGE2. These mice also failed to produce Th1 and Th17 cytokines and CCL-2, but IL-10 levels were similar to those of BaV-injected WT mice. Our results indicate that MyD88 signaling is required for activation of the inflammatory response elicited by BaV, raising the possibility of developing new therapeutic targets to treat Bothrops sp. poisoning. PMID:23474268

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

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    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.

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

    International Nuclear Information System (INIS)

    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. (Author)

  13. 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.

  14. New methodology for the obtainment of antibothropic factors from the South American opossum (Didelphis marsupialis) and jararaca snake (Bothrops jararaca).

    Science.gov (United States)

    Neves-Ferreira, A G; Valente, R H; Sá, P G; Rocha, S L; Moussatché, H; Domont, G B; Perales, J

    1999-10-01

    The antibothropic factor (ABF) from D. marsupialis was collected from perforated hollow plastic golf balls which were surgically implanted subcutaneously in anesthetized opossums, a technique originally described for the production of polyclonal antibodies. Two months after the implantation of the balls, approximately 15 ml of seromatous fluid from D. marsupialis (SFDm-50 mg total protein/ml) could be recovered monthly. Opossum serum as well as SFDm showed similar SDS-PAGE profiles and antihemorrhagic potencies against Bothrops jararaca snake venom (Bjv). The presence of ABF in SFDm was confirmed by immunoblotting, using rabbit polyclonal antibodies raised against ABF isolated from opossum serum. ABF isolated from SFDm or from serum by ion-exchange chromatography showed identical chromatographic and electrophoretic profiles. ABF fromboth sources displayed very similar antihemorrhagic and anticaseinolytic activities against Bjv. In the case of B. jararaca, polyethylene perforated tubes were inserted in the abdominal cavity and two months after implantation, approximately 4 ml of seromatous fluid from B. jararaca (SFBj-23 mg total protein/ml) were recovered. B.jararaca serum and SFBj showed the same native and SDS-PAGE band pattern. Both serum and SFBj inhibited Bjv hemorrhagic activity. We conclude that this new methodology is very suitable for continuously obtaining opossum ABF and SFBj, in large scale and in an easier way, avoiding animal suffering and eventual sacrifice. PMID:10414866

  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. PMID:27152384

  16. Isolation and characterization of two new non-hemorrhagic metalloproteinases with fibrinogenolytic activity from the mapanare (Bothrops colombiensis) venom.

    Science.gov (United States)

    Girón, María E; Rodríguez-Acosta, Alexis; Salazar, Ana María; Sánchez, Elda E; Galán, Jacob; Ibarra, Carlos; Guerrero, Belsy

    2013-01-01

    Colombienases are acidic, low molecular weight metalloproteinases (Mr of 23,074.31 Da colombienase-1 and 23,078.80 Da colombienase-2; pI of 6.0 and 6.2, respectively) isolated from Bothrops colombiensis snake venom. The chromatographic profile in RP-HPLC and its partial sequence confirmed its high homogeneity. Both colombienases present fibrino(geno)lytic activity, but did not show any hemorrhagic, amidolytic, plasminogen activator or coagulant activities, and no effect on platelet aggregation induced by collagen or ADP. Both enzymes were strongly active on fibrinogen Aα chains followed by the Bβ chains, and colombienases-2, at high doses, also degraded the γ chains. This activity was stable at temperatures ranging between 4 and 37 °C, with a maximum activity at 25 °C, and at pHs between 7 and 9. The homology demonstrated by the comparison of sequences, with zinc-dependent metalloproteinases, as well as the metal chelant effects on, confirmed that the colombienases were metalloproteinases, particularly to α-fibrinogenases belonging to the P-I class of SVPMs (20-30 kDa), which contain only the single-domain proteins. The biological characteristics of the colombienases confer a therapeutic potential, since they contain a high fibrino(geno)lytic activity, devoid of hemorrhagic activity. These metalloproteinases might be explored as thrombolytic agents given that they dissolve fibrin clots or prevent their formation.

  17. Identification of protective B-cell epitopes of Atroxlysin-I: A metalloproteinase from Bothrops atrox snake venom.

    Science.gov (United States)

    Schneider, F S; de Almeida Lima, S; Reis de Ávila, G; Castro, K L; Guerra-Duarte, C; Sanchez, E F; Nguyen, C; Granier, C; Molina, F; Chávez-Olortegui, C

    2016-03-29

    Atroxlysin-I (Atr-I) is a hemorrhagic snake venom metalloproteinase (SVMP) from Bothrops atrox venom, the snake responsible for the majority of bites in the north region of South America. SVMPs like Atr-I produce toxic effects in victims including hemorrhage, inflammation, necrosis and blood coagulation deficiency. Mapping of B-cell epitopes in SVMPs might result in the identification of non-toxic molecules capable of inducing neutralizing antibodies and improving the anti-venom therapy. Here, using the SPOT-synthesis technique we identified two epitopes located in the N-ter region of Atr-I (AtrEp1-(22)YNGNSDKIRRRIHQM(36); and AtrEp2-(55)GVEIWSNKDLINVQ(68)). Based on the sequence of AtrEp1 and AtrEp2 a third peptide named Atr-I biepitope (AtrBiEp) was designed and synthesized ((23)NGNSDKIRRRIH(34)GG(55)GVEIWSNKDLINVQ(68)). AtrBiEp was used to immunize BALB/c mice. Anti-AtrBiEp serum cross-reacted against Atr-I in western blot and was able to fully neutralize the hemorrhagic activity of Atr-I. Our results provide a rational basis for the identification of neutralizing epitopes on Atr-I snake venom toxin and show that the use of synthetic peptides could improve the generation of immuno-therapeutics. PMID:26917009

  18. Inflammatory mediators involved in the paw edema and hyperalgesia induced by Batroxase, a metalloproteinase isolated from Bothrops atrox snake venom.

    Science.gov (United States)

    De Toni, Lanuze G B; Menaldo, Danilo L; Cintra, Adélia C O; Figueiredo, Maria J; de Souza, Anderson R; Maximiano, William M A; Jamur, Maria C; Souza, Glória E P; Sampaio, Suely V

    2015-09-01

    Snake venom metalloproteinases have been described as responsible for several inflammatory effects. In this study, we investigated the edema and hyperalgesia induced in rats by Batroxase, a P-I metalloproteinase from Bothrops atrox venom, along with possible inflammatory mediators involved in these responses. Batroxase or sterile saline was injected into rat paws and the edema and hyperalgesic effects were evaluated for 6h by using a plethysmometer and a Von Frey system, respectively. Batroxase induced significant edematogenic and hyperalgesic peak responses in the first hours after administration. The inflammatory mediators involved in these responses were assayed by pretreatment of animals with synthesis inhibitors or receptor antagonists. Peak responses were significantly reduced by administration of the glucocorticoid dexamethasone, the H1 receptor antagonist diphenhydramine and the FLAP inhibitor MK-886. Rat paws injected with compound 48/80, a mast cell degranulating agent, followed by Batroxase injection resulted in significant reduction of the edema and hyperalgesia. However, Batroxase itself induced minor degranulation of RBL-2H3 mast cells in vitro. Additionally, the inflammatory responses did not seem to be related to prostaglandins, bradykinin or nitric oxide. Our results indicate a major involvement of histamine and leukotrienes in the edema and hyperalgesia induced by Batroxase, which could be related, at least in part, to mast cell degranulation. PMID:26072684

  19. Platelet-active substances in the venom of Bothrops moojeni snake-a novel evaluation method using whole blood aggregometry.

    Science.gov (United States)

    Demler, Christine; Bühler, Beatrice; Menin, Laure; Stöcklin, Reto; Wilmer, Marianne; Ernst, Beat; Perchuc, Anna Maria

    2010-01-01

    The objective of the present study was an investigation of the crude Bothrops moojeni venom, aiming at the identification of new compounds with platelet-activating or -inhibiting activity. The venom was separated by gel filtration chromatography into 18 fractions, which were tested by means of whole blood aggregometry for their activities affecting the aggregation of blood platelets. In order to eliminate interferences caused by prothrombin activators or thrombin like-enzymes, which are frequently present in snake venoms, a test method for screening protein mixtures was developed. To avoid clotting of the blood samples, the thrombin inhibitor hirudin and the synthetic inhibitor of fibrin polymerization Pefabloc FG were applied. In the present study, a platelet aggregation activator with an activity resembling thrombocytin from B. atrox was identified in one of the examined venom fractions. In addition, a platelet antagonist-most likely a disintegrin-with broad inhibitory activity against aggregation triggered by collagen, adenosine diphosphate and thrombin receptor activating peptide, was identified. PMID:19938887

  20. On the quaternary structure of a C-type lectin from Bothrops jararacussu venom--BJ-32 (BjcuL).

    Science.gov (United States)

    Silva, F P; Alexandre, G M C; Ramos, C H I; De-Simone, S G

    2008-12-15

    BJ-32 (also known as BjcuL) is a C-type lectin from the venom of Bothrops jararacussu with specificity for beta-galactosides and a remarkable ability to agglutinate several species of trypanosomatids. Our objective was to study the oligomerization state of native BJ-32 by using different biophysical and computational methods. Small-angle X-ray light scattering (SAXS) experiments disclosed a compact, globular protein with a radius of gyration of 36.72+/-0.04A and molecular weight calculated as 147.5+/-2.0kDa. From analytical ultracentrifugation analysis, it was determined that the BJ-32 sedimentation profile fits nicely to a decamer model. The analysis of the intrinsic emitted fluorescence spectra for BJ-32 solutions indicated that association of subunits in the decamer is accompanied by changes in the environment of Tryptophan residues. Both ab initio and comparative models of BJ-32 supported the resemblance of the decamer in the crystallographic structure from a close homologue, the rattlesnake venom lectin (RSL) from Crotalus atrox. PMID:18948130

  1. Evidence of caspase-mediated apoptosis induced by l-amino acid oxidase isolated from Bothrops atrox snake venom.

    Science.gov (United States)

    Alves, Raquel Melo; Antonucci, Gilmara Ausech; Paiva, Helder Henrique; Cintra, Adélia Cristina Oliveira; Franco, João José; Mendonça-Franqueiro, Elaine Paula; Dorta, Daniel Junqueira; Giglio, José Roberto; Rosa, José César; Fuly, André Lopes; Dias-Baruffi, Marcelo; Soares, Andreimar Martins; Sampaio, Suely Vilela

    2008-12-01

    The aim of this work was to investigate the involvement of caspases in apoptosis induced by l-amino acid oxidase isolated from Bothrops atrox snake venom. The isolation of LAAO involved three chromatographic steps: molecular exclusion on a G-75 column; ion exchange column by HPLC and affinity chromatography on a Lentil Lectin column. SDS-PAGE was used to confirm the expected high purity level of BatroxLAAO. It is a glycoprotein with 12% sugar and an acidic character, as confirmed by its amino acid composition, rich in "Asp and Glu" residues. It displays high specificity toward hydrophobic l-amino acids. The N-terminal amino acid sequence and internal peptide sequences showed close structural homology to other snake venom LAAOs. This enzyme induces in vitro platelet aggregation, which may be due to H2O2 production by LAAOs, since the addition of catalase completely inhibited the aggregation effect. It also showed cytotoxicity towards several cancer cell lines: HL60, Jurkat, B16F10 and PC12. The cytotoxicity activity was abolished by catalase. A fluorescence microscopy evaluation revealed a significant increase in the apoptotic index of these cells after BatroxLAAO treatment. This observation was confirmed by phosphatidyl serine exposure and activation of caspases. BatroxLAAO is a protein with various biological functions that can be involved in envenomation. Further investigations of its function will contribute to toxicology advances. PMID:18804547

  2. A tripeptide isolated from Bothrops atrox venom has neuroprotective and neurotrophic effects on a cellular model of Parkinson's disease.

    Science.gov (United States)

    Martins, N M; Santos, N A G; Sartim, M A; Cintra, A C O; Sampaio, S V; Santos, A C

    2015-06-25

    Parkinson's disease (PD) is the second most common neurodegenerative disorder; however, there is no treatment able to prevent the loss of dopaminergic neurons or its consequences. Trophic factors such as NGF and BDNF has positive effects on different disorders of the brain, including neurodegeneration. Additionally, studies have suggested the use of venom peptides as a therapeutic strategy for neurological disorders. Therefore, in the present study, we investigated the neuroprotective activity of a peptide isolated from Bothrops atrox venom and its trophic ability by using a cellular model of dopaminergic neurotoxicity induced by 1-methyl-4-phenylpyridinium (MPP(+)) in PC12 cells. We showed that it decreased the activities of the apoptotic proteases caspase-9 (mitochondrial) and caspase-3 (executor) and increased cell viability and proliferation in this model. Additionally, it increased neuritogenesis in non-treated PC12 cells (neuronal model) as well as in PC12 cells treated with the dopaminergic neurotoxin. The amino acid sequence of the peptide was identified as Glutamic acid-Valine-Tryptophan (Glu-Val-Trp). These findings suggest that this tripeptide has the potential to protect against the dopaminergic neurons loss and that trophic stimulation of neuroplasticity might be involved in its mechanism of neuroprotection. PMID:25868679

  3. Purification from Bothrops lanceolatus (fer de lance) venom of a fibrino(geno)lytic enzyme with esterolytic activity.

    Science.gov (United States)

    Lôbo de Araújo, A; Donato, J L; Bon, C

    1998-05-01

    Bothrops lanceolatus venom has high caseinolytic, phospholipasic, esterolytic and hemorrhagic activities. In spite of having no coagulant effect on plasma, this venom contains a thrombin-like enzyme. Using gel filtration and ion-exchange chromatographies, we have purified an esterolytic fraction (F-II-1a) from this venom with a protein yield of 4% and a 58% recovery in enzyme activity. SDS-PAGE in the presence of beta-mercaptoethanol showed that the enzyme is a single chain polypeptide with a MW=38,100. Immunodiffusion and immunoelectrophoresis of fraction F-II-1a against serum from horses immunized with B. lanceolatus venom and against rabbit antiserum prepared using fraction F-II-1a both showed a single immunoprecipitin line. The Km and Vmax values for TAME hydrolysis were 0.85 mM and 38.6 micromol/min/mg, respectively. The esterolytic activity was completely inhibited by PMSF (10 mM) but not by EDTA (20 mM). Fraction F-II-1a hydrolyzed the alpha and beta chains of fibrinogen. Degradation of the alpha chain occurred within 10 min while that of the beta-chain was slower. The enzyme had no effect on the gamma-chain even after 4 h of hydrolysis. PMID:9655635

  4. Fatal diffuse thrombotic microangiopathy after a bite by the "Fer-de-Lance" pit viper (Bothrops lanceolatus) of Martinique.

    Science.gov (United States)

    Malbranque, Stéphane; Piercecchi-Marti, Marie Dominique; Thomas, Laurent; Barbey, Christophe; Courcier, Dominique; Bucher, Bernard; Ridarch, Alex; Smadja, Didier; Warrell, David A

    2008-06-01

    In Martinique, a man bitten two days earlier by a pit viper (Bothrops lanceolatus) was hospitalized with impaired consciousness and tetraplegia. Investigations confirmed cerebral and myocardial infarctions. Resolving thrombocytopenia was associated with virtually normal blood prothrombin time/activated partial thromboplastin time but increasing hyperfibrinogenemia. Despite specific antivenom treatment, he developed fatal left ventricular failure six days after the bite. At autopsy, multiple cerebral, myocardial and mesenteric infarctions were found. Rupture of mitral chordae tendinae was the likely cause of death. Histopathologic examination showed multi-focal thrombotic microangiopathy with intimal-medial dissection by thrombi extending from foci of endothelial damage in small cerebral, myocardial, pulmonary, mesenteric, and interlobular renal arteries and arterioles. These findings were the causes of infarctions. There was intense angiogenesis in organizing cerebral infarcts. Immunohistochemical analysis showed platelet aggregates and endothelial cells within microthrombi. Viperidae venoms contain vascular endothelial toxins, notably metalloproteinase hemorrhagins, but von Willebrand factor activators or vascular endothelial growth factor-type factors are more likely to have been implicated in this case. PMID:18541759

  5. 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.

  6. 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.

  7. 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.

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

    Science.gov (United States)

    Prado, Nidiane D. R.; Pereira, Soraya S.; da Silva, Michele P.; Morais, Michelle S. S.; Kayano, Anderson M.; Moreira-Dill, Leandro S.; Luiz, Marcos B.; Zanchi, Fernando B.; Fuly, André L.; E. F. Huacca, Maribel; Fernandes, Cleberson F.; Calderon, Leonardo A.; Zuliani, Juliana P.; Soares, Andreimar M.; Stabeli, Rodrigo G.; F. C. Fernandes, Carla

    2016-01-01

    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. PMID:27028872

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

    Science.gov (United States)

    Prado, Nidiane D R; Pereira, Soraya S; da Silva, Michele P; Morais, Michelle S S; Kayano, Anderson M; Moreira-Dill, Leandro S; Luiz, Marcos B; Zanchi, Fernando B; Fuly, André L; Huacca, Maribel E F; Fernandes, Cleberson F; Calderon, Leonardo A; Zuliani, Juliana P; Pereira da Silva, Luiz H; Soares, Andreimar M; Stabeli, Rodrigo G; Fernandes, Carla F C

    2016-01-01

    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. PMID:27028872

  10. Collaborative Calibration of Control Standards of Hemocoagulase Bothrops Atrox%矛头蝮蛇血凝酶工作标准品的协作标定

    Institute of Scientific and Technical Information of China (English)

    李秀琳; 孙东; 丁忠福; 宁静; 于翀; 崔亮亮; 薛雁

    2016-01-01

    目的:标定我公司生产矛头蝮蛇血凝酶工作标准品。方法:以蛇毒血凝酶质量标准WS-349( X-317)-2001中效价测定为基础,改进测定方法,组织多名实验人员协作标定,并对数据进行统计分析。结果:本批矛头蝮蛇血凝酶工作标准品酶活性单位为103 U/支,加权均值标准差为0.8068,95%加权均值可信限的范围为101.0~105.1 U/支。结论:我公司生产的矛头蝮蛇血凝酶工作标准品标化科学、赋值准确,可以用于本公司产品的效价检测。%Objective:To calibrate the control standards of Hemocoagulase Bothrops atrox of our company.Methods:The enzyme ac-tivity was assayed based on the titration of Hemocoagulase Atrox Quality Standard WS-349(X-317)-2001, the method was modified, more than a number of experimenters were organized to collaboratively calibrate and the data was statistically analyzed.Results: The activity of the control standards of Hemocoagulase Bothrops atrox was 103 units per bottle, the weighted average standard deviation was 0.806 8, and the 95%weighted mean confidence range was 101.0-105.1 units per bottle.Conclusion:The calibration of the control standards of Hemocoagulase Bothrops atrox is scientific and accurate, and can be used to detect the titers of our company′s products .

  11. Clinical trial of two antivenoms for the treatment of Bothrops and Lachesis bites in the north eastern Amazon region of Brazil.

    Science.gov (United States)

    Pardal, Pedro Pereira de Oliveira; Souza, Suzana Medeiro; Monteiro, Maria Rita de Cássia da Costa; Fan, Hui Wen; Cardoso, João Luiz Costa; França, Francisco Oscar Siqueira; Tomy, Sandra Corrallo; Sano-Martins, Ida S; de Sousa-e-Silva, Maria Cristina Cirillo; Colombini, Mônica; Kodera, Nancy F; Moura-da-Silva, Ana Maria; Cardoso, Diva Ferreira; Velarde, David Toledo; Kamiguti, Aura S; Theakston, Robert David; Warrell, David A

    2004-01-01

    The efficacies of specific Bothrops atrox-Lachesis and standard Bothrops-Lachesis antivenoms were compared in the north eastern Amazon region of Brazil. The main aim was to investigate whether a specific antivenom raised against the venom of B. atrox, the most important Amazon snake species from a medical point of view, was necessary for the treatment of patients in this region. Seventy-four patients with local and systemic effects of envenoming by Bothrops or Lachesis snakes were randomly allocated to receive either specific (n = 38) or standard (n = 36) antivenoms. In 46 cases (24 in the standard antivenom group, 22 in the other) the snake was identified either by enzyme immunoassay or by examination of the dead snake, as B. atrox in 45, L. muta in one. Patients were similar in all clinical and epidemiological respects before treatment. Results indicated that both antivenoms were equally effective in reversing all signs of envenoming detected both clinically and in the laboratory. Venom-induced haemostatic abnormalities were resolved within 24 h after the start of antivenom therapy in most patients. The extent of local complications, such as local skin necrosis and secondary infection, was similar in both groups. There were no deaths. The incidence of early anaphylactic reactions was 18% and 19%, respectively for specific and standard antivenoms; none was life-threatening. Measurement of serum venom concentrations by enzyme immunoassay (EIA) confirmed that both antivenoms cleared venom antigenaemia effectively. EIA also revealed that one patient had been bitten by Lachesis muta, although the clinical features in this case were not distinctive. PMID:14702836

  12. Combined venomics, venom gland transcriptomics, bioactivities, and antivenomics of two Bothrops jararaca populations from geographic isolated regions within the Brazilian Atlantic rainforest.

    Science.gov (United States)

    Gonçalves-Machado, Larissa; Pla, Davinia; Sanz, Libia; Jorge, Roberta Jeane B; Leitão-De-Araújo, Moema; Alves, Maria Lúcia M; Alvares, Diego Janisch; De Miranda, Joari; Nowatzki, Jenifer; de Morais-Zani, Karen; Fernandes, Wilson; Tanaka-Azevedo, Anita Mitico; Fernández, Julián; Zingali, Russolina B; Gutiérrez, José María; Corrêa-Netto, Carlos; Calvete, Juan J

    2016-03-01

    Bothrops jararaca is a slender and semi-arboreal medically relevant pit viper species endemic to tropical and subtropical forests in southern Brazil, Paraguay, and northern Argentina (Misiones). Within its geographic range, it is often abundant and is an important cause of snakebite. Although no subspecies are currently recognized, geographic analyses have revealed the existence of two well-supported B. jararaca clades that diverged during the Pliocene ~3.8Mya and currently display a southeastern (SE) and a southern (S) Atlantic rainforest (Mata Atlântica) distribution. The spectrum, geographic variability, and ontogenetic changes of the venom proteomes of snakes from these two B. jararaca phylogroups were investigated applying a combined venom gland transcriptomic and venomic analysis. Comparisons of the venom proteomes and transcriptomes of B. jararaca from the SE and S geographic regions revealed notable interpopulational variability that may be due to the different levels of population-specific transcriptional regulation, including, in the case of the southern population, a marked ontogenetic venom compositional change involving the upregulation of the myotoxic PLA2 homolog, bothropstoxin-I. This population-specific marker can be used to estimate the proportion of venom from the southern population present in the B. jararaca venom pool used for the Brazilian soro antibotrópico (SAB) antivenom production. On the other hand, the southeastern population-specific D49-PLA2 molecules, BinTX-I and BinTX-II, lend support to the notion that the mainland ancestor of Bothrops insularis was originated within the same population that gave rise to the current SE B. jararaca phylogroup, and that this insular species endemic to Queimada Grande Island (Brazil) expresses a pedomorphic venom phenotype. Mirroring their compositional divergence, the two geographic B. jararaca venom pools showed distinct bioactivity profiles. However, the SAB antivenom manufactured in Vital Brazil

  13. Combined venomics, venom gland transcriptomics, bioactivities, and antivenomics of two Bothrops jararaca populations from geographic isolated regions within the Brazilian Atlantic rainforest.

    Science.gov (United States)

    Gonçalves-Machado, Larissa; Pla, Davinia; Sanz, Libia; Jorge, Roberta Jeane B; Leitão-De-Araújo, Moema; Alves, Maria Lúcia M; Alvares, Diego Janisch; De Miranda, Joari; Nowatzki, Jenifer; de Morais-Zani, Karen; Fernandes, Wilson; Tanaka-Azevedo, Anita Mitico; Fernández, Julián; Zingali, Russolina B; Gutiérrez, José María; Corrêa-Netto, Carlos; Calvete, Juan J

    2016-03-01

    Bothrops jararaca is a slender and semi-arboreal medically relevant pit viper species endemic to tropical and subtropical forests in southern Brazil, Paraguay, and northern Argentina (Misiones). Within its geographic range, it is often abundant and is an important cause of snakebite. Although no subspecies are currently recognized, geographic analyses have revealed the existence of two well-supported B. jararaca clades that diverged during the Pliocene ~3.8Mya and currently display a southeastern (SE) and a southern (S) Atlantic rainforest (Mata Atlântica) distribution. The spectrum, geographic variability, and ontogenetic changes of the venom proteomes of snakes from these two B. jararaca phylogroups were investigated applying a combined venom gland transcriptomic and venomic analysis. Comparisons of the venom proteomes and transcriptomes of B. jararaca from the SE and S geographic regions revealed notable interpopulational variability that may be due to the different levels of population-specific transcriptional regulation, including, in the case of the southern population, a marked ontogenetic venom compositional change involving the upregulation of the myotoxic PLA2 homolog, bothropstoxin-I. This population-specific marker can be used to estimate the proportion of venom from the southern population present in the B. jararaca venom pool used for the Brazilian soro antibotrópico (SAB) antivenom production. On the other hand, the southeastern population-specific D49-PLA2 molecules, BinTX-I and BinTX-II, lend support to the notion that the mainland ancestor of Bothrops insularis was originated within the same population that gave rise to the current SE B. jararaca phylogroup, and that this insular species endemic to Queimada Grande Island (Brazil) expresses a pedomorphic venom phenotype. Mirroring their compositional divergence, the two geographic B. jararaca venom pools showed distinct bioactivity profiles. However, the SAB antivenom manufactured in Vital Brazil

  14. Pharmacological characterisation of arthritis induced by Bothrops jararaca venom in rabbits: a positive cross talk between bradykinin, nitric oxide and prostaglandin E2.

    OpenAIRE

    Suzana B. V. Mello; Maria Luiza Guzzo; Luiz Filipe Santiago Lisboa; Farsky, Sandra H P

    2002-01-01

    BACKGROUND: Our previous results showed that nitric oxide (NO) and bradykinin (BK) mediate the arthritis induced by Bothrops jararaca venom (BjV) in rabbits. In this study, we investigated the contribution of each receptor of BK as well as the inter-relationship between NO and eicosanoids in BjV-induced arthritis. METHODS: The arthritis was induced in rabbits with 16 microg of BjV injected intra-articularly. Prostaglandin E2 (PGE2), thromboxane B2 (TxB2), leukotriene B4 (LTB4) (radioimmunoass...

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

    OpenAIRE

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

    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). D e 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 si...

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

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    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.

  17. 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)

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

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    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.

  19. 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.

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

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    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.

  1. 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

    2016-01-01

    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. PMID:26931059

  2. 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

  3. Inhibition of toxic activities of Bothrops asper venom and other crotalid snake venoms by a novel neutralizing mixture.

    Science.gov (United States)

    Borkow, G; Gutierrez, J M; Ovadia, M

    1997-12-01

    The majority of snake bites in Central America are caused by Bothrops asper, whose venom induce complex local effects such as myonecrosis, edema and especially hemorrhage. These effects are only partially neutralized by the clinically used antivenom, even when administered rapidly after envenomation. Recently we screened 49 substances for antihemorrhagic activity and found that a mixture composed of CaNa2, EDTA, a B. asper serum fraction (natural antidote), and the currently used horse polyvalent antiserum is highly effective in the neutralization of local and systemic hemorrhage developing after B. asper envenomation (Borkow et al., Toxicon 35, 865-877, 1997). In the present study we screened the best six antihemorrhagic compounds for their capacity to neutralize the lethal activity in mice and the proteolytic, hemolytic, and antiattachment activities in vitro of the venom. The compounds tested included the currently used horse antivenom, rabbit antiserum against whole B. asper venom or against heated venom, B. asper and Natrix tessellata serum fractions, and CaNa2 EDTA. The constituents of the antihemorrhagic mixture were also the best inhibitors of the other examined toxic activities. Importantly, the mixture effectively neutralized toxic activities of an additional nine venoms from snakes abundant in Central America. This work suggests that the polyvalent antivenom used in Central America could be enriched with a B. asper serum fraction producing a more effective antivenom. In addition, the local application of CaNa2 EDTA to neutralize hemorrhagic toxins, immediately after a snake bite, may provide rapid inhibition of local damage caused by the venoms. PMID:9439739

  4. 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

  5. Hemorrhagic, coagulant and fibrino(geno)lytic activities of crude venom and fractions from mapanare (Bothrops colombiensis) snakes.

    Science.gov (United States)

    Girón, María E; Salazar, Ana M; Aguilar, Irma; Pérez, John C; Sánchez, Elda E; Arocha-Piñango, Carmen L; Rodríguez-Acosta, Alexis; Guerrero, Belsy

    2008-01-01

    Bothrops colombiensis venom from two similar geographical locations were tested for their hemostatic functions and characterized by gel-filtration chromatography and SDS-PAGE electrophoresis. The snakes were from Caucagua and El Guapo towns of the Venezuelan state of Miranda. Fibrino(geno)lytic, procoagulant, hemorrhagic, lethal activities, gel-filtration chromatography and SDS-PAGE profiles were analyzed and compared for both venoms. The highest hemorrhagic activity of 5.3 mug was seen in El Guapo venom while Caucagua venom had the lowest LD(50) of 5.8 mg/kg. Both venoms presented similar thrombin-like activity. El Guapo showed a factor Xa-like activity two times higher than Caucagua. Differences were observed in kallikrein-like and t-PA activities, being highest in El Guapo. Caucagua venom showed the maximum fibrin lysis. Both crude venom runs on Sephadex G-100 chromatography gave fraction SII with the high fibrinolytic activity. Proteases presented in SII fractions and eluted from Benzamidine-Sepharose (not bound to the column) provoked a fast degradation of fibrinogen alpha chains and a slower degradation of beta chains, which could possibly be due to a higher content of alpha fibrinogenases in these venoms. The fibrinogenolytic activity was decreased by metalloprotease inhibitors. The results suggested that metalloproteases in SII fractions were responsible for the fibrinolytic activity. The analysis of samples for fibrin-zymography of SII fractions showed an active band with a molecular mass of approximately 30 kDa. These results reiterate the importance of using pools of venoms for antivenom immunization, to facilitate the neutralization of the maximum potential number of toxins.

  6. Local inflammatory events induced by Bothrops atrox snake venom and the release of distinct classes of inflammatory mediators.

    Science.gov (United States)

    Moreira, Vanessa; Dos-Santos, Maria Cristina; Nascimento, Neide Galvão; Borges da Silva, Henrique; Fernandes, Cristina Maria; D'Império Lima, Maria Regina; Teixeira, Catarina

    2012-07-01

    Bothrops atrox is responsible for most accidents involving snakes in the Brazilian Amazon and its venom induces serious systemic and local effects. The local effects are not neutralized effectively by commercial antivenoms, resulting in serious sequelae in individuals bitten by this species. This study investigates the local inflammatory events induced in mice by B. atrox venom (BaV), such as vascular permeability, leukocyte influx and the release of important inflammatory mediators such as cytokines, eicosanoids and the chemokine CCL-2, at the injection site. The effect of BaV on cyclooxygenase (COX-1 and COX-2) expression was also investigated. The results showed that intraperitoneal (i.p.) injection of BaV promoted a rapid and significant increase in vascular permeability, which reached a peak 1 h after venom administration. Furthermore, BaV caused leukocyte infiltration into the peritoneal cavity between 1 and 8 h after i.p. injection, with mononuclear leukocytes (MNs) predominating in the first 4 h, and polymorphonuclear leukocytes (PMNs) in the last 4 h. Increased protein expression of COX-2, but not of COX-1, was detected in leukocytes recruited in the first and fourth hours after injection of BaV. The venom caused the release of eicosanoids PGD₂, PGE₂, TXA₂ and LTB₄, cytokines TNF-α, IL-6, IL-10 and IL-12p70, but not IFN-γ, and chemokine CCL-2 at different times. The results show that BaV is able to induce an early increase in vascular permeability and a leukocyte influx to the injection site consisting mainly of MNs initially and PMNs during the later stages. These phenomena are associated with the production of cytokines, the chemokine CCL-2 and eicosanoids derived from COX-1 and COX-2. PMID:22465491

  7. Cell cycle arrest evidence, parasiticidal and bactericidal properties induced by L-amino acid oxidase from Bothrops atrox snake venom.

    Science.gov (United States)

    de Melo Alves Paiva, Raquel; de Freitas Figueiredo, Raquel; Antonucci, Gilmara Ausech; Paiva, Helder Henrique; de Lourdes Pires Bianchi, Maria; Rodrigues, Kelly C; Lucarini, Rodrigo; Caetano, Renato Cesar; Linhari Rodrigues Pietro, Rosemeire Cristina; Gomes Martins, Carlos Henrique; de Albuquerque, Sérgio; Sampaio, Suely Vilela

    2011-05-01

    The present article describes an l-amino acid oxidase from Bothrops atrox snake venom as with antiprotozoal activities in Trypanosoma cruzi and in different species of Leishmania (Leishmania braziliensis, Leishmania donovani and Leishmania major). Leishmanicidal effects were inhibited by catalase, suggesting that they are mediated by H(2)O(2) production. Leishmania spp. cause a spectrum of diseases, ranging from self-healing ulcers to disseminated and often fatal infections, depending on the species involved and the host's immune response. BatroxLAAO also displays bactericidal activity against both Gram-positive and Gram-negative bacteria. The apoptosis induced by BatroxLAAO on HL-60 cell lines and PBMC cells was determined by morphological cell evaluation using a mix of fluorescent dyes. As revealed by flow cytometry analysis, suppression of cell proliferation with BatroxLAAO was accompanied by the significant accumulation of cells in the G0/G1 phase boundary in HL-60 cells. BatroxLAAO at 25 μg/mL and 50 μg/mL blocked G0-G1 transition, resulting in G0/G1 phase cell cycle arrest, thereby delaying the progression of cells through S and G2/M phase in HL-60 cells. This was shown by an accentuated decrease in the proportion of cells in S phase, and the almost absence of G2/M phase cell population. BatroxLAAO is an interesting enzyme that provides a better understanding of the ophidian envenomation mechanism, and has biotechnological potential as a model for therapeutic agents. PMID:21300133

  8. 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.

  9. Ontogenetic variation of metalloproteinases and plasma coagulant activity in venoms of wild Bothrops atrox specimens from Amazonian rain forest.

    Science.gov (United States)

    López-Lozano, Jorge Luis; de Sousa, Marcelo Valle; Ricart, Carlos André O; Chávez-Olortegui, Carlos; Flores Sanchez, Eladio; Muniz, Emiro G; Bührnheim, Paulo F; Morhy, Lauro

    2002-07-01

    A comparative study of venoms from juvenile, sub-adult and adult wild Bothrops atrox specimens captured in Manaus region (Brazil) was performed. All venoms tested had acidic pH (5.5) and the human plasma coagulant activity was higher in venoms from juvenile and sub-adult specimens than in adults. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that the most intense bands in adult venoms corresponded to polypeptides of 23 and 50kDa. The 23kDa protein was not detected in juvenile venoms. The 23 and 50kDa proteins were purified by two steps of reversed phase-HPLC followed by size exclusion HPLC. Partial amino acid sequence of the 23kDa protein showed homology to metalloproteinases from other snake venoms. Electrospray ionization mass spectrometric analysis (ESI-MS) showed that the 23kDa band contained at least three isoforms of 23030, 23300 and 23645Da. The 50kDa polypeptide was N-terminally blocked for Edman degradation and presented molecular masses ranging from 46.8 to 49.4kDa by ESI-MS. Both proteins were detected by anti-mutalysin II antibodies in immunoblotting assay indicating that they belong to the metalloproteinase family. Immunoblotting analysis also showed that the 23kDa band increased in intensity from juvenile to adult specimens.SDS-PAGE analysis of juvenile and adult venoms following autoproteolysis in pH 7.4 suggested that endogenous venom metalloproteinases can digest the 50kDa metalloproteinase, originating a new protein band of 27kDa. It was also demonstrated in juvenile venoms that the 23kDa band was not the result of proteolytic processing of the 50kDa metalloproteinase. PMID:12076654

  10. Activation of J77A.1 macrophages by three phospholipases A2 isolated from Bothrops atrox snake venom.

    Science.gov (United States)

    Furtado, Juliana L; Oliveira, George A; Pontes, Adriana S; Setúbal, Sulamita da S; Xavier, Caroline V; Lacouth-Silva, Fabianne; Lima, Beatriz F; Zaqueo, Kayena D; Kayano, Anderson M; Calderon, Leonardo A; Stábeli, Rodrigo G; Soares, Andreimar M; Zuliani, Juliana P

    2014-01-01

    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. PMID:24592395

  11. Bothrops lanceolatus (Fer de lance) venom induces oedema formation and increases vascular permeability in the mouse hind paw.

    Science.gov (United States)

    de Araújo, A L; de Souza, A O; da Cruz-Höfling, M A; Flores, C A; Bon, C

    2000-02-01

    The ability of snake venoms to increase vascular permeability and to induce oedema through the release of pharmacologically active substances is well known. We have studied the oedema and vascular permeability induced by Bothrops lanceolatus venom in male Swiss white mice. Paw oedema was induced by the subplantar injection of B. lanceolatus venom (125-1000 ng/paw) and was quantified as the increase in paw weight. Changes in vascular permeability were assessed by measuring the amount of Evans blue dye extravasation. The oedema and the increase in vascular permeability were maximal within 2 h and had resolved after 24 h. The administration of the vasodilator iloprost (20 ng/paw) immediately after B. lanceolatus venom potentiated the oedema and the increase in vascular permeability by approximately four-fold. Pretreating the mice with indomethacin, dexamethasone, NDGA or BW A4C inhibited the venom-induced oedema and the increase in vascular permeability. In contrast, histamine, serotonin and PAF-acether antagonists (mepyramine, cyproheptadine and WEB 2086, respectively) were ineffective. Histological examination showed that B. lanceolatus venom (250 ng and 500 ng/paw) caused thickening of the inner dermal layers which was accompanied by extensive intercellular spaces indicative of oedema. In addition, there was a marked infiltration of inflammatory cells, particularly neutrophils, into the underlying muscle layer. The latter, however, remained morphologically unaffected during the 3 h of observation. Venom doses larger than 500 ng/paw produced intense haemorrhage. These results indicate that B. lanceolatus venom induces oedema and increases vascular permeability in the mouse hind paw. The principal mediators of this inflammatory response are cyclooxygenase and lipoxygenase products. PMID:10665802

  12. 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

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

    International Nuclear Information System (INIS)

    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 IC50 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

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

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    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

  15. Heterologous expression and biochemical and functional characterization of a recombinant alpha-type myotoxin inhibitor from Bothrops alternatus snake.

    Science.gov (United States)

    Santos-Filho, Norival A; Boldrini-França, Johara; Santos-Silva, Ludier K; Menaldo, Danilo L; Henrique-Silva, Flávio; Sousa, Tiago S; Cintra, Adélia C O; Mamede, Carla C N; Oliveira, Fábio; Arantes, Eliane C; Antunes, Lusânia M Greggi; Cilli, Eduardo M; Sampaio, Suely V

    2014-10-01

    Venomous and non-venomous snakes possess phospholipase A2 (PLA2) inhibitory proteins (PLIs) in their blood serum. This study shows the expression and biochemical and functional characterization of a recombinant alpha inhibitor from Bothrops alternatus snake, named rBaltMIP. Its expression was performed in Pichia pastoris heterologous system, resulting in an active recombinant protein. The expressed inhibitor was tested regarding its ability to inhibit the phospholipase activity of different PLA2s, showing slight inhibitions especially at the molar ratios of 1:1 and 1:3 (PLA2:PLI). rBaltMIP was also effective in decreasing the myotoxic activity of the tested toxins at molar ratios greater than 1:0.4 (myotoxin:PLI). The inhibition of the myotoxic activity of different Asp49 (BthTX-II and PrTX-III) and Lys49 (BthTX-I and PrTX-I) myotoxins was also performed without the prior incubation of myotoxins/inhibitor in order to analyze the real possibility of using snake plasma inhibitors or recombinant inhibitors as therapeutic agents for treating envenomations. As a result, rBaltMIP was able to significantly inhibit the myotoxicity of Lys49 myotoxins. Histopathological analysis of the gastrocnemius muscles of mice showed that the myotoxins are able to induce severe damage to the muscle fibers of experimental animals by recruiting a large number of leukocyte infiltrates, besides forming an intense accumulation of intercellular fluid, leading to local edema. When those myotoxins were incubated with rBaltMIP, a reduction of the damage site could be observed. Furthermore, the cytotoxic activity of Asp49 PLA2s and Lys49 PLA2-like enzymes on C2C12 cell lines was decreased, as shown by the higher cell viabilities after preincubation with rBaltMIP. Heterologous expression would enable large-scale obtainment of rBaltMIP, thus allowing further investigations for the elucidation of possible mechanisms of inhibition of snake PLA2s, which have not yet been fully clarified. PMID:25047442

  16. 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

  17. 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

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

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    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.

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

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    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.

  20. Clinical indicators of envenoming and serum levels of venom antigens in patients bitten by Bothrops lanceolatus in Martinique. Research Group on Snake Bites in Martinique.

    Science.gov (United States)

    Bucher, B; Canonge, D; Thomas, L; Tyburn, B; Robbe-Vincent, A; Choumet, V; Bon, C; Ketterlé, J; Lang, J

    1997-01-01

    An enzyme-linked immunosorbent assay was developed to measure venom antigen levels in the serum of 40 patients bitten by Bothrops lanceolatus. The grading system used for the severity of envenomation (grades 1 to 4, minor to major) was predominantly based on the presence of local signs. Serum venom levels increased with the grade of severity (P or = 15 ng/mL were observed in all patients with progressive aggravation of swelling despite the use of early antivenom therapy. No venom was detectable in blood samples taken after completion of serotherapy. All patients recovered. These results confirm the efficacy of both the clinical severity scoring system used and the therapeutic regimen. PMID:9196765

  1. An Isoflavone from Dipteryx alata Vogel is Active against the in Vitro Neuromuscular Paralysis of Bothrops jararacussu Snake Venom and Bothropstoxin I, and Prevents Venom-Induced Myonecrosis

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    Miriéle C. Ferraz

    2014-05-01

    Full Text Available Snakebite is a neglected disease and serious health problem in Brazil, with most bites being caused by snakes of the genus Bothrops. Although serum therapy is the primary treatment for systemic envenomation, it is generally ineffective in neutralizing the local effects of these venoms. In this work, we examined the ability of 7,8,3'-trihydroxy-4'-methoxyisoflavone (TM, an isoflavone from Dipteryx alata, to neutralize the neurotoxicity (in mouse phrenic nerve-diaphragm preparations and myotoxicity (assessed by light microscopy of Bothrops jararacussu snake venom in vitro. The toxicity of TM was assessed using the Salmonella microsome assay (Ames test. Incubation with TM alone (200 μg/mL did not alter the muscle twitch tension whereas incubation with venom (40 μg/mL caused irreversible paralysis. Preincubation of TM (200 μg/mL with venom attenuated the venom-induced neuromuscular blockade by 84% ± 5% (mean ± SEM; n = 4. The neuromuscular blockade caused by bothropstoxin-I (BthTX-I, the major myotoxic PLA2 of this venom, was also attenuated by TM. Histological analysis of diaphragm muscle incubated with TM showed that most fibers were preserved (only 9.2% ± 1.7% were damaged; n = 4 compared to venom alone (50.3% ± 5.4% of fibers damaged; n = 3, and preincubation of TM with venom significantly attenuated the venom-induced damage (only 17% ± 3.4% of fibers damaged; n = 3; p < 0.05 compared to venom alone. TM showed no mutagenicity in the Ames test using Salmonella strains TA98 and TA97a with (+S9 and without (−S9 metabolic activation. These findings indicate that TM is a potentially useful compound for antagonizing the neuromuscular effects (neurotoxicity and myotoxicity of B. jararacussu venom.

  2. Caracterización de la enzima similar a trombina del veneno de Bothrops pictus "jergón de costa

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    Dan Vivas-Ruiz

    2015-12-01

    Full Text Available Objetivos. Realizar una caracterización bioquímica y molecular del principio coagulante del veneno de Bothrops pictus. Materiales y métodos. Se realizó la amplificación del gen a partir de cDNA, se analizó la homología de la secuencia nucleotídica y de la proteína deducida. Se procedió a purificar la enzima para los análisis de secuenciación directa N terminal de los primeros 20 aminoácidos y los ensayos de coagulación sobre plasma humano y fibrinógeno humano, por otro lado, se evaluó el patrón de corte del fibrinógeno por medio de PAGE SDS y la actividad defibrinogenante en roedores albinos (18-22 g. Se determinó el contenido de carbohidratos asociados, el efecto de inhibidores clásicos de proteasas y el efecto de iones bajo la forma de cloruros. Resultados. La enzima mostró homología en la estructura primaria con otras TLEs reportadas para la familia Viperidae, la dosis coagulante mínima (DCM sobre plasma y fibrinógeno humano fue de 18 y 6 µg respectivamente y su potencia coagulante fue de 131,1 NHI unidades de trombina. La enzima se mostró estable a condiciones fisiológicas y prescinde de iones para su actividad. Los carbohidratos asociados detectados fueron hexosas (25,76%, hexosaminas (13,1% y ácido siálico (0,76%. Los agentes fluoruro de fenil metil sulfonil floruro (PMSF ditiotreitol (DTT fueron los principales inhibidores de la actividad enzimática en tanto que la heparina no tuvo efecto inhibidor. Conclusiones. El principio coagulante del veneno de Bothrops pictus es una enzima similar a trombina

  3. Caracterización de la enzima similar a trombina del veneno de Bothrops pictus "jergón de costa

    Directory of Open Access Journals (Sweden)

    Dan Vivas-Ruiz

    2015-10-01

    Full Text Available Objetivos. Realizar una caracterización bioquímica y molecular del principio coagulante del veneno de Bothrops pictus. Materiales y métodos. Se realizó la amplificación del gen a partir de cDNA, se analizó la homología de la secuencia nucleotídica y de la proteína deducida. Se procedió a purificar la enzima para los análisis de secuenciación directa N terminal de los primeros 20 aminoácidos y los ensayos de coagulación sobre plasma humano y fibrinógeno humano, por otro lado, se evaluó el patrón de corte del fibrinógeno por medio de PAGE SDS y la actividad defibrinogenante en roedores albinos (18-22 g. Se determinó el contenido de carbohidratos asociados, el efecto de inhibidores clásicos de proteasas y el efecto de iones bajo la forma de cloruros. Resultados. La enzima mostró homología en la estructura primaria con otras TLEs reportadas para la familia Viperidae, la dosis coagulante mínima (DCM sobre plasma y fibrinógeno humano fue de 18 y 6 µg respectivamente y su potencia coagulante fue de 131,1 NHI unidades de trombina. La enzima se mostró estable a condiciones fisiológicas y prescinde de iones para su actividad. Los carbohidratos asociados detectados fueron hexosas (25,76%, hexosaminas (13,1% y ácido siálico (0,76%. Los agentes fluoruro de fenil metil sulfonil floruro (PMSF ditiotreitol (DTT fueron los principales inhibidores de la actividad enzimática en tanto que la heparina no tuvo efecto inhibidor. Conclusiones. El principio coagulante del veneno de Bothrops pictus es una enzima similar a trombina

  4. 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. PMID:21278006

  5. Campanha Antiofídica em Minas Gerais

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    Octávio de Magalhães

    1958-12-01

    Full Text Available O A. descreve as fases da campnha antiofídica em Minas Gerais (1918 a 1940. Registra as seguintes espécies de ofídios venenosos encontrados: Bothrops cotiara, Bothrops alternata, Bothrops atrox, Bothrops neuwiedii, Bothrops jararaca, Bothrops inaequalis, Bothrops jararacussú, Crotalus terrificus terrificus, Lachesis mutus, Micrurus lemniscatus, Micrurus frontalis, Micrurus corallinus corallinus e numerosas espécies não venenosas, fazendo um mapa para a distribuição geográfica dos ofídios venenosos em Minas Gerais. Apurou que dominam no Estado as espécies do gênero Bothrops e não do gênero Crotalus. Registrou diferenças morfológicas, principalmente de tamanho dos Crotalus terrificus terrificus do Norte do Estado e do Centro e Sul. Relata 2 casos de alergia, de certa gravidade, pelo veneno ofídico, líquido ou sêco, curando-se os pacientes pelo afastamento dos trabalhos da seção antiofídica do Instituto Ezequiel Dias. Registra, ainda, 22 anos de trabalhos, numerosas observações pelas picadas de ofídios venenosos e não venenosos, sendo que estas últimas, em gravidade. Das 980 primeiras observações, 921 foram de acidentes humanos e, destas, 531 tomaram sôro, com a mortalidade de 1,30%; 390 não tomaram sôro e tiveram mortalidade variável, conforme a dose, a época do emprêgo do sôro e a espécie do ofídio, entre 7,6 e 100% dos casos, sendo a média geral de 33%. nenhum acidentado pela Lachesis mutus se salvou.The author describes the phases of the antiophidian campaign in Minas Gerais (1918 to 1940. He registers the following species of poisonous snakes found: Bothrops cotiara, Bothrops alternata, Bothrops atrox, Bothrops neuwiedii, Bothrops jararaca, Bothrops inaequalis, Bothrops jararacussú, Crotalus terrificus terrificus, Lachesis mutus, Micrurus lemniscatus, Micrurus frontalis, Micrurus corallinus and numerous species not poisonous, making a map for the geographical distribution of the poisonous snakes in Minas

  6. 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

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    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

  7. Comparación de caracteres corporales y del veneno de Bothrops alternatus entre poblaciones de las provincias de Buenos Aires y Entre Ríos, Argentina

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    Rocco, Daniela Marisa

    2011-11-01

    Full Text Available Comparamos caracteres corporales y producción de veneno de ejemplares de Bothrops alternatus de una población aislada geográficamente (Olavarría, región de Tandilia, Buenos Aires con otra en su área de distribución continua de Concordia (Entre Ríos. Estudiamos el largo corporal, peso, separación entre dientes inoculadores, cantidad de veneno y de proteínas en el veneno por ejemplar. No se hallaron diferencias en los caracteres estudiados entre ambas poblaciones (p > 0.05. Las hembras fueron mayores que los machos en ambas muestras, entre un 12-18% (p 0.5; Olavarría: 142 ± 65 mg/animal, Concordia: 160 ± 80 mg/animal, aún ajustando la cantidad de veneno producida respecto al tamaño, mediante el cociente veneno/largo corporal (p >0.6. Tampoco hubo diferencias en el contenido proteico, siendo para ambas muestras de 0.697 ± 0.096 mg de proteínas/mg de veneno seco. Nuestros datos sugieren que los ejemplares de la población aislada de Tandilia no presentan variaciones en el tamaño corporal o en la cantidad de veneno producida, respecto a los ejemplares de Concordia. Some corporal characters and venom yield of adult specimens of Bothrops alternatus from the isolated region of Olavarría (Tandilian Region, Buenos Aires and the region of continuous distribution of Concordia (Entre Ríos were compared. Corporal length, weight, separation between fangs, venom yield by snake and the amount of protein in the venom were determined. No differences in the distinct characters from specimens from both regions were found (p > 0.05. Females were greater than males in both samples, 12-18% (p 0.5; Olavarría= 142 ± 65 mg / animal, Concordia= 161 ± 80 mg / animal even adjusting the venom yield with the corporal length by the relation mg of venom / corporal length (p > 0.6. No differences were observed in the protein content of the dry venom which was of 0.697 ± 0.096 mg of protein / mg of dry venom. From the study of these samples, it could be

  8. Occurrence of sulfated fucose branches in fucosylated chondroitin sulfate are essential for the polysaccharide effect preventing muscle damage induced by toxins and crude venom from Bothrops jararacussu snake.

    Science.gov (United States)

    Monteiro-Machado, Marcos; Tomaz, Marcelo A; Fonseca, Roberto J C; Strauch, Marcelo A; Cons, Bruno L; Borges, Paula A; Patrão-Neto, Fernando C; Tavares-Henriques, Matheus S; Teixeira-Cruz, Jhonatha M; Calil-Elias, Sabrina; Cintra, Adélia C O; Martinez, Ana Maria B; Mourão, Paulo A S; Melo, Paulo A

    2015-05-01

    Snake envenoming is an important public health problem around the world, particularly in tropics. Beyond deaths, morbidity induced by snake venoms, such as myotoxicity, is of pivotal consequence to population. Bothrops jararacussu is the main venomous snake in southeast region of Brazil, and particularly presents strong myotoxic effect. The only available therapy, antibothropic antivenom, poorly affects venom-induced myotoxicity. The aim of this study is to assess the ability of fucosylated chondroitin sulfate (fucCS), a glycosaminoglycan with anticoagulant and antithrombotic properties, and its derivatives to inhibit toxic activities of B. jararacussu crude venom and its isolated toxins, named bothropstoxins (BthTX-I and BthTX-II). The in vitro myotoxic activities induced by crude venom, by BthTX-I alone and by toxins together were abolished by fucCS. Carboxyl reduction (fucCS-CR) kept this ability whereas defucosilation (defucCS) abrogates myoprotection. We observed the same pattern in the response of these polysaccharides in antagonizing the increase in plasma creatine kinase (CK) levels, the reduction of skeletal muscle CK content and the rise of myeloperoxidase (MPO) activity induced by crude venom and isolated toxins. FucCS inhibited edematogenic activity and partially prevented the reduction of total leukocytes in blood when pre-incubated with crude venom. Furthermore, the venom procoagulant effect was completely antagonized by increasing concentrations of fucCS, although this polyanion could stop neither the tail bleeding nor the skin hemorrhage induced by Bothrops jararaca venom. The B. jararacussu phospholipase, hyaluronidase, proteolytic and collagenase activities were inhibited in vitro. The results suggest that fucCS could be able to interact with both toxins, and it is able to inhibit BthTX-II phospholipase activity. Light microscopy of extensor digitorum longus muscle (EDL) muscle showed myoprotection by fucCS, once necrotic areas, edema and

  9. 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

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    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.

  10. Comparação entre a epidemiologia do acidente e a clínica do envenenamento por serpentes do gênero Bothrops, em adultos idosos e não idosos Comparison between the epidemiology of accidents and the clinical features of envenoming by snakes of the genus Bothrops, among elderly and non-elderly adults

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    Lindioneza Adriano Ribeiro

    2008-02-01

    Full Text Available O objetivo do presente estudo foi conhecer diferenças epidemiológicas e clínicas do envenenamento por Bothrops spp em adultos idosos (>60 anos e não idosos (20 a 59 anos. Os dados foram obtidos de 1.930 prontuários de pacientes atendidos no Instituto Butantan de 1981 a 1992. Quanto maior a idade do paciente maior a freqüência do acometimento das mãos em relação aos pés (p12 horas após a picada (pThis study had the aim of ascertaining epidemiological and clinical differences in envenoming caused by Bothrops spp between elderly adults (>60 years and non-elderly adults (20 to 59 years. The data were obtained from 1,930 medical records of patients attended at the Butantan Institute between 1981 and 1992. The greater the patient’s age was, the higher the frequency of bites on the hands rather than on the feet was (p 12 hours after the bite (p < 0.05. Necrosis was more common among the elderly patients (p < 0.05 and renal failure was more common among patients aged 50 years or over (p < 0.05, in relation to younger patients. It was concluded that elderly individuals are more often bitten on the hands and less often on the feet, and that they develop local necrosis and renal failure more frequently than do younger individuals.

  11. Alkylation of Histidine Residues of Bothrops jararacussu Venom Proteins and Isolated Phospholipases A2: A Biotechnological Tool to Improve the Production of Antibodies

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    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.

  12. Purification and partial characterization of phospholipases A2 from Bothrops asper (barba amarilla snake venom from Chiriguaná (Cesar, Colombia

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    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.

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

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    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.

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

    International Nuclear Information System (INIS)

    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)

  15. Sexual dimorphism in development and venom production of the insular threatened pit viper Bothrops insularism (Serpentes: Viperidae of Queimada Grande Island, Brazil

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    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.

  16. Prognostic significance of clinical grading of patients envenomed by Bothrops lanceolatus in Martinique. Members of the Research Group on Snake Bite in Martinique.

    Science.gov (United States)

    Thomas, L; Tyburn, B; Ketterlé, J; Biao, T; Mehdaoui, H; Moravie, V; Rouvel, C; Plumelle, Y; Bucher, B; Canonge, D; Marie-Nelly, C A; Lang, J

    1998-01-01

    The correlation between clinical grading of patients bitten by Bothrops lanceolatus and the subsequent development of their envenoming was examined. Severity of envenoming was graded using a 1-4 scale (minor to major). Patients were classified into 2 groups according to the time elapsed between bite and treatment with a specific purified equine F(ab')2 antivenom. The late/no treatment group (n = 33) was characterized by a systemic thrombotic complication rate of 14/33 (42.4%) leading to 4 deaths, which increased with the maximum severity assessed on the first day following the bite (P = 0.003). However, infarctions could develop in patients who presented initially with signs of moderate envenoming, normal blood clotting and low serum levels of venom antigens. No such complication of fatality occurred in the early (0.5-6 h) treatment group (n = 70). Multiple regression analysis showed that duration of stay in hospital in this group increased with the length of the snake (P = 0.017), venom antigenaemia (P = 0.016), initial grading (P < 0.001), and with the need for surgical debridement (n = 10/70, P < 0.001). Outcome was correlated with initial severity of envenoming. However, the only factor with a positive prognostic significance for the individual envenomed patient was the early infusion of specific antivenom, which led to 100% recovery in our series. PMID:9861375

  17. 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

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    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.

  18. Structural and functional characterization of myotoxin I, a Lys49 phospholipase A2 homologue from the venom of the snake Bothrops atrox.

    Science.gov (United States)

    Núñez, Vitelbina; Arce, Viviana; Gutiérrez, José María; Lomonte, Bruno

    2004-07-01

    A new myotoxin was isolated from the venom of Bothrops atrox from Colombia. B. atrox myotoxin I is a homodimer, with a subunit molecular mass of 13,826, and a pI of 8.9. Its complete nucleotide sequence was obtained by cDNA cloning, indicating a mature product of 122 residues that belongs to the family of Lys49 phospholipase A(2) (PLA(2)) homologues, a subgroup of catalytically inactive proteins within the group IIA. Accordingly, the toxin was devoid of phospholipase and anticoagulant activities, in vitro. In mice, it induced conspicuous local myonecrosis, edema, and a systemic interleukin-6 response. In vitro, it was cytolytic upon myoblasts, and weakly bactericidal. The toxin showed highest homology with other Lys49 PLA(2)s, both in its primary and three-dimensional modeled structure, although with an evident difference in the C-terminal region. Unlike Lys49 proteins of American crotalids having 121 residues, this toxin presents an insertion (Asn) between positions 118 and 119. Despite several substitutions within the C-terminal region 115-129 between B. atrox myotoxin I and B. asper myotoxin II, antibodies against synthetic peptide 115-129 of the latter were strongly cross-reactive to the former, indicating the antigenic conservation of this site, known to be critical for the membrane-damaging activities of Lys49 myotoxins. PMID:15225567

  19. Low-molecular-mass peptides from the venom of the Amazonian viper Bothrops atrox protect against brain mitochondrial swelling in rat: potential for neuroprotection.

    Science.gov (United States)

    Martins, N M; Ferreira, D A S; Carvalho Rodrigues, M A; Cintra, A C O; Santos, N A G; Sampaio, S V; Santos, A C

    2010-08-01

    The neurodegenerative diseases are important causes of morbidity and mortality in Western countries. Common mechanisms of toxicity involving mitochondrial damage have been suggested; however, a definitive treatment has not yet been found. Therefore, there has been great interest in the development of mitochondria-targeted protective compounds for the treatment of neuropathies. Animal toxins represent a promising source of new molecules with neuroprotective activity and potential to originate new drugs. We present here the effects of a low-molecular-mass peptides fraction (Ba-V) from Bothrops atrox snake venom, on rat brain mitochondrial function. Ba-V did not induce the mitochondrial swelling and moreover, was as effective as cyclosporin A (CsA) to inhibit the calcium/phosphate-induced swelling, which indicates its potential to prevent the mitochondrial permeability transition (MPT). The membrane electrochemical potential, the oxygen consumption during states-3 and -4 respirations as well as the respiratory control ratio (RCR) were not affected by Ba-V. Additionally, Ba-V did not induce reactive oxygen species (ROS) generation. Interestingly, Ba-V did not protect against the generation of ROS induced by t-BOH, which suggests a protection mechanism other than ROS scavenging. Given the important role of the mitochondrial damage and, more specifically, of MPT, in the development of neuropathies, Ba-V might be useful in the future strategies for the treatment of these diseases. PMID:20338188

  20. The novel metalloproteinase atroxlysin-I from Peruvian Bothrops atrox (Jergón) snake venom acts both on blood vessel ECM and platelets.

    Science.gov (United States)

    Sanchez, Eladio F; Schneider, Francisco S; Yarleque, Armando; Borges, Marcia H; Richardson, Michael; Figueiredo, Suely G; Evangelista, Karla S; Eble, Johannes A

    2010-04-01

    We report the isolation and structure-function relationship of a 23kDa metalloproteinase named atroxlysin-I from the venom of the Peruvian Bothrops atrox (Jergón). Atroxlysin is a P-I metalloproteinase and contains 204 residues. Its proteolytic activity towards dimethylcasein is enhanced by Ca2+ but inhibited by EDTA, dithiothreitol, excessive Zn2+ and alpha2-macroglobulin. Unlike other structurally homologous P-I metalloproteinases, atroxlysin-I causes hemorrhages. To examine its hemorrhagic activity mechanistically, we studied its function in vitro and in vivo. It cleaved the Ala14-Leu15 and Tyr16-Leu17 bonds in oxidized insulin B-chain and specifically hydrolyzed the alpha-chains of fibrin(ogen) in a dose- and time-dependent manner. Atroxlysin-I cleaved plasma fibronectin and other extracellular matrix proteins (collagens I and IV) and the triple-helical fragment CB3 of collagen IV, but did not degrade laminin-111. Complementarily, the laminin and collagen binding integrins alpha7beta1 and alpha1beta1 were cleaved by atroxlysin. Even without catalytic activity atroxlysin-I inhibited collagen- and ADP-triggered platelet aggregation. PMID:20102699

  1. Osteopontin, a chemotactic protein with cytokine-like properties, is up-regulated in muscle injury caused by Bothrops lanceolatus (fer-de-lance) snake venom.

    Science.gov (United States)

    Barbosa-Souza, Valéria; Contin, Daniel Kiss; Filho, Waldemar Bonventi; de Araújo, Albetiza Lôbo; Irazusta, Silvia Pierre; da Cruz-Höfling, Maria Alice

    2011-10-01

    Osteopontin (OPN) is a chemotactic, adhesive protein whose receptors include some integrins and matrix proteins known to have role in inflammatory and repair processes. We examined the time course of OPN expression at acute and chronic stages after intramuscular injection of Bothrops lanceolatus venom in rats. Additionally, we examined the expression of CD68 (a marker for phagocytic macrophages) and the myogenic factors, myoD and myogenin. There was a biphasic upregulation of OPN (6-48 h and 3-14 days post-venom), i.e., during acute inflammation and myogenic cell proliferation and differentiation phases. OPN was detected in CD68 + macrophages, fibroblasts, normal and damaged myofibers, myoblasts and myotubes. Myogenin was expressed in the cytoplasm (atypical pattern) and nucleus of myoblasts and myotubes from 18 h to 7 days, after which it was expressed only in nuclei. Macrophage numbers, OPN and myogenin expression were still elevated at 7, 14 and 7 days. At 3 days, when OPN achieved the peak, some clusters of myoblasts were within regions of intense collagen deposition. Fibrosis may represent limitation for repairing processes and may explain the small diameter of regenerated fibers at 21 days post-venom. The expression of OPN in the course of venom-induced damage and regeneration suggests stages-specific mediation role along the whole process. PMID:21839764

  2. Neutralization, by a monospecific Bothrops lanceolatus antivenom, of toxic activities induced by homologous and heterologous Bothírops snake venoms.

    Science.gov (United States)

    Bogarín, G; Romero, M; Rojas, G; Lutsch, C; Casadamont, M; Lang, J; Otero, R; Gutiérrez, J M

    1999-03-01

    A monospecific Bothrops lanceolatus antivenom, currently used in Martinique, was tested for its efficacy in the neutralization of several toxic and enzymatic activities of the venoms of B. lanceolatus, B. atrox and B. asper. When tested by the i.p. route in mice, B. lanceolatus venom had an LD50 of 12.8 microg/g. In addition, it induced local tissue damage (hemorrhage, edema and myotoxicity) and showed indirect hemolytic activity, but was devoid of coagulant effect on human plasma in vitro and of defibrinating activity in mice. Antivenom was fully effective in the neutralization of lethal, hemorrhagic, edema-forming, myotoxic and indirect hemolytic effects of B. lanceolatus venom in assays involving preincubation of venom and antivenom. When tested against the venoms of B. asper and B. atrox, the antivenom completely neutralized the lethal, hemorrhagic, myotoxic and indirect hemolytic effects, and was partially effective in neutralizing edema-forming activity. In contrast, the antivenom was ineffective in the neutralization of in vitro coagulant and in vivo defibrinating effects induced by these two venoms. PMID:10080358

  3. Quadros clínico-patológicos do envenenamento ofídico por Crotalus durissus terrificus e Bothrops spp. em animais de produção

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    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.

  4. Evaluación biológica preliminar de extractos vegetales utilizados en la medicina tradicional de la Sierra Nevada deSanta Marta contra el veneno de la Bothrops asper

    OpenAIRE

    Willinton Barranco Pérez; Vitelbina Nuñez; Mauricio Sanchez

    2013-01-01

    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 ...

  5. Ineficácia do antiveneno botrópico na neutralização da atividade coagulante do veneno de Lachesis muta muta: relato de caso e comprovação experimental Inefficacy of bothropic antivenom on the neutralization of the coagulant activity of Lachesis muta muta venom: case report and experimental evidence

    Directory of Open Access Journals (Sweden)

    Ramza Bard

    1994-02-01

    Full Text Available No Estado do Amazonas, nas regiões circunvizinhas à cidade de Manaus, as principais espécies de serpentes causadoras de acidentes são Bothrops atrox e Lachesis muta muta com um percentual de ocorrência, dos acidentes confirmados, de 76% e 17%, respectivamente. Rotineiramente, na ausência dos soros antilaquético e antibotrópico-laquético o Instituto de Medicina Tropical de Manaus (IMTM, utiliza-se do soro antibotrópico no tratamento do acidente laquético. Neste trabalho relatamos um caso de acidente por L m. muta, onde o paciente foi tratado com 20 ampolas do soro antibotrópico e permaneceu com o sangue incoagulável até o 13º dia após o acidente. Experimentos foram realizados para obtenção das potências do soro antibotrópico para as atividades coagulante e hemorrágica dos venenos de L. m. muta e de B. atrox. Os resultados mostram que as potências do soro para a atividade hemorrágica dos venenos de L. m. muta e de B. atrox foram similares enquanto que a potência, para a atividade coagulante do veneno de L. m. muta, foi 9,2 vezes menor. Os títulos de anticorpos de três diferentes lotes de soro antibotrópico variaram para o veneno de L. m. muta e foram constantes para o veneno de B. atrox. Devido a ineficácia do soro antibotrópico em neutralizar, principalmente, a atividade coagulante do veneno de L. m. muta, sugerimos a não utilização do antibotrópico no tratamento dos acidentes por L. m. muta.In the State of Amazonas and in areas nearby Manaus, the snakes responsable for the majority of accidents are Bothrops atrox and Lachesis muta muta, with a percentage of confirmed species of 76% and 17%, respectively. Frequently, in the absence of the laquetic and bothropic-laquetic antivenoms, the Instituto de Medicina Tropical de Manaus (IMTM has been using bothropic antivenom in the treatment of laquetic accident. In this paper is related a case of accident caused by L. muta muta; the patient was treated with bothropic

  6. 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

  7. 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

    2009-11-01

    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. PMID:19665598

  8. Anti-parasitic effect on Toxoplasma gondii induced by BnSP-7, a Lys49-phospholipase A2 homologue from Bothrops pauloensis venom.

    Science.gov (United States)

    Borges, Isabela Pacheco; Castanheira, Letícia Eulalio; Barbosa, Bellisa Freitas; de Souza, Dayane Lorena Naves; da Silva, Rafaela José; Mineo, José Roberto; Tudini, Kelly Aparecida Yoneyama; Rodrigues, Renata Santos; Ferro, Eloísa Amália Vieira; de Melo Rodrigues, Veridiana

    2016-09-01

    Toxoplasmosis affects a third of the global population and presents high incidence in tropical areas. Its great relevance in public health has led to a search for new therapeutic approaches. Herein, we report the antiparasitic effects of BnSP-7 toxin, a Lys49 phospholipase A2 (PLA2) homologue from Bothrops pauloensis snake venom, on Toxoplasma gondii. In an MTT assay, BnSP-7 presented significant cytotoxicity against host HeLa cells at higher doses (200 μg/mL to 50 μg/mL), whereas lower doses (25 μg/mL to 1.56 μg/mL) produced low cytotoxicity. Furthermore, the toxin showed no effect on T. gondii tachyzoite viability when evaluated by trypan blue exclusion, but decreased both adhesion and parasite proliferation when tachyzoites were treated before infection. We also measured cytokines in supernatants collected from HeLa cells infected with T. gondii tachyzoites previously treated with RPMI or BnSP-7, which revealed enhancement of only MIF and IL-6 cytokines levels in supernatants of HeLa cells after BnSP-7 treatment. Our results showed that the BnSP-7 PLA2 exerts an anti-Toxoplasma effect at a lower dose than that required to induce cytotoxicity in HeLa cells, and also modulates the immune response of host cells. In this sense, the anti-parasitic effect of BnSP-7 PLA2 demonstrated in the present study opens perspectives for use of this toxin as a tool for future studies on toxoplasmosis. PMID:27212627

  9. 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)

    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 (EC50), 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 EC50 of 11|μg/ml. Irradiation of LAO with 60Co 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 EC50 of 0.67mM. These data could be of help in the development of alternative therapeutic approaches to the treatment of leishmaniasis. (author)

  10. Alternagin-C, a disintegrin-like protein from the venom of Bothrops alternatus, modulates a2ß1 integrin-mediated cell adhesion, migration and proliferation

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    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.

  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. PMID:15264003

  12. Snakebites and ethnobotany in the northwest region of Colombia: Part II: neutralization of lethal and enzymatic effects of Bothrops atrox venom.

    Science.gov (United States)

    Otero, R; Núñez, V; Jiménez, S L; Fonnegra, R; Osorio, R G; García, M E; Díaz, A

    2000-08-01

    Twelve of 74 ethanolic extracts of plants used by traditional healers for snakebites in the northwest region of Colombia, were active against lethal effect of Bothrops atrox venom when they were i.p. injected into mice (18-20 g). After preincubation of sublethal doses of every extract (0.5-4.0 mg/mouse) with 1.5 i.p. lethal dose 50% (LD50) (99.3 microg) of venom, seven of them demonstrated 100% neutralizing capacity within 48 h. These were the stem barks of Brownea rosademonte (Caesalpiniaceae) and Tabebuia rosea (Bignoniaceae); rhizomes of Renealmia alpinia (Zingiberaceae) and Heliconia curtispatha (Heliconiaceae); the whole plants of Pleopeltis percussa (Polypodiaceae) and Trichomanes elegans (Hymenophyllaceae); and the ripe fruits of Citrus limon (Rutaceae). The other five extracts showing partial neutralization (45-80%; 10-30% survival rate in the control group receiving the venom alone; PBixa orellana (Bixaceae) and Struthanthus orbicularis (Loranthaceae). When the extracts were independently administered per oral or i.p. route 60 min before an i.m. venom injection (204 microg=1.5 i.m. LD50), C. limon, T. elegans, B. orellana and T. rosea extracts had partial and significant neutralizing capacity against B. atrox venom lethal effect. C. limon extract was also partially effective when it was administered either i.v. 15 min before or i.p. 5 min after an i.m. venom injection. Three of the 12 extracts with anti-lethal effect (C. limon, D. croatii and S. acuta) were devoid of antiphospholipase A2 activity, when they were tested against one minimum indirect hemolytic dose of B. atrox venom (2 microg) in agarose-erythrocyte-egg yolk gels. PMID:10940590

  13. Anti-parasitic effect on Toxoplasma gondii induced by BnSP-7, a Lys49-phospholipase A2 homologue from Bothrops pauloensis venom.

    Science.gov (United States)

    Borges, Isabela Pacheco; Castanheira, Letícia Eulalio; Barbosa, Bellisa Freitas; de Souza, Dayane Lorena Naves; da Silva, Rafaela José; Mineo, José Roberto; Tudini, Kelly Aparecida Yoneyama; Rodrigues, Renata Santos; Ferro, Eloísa Amália Vieira; de Melo Rodrigues, Veridiana

    2016-09-01

    Toxoplasmosis affects a third of the global population and presents high incidence in tropical areas. Its great relevance in public health has led to a search for new therapeutic approaches. Herein, we report the antiparasitic effects of BnSP-7 toxin, a Lys49 phospholipase A2 (PLA2) homologue from Bothrops pauloensis snake venom, on Toxoplasma gondii. In an MTT assay, BnSP-7 presented significant cytotoxicity against host HeLa cells at higher doses (200 μg/mL to 50 μg/mL), whereas lower doses (25 μg/mL to 1.56 μg/mL) produced low cytotoxicity. Furthermore, the toxin showed no effect on T. gondii tachyzoite viability when evaluated by trypan blue exclusion, but decreased both adhesion and parasite proliferation when tachyzoites were treated before infection. We also measured cytokines in supernatants collected from HeLa cells infected with T. gondii tachyzoites previously treated with RPMI or BnSP-7, which revealed enhancement of only MIF and IL-6 cytokines levels in supernatants of HeLa cells after BnSP-7 treatment. Our results showed that the BnSP-7 PLA2 exerts an anti-Toxoplasma effect at a lower dose than that required to induce cytotoxicity in HeLa cells, and also modulates the immune response of host cells. In this sense, the anti-parasitic effect of BnSP-7 PLA2 demonstrated in the present study opens perspectives for use of this toxin as a tool for future studies on toxoplasmosis.

  14. 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.

  15. Neutralizing activities of ethanolic extracts of six plants traditionally used in Guatemala as antidotes for the envenomation caused by the snake Bothrops asper

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    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.

  16. Isolation and characterization of a new serine protease with thrombin-like activity (TLBm) from the venom of the snake Bothrops marajoensis.

    Science.gov (United States)

    Vilca-Quispe, Augusto; Ponce-Soto, Luis Alberto; Winck, Flavia Vischi; Marangoni, Sergio

    2010-04-01

    The thrombin-like serine protease TLBm from Bothrops marajoensis was isolated in one chromatographic step in reverse phase HPLC. Its molecular mass was 33239.95 Da, as based on the determined primary structure and confirmed experimentally by MALDI-TOF mass spectrometry (33332.5 Da) and it contains 12 half-cysteine residues. This TLBm exhibited high specificity for BArhoNA, Michaelis-Menten behavior with K(m) 2.3x10(-1)M and the V(max) 0.52x10(-1) nmoles rho-NA/lt/min for this substrate. TLBm also showed ability to coagulate bovine fibrinogen and was inhibited by soybean trypsin inhibitor, EDTA and S(Dm) from the serum of the species Didelphis marsupialis. The primary structure of TLBm showed the presence of His(45), Asp(103) and Ser(228) residues in the corresponding positions of the catalytic triad established in the serine proteases and Ser(228) are inhibited by phenylmethylsulfonyl fluoride (PMSF). Amino acid analysis showed a high content of Asp, Glu, Gly, Ser, Ala and Pro as well as 12 half-cysteine residues and calculated pI of 6.47; TLBm presented 285 amino acid residues. In this work, we investigated the ability of TLBm to degrade fibrinogen and we observed that it is able to cause alpha- and beta-chain cleavage. Enzymatic as well as the platelet aggregation activities were strongly inhibited when incubated with PMSF, a specific inhibitor of serine protease. Also, TLBm induced platelet aggregation in washed and platelet-rich plasma, and in both cases, PMSF inhibited its activity. PMID:19931298

  17. 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.

  18. 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. PMID:16529786

  19. Prevention of thromboses in human patients with Bothrops lanceolatus envenoming in Martinique: failure of anticoagulants and efficacy of a monospecific antivenom. Research Group on Snake Bites in Martinique.

    Science.gov (United States)

    Thomas, L; Tyburn, B; Bucher, B; Pecout, F; Ketterle, J; Rieux, D; Smadja, D; Garnier, D; Plumelle, Y

    1995-05-01

    Envenomation by the Bothrops lanceolatus, a snake found only in Martinique, leads to swelling and pain, and occasionally to systemic signs and/or coagulopathy. Severe thromboses at some distance from the site of the bite may appear within 48 hr. Uncertainties as to the actual development of thrombotic complications in patients appearing to be suffering from moderate poisoning and as to the availability and the toxicity of a monospecific antivenom (AVS) initially led us to reserve antivenom for the most severe cases, and to use anticoagulants to prevent thromboses in all patients. This approach was modified after we observed serious thromboses in patients with moderate poisoning. Of 50 adult snake bite cases hospitalized between June 1991 and August 1994, 11 developed serious thrombotic complications at 36 /+- 27 hr (mean +/- SD) (range 12-96) following envenomation, despite early preventive anticoagulant therapy. Those included pulmonary embolism (two cases), cerebral infarction (six cases), myocardial infarction (one case), and cerebral and myocardial infarctions (two cases). Sixteen patients were not treated with AVS: 10 of these recovered without complications and six developed systemic thrombosis causing permanent disability in three cases. Thirty were treated with an intravenous infusion of 2-6 vials of AVS given 2-48 hr after the bite. Of these, three died of cerebral infarction that developed before the initiation of serotherapy. All others recovered. Among patients treated with AVS, three presented with mild anaphylactic reactions, while one developed serum sickness that responded to steroids. These data indicate that preventive anticoagulant therapy is of limited efficacy in Martinique.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7771608

  20. Snakebites and ethnobotany in the northwest region of Colombia: Part II: neutralization of lethal and enzymatic effects of Bothrops atrox venom.

    Science.gov (United States)

    Otero, R; Núñez, V; Jiménez, S L; Fonnegra, R; Osorio, R G; García, M E; Díaz, A

    2000-08-01

    Twelve of 74 ethanolic extracts of plants used by traditional healers for snakebites in the northwest region of Colombia, were active against lethal effect of Bothrops atrox venom when they were i.p. injected into mice (18-20 g). After preincubation of sublethal doses of every extract (0.5-4.0 mg/mouse) with 1.5 i.p. lethal dose 50% (LD50) (99.3 microg) of venom, seven of them demonstrated 100% neutralizing capacity within 48 h. These were the stem barks of Brownea rosademonte (Caesalpiniaceae) and Tabebuia rosea (Bignoniaceae); rhizomes of Renealmia alpinia (Zingiberaceae) and Heliconia curtispatha (Heliconiaceae); the whole plants of Pleopeltis percussa (Polypodiaceae) and Trichomanes elegans (Hymenophyllaceae); and the ripe fruits of Citrus limon (Rutaceae). The other five extracts showing partial neutralization (45-80%; 10-30% survival rate in the control group receiving the venom alone; P<0.05) were: leaves, branches and stem of Costus lasius (Costaceae); the whole plant of Sida acuta (Malvaceae); rhizomes of Dracontium croatii (Araceae); leaves and branches of Bixa orellana (Bixaceae) and Struthanthus orbicularis (Loranthaceae). When the extracts were independently administered per oral or i.p. route 60 min before an i.m. venom injection (204 microg=1.5 i.m. LD50), C. limon, T. elegans, B. orellana and T. rosea extracts had partial and significant neutralizing capacity against B. atrox venom lethal effect. C. limon extract was also partially effective when it was administered either i.v. 15 min before or i.p. 5 min after an i.m. venom injection. Three of the 12 extracts with anti-lethal effect (C. limon, D. croatii and S. acuta) were devoid of antiphospholipase A2 activity, when they were tested against one minimum indirect hemolytic dose of B. atrox venom (2 microg) in agarose-erythrocyte-egg yolk gels.

  1. Neutralization of the edema-forming, defibrinating and coagulant effects of Bothrops asper venom by extracts of plants used by healers in Colombia.

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    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.

  2. Neutralization of the edema-forming, defibrinating and coagulant effects of Bothrops asper venom by extracts of plants used by healers in Colombia

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    Núñez V.

    2004-01-01

    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.

  3. A comparative analysis of the clotting and fibrinolytic activities of the snake venom (Bothrops atrox) from different geographical areas in Venezuela.

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    Salazar, Ana Maria; Rodriguez-Acosta, Alexis; Girón, Maria E; Aguilar, Irma; Guerrero, Belsy

    2007-01-01

    Venom constitution within the same snake species can present considerable geographical variations. Bothrops atrox venoms were obtained from adult snakes captured at different geographical locations: Parguasa (Bolívar state); Puerto Ayacucho 1, Serranía del Cuao and Puerto Ayacucho 2 (Amazon state). The coagulant and fibrinolytic activities of these venoms were compared. Amidolytic activity of crude snake venom was measured by a micromethod designed in our laboratory. Coagulant activity on plasma and fibrinogen due to thrombin-like activity in venoms was also determined. Crude snake venom fibrinolytic activity by the fibrin plate method was assayed. Chromatographic studies were developed on Protein-Pack 300 column. Polyacrylamide gel electrophoresis was carried out under reduced conditions. After SDS-PAGE of samples, the fibrin-zymography was tested on agarose-fibrin plates. The results demonstrated several differences among B. atrox venoms from different geographical areas. Chromatograms and SDS-PAGE profiles indicated that venoms from the same species presented differences in the molecular mass of their components. The procoagulant activity depended on the utilized method (amidolytic versus clotting). Parguasa and Puerto Ayacucho 2 venoms presented procoagulant activity for both methods. Furthermore, Parguasa venom had also the highest hemorrhagic activity and the lowest LD50. In relation to the fibrinolytic activity, Puerto Ayacucho 1 venom was the most active, equally for fibrin plates as for the amidolytic method (t-PA like). This venom had the lowest coagulant activity, which induced us to think that probably its procoagulant activity was interfered by its fibrinolytic activity.

  4. 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.

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    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. PMID:26276061

  5. A comparative analysis of the clotting and fibrinolytic activities of the snake venom (Bothrops atrox) from different geographical areas in Venezuela.

    Science.gov (United States)

    Salazar, Ana Maria; Rodriguez-Acosta, Alexis; Girón, Maria E; Aguilar, Irma; Guerrero, Belsy

    2007-01-01

    Venom constitution within the same snake species can present considerable geographical variations. Bothrops atrox venoms were obtained from adult snakes captured at different geographical locations: Parguasa (Bolívar state); Puerto Ayacucho 1, Serranía del Cuao and Puerto Ayacucho 2 (Amazon state). The coagulant and fibrinolytic activities of these venoms were compared. Amidolytic activity of crude snake venom was measured by a micromethod designed in our laboratory. Coagulant activity on plasma and fibrinogen due to thrombin-like activity in venoms was also determined. Crude snake venom fibrinolytic activity by the fibrin plate method was assayed. Chromatographic studies were developed on Protein-Pack 300 column. Polyacrylamide gel electrophoresis was carried out under reduced conditions. After SDS-PAGE of samples, the fibrin-zymography was tested on agarose-fibrin plates. The results demonstrated several differences among B. atrox venoms from different geographical areas. Chromatograms and SDS-PAGE profiles indicated that venoms from the same species presented differences in the molecular mass of their components. The procoagulant activity depended on the utilized method (amidolytic versus clotting). Parguasa and Puerto Ayacucho 2 venoms presented procoagulant activity for both methods. Furthermore, Parguasa venom had also the highest hemorrhagic activity and the lowest LD50. In relation to the fibrinolytic activity, Puerto Ayacucho 1 venom was the most active, equally for fibrin plates as for the amidolytic method (t-PA like). This venom had the lowest coagulant activity, which induced us to think that probably its procoagulant activity was interfered by its fibrinolytic activity. PMID:17045631

  6. Galatrox is a C-type lectin in Bothrops atrox snake venom that selectively binds LacNAc-terminated glycans and can induce acute inflammation.

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    Sartim, Marco A; Riul, Thalita B; Del Cistia-Andrade, Camillo; Stowell, Sean R; Arthur, Connie M; Sorgi, Carlos A; Faccioli, Lucia H; Cummings, Richard D; Dias-Baruffi, Marcelo; Sampaio, Suely V

    2014-11-01

    Previous studies indicate that snake venom contains glycan-binding proteins (GBPs), although the binding specificity and biological activities of many of these GBPs is unclear. Here we report our studies on the glycan binding specificity and activities of galatrox, a Bothrops atrox snake venom-derived GBP. Glycan microarray analysis indicates that galatrox binds most strongly to glycans expressing N-acetyllactosamine (LacNAc), with a significant preference for Galβ1-4GlcNAcβ over Galβ1-3GlcNAcβ compounds. Galatrox also bound immobilized laminin, a LacNAc-dense extracellular matrix component, suggesting that this GBP can bind LacNAc-bearing glycoproteins. As several endogenous mammalian GBPs utilize a similar binding LacNAc binding preference to regulate neutrophil and monocyte activity, we hypothesized that galatrox may mediate B. atrox toxicity through regulation of leukocyte activity. Indeed, galatrox bound neutrophils and promoted leukocyte chemotaxis in a carbohydrate-dependent manner. Similarly, galatrox administration into the mouse peritoneal cavity induced significant neutrophil migration and the release of pro-inflammatory cytokines IL-1α and IL-6. Exposure of bone marrow-derived macrophages to galatrox induced generation of pro-inflammatory mediators IL-6, TNF-α, and keratinocyte-derived chemokine. This signaling by galatrox was mediated via its carbohydrate recognition domain by activation of the TLR4-mediated MyD88-dependent signaling pathway. These results indicate that galatrox has pro-inflammatory activity through its interaction with LacNAc-bearing glycans on neutrophils, macrophages and extracellular matrix proteins and induce the release of pro-inflammatory mediators. PMID:24973254

  7. The complement system is involved in acute inflammation but not in the hemorrhage produced by a Bothrops atrox snake venom low molecular mass proteinase.

    Science.gov (United States)

    Rodrigues, Flávia G; Petretski, Jorge H; Kanashiro, Milton M; Lemos, Luciana; da Silva, Wilmar Dias; Kipnis, Thereza L

    2004-03-01

    Low molecular weight hemorrhagins were purified from crude Bothrops atrox snake venom by gel filtration followed by ionic strength chromatography. The protein fractions obtained, designated HI-1 to HI-8, contained proteins with molecular masses lower than 30 kDa. HI-5, the most representative among of these fractions, exhibited, in vitro, proteolytic and C inactivating properties, as analyzed by proteolysis of a protein substrate, and C system consumptive activities as assayed by reduction of the hemolytic C activity in normal human serum and by cleavage of partially purified component C3. HI-5 hemorrhagin injected i.m. into C-sufficient BALB/c mice induced a local inflammation characterized by edema, accumulation of polymorphonuclear leucocytes (PMN) and hemorrhage. In contrast, when injected into BALB/c mice previously C-depleted, the number of PMN per tissue section, but not hemorrhage, was significantly reduced (129.668 +/- 31.341 cells per microscopic field) as compared with the control C-sufficient mice (812.168 +/- 111.194 cells per microscopic field). The observations were confirmed by using C5-deficient mice instead of C-depleted mice. The average number of PMN per tissue section in C5-defficient A/J mice was 72.666 +/- 19.416 cells per microscopic field. These data indicate that the C system is involved in PMN accumulation, but not in the hemorrhage, at the local induced lesions by low molecular mass B. atrox hemorrhagins. HI-5 apparently is not contaminated with other direct or indirect inflammation mediators, PMN accumulation and hemorrhage, however, an independent phenomenon, could be mediated by the same hemorrhagin proteinase domain. PMID:15104120

  8. Use of a synthetic biosensor for neutralizing activity-biased selection of monoclonal antibodies against atroxlysin-I, an hemorrhagic metalloproteinase from Bothrops atrox snake venom.

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    Francisco Santos Schneider

    2014-04-01

    Full Text Available BACKGROUND: The snake Bothrops atrox is responsible for the majority of envenomings in the northern region of South America. Severe local effects, including hemorrhage, which are mainly caused by snake venom metalloproteinases (SVMPs, are not fully neutralized by conventional serum therapy. Little is known about the immunochemistry of the P-I SVMPs since few monoclonal antibodies (mAbs against these molecules have been obtained. In addition, producing toxin-neutralizing mAbs remains very challenging. METHODOLOGY/PRINCIPAL FINDINGS: Here, we report on the set-up of a functional screening based on a synthetic peptide used as a biosensor to select neutralizing mAbs against SVMPs and the successful production of neutralizing mAbs against Atroxlysin-I (Atr-I, a P-I SVMP from B. atrox. Hybridomas producing supernatants with inhibitory effect against the proteolytic activity of Atr-I towards the FRET peptide Abz-LVEALYQ-EDDnp were selected. Six IgG1 Mabs were obtained (named mAbatr1 to mAbatr6 and also two IgM. mAbatrs1, 2, 3 and 6 were purified. All showed a high specific reactivity, recognizing only Atr-I and B. atrox venom in ELISA and a high affinity, showing equilibrium constants in the nM range for Atr-I. These mAbatrs were not able to bind to Atr-I overlapping peptides, suggesting that they recognize conformational epitopes. CONCLUSIONS/SIGNIFICANCE: For the first time a functional screening based on a synthetic biosensor was successfully used for the selection of neutralizing mAbs against SVMPs.

  9. Estudo histopatológico das lesões causadas pelo veneno de urutu (Bothrops alternatus em músculo esquelético de camundongos

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    L. S. Queiroz

    1984-10-01

    Full Text Available Veneno bruto de urutu (Bothrops alternatus dissolvido em solução salina fisiológica foi injetado no músculo tibial anterior direito de camundongos adultos na dose de 80 μg. Os músculos foram examinados em cortes de parafina, corados por Hematoxilina e Eosina. Aos 10 minutos já havia intensa hemorragia difusa no M. tibial anterior, mas apenas raras fibras musculares estavam necróticas. Nas horas seguintes, contudo, observou-se rápido aumento do número de fibras afetadas, sendo que às 24 hs o músculo apresentava-se totalmente necrótico. Vasos sangüíneos intramusculares e nas proximidades do M. tibial anterior mostravam necrose hialina da camada média e por vezes trombose. A fagocitose dos restos celulares ocorreu da periferia para o centro e acompanhou-se de regeneração muscular. Após 1 a 2 meses, em vários animais houve recuperação considerável do músculo, embora com persistência de cicatriz. As fibras regeneradas possuiam núcleos centrais e variavam em diâmetro, estando muitas atróficas. Em outros camundongos a regeneração do M. tibial anterior foi muito precária, tendo este sido substituído por tecido fibroadiposo com apenas raras fibras musculares. Os resultados mostram que, apesar da gravidade das lesões iniciais devidas ao veneno, ocorre regeneração muscular em grau variável de animal para animal. Sugere-se que a má regeneração observada em alguns casos poderia ser devida, ao menos em parte, a dano vascular permanente.

  10. 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

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    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

  11. Identificación molecular y actividad sobre sustratos cromogénicos de la venombina A del veneno de la serpiente peruana Bothrops atrox

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    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

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    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. Neutrophils regulate the expression of cytokines, chemokines and nitric oxide synthase/nitric oxide in mice injected with Bothrops atrox venom.

    Science.gov (United States)

    Escocard, Rita de Cássia Mothé; Kanashiro, Milton Masahiko; Petretski, Jorge Hudson; Azevedo-Silva, Juliana; Queiroz de Carvalho, Eulógio Carlos; Dias da Silva, Wilmar; Kipnis, Thereza Liberman

    2006-01-01

    Bothrops atrox crude venom injected intraperitoneal (i.p.) into BALB/c mice induced local afflux of inflammatory cells, one neutrophil-rich peak after 6h and another macrophage-rich peak after 48 h. A similar pattern of local cell afflux plus edema, Delta lesions of some skeletal muscle cells, and hemorrhage were observed in mice intramuscular (i.m.) injected with the venom. Measurement of serum cytokines in neutrophil-depleted (by anti-mouse rat monoclonal antibody (mAb) RB6-8C5) and non-depleted BALB/c mice was performed by ELISA. With the exception of IL-1beta (78 pg/ml), higher levels of IL-6 (1348 pg/ml), MIP-1beta (437 pg/ml) and MIP-2 (904 pg/ml) were observed in neutrophil-depleted mice, in comparison to the values found in non-neutrophil depleted mice: IL-1beta (437 pg/ml), IL-6 (750 pg/ml), MIP-1beta (165 pg/ml) and MIP-2 (90 pg/ml). TNF-alpha was not detected. NO was detected (18 microM) 24h after venom injection in neutrophil-depleted mice. RT-PCR using representative primers detected expression of mRNA in cells from BALB/c mice injected with B. atrox venom: (a) for IL-1beta, IL-6, inducible nitric oxide synthase (iNOS), CXCR2, MIP-2 and RANTES in cells from mice that were neutrophil-depleted or not; (b) for CCR1, CCR5 and MIP-1beta in cells from neutrophil-depleted mice; (c) for MIP-1alpha in cells from non-neutrophil-depleted mice; (d) TNF-alpha and TGF-beta were not detected in either of the mice. These results indicate that neutrophils play a role in regulating the production of some cytokines and chemokines as well as locally expressed or liberated iNOS/NO in tissues injected with B. atrox crude venom. PMID:16446169

  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

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    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

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    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. Identificación de Bacterias en Cavidad Oral y Valores Hematológicos en Bothrops asper en el Serpentario de la Universidad de Antioquia -resumen-

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    C Martínez-Mejía

    2014-12-01

    Full Text Available La flora bacteriana de la cavidad oral de las serpientes en cautiverio ha sido asociada a infecciones por estomatitis y abscesos secundarios por auto-mordedura. Igualmente‚ su identificación es importante debido a las infecciones secundarias que pueden causar los accidentes ofídicos en humanos y animales. En particular el veneno de la serpiente mapaná‚ Bothrops asper‚ responsable del 70% de los accidentes ofídicos en Colombia‚ genera efectos sistémicos y locales incluyendo dermonecrosis y mionecrosis que puede complicarse con abscesos‚ celulitis y fascitis hasta en el 30% de los casos. En este estudio se realizaron cultivos bacteriológicos y antibiogramas para identificar las bacterias presentes en la cavidad oral‚ y se determinaron los valores hematológicos no reportados aún para B. asper‚ mantenidas en el serpentario de la Universidad de Antioquia. Mediante hisopados de cavidad oral (n=16 se aislaron 9 especies de bacterias pertenecientes a 4 familias: Enterobacteriaceae (Escherichia coli‚ Citrobacter sp.‚ Proteus sp.‚ Salmonella sp. y Enterobacter sp.‚ Enterococcaceae (Enterococcus sp.‚ Staphylococcaceae (Staphylococcus epidermidis‚ Staphylococcus haemolyticus y Bacillaceae (Bacillus licheniformis. De este total‚ la mayoría (55,5% resultaron Gram negativas y el resto (45,5% fueron Gram positivas. Se observó además una asociación entre los individuos con más tiempo en cautiverio (+ 60 meses‚ 4 ♀ y 3♂ y cultivos simultáneos de 2 bacterias‚ mientras que de los individuos con menos tiempo (- 50 meses‚ 4 ♀ y 5♂ se aisló solo una especie de bacteria. En acuerdo con la literatura‚ todas las bacterias aisladas fueron sensibles a la Amikacina y Enrofloxacina‚ mientras que el 66‚6% (n=9 de las bacterias fue resistente a la Ampicilina Sulbactam. Los parámetros hematológicos se determinaron a partir de 24 serpientes en buen estado de salud. El promedio del hematocrito (Hto‚ % fue de 27

  17. Alteraciones histológicas en riñones de ratas, inducidas por dosis bajas del veneno de la serpiente Bothrops asper. - Histological injuries in rat’s kidneys induced to low dose of snake’s poison Bothrops asper.

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    Reyes, Adriana

    2009-09-01

    Full Text Available ResumenEl objetivo del trabajo fue demostrar que dosis bajas del veneno deBothrops asper pueden causar patologías renales que se acentúancon el tiempo de exposición al veneno, especialmente a nivelglomérular. Se emplearon cinco grupos de ratas de cinco individuoscada uno, los cuales fueron inoculados con una dosis de veneno de2.1µg/g de peso (por vía intraperitoneal, con adición de un grupocontrol, al cual se le inyectó solución salina. A los animales se lespracticó la eutanasia secuencialmente (1, 48, 96, 144 y 288 horas.En cada tiempo se extrajeron los riñones del grupo seleccionado, paraprocesarlos con técnicas de coloración (HE – PAS. Con el diagnósticohistopatológico se evidenció en la primera hora gloméruloscongestionados con tumefacción, después de 48 horas se presentóuna glomérulonefritis, seguida de un colapso glomerular a las 96horas. Las patologías se evidenciaron mas en los últimos períodos deexperimentación; a las 144 horas se observó glomérulonefritismembranosa y proliferativa endocapilar culminando con la desaparición de la cápsula de Bowman y la estructura total del glomérulo a las 288 horas. Se concluye que cantidades bajas del veneno pueden causar patologías renales progresivas con el paso del tiempo.SummaryAim of this work was to demonstrate that low doses of Bothropsasper venom can cause pathologies that progress over time on therenal tissue, at the glomerular level especially. Five groups of ratsconsisting of five individuals were used and a single dose of poison2.1µg/g in weight was injected. Control group was injected withsaline solution. Euthanasia sequentially was practiced (1, 48, 96, 144and 288 hours. In each time was extracted the kidneys of groupselection, to procession with color’s techniques (HE – PAS.Histopathological diagnostic evidenced congested glomeruli withswelling in the first hour, after 48 hours a glomerulonephritis waspresented, followed by a collapsed glomerular at

  18. Purification and Characterization of F Ⅱ A, a Novel Prothrombin-activator from Bothrops atrox Venom%一种新的蛇毒凝血酶原激活物的分离纯化及特征

    Institute of Scientific and Technical Information of China (English)

    孙东; 石皎; 丁忠福; 李秀琳; 李秀娜; 崔亮亮; 薛雁

    2011-01-01

    In this study, a novel prothrombin-activator from the venom of Bothrops atrox designated F II A was isolated. F H A was purified using a SP Sepharose Fast Flow anion-exchange chromatography, DEAE-Sepharose Fast Flow cation-exchange chromatography followed by SP Sepharose High Performance anion-exchange chromatography. F II A was a single-chain polypeptide with a molecular mass 72 kD as measured by reduced SDS-PAGE. Its pi was 6. 67 as measured by IEF. Its N-terminal amino acid sequence was ALVLIAFAQVLQQCP and the accession number was B3A0N1 in UniProtKB Database. The enzymatic activity of F II A was inhibited by the chelating EDTA, whereas the serine protease inhibitor PMSF had no effect on its activity. Interestingly, its enzymatic activity is not requiring cofactors such as Ca2+ , Fva or phospholipid. The results suggest that F II A is a P- I metalloproteinase and its pathways for prothrombin activation is similar to F X a.%为获得矛头蝮蛇(Bothrops atrox)蛇毒凝血酶原激活物并研究其基本性质,采用SP Sepharose Fast Flow,DEAE-Sepharose Fast Flow和SP Sepharose High Performance等层析方法从巴西矛头蝮蛇蛇毒中分离纯化得到1种单一组分的凝血酶原激活物(prothrombin activator,FⅡA).还原性SDSPAGE结果显示,其分子质量约为72 kD,等电点为6.67.HPSEC显示纯度大于95%.该酶是1种N连接的糖蛋白,N末端氨基酸序列为ALVLIAFAQYLQQCP,获得登录号为:B3A0N1.其活性可被EDTA-Na2抑制,PMSF对其活性无影响,对凝血酶原的激活过程无需Ca2+、FVa、磷脂的参与,为P-I金属蛋白酶,对凝血酶原的激活方式与FXa相似.本研究纯化与鉴定的新凝血酶原激活物为其药学研究及临床应用提供参考.

  19. Purificación y caracterización de la l-amino ácido oxidasa del veneno de la serpiente Bothrops brazili "jergón shushupe"

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    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.

  20. Bothrops jararaca venom (BjV) induces differential leukocyte accumulation in mice genetically selected for acute inflammatory reaction: the role of host genetic background on expression of adhesion molecules and release of endogenous mediators.

    Science.gov (United States)

    Carneiro, Adriana S; Ribeiro, Orlando G; Cabrera, Wafa H K; Vorraro, Francisca; De Franco, Marcelo; Ibañez, Olga M; Starobinas, Nancy

    2008-10-01

    The dynamics of the local inflammatory events induced by Bothrops jararaca venom (BjV) inoculation in footpad of mice genetically selected for maximal (AIRmax) and minimal (AIRmin) acute inflammatory reactivity (AIR) was investigated. The BjV injection induced a marked inflammatory cell infiltrate with predominance of neutrophils, with increased blood cell numbers before its accumulation, suggesting a stimulatory action of BjV on mechanisms of cell mobilization from bone marrow. The process of cell migration is regulated by different cell-adhesion molecules (CAM). Our results showed that neutrophil cells from both lines had the same pattern of response concerning CAMs expression, presenting the involvement of l-selectin, Mac-1 and PECAM-1 adhesion molecules in BjV-induced neutrophil accumulation. The effect of BjV on the release of pro-inflammatory cytokines and chemokines related with cellular migration was also studied and IL-1beta, IL-6, TNF-alpha and MIP-2 levels could be detected after venom injection. The AIRmax mice were shown to be more responsive than AIRmin with respect to leukocyte influx, expression of MIP-2 and release of IL-1beta and IL-6. These results demonstrate the importance of host genetic background in the local response and the involvement of alleles accumulated in AIRmax mice in the inflammatory events induced by BjV. PMID:18723041

  1. Interaction between TNF and BmooMP-Alpha-I, a Zinc Metalloprotease Derived from Bothrops moojeni Snake Venom, Promotes Direct Proteolysis of This Cytokine: Molecular Modeling and Docking at a Glance

    Science.gov (United States)

    Silva, Maraisa Cristina; Lopes Silva, Tamires; Silva, Murilo Vieira; Mota, Caroline Martins; Santiago, Fernanda Maria; Fonseca, Kelly Cortes; Oliveira, Fábio; Mineo, Tiago Wilson Patriarca; Mineo, José Roberto

    2016-01-01

    Tumor necrosis factor (TNF) is a major cytokine in inflammatory processes and its deregulation plays a pivotal role in several diseases. Here, we report that a zinc metalloprotease extracted from Bothrops moojeni venom (BmooMP-alpha-I) inhibits TNF directly by promoting its degradation. This inhibition was demonstrated by both in vitro and in vivo assays, using known TLR ligands. These findings are supported by molecular docking results, which reveal interaction between BmooMP-alpha-I and TNF. The major cluster of interaction between BmooMP-alpha-I and TNF was confirmed by the structural alignment presenting Ligand Root Mean Square Deviation LRMS = 1.05 Å and Interactive Root Mean Square Deviation IRMS = 1.01 Å, this result being compatible with an accurate complex. Additionally, we demonstrated that the effect of this metalloprotease on TNF is independent of cell cytotoxicity and it does not affect other TLR-triggered cytokines, such as IL-12. Together, these results indicate that this zinc metalloprotease is a potential tool to be further investigated for the treatment of inflammatory disorders involving TNF deregulation. PMID:27447669

  2. Two phospholipase A2 inhibitors from the plasma of Cerrophidion (Bothrops) godmani which selectively inhibit two different group-II phospholipase A2 myotoxins from its own venom: isolation, molecular cloning and biological properties.

    Science.gov (United States)

    Lizano, S; Angulo, Y; Lomonte, B; Fox, J W; Lambeau, G; Lazdunski, M; Gutiérrez, J M

    2000-01-01

    Myotoxic phospholipases A(2) (PLA(2)s; group II) account for most of the muscle-tissue damage that results from envenomation by viperid snakes. In the venom of the Godman's viper (Cerrophidion godmani, formerly Bothrops godmani), an enzymically active PLA(2) (myotoxin I) and an inactive, Lys-49 variant (myotoxin II) induce extensive muscle damage and oedema. In this study, two distinct myotoxin inhibitor proteins of C. godmani, CgMIP-I and CgMIP-II, were purified directly from blood plasma by selective binding to affinity columns containing either myotoxin I or myotoxin II, respectively. Both proteins are glycosylated, acidic (pI=4) and composed of 20-25-kDa subunits that form oligomers of 110 kDa (CgMIP-I) or 180 kDa (CgMIP-II). In inhibition studies, CgMIP-I specifically neutralized the PLA(2) and the myotoxic, oedema-forming and cytolytic activities of myotoxins I, whereas CgMIP-II selectively inhibited the toxic properties of myotoxin II. N-terminal amino acid sequence analysis and sequencing of cDNAs encoding the two inhibitors revealed that CgMIP-I is similar to gamma-type inhibitors, which share a pattern of cysteine residues present in the Ly-6 superfamily of proteins, whereas CgMIP-II shares sequence identity with alpha-type inhibitors that contain carbohydrate-recognition-like domains, also found in C-type lectins and mammalian PLA(2) receptors. N-terminal sequencing of myotoxin I revealed a different primary structure from myotoxin II [De Sousa, Morhy, Arni, Ward, Díaz and Gutiérrez (1998) Biochim. Biophys. Acta 1384, 204-208], which provides insight into the nature of such pharmacological specificity. PMID:10698689

  3. 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.

  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. 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

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    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

  6. 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

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    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

  7. Seasonal, daily activity, and habitat use by three sympatric pit vipers (Serpentes, Viperidae) from southern Brazil.

    Science.gov (United States)

    Rocha, Marcelo C; Hartmann, Paulo A; Winck, Gisele R; Cechin, Sonia Z

    2014-04-25

    Viperid snakes are widely distributed in the South America and the greater distribution range of the family is found at the Crotalinae subfamily. Despite the abundance of this snakes along their geographic distribution, some ecological aspects remain unknown, principally at subtropical areas. In the present study, we evaluated the activity (daily and seasonal) and the use of the habitat by Bothrops diporus, B. jararaca and B. jararacussu, in an Atlantic Forest area at southern Brazil. We observed higher incidence of viperid snakes during the months with higher temperatures, while no snakes were found during the months with lower temperatures. The data suggest the minimum temperature as environmental variable with the greatest influence on the seasonal activity of this species. Considering the daily activity, we observed a tendency of snakes to avoid the warmest hours. Bothrops jararacussu tend to avoid open areas, being registered only inside and at the edges of the forest. We compared our results with previous studies realized at tropical areas and we suggest the observed seasonal activity as an evolutive response, despite the influence of the different environmental variables, according to the occurence region.

  8. 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

  9. A comparison of the ability of Bellucia dichotoma Cogn. (Melastomataceae) extract to inhibit the local effects of Bothrops atrox venom when pre-incubated and when used according to traditional methods.

    Science.gov (United States)

    Mourão de Moura, Valéria; Serra Bezerra, Adrielle N; Veras Mourão, Rosa Helena; Varjão Lameiras, Juliana L; Almeida Raposo, Juliana D; Luckwu de Sousa, Rafael; Boechat, Antônio Luiz; Bezerra de Oliveira, Ricardo; de Menezes Chalkidis, Hipocrátes; Dos-Santos, Maria Cristina

    2014-07-01

    Bellucia dichotoma Cogn. (Melastomataceae) is one of various plant species used in folk medicine in the west of the state of Pará, Brazil, to treat snake bites. Many studies have been carried out to evaluate the effectiveness of anti-snake bite plants, but few of these use the same preparation methods and doses as those traditionally used by the local populations. This study therefore compared inhibition of the main local effects of B. atrox venom (BaV) by aqueous extract of B. dichotoma (AEBd) administered according to traditional methods and pre-incubated with BaV). The concentrations of phenolic compounds (tannins and flavonoids) in AEBd were determined by colorimetric assays. The effectiveness of AEBd in inhibiting the hemorrhagic and edematogenic activities of BaV was evaluated in mice in four different experimental in vivo protocols: (1) pre-incubation (venom:extract, w/w); (2) pre-treatment (p.o.); (3) post-treatment (p.o.); and (4) AEBd (p.o.) in combination with Bothrops antivenom (BA) (i.v.). To assess in vitro inhibition of BaV phospholipase A₂ activity, the pre-incubation method or incorporation of AEBd or BA in agarose gels were used. The effect of AEBd on BaV was determined by SDS-PAGE, zymography and Western blot. Colorimetric assays revealed higher concentrations of (condensed and hydrolyzable) tannins than flavonoids in AEBd. Hemorrhagic activity was completely inhibited using the pre-incubation protocol. However, with pre-treatment there was no significant inhibition for the concentrations tested, and with the post-treatment only the 725 mg/kg dose of AEBd was able to inhibit 40.5% (p = 0.001) of the hemorrhagic activity of BaV. Phospholipase A₂ activity was only inhibited when AEBd was pre-incubated with BaV. BaV-induced edema was completely inhibited with pre-incubation (p < 0.05) and significantly reduced (p < 0.05) with pre- and post-treatment (p.o.) for the concentrations tested. The reduction in local edema was even greater when AEBd

  10. A comparison of the ability of Bellucia dichotoma Cogn. (Melastomataceae) extract to inhibit the local effects of Bothrops atrox venom when pre-incubated and when used according to traditional methods.

    Science.gov (United States)

    Mourão de Moura, Valéria; Serra Bezerra, Adrielle N; Veras Mourão, Rosa Helena; Varjão Lameiras, Juliana L; Almeida Raposo, Juliana D; Luckwu de Sousa, Rafael; Boechat, Antônio Luiz; Bezerra de Oliveira, Ricardo; de Menezes Chalkidis, Hipocrátes; Dos-Santos, Maria Cristina

    2014-07-01

    Bellucia dichotoma Cogn. (Melastomataceae) is one of various plant species used in folk medicine in the west of the state of Pará, Brazil, to treat snake bites. Many studies have been carried out to evaluate the effectiveness of anti-snake bite plants, but few of these use the same preparation methods and doses as those traditionally used by the local populations. This study therefore compared inhibition of the main local effects of B. atrox venom (BaV) by aqueous extract of B. dichotoma (AEBd) administered according to traditional methods and pre-incubated with BaV). The concentrations of phenolic compounds (tannins and flavonoids) in AEBd were determined by colorimetric assays. The effectiveness of AEBd in inhibiting the hemorrhagic and edematogenic activities of BaV was evaluated in mice in four different experimental in vivo protocols: (1) pre-incubation (venom:extract, w/w); (2) pre-treatment (p.o.); (3) post-treatment (p.o.); and (4) AEBd (p.o.) in combination with Bothrops antivenom (BA) (i.v.). To assess in vitro inhibition of BaV phospholipase A₂ activity, the pre-incubation method or incorporation of AEBd or BA in agarose gels were used. The effect of AEBd on BaV was determined by SDS-PAGE, zymography and Western blot. Colorimetric assays revealed higher concentrations of (condensed and hydrolyzable) tannins than flavonoids in AEBd. Hemorrhagic activity was completely inhibited using the pre-incubation protocol. However, with pre-treatment there was no significant inhibition for the concentrations tested, and with the post-treatment only the 725 mg/kg dose of AEBd was able to inhibit 40.5% (p = 0.001) of the hemorrhagic activity of BaV. Phospholipase A₂ activity was only inhibited when AEBd was pre-incubated with BaV. BaV-induced edema was completely inhibited with pre-incubation (p < 0.05) and significantly reduced (p < 0.05) with pre- and post-treatment (p.o.) for the concentrations tested. The reduction in local edema was even greater when AEBd

  11. Molecular cloning of a hyaluronidase from Bothrops pauloensis venom gland

    OpenAIRE

    Castanheira, Letícia Eulalio; Rodrigues, Renata Santos; Boldrini-França, Johara; Fonseca, Fernando PP; Henrique-Silva, Flávio; Homsi-Brandeburgo, Maria I; Rodrigues, Veridiana M.

    2014-01-01

    Background Hyaluronate is one of the major components of extracellular matrix from vertebrates whose breakdown is catalyzed by the enzyme hyaluronidase. These enzymes are widely described in snake venoms, in which they facilitate the spreading of the main toxins in the victim’s body during the envenoming. Snake venoms also present some variants (hyaluronidases-like substances) that are probably originated by alternative splicing, even though their relevance in envenomation is still under inve...

  12. Clinical and immunological aspects of envenomations by Bothrops snakes

    OpenAIRE

    KPO Luna; MB da Silva; VRA Pereira

    2011-01-01

    Accidents caused by snakes, especially in tropical and subtropical countries, still constitute a serious public health problem due to the lack of knowledge of health professionals and the precariousness of health systems in the regions where most accidents occur. Snake venoms contain a range of molecules that may provoke local swelling, pain, renal and respiratory insufficiencies. The study of the effects of each molecule on humans can help the development of complementary therapy. Similarly,...

  13. Peptidomics of Three Bothrops Snake Venoms: Insights Into the Molecular Diversification of Proteomes and Peptidomes*

    OpenAIRE

    Tashima, Alexandre K.; Zelanis, André; Kitano, Eduardo S.; Ianzer, Danielle; Melo, Robson L.; Rioli, Vanessa; Sant'anna, Sávio S.; Schenberg, Ana C. G.; Camargo, Antônio C. M.; Serrano, Solange M. T.

    2012-01-01

    Snake venom proteomes/peptidomes are highly complex and maintenance of their integrity within the gland lumen is crucial for the expression of toxin activities. There has been considerable progress in the field of venom proteomics, however, peptidomics does not progress as fast, because of the lack of comprehensive venom sequence databases for analysis of MS data. Therefore, in many cases venom peptides have to be sequenced manually by MS/MS analysis or Edman degradation. This is critical for...

  14. A rational protocol for the successful crystallization of l-amino-acid oxidase from Bothrops atrox

    OpenAIRE

    Alves, Raquel Melo; Feliciano, Patricia Rosa; Sampaio, Suely Vilela; Nonato, Maria Cristina

    2011-01-01

    l-Amino-acid oxidase from B. atrox has been crystallized by combining seeding with oil modulation of vapour equilibration. A complete data set has been collected to 2.3 Å resolution from a native crystal.

  15. Preliminary assessment of Hedychium coronarium essential oil on fibrinogenolytic and coagulant activity induced by Bothrops and Lachesis snake venoms

    OpenAIRE

    Miranda, Cíntia A SF; Cardoso, Maria G; Mansanares, Mariana E; Gomes, Marcos S; Marcussi, Silvana

    2014-01-01

    Background The search for new inhibitors of snake venom toxins is essential to complement or even replace traditional antivenom therapy, especially in relation to compounds that neutralize the local effects of envenomations. Besides their possible use as alternative to traditional antivenom therapy, some plant species possess bioactive secondary metabolites including essential oils, which can be extracted from weeds that are considered substantial problems for agriculture, such as Hedychium c...

  16. Action of neuwiedase, a metalloproteinase isolated from Bothrops neuwiedi venom, on skeletal muscle: an ultrastructural and immunocytochemistry study

    Directory of Open Access Journals (Sweden)

    C Baldo

    2010-01-01

    Full Text Available The damaging effects of neuwiedase, a non-hemorrhagic snake venom metalloproteinase from P-I class, on gastrocnemius muscle are studied herein. Following neuwiedase injection, ultrastructural alterations were detected early showing disarrangement of skeletal muscle fibers (characterized by discontinuity of Z lines, mitochondrial swelling, and disruption of plasma membrane and basal lamina. Degradation of skeletal muscle and the appearance of an amorphous substance, primarily composed of cellular debris, were noted after 24 hours. The presence of neuwiedase at the injection site (detected by immunocytochemistry revealed highly specific labeling of myofibril components of damaged myocytes. In addition, proteolysis of muscle proteins assayed through myofibrils extracted from gastrocnemius muscle indicated that neuwiedase provoked degradation of myofibrils, especially myosin. These results suggest that skeletal muscle damage, induced by neuwiedase, is probably due to its proteolytic action on myofibrils, which are responsible for the maintenance of the cellular architecture.

  17. Isolation of a galactose-binding lectin from the venom of the snake Bothrops godmani (Godmann's pit viper).

    Science.gov (United States)

    Lomonte, B; Rojas, G; Gutiérrez, J M; Ramírez, G

    1990-01-01

    A galactose-binding lectin, isolated from the venom of B. godmani by affinity chromatography, is an acidic protein (pI 4.9) with a subunit mol. wt of about 14,000, occurring mostly as a disulfide-linked dimer of 28,000. A small proportion of lectin appears as a monomer and as a tetramer. The lectin agglutinates erythrocytes from mice, rabbit, cow and human (all ABO types, either Rh positive or negative), but does not agglutinate horse, sheep, goat and snake (Oxybelis aeneus, Colubridae) erythrocytes. The agglutinating activity is inhibited by 1 mM EDTA. The lectin is devoid of lethal, hemorrhagic, myotoxic, proteolytic and phospholipase A2 activities. It is not mitogenic for human peripheral blood mononuclear cells. The only effect observed was a moderate induction of edema in the footpad of mice, with a minimal edema-forming dose of 22 micrograms. This effect developed rapidly, and was significantly inhibited by i.p. administration of cyproheptadine, a histamine and serotonin antagonist, before injection of the lectin. Despite the edema-forming activity observed, the low concentration of lectin in crude venom, together with its relatively low potency, suggest that this lectin is not a key component in the development of edema following envenomations by B. godmani.

  18. Partial in vitro analysis of toxic and antigenic activities of eleven Peruvian pitviper snake venoms.

    Science.gov (United States)

    Guerra-Duarte, C; Lopes-Peixoto, J; Fonseca-de-Souza, B R; Stransky, S; Oliveira, D; Schneider, F S; Lopes-de-Souza, L; Bonilla, C; Silva, W; Tintaya, B; Yarleque, A; Chávez-Olórtegui, C

    2015-12-15

    This work used eleven Peruvian snake venoms (Bothrops andianus, Bothrops atrox, Bothrops barnetti, Bothrops castelnaudi, Bothriopsis chloromelas, Bothrocophias microphthalmus, Bothrops neuwiedi, Bothriopsis oligolepis, Bothriopsis peruviana, Bothrops pictus and Bothriopsis taeniata) to perform in vitro experimentation and determine its main characteristics. Hyaluronidase (HYAL), phospholipase A2 (PLA2), snake venom metalloproteinase (SVMP), snake venom serine protease (SVSP) and L-amino acid oxidase (LAAO) activities; toxicity by cell viability assays using MGSO3, VERO and HeLa cell lineages; and crossed immunoreactivity with Peruvian (PAV) and Brazilian (BAV) antibothropic polyvalent antivenoms, through ELISA and Western Blotting assays, were determined. Results show that the activities tested in this study were not similar amongst the venoms and each species present their own peculiarities, highlighting the diversity within Bothrops complex. All venoms were capable of reducing cell viability of all tested lineages. It was also demonstrated the crossed recognition of all tested venoms by both antivenoms. PMID:26365916

  19. Presynaptic Proteins as Markers of the Neurotoxic Activity of BmjeTX-I and BmjeTX-II Toxins from Bothrops marajoensis (Marajó Lancehead) Snake Venom

    Science.gov (United States)

    Lisboa, Antonio; Melaré, Rodolfo; Franco, Junia R. B.; Bis, Carolina V.; Gracia, Marta; Ponce-Soto, Luis A.; Marangoni, Sérgio; Rodrigues-Simioni, Léa; da Cruz-Höfling, Maria Alice

    2016-01-01

    Neuromuscular preparations exposed to B. marajoensis venom show increases in the frequency of miniature end-plate potentials and twitch tension facilitation followed by presynaptic neuromuscular paralysis, without evidences of muscle damage. Considering that presynaptic toxins interfere into the machinery involved in neurotransmitter release (synaptophysin, synaptobrevin, and SNAP25 proteins), the main objective of this communication is to analyze, by immunofluorescence and western blotting, the expression of the synaptic proteins, synaptophysin, synaptobrevin, and SNAP25 and by myography, light, and transmission electron microscopy the pathology of motor nerve terminals and skeletal muscle fibres of chick biventer cervicis preparations (CBC) exposed in vitro to BmjeTX-I and BmjeTX-II toxins from B. marajoensis venom. CBC incubated with toxins showed irreversible twitch tension blockade and unaffected KCl- and ACh-evoked contractures, and the positive colabelling of acetylcholine receptors confirmed that their action was primarily at the motor nerve terminal. Hypercontraction and loose myofilaments and synaptic vesicle depletion and motor nerve damage indicated that the toxins displayed both myotoxic and neurotoxic effect. The blockade resulted from interference on synaptophysin, synaptobrevin, and SNAP25 proteins leading to the conclusion that BmjeTX-I and BmjeTX-II affected neurotransmitter release machinery by preventing the docking of synaptic vesicles to the axolemma of the nerve terminal. PMID:27635261

  20. 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.

  1. 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.

  2. Hemocoagulase from Agkistrodon halys (pallas) vs hemocoagulase from Bothrops atrox%白眉蛇毒血凝酶与血凝酶的对比

    Institute of Scientific and Technical Information of China (English)

    黄仲义

    2005-01-01

    通过对血凝酶和白眉蛇毒血凝酶的组成成分、药理作用、质量标准等方面的研究,提示白眉蛇毒血凝酶与血凝酶不同,是另外一种从蛇毒提取的含有类凝血酶和类凝血激酶的具有止血作用的多种组分复合制剂,其临床疗效和安全性需进一步研究和证明.

  3. Gene : CBRC-ACAR-01-0620 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available AAO72730.1| follicle-stimulating hormone receptor [Bothrops jararaca] 0.0 85% MALVYLCLLLLLLPSGTCSGCQHPLCQCSD...GANGPSFLDISETRISQLPAIGLESISKLRARSAYSLKKLPPLDKFRALIEANLTYPSHCCAFANWTKQNSMLHPLCNKSSTKQDPEGLYFELDEDDYYQSLCKEEVQ

  4. Drug: D06076 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available D06076 Drug Teprotide (USAN/INN) C53H76N14O12 1100.5767 1101.2571 D06076.gif Bothrops...ing] Same as: C11712 venom of the Bothrops jararaca angiotensin-converting enzyme (ACE) inhibitor [HSA:1636

  5. AcEST: BP914257 [AcEST

    Lifescience Database Archive (English)

    Full Text Available .4 sp|Q9DGI0|FTE46_BOTJA Antihemorragic factor BJ46a OS=Bothrops ja... 30 4.4 sp|Q3TIT8|SPX3_MOUSE Sugar pho...JA Antihemorragic factor BJ46a OS=Bothrops jararaca PE=1 SV=1 Length = 341 Score = 30.4 bits (67), Expect =

  6. AcEST: BP914004 [AcEST

    Lifescience Database Archive (English)

    Full Text Available _BOTIN Phospholipase A2 2 (Fragments) OS=Bothrops ... 29 8.2 >sp|Q9SS60|PP210_ARATH Pentatricopeptide repeat...CFENGQCS 686 >sp|P84397|PA22_BOTIN Phospholipase A2 2 (Fragments) OS=Bothrops ins

  7. AcEST: DK960586 [AcEST

    Lifescience Database Archive (English)

    Full Text Available 8UF9 Definition sp|Q98UF9|VMHF3_BOTJA Zinc metalloproteinase-disintegrin HF3 OS=Bothrops jararaca Align leng...rinogen decarboxylase OS=Agrobacte... 30 8.9 >sp|Q98UF9|VMHF3_BOTJA Zinc metalloproteinase-disintegrin HF3 OS=Bothrops

  8. Dicty_cDB: Contig-U14983-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 37 2.3 EU733640_1( EU733640 |pid:none) Bothrops atrox batroxstatin-2 mRNA... 37 2.3 EU733643_1( EU733643 |p...7_6( BX649457 |pid:none) Zebrafish DNA sequence from clone ... 35 8.7 AY255004_1( AY255004 |pid:none) Bothrops

  9. Dicty_cDB: Contig-U16376-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 2 ( DW713082 ) JARO001014H04 001 Bothrops jararaca cDNA, mRNA se... 60 9e-11 2 ( EL821265 ) CBWN5685.g1 NIC...Hawdon Ancylostoma caninum L3 Ancylost... 58 1e-08 2 ( BM401720 ) GH077R Snake Bothrops insularis library IL

  10. AcEST: BP913809 [AcEST

    Lifescience Database Archive (English)

    Full Text Available OTJA Bothrojaracin subunit beta OS=Bothrops jara... 30 4.9 sp|Q46915|GUDX_ECOLI Glucarate dehydratase-relate...A Bothrojaracin subunit beta OS=Bothrops jararaca PE=1 SV=1 Length = 150 Score = 29.6 bits (65), Expect = 4.

  11. Toxicity of South American snake venoms measured by an in vitro cell culture assay.

    Science.gov (United States)

    Oliveira, J C R; de Oca, H M; Duarte, M M; Diniz, C R; Fortes-Dias, C L

    2002-03-01

    Cytotoxicity of venoms from eight medically important South American Crotalidae snakes (Bothrops and Lachesis genera) was determined, based on a procedure originally described for the screening of cytotoxic agents in general. The assay, the conditions of which were adapted to snake venoms, determines the survival of viable cells in monolayer culture upon exposure to the toxic agent. Snake venom toxicity was expressed as the venom dose that killed 50% of the cells (CT(50)) under the assay conditions. Bothrops neuwieddi mattogrossensis (CT(50)=4.74+/-0.35 microg/ml) and Bothrops leucurus (CT(50)=4.95+/-0.51 microg/ml) were the most cytotoxic whereas Bothrops atrox (CT(50)=34.64+/-2.38 microg/ml) and Bothrops sp. (CT(50)=33.89+/-3.89 microg/ml) were the least cytotoxic venoms, respectively. The relationship between CT(50) and other biological activities of these snake venoms was evaluated. PMID:11711131

  12. AcEST: DK951785 [AcEST

    Lifescience Database Archive (English)

    Full Text Available homolog OS=Caeno... 32 2.1 sp|P83512|VMBP1_BOTAS Zinc metalloproteinase Bap1 OS=Bothrops as... 32 2.8 sp|Q0...SLFSAEGKAKTESAV 647 >sp|Q7T1T4|VMMP2_BOTJR Zinc metalloproteinase BjussuMP-2 (Fragment) OS=Bothrops jararacu...talloproteinase/disintegrin OS=Bothrops insularis PE=1 SV=1 Length = 476 Score = 32.3 bits (72), Expect = 2....VMBP1_BOTAS Zinc metalloproteinase Bap1 OS=Bothrops asper PE=1 SV=2 Length = 408 Score = 32.0 bits (71), Exp...PEQQRFSPRYIELAVVADH 208 >sp|Q072L5|VM2_BOTAS Zinc metalloproteinase/disintegrin OS=Bothrops asper PE=2 SV=1

  13. 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.

  14. 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.

  15. 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)

    Cristina Giannotti, Karina; 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 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. PMID:23509782

  16. AcEST: DK960559 [AcEST

    Lifescience Database Archive (English)

    Full Text Available id Q9I8F8 Definition sp|Q9I8F8|PA2_BOTPC Phospholipase A2 OS=Bothrops pictus Align length 80 Score (bit) 32....its) Value sp|Q9I8F8|PA2_BOTPC Phospholipase A2 OS=Bothrops pictus PE=2 SV=1 33 1...ke protein 18 OS=Arabidopsis thali... 30 8.5 >sp|Q9I8F8|PA2_BOTPC Phospholipase A2 OS=Bothrops pictus PE=2 S

  17. 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...

  18. 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.

  19. Management of venomous snakebites in dogs and cats in Brazil

    OpenAIRE

    R. S. FERREIRA JÚNIOR; B. Barravieira

    2004-01-01

    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, di...

  20. AcEST: DK950256 [AcEST

    Lifescience Database Archive (English)

    Full Text Available catalytic sub... 33 1.0 sp|Q9I8F8|PA2_BOTPC Phospholipase A2 OS=Bothrops pictus PE=2 SV=1 33 1.8 sp|Q02957|K...ESASRSSSQSSHQRHSRDGRGELG-SEHGERRS 159 >sp|Q9I8F8|PA2_BOTPC Phospholipase A2 OS=Bothrops pictus PE=2 SV=1 Len

  1. [Study on the venoms of the principal venomous snakes from French Guiana and the neutralization].

    Science.gov (United States)

    Estévez, J; Magaña, P; Chippaux, J P; Vidal, N; Mancilla, R; Paniagua, J F; de Roodt, A R

    2008-10-01

    We studied some biochemical, toxic and immunological characteristics of the venoms of Bothrops atrox, Bothrops brazili and Lachesis muta, Viperidae responsible for most of the bites of venomous snakes in French Guiana. Chromatographic (HPLC) and electrophoretical profiles (SDS-PAGE), lethal, hemorrhagic, defibrinogenating, coagulant, thrombin like, proteolytic, fibrino(geno)lytic and phospholipase activities were studied. In addition, the neutralization of some toxic activities conferred by four antivenins was compared. The chromatographic and electrophoretic profiles were different for the three venoms, showing differences between Bothrops and L. muta venoms. In general, bothropic venoms showed the highest toxic and enzymatic activities, while the venom of L. muta showed the lowest lethal, hemorrhagic and coagulant activities. The enzymes of bothropic venoms responsible for gelatinolytic activity were around 50-90 kDa. All the venoms were able to hydrolyze a and beta chains of the fibrinogen, showing different patterns of degradation. Although all the antivenoms tested were effective to various degrees in neutralizing the venom of B. brazili and B. atrox, neutralization of L. muta venom was significantly better achieved using the antivenom including this venom in its immunogenic mixture. For the neutralization of L. muta venom, homologous or polyvalent antivenoms that include the "bushmaster" venom in their immunogenic mixture should be preferred. PMID:18956820

  2. 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.

  3. [Snakes from the urban area of Cuiabá, Mato Grosso: ecological aspects and associated snakebites].

    Science.gov (United States)

    de Carvalho, M A; Nogueira, F N

    1998-01-01

    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 snake bites (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 snake bites 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 probably responsible for most of the non-venomous snake bites. PMID:9878908

  4. 两种巴曲酶制剂的区别

    Institute of Scientific and Technical Information of China (English)

    张毅

    2008-01-01

    @@ 巴曲酶制剂在神经科的应用十分广泛.矛头蝮蛇属(bothrops)是南美两大毒蛇家族之一,种类繁多,从巴西矛头蝮蛇(bothrops atrox)蛇毒中分离获得的巴曲酶具有促凝血作用,称为血凝酶(hemopexin),临床药物有立止血和巴曲亭.而从另一亚种bothrops moojeni蛇毒分离而来的巴曲酶则具有去纤维蛋白原作用,称为去纤酶(defibrinogenase),临床药物有东菱精纯克栓酶.

  5. The application of indirect ELISA on snake venom species identification in Hemocoagulase Bothrops atrox for Injection%间接ELISA法在注射用矛头蝮蛇血凝酶蛇毒种属鉴别中的应用

    Institute of Scientific and Technical Information of China (English)

    李秀娜; 孙东; 李秀琳; 崔亮亮; 丁忠福; 宁静

    2016-01-01

    目的 建立间接ELISA法定性检测矛头蝮蛇血凝酶,用于巴曲亭蛇毒种属来源检测.方法 将矛头蝮蛇血凝酶为抗原免疫小鼠,与尖吻蝮蛇血凝酶、白眉蝮蛇血凝酶交叉筛选获得特异性单克隆抗体.优化矛头蝮蛇血凝酶单抗(一抗)、HRP标记的羊抗鼠(二抗)IgG、矛头蝮蛇血凝酶(抗原)工作浓度,建立间接ELISA检测方法.考察方法的重复性、中间精密度,确定方法的适用性.采用膜超滤法将巴曲亭中辅料去除,按照建立的ELISA法检测,判定种属来源.结果 矛头蝮蛇血凝酶单抗具有强特异性,一抗的最佳工作浓度1∶4000(250 ng),二抗的最佳工作浓度1∶4000(250 ng),抗原浓度2μg/孔,该法重复性和中间精密度较好.巴曲亭与矛头蝮蛇血凝酶单抗有特异性反应,与其它蛇毒来源血凝酶单抗无交叉反应,确定巴曲亭来源于矛头蝮蛇蛇毒.结论 本研究建立的ELISA法具有较好的特异性,可作为巴曲亭蛇毒种属来源判断的方法.

  6. 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.

  7. 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. PMID:9428185

  8. Drug: D03352 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available D03352 Drug Batroxobin (JAN/INN); Defibrase (TN) Bothrops atrox [TAX:8725] Enzyme Therapeutic... category: 3959 ATC code: B02BX03 thrombin-like proteolytic enzyme Therapeutic category of drugs i...ons 3959 Others D03352 Batroxobin (JAN/INN) Anatomical Therapeutic Chemical (ATC)

  9. Characterization of allergens in four South American snake species.

    Science.gov (United States)

    Madero, Mauro F; Gámez, Cristina; Madero, Mauro A; Fernández-Nieto, Mar; Sastre, Joaquín; del Pozo, Victoria

    2009-01-01

    A 55-year-old herpetologist developed rhinitis, asthma, urticaria and anaphylaxis when handling 4 different viper snake venoms. Allergen characterizations were done using SDS-PAGE, IgE immunoblotting and IgE inhibition experiments. The most prominent immunoreactive proteins were analyzed by MALDI-TOF mass spectrometry, and peptide identity was demonstrated by homology with known peptide sequences. SDS-PAGE showed several protein bands ranging from 5 to 99 kDa in each of the 4 snake venoms. Immunoblotting demonstrated 4 IgE-binding bands in the Bothrops extract of about 60, 28, 14 and 7 kDa. The bands of 28 and 14 kDa were also present in Lachesis muta. Two IgE-binding proteins of about 50 and 35 kDa were found in Bothrops atrox and L. muta, respectively. A strong inhibition of IgE binding to immobilize Bothrops asper proteins was observed after preabsorption of sera with B. asper, B. atrox,Bothrops xanthograma and L. muta extracts. MALDI-TOF analysis showed a 14-kDa phospholipase and the 60- and 28-kDa proteins showed significant similarity with metalloproteinases. In this report we have characterized the snake venom allergens that can elicit IgE-mediated symptoms. PMID:19494529

  10. Fluorometric assay using naphthylamide substrates for assessing novel venom peptidase activities.

    Science.gov (United States)

    Gasparello-Clemente, Elaine; Silveira, Paulo Flávio

    2002-11-01

    In the present study we examined the feasibility of using the fluorometry of naphthylamine derivatives for revealing peptidase activities in venoms of the snakes Bothrops jararaca, Bothrops alternatus, Bothrops atrox, Bothrops moojeni, Bothrops insularis, Crotalus durissus terrificus and Bitis arietans, of the scorpions Tityus serrulatus and Tityus bahiensis, and of the spiders Phoneutria nigriventer and Loxosceles intermedia. Neutral aminopeptidase (APN) and prolyl-dipeptidyl aminopeptidase IV (DPP IV) activities were presented in all snake venoms, with the highest levels in B. alternatus. Although all examined peptidase activities showed relatively low levels in arthropod venoms, basic aminopeptidase (APB) activity from P. nigriventer venom was the exception. Compared to the other peptidase activities, relatively high levels of acid aminopeptidase (APA) activity were restricted to B. arietans venom. B. arietans also exhibited a prominent content of APB activity which was lower in other venoms. Relatively low prolyl endopeptidase and proline iminopeptidase activities were, respectively, detectable only in T. bahiensis and B. insularis. Pyroglutamate aminopeptidase activity was undetectable in all venoms. All examined peptidase activities were undetectable in T. serrulatus venom. In this study, the specificities of a diverse array of peptidase activities from representative venoms were demonstrated for the first time, with a description of their distribution which may contribute to guiding further investigations. The expressive difference between snake and arthropod venoms was indicated by APN and DPP IV activities while APA and APB activities distinguished the venom of B. arietans from those of Brazilian snakes. The data reflected the relatively uniform qualitative distribution of the peptidase activities investigated, together with their unequal quantitative distribution, indicating the evolutionary divergence in the processing of peptides in these different

  11. 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

  12. 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.

  13. Inhibition of Crotalidae venom hemorrhagic activities by Didelphis marsupialis liver spheroids culture supernatants.

    Science.gov (United States)

    Salgueiro, L M; Rodríguez-Acosta, A; Rivas-Vetencourt, P; Zerpa, M; Carillo, G; Aguilar, I; Girón, M E; Acevedo, C E; Gendzekhadze, K

    2001-05-01

    The main aim of this work was the development of a primary hepatocyte culture from Didelphis marsupialis, to determine the possible use of culture medium supernatants as a source of inhibitors of the Bothrops lanceolatus venom hemorrhagic activity. The cellular culture was carried out from isolated hepatocytes by the double perfusion technique, and digestion of the liver with collagenase and culturing the hepatocytes in a liquid media under continuous agitation at 37 degrees C in 5% CO2. The hemorrhagic activity inhibition assays were performed inoculating intradermically, a mixture of Bothrops lanceolatus venom plus a pool of liver spheroids culture supernatants, in mice. These liver Didelphis marsupialis spheroid cultures were adequate to obtain large supernatant volumes with inhibitors of hemorrhagic activity. PMID:11405280

  14. 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.

  15. Functional analysis of DM64, an antimyotoxic protein with immunoglobulin-like structure from Didelphis marsupialis serum.

    Science.gov (United States)

    Rocha, Surza L G; Lomonte, Bruno; Neves-Ferreira, Ana G C; Trugilho, Monique R O; Junqueira-de-Azevedo, Inácio de L M; Ho, Paulo L; Domont, Gilberto B; Gutiérrez, José M; Perales, Jonas

    2002-12-01

    Bothrops snake venoms are known to induce local tissue damage such as hemorrhage and myonecrosis. The opossum Didelphis marsupialis is resistant to these snake venoms and has natural venom inhibitors in its plasma. The aim of this work was to clone and study the chemical, physicochemical and biological properties of DM64, an antimyotoxic protein from opossum serum. DM64 is an acidic protein showing 15% glycosylation and with a molecular mass of 63 659 Da when analysed by MALDI-TOF MS. It was cloned and the amino acid sequence was found to be homologous to DM43, a metalloproteinase inhibitor from D. marsupialis serum, and to human alpha1B-glycoprotein, indicating the presence of five immunoglobulin-like domains. DM64 neutralized both the in vivo myotoxicity and the in vitro cytotoxicity of myotoxins I (mt-I/Asp49) and II (mt-II/Lys49) from Bothrops asper venom. The inhibitor formed noncovalent complexes with both toxins, but did not inhibit the PLA2 activity of mt-I. Accordingly, DM64 did not neutralize the anticoagulant effect of mt-I nor its intracerebroventricular lethality, effects that depend on its enzymatic activity, and which demonstrate the dissociation between the catalytic and toxic activities of this Asp49 myotoxic PLA2. Furthermore, despite its similarity with metalloproteinase inhibitors, DM64 presented no antihemorrhagic activity against Bothrops jararaca or Bothrops asper crude venoms, and did not inhibit the fibrinogenolytic activity of jararhagin or bothrolysin. This is the first report of a myotoxin inhibitor with an immunoglobulin-like structure isolated and characterized from animal blood. PMID:12473101

  16. Philodryas patagoniensis bite and local envenoming Picada por Philodryas patagoniensis e envenenamento local

    OpenAIRE

    Sérgio de Andrade Nishioka; Paulo Vitor Portella Silveira

    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...

  17. 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.

  18. Protective effect of snake venom antibodies in sera of previous snake bite victims.

    Science.gov (United States)

    Theakston, R D; Reid, H A; Iddon, D; Larrick, J W

    1983-06-01

    Three out of seven serum samples from Ecuadorian Indians had very high antibody levels against Bothrops nasutus venom, and IgG concentrates of these sera effectively neutralized this venom when subsequently injected into mice. It is concluded that the high mortality rate among these Indians would be even higher if there were not such natural protection. Further research into active immunization of humans should be encouraged. PMID:6625731

  19. 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. PMID:18455482

  20. Serpentes da área urbana de Cuiabá, Mato Grosso: aspectos ecológicos e acidentes ofídicos associados

    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.

  1. 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.

  2. IgE antibodies against snake venoms.

    Science.gov (United States)

    Alonso, A; Scavini, L M; Marino, G A; Rodríguez, S M

    1995-01-01

    A similar event was detected in the clinical records of a small group of atopic patients living in the northern provinces of Argentina, i.e., they were bitten by a snake of the Bothrops species (or yarará) during their rural activities (woodcutters, cattle-drivers and farmers). Those who were bitten twice suffered an acute episode of hives and angioedema within 15 minutes after the snake bite. The presence of specific antibodies against Bothrops alternata (Ba) extract was detected by means of RAST for IgE and Ouchterlony and Boyden for IgG. The Ouchterlony also demonstrated crossreactivity among the venoms of the Bothrops species and the positivity of the six fractions obtained by DEAE-cellulose column fractionation against the horse anti-Ba serum. The Ba antigen induced a definite inhibition of the RAST. We presume that hives and angioedema in atopic patients immediately after a second snake bite could be attributed to the presence of a specific IgE antibody against the venom, and must not be misinterpreted with the toxic effects that appear later. PMID:7551202

  3. Specific identification of Lachesis muta muta snake venom using antibodies against the plasminogen activator enzyme, LV-PA.

    Science.gov (United States)

    Felicori, Liza F; Chávez-Olórtegui, Carlos; Sánchez, Eladio F

    2005-05-01

    Sandwich-type enzyme linked immunosorbent assays (ELISA) were developed to detect Lachesis muta muta (bushmaster) snake venom using antibodies against the plasminogen activator enzyme (LV-PA). Antibodies to LV-PA were obtained by immunization of one rabbit with the purified enzyme. The IgG fraction was purified from rabbit blood in a single step on a column of Sepharose-L. m. muta venom and used to coat the microtiter plates. The specificity of the assay was demonstrated by its capacity to correctly discriminate between the circulating antigens in mice that were experimentally inoculated with L. m. muta venom from those in mice inoculated with venoms from Bothrops atrox, B. brazili, B. castelnaudi, Bothriopsis taeniata, B. bilineata, Crotalus durissus ruruima and the antigenic Bothrops (AgB) and Crotalus (AgC) pools venoms used to produce Bothropic and Crotalic antivenoms at Fundacao Ezequiel Dias (FUNED). Measurable absorbance signals were obtained with 1.5 ng of venom per assay. The ELISA was used to follow the kinetic distribution of antigens in experimentally envenomed mice. PMID:15804530

  4. Crotalid snake venom subproteomes unraveled by the antiophidic protein DM43.

    Science.gov (United States)

    Rocha, Surza L G; Neves-Ferreira, Ana G C; Trugilho, Monique R O; Chapeaurouge, Alex; León, Ileana R; Valente, Richard H; Domont, Gilberto B; Perales, Jonas

    2009-05-01

    Snake venoms are mixtures of proteins and peptides with different biological activities, many of which are very toxic. Several animals, including the opossum Didelphis aurita, are resistant to snake venoms due to the presence of neutralizing factors in their blood. An antihemorrhagic protein named DM43 was isolated from opossum serum. It inhibits snake venom metalloproteinases through noncovalent complex formation with these enzymes. In this study, we have used DM43 and proteomic techniques to explore snake venom subproteomes. Four crotalid venoms were chromatographed through an affinity column containing immobilized DM43. Bound fractions were analyzed by one- and two-dimensional gel electrophoresis, followed by identification by MALDI-TOF/TOF mass spectrometry. With this approach, we could easily visualize and compare the metalloproteinase compositions of Bothrops atrox, Bothrops jararaca, Bothrops insularis, and Crotalus atrox snake venoms. The important contribution of proteolytic processing to the complexity of this particular subproteome was demonstrated. Fractions not bound to DM43 column were similarly analyzed and were composed mainly of serine proteinases, C-type lectins, C-type lectin-like proteins, l-amino acid oxidases, nerve growth factor, cysteine-rich secretory protein, a few metalloproteinases (and their fragments), and some unidentified spots. Although very few toxin families were represented in the crotalid venoms analyzed, the number of protein spots detected was in the hundreds, indicating an important protein variability in these natural secretions. DM43 affinity chromatography and associated proteomic techniques proved to be useful tools to separate and identify proteins from snake venoms, contributing to a better comprehension of venom heterogeneity. PMID:19267469

  5. Contribuição à biologia de serpentes da Bahia, Brasil: I. vivíparas Contribution to reproductive biology of snakes in Bahia, Brazil: I. viviparous

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    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.

  6. 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)

    OpenAIRE

    Marisa M. T. da Rocha; Maria de F. D. Furtado

    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...

  7. Snake venom antibodies in Ecuadorian Indians.

    Science.gov (United States)

    Theakston, R D; Reid, H A; Larrick, J W; Kaplan, J; Yost, J A

    1981-10-01

    Serum samples from 223 Waorani Indians, a tribe in eastern Ecuador, were investigated by enzyme-linked immunosorbent assay for antibodies to snake venom. Seventy-eight per cent were positive, confirming the highest incidence and mortality from snake bite poisoning yet recorded in the world. Most samples were positive for more than one venom antibody. Antibodies were found to venoms of Bothrops viper in 60% of positive cases, of Micrurus coral snake in 21%, and of the bushmaster, Lachesis muta, in 18%. Further studies are needed to determine whether high venom-antibody levels afford protection against further snake envenoming. PMID:7299877

  8. Biochemical and biological activities of the venom of a new species of pitviper from Vietnam, Triceratolepidophis sieversorum.

    Science.gov (United States)

    Mebs, Dietrich; Kuch, Ulrich; Herrmann, Hans Werner; Ziegler, Thomas

    2003-02-01

    Biochemical and biological activities of a venom sample from a recently discovered new genus and species of pitviper from Vietnam, Triceratolepidophis sieversorum, were assayed and compared with those of five other viperid snakes (Bothrops asper, Crotalus atrox, Protobothrops flavoviridis, Trimeresurus insularis, and Vipera ammodytes). The venom had high casein hydrolysis, arginine ester hydrolysis and haemorrhagic activities, lacked measurable phosphodiesterase and L-amino acid oxidase activities, and had no procoagulant activity on either bovine fibrinogen or human plasma. Other enzymatic activities (phospholipase A(2), kallikrein) were moderate. The approximate i.p. LD(50) (mice) of the venom is about 5-6 mg/kg. PMID:12565732

  9. Proteomics application exercise of the Swiss Proteomics Society: report of the SPS'02 session.

    Science.gov (United States)

    Binz, Pierre-Alain; Abdi, Fadi; Affolter, Michael; Allard, Laure; Barblan, Jachen; Bhardwaj, Sanjeev; Bienvenut, Willy V; Bulet, Philippe; Burgess, Jennifer; Carrette, Odile; Corthals, Garry; Delalande, François; Diemer, Hélène; Favreau, Philippe; Giuliano, Elia; Gueguen, Yannick; Guillaume, Elisabeth; Hahner, Stephanie; Man, Petr; Michalet, Sophie; Neri, Dario; Noukakis, Dimitrios; Palagi, Patricia; Paroutaud, Pierre; Pimenta, Daniel Carvalho; Quadroni, Manfredo; Resemann, Anja; Richert, Sophie; Rybak, Jascha; Sanchez, Jean-Charles; Scherl, Alexander; Scheurer, Simone; Schweiger Hufnagel, Ulrike; Siethoff, Christoph; Suckau, Detlev; van Dorsselaer, Alain; Wagner Redeker, Winfried; Walter, Nadia; Stöcklin, Reto

    2003-08-01

    After the success of the mass spectrometry (MS) round table that was held at the first Swiss Proteomics Society congress (SPS'01) in Geneva, the SPS has organized a proteomics application exercise and allocated a full session at the SPS'02 congress. The main objective was to encourage the exchange of expertise in protein identification, with a focus on the use of mass spectrometry, and to create a bridge between the users' questions and the instrument providers' solutions. Two samples were sent to fifteen interested labs, including academic groups and MS hardware providers. Participants were asked to identify and partially characterize the samples. They consisted of a complex mixture of peptide/proteins (sample A) and an almost pure recombinant peptide carrying post-translational modifications (sample B). Sample A was an extract of snake venom from the species Bothrops jararaca. Sample B was a recombinant and modified peptide derived from the shrimp Penaeus vannamei penaeidin 3a. The eight labs that returned results reported the use of a wide range of MS instrumentation and techniques. They mentioned a variety of time and manpower allocations. The origin of sample A was generally identified together with a number of database protein entries. The difficulty of the sample identification lay in the incomplete knowledge of the Bothrops species genome sequence and is discussed. Sample B was generally and correctly identified as penaeidin. However, only one group reported the full primary structure. Interestingly, the approaches were again varied and are discussed in the text.

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

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    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.

  11. 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

  12. Venomous Frogs Use Heads as Weapons.

    Science.gov (United States)

    Jared, Carlos; Mailho-Fontana, Pedro Luiz; Antoniazzi, Marta Maria; Mendes, Vanessa Aparecida; Barbaro, Katia Cristina; Rodrigues, Miguel Trefaut; Brodie, Edmund D

    2015-08-17

    Venomous animals have toxins associated with delivery mechanisms that can introduce the toxins into another animal. Although most amphibian species produce or sequester noxious or toxic secretions in the granular glands of the skin to use as antipredator mechanisms, amphibians have been considered poisonous rather than venomous because delivery mechanisms are absent. The skin secretions of two Brazilian hylid frogs (Corythomantis greening and Aparasphenodon brunoi) are more toxic than the venoms of deadly venomous Brazilian pitvipers, genus Bothrops; C. greeningi secretion is 2-fold and A. brunoi secretion is 25-fold as lethal as Bothrops venom. Like the venoms of other animals, the skin secretions of these frogs show proteolytic and fibrinolytic activity and have hyaluronidase, which is nontoxic and nonproteolytic but promotes diffusion of toxins. These frogs have well-developed delivery mechanisms, utilizing bony spines on the skull that pierce the skin in areas with concentrations of skin glands. C. greeningi has greater development of head spines and enlarged skin glands producing a greater volume of secretion, while A. brunoi has more lethal venom. C. greeningi and A. brunoi have highly toxic skin secretions and an associated delivery mechanism; they are therefore venomous. Because even tiny amounts of these secretions introduced into a wound caused by the head spines could be dangerous, these frogs are capable of using their skin toxins as venoms against would-be predators. PMID:26255851

  13. Ethnobotanic study of Randia aculeata (Rubiaceae in Jamapa, Veracruz, Mexico, and its anti-snake venom effects on mouse tissue

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    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.

  14. 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)

  15. Non-venomous snake bite and snake bite without envenoming in a Brazilian teaching hospital. Analysis of 91 cases.

    Science.gov (United States)

    Silveria, P V; Nishioka, S de A

    1992-01-01

    A retrospective survey of 473 cases of snake bite admitted to a Brazilian teaching hospital from 1984 to 1990 revealed 91 cases of bite without envenoming and/or caused by non-venomous snakes. In 17 of these cases the snake was identified, and one patient was bitten by a snake-like reptile (Amphisbaena mertensii). In 43 cases diagnosis was made on clinical grounds (fang marks in the absence of signs of envenoming). The other 30 cases were of patients who complained of being bitten but who did not show any sign of envenoming or fang mark. Most cases occurred in men (66;73%), in the 10-19 years age group (26;29%), in the lower limbs (51/74;69%), between 6 A. M. and 2 P.M. (49;61%) and in the month of April (16;18%). One patient bitten by Philodryas olfersii developed severe local pain, swelling and redness at the site of the bite, with normal clotting time. The patient bitten by Drymarcon corais was misdiagnosed as being bitten by a snake of the genus Bothrops, was given the specific antivenom, and developed anaphylaxis. One patient bitten by Sibynomorphus mikanii presented prolonged clotting time, and was also given antivenom as a case of Bothrops bite. Correct identification of venomous snakes by physicians is necessary to provide correct treatment to victims of snake bite, avoiding unnecessary distress to the patient, and overprescription of antivenom, which may eventually cause severe untoward effects. PMID:1342117

  16. Nucleotidase and DNase activities in Brazilian snake venoms.

    Science.gov (United States)

    Sales, Paulo Bruno Valadão; Santoro, Marcelo L

    2008-01-01

    Among the myriad of enzymes present in animal venoms, nucleotidases and nucleases are poorly investigated. Herein, we studied such enzymes in 28 crude venoms of animals found in Brazil. Higher levels of ATPase, 5'-nucleotidase, ADPase, phosphodiesterase and DNase activities were observed in snake venoms belonging to Bothrops, Crotalus and Lachesis genera than to Micrurus genus. The venom of Bothrops brazili snake showed the highest nucleotidase and DNase activities, whereas that of Micrurus frontalis snake the highest alkaline phosphatase activity. On the other hand, the venoms of the snake Philodryas olfersii and the spider Loxosceles gaucho were devoid of most nucleotidase and DNase activities. Species that exhibited similar nucleotidase activities by colorimetric assays showed different banding pattern by zymography, suggesting the occurrence of structural differences among them. Hydrolysis of nucleotides showed that 1 mol of ATP is cleaved in 1 mol of pyrophosphate and 1 mol of orthophosphate, whereas 1 mol of ADP is cleaved exclusively in 2 mol of orthophosphates. Pyrophosphate is barely hydrolyzed by snake venoms. Phosphodiesterase activity was better correlated with 5'-nucleotidase, ADPase and ATPase activities than with DNase activity, evidencing that phosphodiesterases are not the main agent of DNA hydrolysis in animal venoms. The omnipresence of nucleotidase and DNase activities in viperid venoms implies a role for them within the repertoire of enzymes involved in immobilization and death of preys. PMID:17904425

  17. Hemorrhagic, fibrinogenolytic and edema-forming activities of the venom of the colubrid snake Philodryas olfersii (green snake).

    Science.gov (United States)

    Assakura, M T; Salomão, M G; Puorto, G; Mandelbaum, F R

    1992-04-01

    The venom of P. olfersii has high hemorrhagic, edema-inducing and fibrin(ogen)olytic activities. It is devoid of thrombin-like, procoagulant, phospholipase A2 and platelet aggregating enzymes. The main activities are metalloproteinases inhibited by metal chelators (EDTA and 1,10-phenanthroline) and sulfhydryl compounds (DTT and cysteine). The hemorrhagic and fibrinogenolytic enzymes were partially purified by gel filtration on HPLC. The hemorrhagic activity of the venom was neutralized by commercial horse antivenoms to Bothrops species, as well as by rabbit antisera specific for hemorrhagic factors isolated from these Bothrops venoms. No immunoprecipitin reactions were obtained, indicating that the few epitopes of the P. olfersii hemorrhagin are involved in these neutralization reactions. The fibrinogenolytic enzyme cleaves A alpha-chain more quickly than the B beta-chain of human fibrinogen. The venom also solubilizes fibrin. This solubilization appears to occur from the hydrolysis of unpolymerized alpha-chain and cross-linked gamma-gamma dimer. The fibrin peptide products are distinct from those produced by plasmin. PMID:1626324

  18. 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

  19. [Toxicity of venoms from snakes of medical importance in México].

    Science.gov (United States)

    de Roodt, Adolfo R; Estévez-Ramírez, Judith; Paniagua-Solís, Jorge F; Litwin, Silvana; Carvajal-Saucedo, Alejandro; Dolab, Jorge A; Robles-Ortiz, Luis E; Alagón, Alejandro

    2005-01-01

    The characterization of the toxic activities of snake venoms is necessary to understand the physiopathology of the envenomation and to test the potency of the antivenoms used to treat this pathology. Because of the lack of data on the toxic activities of venoms from Mexican snakes of medical importance, we studied the venoms from Bothrops asper, Athropoides nummifrr, Agkistrodon billineatus, Crotalus durissus durissus, Crotalus basiliscus, Crotalus scutulatus, Crotalus atrox and Micrurus nigrocinctus. The studies performed were: SDS-PAOE, determination of lethal potency, hemorrhagic, necrotizing, coagulation on plasma and fibrinogen, phospholipasic and fibri(noge)nolytic activities. In addition we studied the neutralizing capacity of the toxic activities of an antivenom currently used for the treatment of snakebites in Mexico. The venom from viperids showed important hemorrhagic, necrotizing, coagulative on plasma, prothrombinic, fibrinogenolytic and phospholipase activities. The venoms with the highest lethal potency were those of Micrurus nigrocinctus and Crotalus scutulatus; however, the viperine venom that globally displayed the most potent toxic activities was from Bothrops asper. All the venoms showed toxic activities of similar range to those described for other American venomous snakes. The activity on plasma or fibrinogen varied widely among the different venoms but all displayed capacity to act on the coagulation system. The antivenom tested not only neutralized the lethality B. asper venom but also its other toxic activities. PMID:15754746

  20. 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. PMID:26676823

  1. 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. PMID:20677373

  2. 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.

  3. Atividade antiinflamatória do extrato aquoso de Arrabidaea chica (Humb. & Bonpl. B. Verl. sobre o edema induzido por venenos de serpentes amazônicas Anti-inflammatory activity of the aqueous extract of Arrabidaea chica (Humb. & Bonpl. B. Verl. on the self-induced inflammatory process from venoms amazonians snakes

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    Denys Paixão Costa de Oliveira

    2009-06-01

    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.The study investigated the anti-inflammatory effects of the aqueous extract (EAq, the Arrabidaea chica (Humb. & Bonpl. B. Verl., Bignoniaceae, known as the "crajiru", tested on the paw oedema induced in inflammatory process by venoms Amazon snakes from the Bothrops and Crotalus species, on albino mice. This process was done by an oral, intraperitoneal and subcutaneous way. The cushions thickness was measured at time breaks of 1, 3, 6, 12 and 24 hours, Bothrops genus and 1, 3 and 6 hours, Crotalus genus, and a histopathologic analysis was made. The Bothrops genus, was used as an inhibitory effect for the subcutaneous and intraperitoneal way (12 hours, and it

  4. 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

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    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

  5. Photobiomodulation Protects and Promotes Differentiation of C2C12 Myoblast Cells Exposed to Snake Venom

    Science.gov (United States)

    da Silva, Aline; Vieira, Rodolfo Paula; Mesquita-Ferrari, Raquel Agnelli; Cogo, José Carlos; Zamuner, Stella Regina

    2016-01-01

    Background 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. Methodology 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. Results 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. Conclusion 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

  6. Trimeresurus venom inhibition of anti-HPA-1a and anti-HPA-1b antibody binding to human platelets.

    Science.gov (United States)

    Wlodar, S J; Stone, D L; Sinor, L T

    1995-01-01

    A solid-phase red cell adherence assay was used to demonstrate the specific inhibitory effect of seven species of Trimeresurus snake venom on the binding of HPA-1a- and HPA-1b-specific platelet antibodies. Trimeresurus venom did not inhibit the binding of HLA-, HPA-3a-, HPA-3b-, HPA-4a-, HPA-5a-, and HPA-5b-specific platelet antibodies. Venom from other genera of snakes, including representatives from Agkistrodon, Ancistrodon, Bitis, Bothrops, Bungarus, Causus, Crotalus, Dendroaspis, Ecis, Micrurus, Naja, Notechis, Ophiophagus, Pseudechis, Sepedon (Hemachatus), and Vipera, all failed to specifically inhibit anti-HPA-1a and HPA-1b binding. These results may indicate that the component in Trimeresurus snake venom previously reported to bind to the platelet GPIIb-IIIa complex, inhibiting fibrinogen binding, binds close to the HPA-1a and HPA-1b epitopes.

  7. Faculdade de Saúde e Desenvolvimento Humano Santo Agostinho

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    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.

  8. Ochetosoma heterocoelium (Digenea: Plagiorchiidae in snakes from Colombia Ochetosoma heterocoelium (Digenea: Plagiorchiidae en ofidios de Colombia

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    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

  9. Influence of gamma-radiation on the biological activity of snake venoms in Peru

    International Nuclear Information System (INIS)

    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

  10. Distribution of the herpetofauna community associated to four areas with different interference degree in Gorgona Island, Colombian pacific

    International Nuclear Information System (INIS)

    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

  11. 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)

  12. Influenza em animais heterotérmicos Influenza in heterothermics

    OpenAIRE

    Dalva Assunção Portari Mancini; Rita Maria Zucatelli Mendonça; Aurora Marques Cianciarullo; Leonardo Setsuo Kobashi; Hermínio Gomes Trindade; Wilson Fernandes; José Ricardo Pinto

    2004-01-01

    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 d...

  13. Some new species of Caryospora (Apicomplexa: Eimeriidae from brazilian snakes, and a re-description of C. jararacae Carini, 1939

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    Ralph Lainson

    1991-09-01

    Full Text Available The mature ooxysts of six new species of Caryospora are described from the faeces of Brazilian snakes. They are differentiated from other species previously recorded from reptiles, largely on the size and shape of the oocyst and sporocyst, structure of the oocyst wall, and presence or absence of a polar body. C. paraensis n. sp., and C. carajasensis n. sp., are from the "false coral", Oxyrhopus petola digitalis; C. pseustesi n. sp., from the "egg-eater", Pseustes sulphureus sulphureus; C. epicratesi n. sp., from the "red boa", Epicrates cenchria cenchria; and C. micruri n. sp., and C. constancieae n. sp., from the "coral snake", Micrurus spixii spixii. A re-description is given of C. jararacae Carini, 1939, from the "jararaca" Bothrops atrox, embodying some additional morphological features.

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

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    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.

  15. 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

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    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

  16. Software system for three-dimensional volumetric reconstruction of histological sections: a case study for the snake chondrocranium.

    Science.gov (United States)

    Hofstadler-Deiques, Clarice; Walter, Marcelo; Mierlo, Fábio; Ruduit, Rodrigo

    2005-10-01

    Volumetric digital computer-assisted reconstruction of histological sections is an attractive possibility for developmental studies. Commercial solutions are very expensive for many educational institutions. Therefore, we developed a software system for three-dimensional reconstruction of anatomical virtual models. The input data for the system are the digitized images from the histological samples of the chondrocranium of two crotalines, Bothrops jararaca and Crotalus durissus terrificus, and one colubrid, Philodryas olfersii, using a stereomicroscope connected to a digital camera. These images are then manually registered and segmented. We use computer graphics visualization algorithms such as marching cubes and ray casting to generate three-dimensional visualizations of the volumes. The results show that the digital reconstruction is as good as the manual reconstruction with the advantages of speed of reconstruction, accuracy, and flexibility to handle and study the volume. Compared with commercial options, our system has approximately the same features, and it is available free for the scientific community. PMID:16114067

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

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    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.

  18. 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. PMID:7855493

  19. Noteworthy records of reptiles from natural open vegetation areas in plateau and coastal areas of the state of Santa Catarina, southern Brazil

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    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.

  20. Constituintes químicos e atividade antiedematogênica de Peltodon radicans (Lamiaceae Chemical constituents and antiedematogenic activity of Peltodon radicans (Lamiaceae

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    Habdel Nasser Rocha da Costa

    2008-01-01

    Full Text Available 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, 3beta-OH,beta-amirin (1, 3beta-OH,alpha-amirin (2, beta-sitosterol (3, stigmasterol (4, ursolic acid (5, 2alpha,3beta,19alpha-trihydroxy-urs-12-en-28-oic acid (tormentic acid, 6, methyl 3beta-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.

  1. Plant natural products active against snake bite--the molecular approach.

    Science.gov (United States)

    Mors, W B; Nascimento, M C; Pereira, B M; Pereira, N A

    2000-11-01

    The article surveys the substances identified in plants reputed to neutralize the effects of snake venoms. Protective activity of many of them against the lethal action of the venom of the jararaca (Bothrops jararaca) snake was confirmed by biological assays. It was shown that all belong to chemical classes capable of interacting with macromolecular targets--receptors and enzymes. In a few cases it has been shown that exogenous natural micromolecules can mimic the biological activity of endogenous macromolecules. From the evidence presented, it can be inferred that micromolecules which neutralize the action of snake venoms mechanistically replace endogenous antitoxic serum proteins with venom neutralizing capacity such as produced by some animals. PMID:11130675

  2. An alternative method to isolate protease and phospholipase A2 toxins from snake venoms based on partitioning of aqueous two-phase systems

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    GN Gómez

    2012-01-01

    Full Text Available Snake venoms are rich sources of active proteins that have been employed in the diagnosis and treatment of health disorders and antivenom therapy. Developing countries demand fast economical downstream processes for the purification of this biomolecule type without requiring sophisticated equipment. We developed an alternative, simple and easy to scale-up method, able to purify simultaneously protease and phospholipase A2 toxins from Bothrops alternatus venom. It comprises a multiple-step partition procedure with polyethylene-glycol/phosphate aqueous two-phase systems followed by a gel filtration chromatographic step. Two single bands in SDS-polyacrylamide gel electrophoresis and increased proteolytic and phospholipase A2 specific activities evidence the homogeneity of the isolated proteins.

  3. 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.

  4. Application of ultrasound in the dissolution of potential antiophidian compounds from two ethanolics extracts of two species of Heliconias.

    Science.gov (United States)

    Estrada, G Sebastián; Jiménez, Silvia L; Alarcon, P Juan C; Vargas, Leidy J

    2010-06-01

    This study evaluates the favorable effects of using ultrasound during the dissolution process of ethanolic extracts of Heliconia psittacorum and Heliconia rostrata (Heliconiaceae), a family of plants reported to have antiophidic activities. The extracts were subjected to an ultrasound treatment before incubation with venom, and carbohydrate and protein contents were calculated. The ultrasound-treated extracts delayed the clotting effect of venom by up to 45.59 s compared to the positive control (venom). The metabolites content increased to 296.6% and 61.6% in protein and carbohydrate content, respectively. Ultrasound increases the amount of primary and secondary metabolites (potentially antiophidics) released from the extract into the media, and thereby enhances the anticoagulant activity of these plants against the Bothrops asper (mapaná X) venom.

  5. Application of ultrasound in the dissolution of potential antiophidian compounds from two ethanolics extracts of two species of Heliconias.

    Science.gov (United States)

    Estrada, G Sebastián; Jiménez, Silvia L; Alarcon, P Juan C; Vargas, Leidy J

    2010-06-01

    This study evaluates the favorable effects of using ultrasound during the dissolution process of ethanolic extracts of Heliconia psittacorum and Heliconia rostrata (Heliconiaceae), a family of plants reported to have antiophidic activities. The extracts were subjected to an ultrasound treatment before incubation with venom, and carbohydrate and protein contents were calculated. The ultrasound-treated extracts delayed the clotting effect of venom by up to 45.59 s compared to the positive control (venom). The metabolites content increased to 296.6% and 61.6% in protein and carbohydrate content, respectively. Ultrasound increases the amount of primary and secondary metabolites (potentially antiophidics) released from the extract into the media, and thereby enhances the anticoagulant activity of these plants against the Bothrops asper (mapaná X) venom. PMID:20189865

  6. 介入超声:蝮(蝰)蛇毒酶(立芷雪)过敏1例%Interventional ultrasound:reptilase from viper venom induced anaphylaxis (a case report)

    Institute of Scientific and Technical Information of China (English)

    侯庆峰; 王晴文

    2009-01-01

    立芷雪,商名Reptilase,(源于reptile爬行类+ase酶),临床应用作为止血或减少出血的辅助药物,一般甚为安全。其2000年的通行定义,为大具窍矛头蝮蛇毒液中的酶,它分解纤维蛋白多肽使纤维蛋白原凝固venom of Bothrops atrox that clots fibrinogen by splitting off its fibronopeptide)。本文介绍超声介入1例患者注射该血凝酶(haemocoagulabe)后发生过敏反应,因较少见,现报告如下。

  7. Cross reactivity of three antivenoms against North American snake venoms.

    Science.gov (United States)

    Sánchez, Elda E; Ramírez, María Susana; Galán, Jacob A; López, Gonzalo; Rodríguez-Acosta, Alexis; Pérez, John C

    2003-03-01

    The antivenom in the United States today is in short supply, expensive and may not even be the most effective in neutralizing venoms from snakes in certain geographical locations. The ED(50) is considered to be the best indicator of antivenom efficacy, however, other tests are needed. In this study, three antivenoms (Antivipmyn (Fab(2)H), Crotalidae Polyvalent Immune Fab (Ovine) (FabO) and UCV (FabV) were used to test the effectiveness of neutralization of eight venoms (Agkistrodon piscivorus piscivorus, Bothrops asper, Crotalus adamanteus, C. durissus durissus, C. horridus atricaudatus, C. h. horridus, C. atrox, and C. molossus molossus). Four different assays were used to study the efficacy of the antivenoms: the antihemorrhagic, antigelatinase, antifibrinolytic and antihide powder azure. Fab(2)H antivenom was more effective in neutralizing the enzymatic activities of these eight venoms than FabO and FabV antivenoms. PMID:12565754

  8. 蛇毒类凝血酶基因工程的研究进展%Research Progress on Cloning of Snake Venom Thrombin-like Enzymes

    Institute of Scientific and Technical Information of China (English)

    李家祺; 李谦; 唐松山; 李红枝

    2013-01-01

    蛇毒类凝血酶(Thrombin-like enzyme,TLE)是蛇毒中与血浆凝血酶性质相似的一类丝氨酸蛋白酶,所不同的是在蛇毒类凝血酶结构中已经没有纤维蛋白稳定因子激活组分.由于蛇毒类凝血酶在新药研究中扮演重要角色,比如来自Bothrops jararaca和Bothrops atrox蛇毒的立芷雪、来自东北白眉蝮蛇毒的邦停和来自尖吻蝮蛇毒的苏灵,它们是具有止血作用的新药;来自Gloydius shedaoensis和Gloydius ussuriensis蛇毒的东菱克栓酶、来自白眉蝮蛇和尖吻蝮蛇毒的降纤酶、来自Calloselasma rhodostoma蛇毒的安克洛酶和来自Crotalus adamanteus蛇毒的Crotalase,它们是具有溶栓作用的新药.这些药用蛋白质的来源和产量因有限的蛇毒原料而有很大限制,通过基因克隆可解决资源问题.文章综述了蛇毒类凝血酶的基因结构、糖基化特点和各种重组表达体系,为大量制备供临床和基础研究使用奠定基础.%Snake venom thrombin-like enzymes (snTLE) are one kind of serine proteases similar to thrombin from blood, but the basic difference between them is that snTLE has no XIII-activated domain to lead thrombosis in vivo or in vitro. Because snake venoms play very important role in pharmaceutics,for example Reptilase from Bothrops jararaca or Bothrops atrox snake venom, Bangtin from Agkistrodon halys snake venom,or Siding from Agkistrodon acutus snake venom,are hemostatic drugs. Defibrase from Gloydius shedaoensis ,Cloydius ussuriensis,Agkistrodon halys,or Agkistrodon acutus snake venom,Ancrod from Calloselasma rhodostoma snake venom, or Crotalase from Crotalus adamanteus snake venom, is thrombolytic drug. So the shortage of snake venom source can be overcome by using gene recombination method. The gene structures, protein glycosylation, and all kinds of cloning expression systems for snTLE protein are summarized in the paper,which will provide a basis for large-scale production of some snTLE proteins.

  9. 注射用血凝酶在耳鼻咽喉头颈部术中的应用研究

    Institute of Scientific and Technical Information of China (English)

    贾笑芳

    2006-01-01

    注射用血凝酶(Reptilase,商品名:立芷雪/立止血)是由巴西矛头蝮蛇(Bothrops atrox)蛇毒经分离、提纯而得的酶性止血药.其含巴曲酶(Batroxobin)和磷脂依赖性凝血因子X激活物(FXA)2种成分,具有安全、高效、速效、使用方便等特点.本文拟就近年来注射用血凝酶在耳鼻咽喉头颈部术中的临床应用综述如下.

  10. Functional characterization of recombinant batroxobin, a snake venom thrombin-like enzyme, expressed from Pichia pastoris.

    Science.gov (United States)

    You, Weon-Kyoo; Choi, Won-Seok; Koh, You-Seok; Shin, Hang-Cheol; Jang, Yangsoo; Chung, Kwang-Hoe

    2004-07-30

    A thrombin-like enzyme of Bothrops atrox moojeni venom, batroxobin, specifically cleaves fibrinogen alpha chain, resulting in the formation of non-crosslinked fibrin clots. The cDNA encoding batroxobin was cloned, expressed in Pichia pastoris and the molecular function of purified recombinant protein was also characterized. The recombinant batroxobin had an apparent molecular weight of 33 kDa by SDS-PAGE analysis and biochemical activities similar to those of native batroxobin. The purified recombinant protein strongly converted fibrinogen into fibrin clot in vitro, and shortened bleeding time and whole blood coagulation time in vivo. However, it did not make any considerable alterations on other blood coagulation factors. Several lines of experimental evidence in this study suggest that the recombinant batroxobin is a potent pro-coagulant agent. PMID:15280019

  11. Snake venom galactoside-binding lectins: a structural and functional overview.

    Science.gov (United States)

    Sartim, Marco A; Sampaio, Suely V

    2015-01-01

    Snake venom galactoside-binding lectins (SVgalLs) comprise a class of toxins capable of recognizing and interacting with terminal galactoside residues of glycans. In the past 35 years, since the first report on the purification of thrombolectin from Bothrops atrox snake venom, several SVgalLs from Viperidae and Elapidae snake families have been described, as has progressive improvement in the investigation of structural/functional aspects of these lectins. Moreover, the advances of techniques applied in protein-carbohydrate recognition have provided important approaches in order to screen for possible biological targets. The present review describes the efforts over the past 35 years to elucidate SVgalLs, highlighting their structure and carbohydrate recognition function involved in envenomation pathophysiology and potential biomedical applications. PMID:26413085

  12. Low-Level Laser Therapy (904 nm) Counteracts Motor Deficit of Mice Hind Limb following Skeletal Muscle Injury Caused by Snakebite-Mimicking Intramuscular Venom Injection.

    Science.gov (United States)

    Vieira, Willians Fernando; Kenzo-Kagawa, Bruno; Cogo, José Carlos; Baranauskas, Vitor; Cruz-Höfling, Maria Alice da

    2016-01-01

    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 (plaser 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.

  13. 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.

  14. Antihemorrhagic, antinucleolytic and other antiophidian properties of the aqueous extract from Pentaclethra macroloba.

    Science.gov (United States)

    da Silva, Jocivânia O; Coppede, Juliana S; Fernandes, Vanessa C; Sant'ana, Carolina D; Ticli, Fábio K; Mazzi, Maurício V; Giglio, José R; Pereira, Paulo S; Soares, Andreimar M; Sampaio, Suely V

    2005-08-22

    Several Brazilian plants have been utilized in folk medicine as active agents against various effects induced by snake venoms. The inhabitants of the Amazon region use, among others, the macerated bark of a plant popularly named "Pracaxi" (Pentaclethra macroloba Willd) to combat these effects. We report now the antihemorrhagic properties against snake venoms of the aqueous extract of Pentaclethra macroloba (EPema). EPema exhibited full inhibition of hemorrhagic and nucleolytic activities induced by several snake venoms. Additionally, partial inhibition of myotoxic, lethal, phospholipase and edema activities of snake venoms and its isolated PLA(2)s by EPema is reported. In vivo tests showed that EPema is able to totally inhibit a Bothrops jararacussu metalloprotease (BjussuMP-I) induced hemorrhage, suggesting interaction of the extract compounds with this high molecular weight protein. The extract did induce neither hemorrhage nor death in mice when administered alone by i.m. route. When administered separately by i.m. route, the extract did not induce death in mice at 12.5--300 mg/kg doses. Other assays demonstrated that EPema was unable to inhibit fibrinogenolytic and coagulant activities of Bothrops atrox venom. Although the mechanism of action of EPema is still unknown, the finding that no visible change was detected in the electrophoretic pattern of snake venom after incubation with the extract excludes proteolytic degradation as a potential mechanism. The search for new inhibitors of venom metalloproteases and DNAases are a relevant task. Investigation of snake venom inhibitors can provide useful tools for the elucidation of the action mechanisms of purified toxins. Furthermore, these inhibitors can be used as molecular models for development of new therapeutical agents in the treatment of ophidian accidents. PMID:16054531

  15. Vipericidins: a novel family of cathelicidin-related peptides from the venom gland of South American pit vipers.

    Science.gov (United States)

    Falcao, C B; de La Torre, B G; Pérez-Peinado, C; Barron, A E; Andreu, D; Rádis-Baptista, G

    2014-11-01

    Cathelicidins are phylogenetically ancient, pleiotropic host defense peptides-also called antimicrobial peptides (AMPs)-expressed in numerous life forms for innate immunity. Since even the jawless hagfish expresses cathelicidins, these genetically encoded host defense peptides are at least 400 million years old. More recently, cathelicidins with varying antipathogenic activities and cytotoxicities were discovered in the venoms of poisonous snakes; for these creatures, cathelicidins may also serve as weapons against prey and predators, as well as for innate immunity. We report herein the expression of orthologous cathelicidin genes in the venoms of four different South American pit vipers (Bothrops atrox, Bothrops lutzi, Crotalus durissus terrificus, and Lachesis muta rhombeata)-distant relatives of Asian cobras and kraits, previously shown to express cathelicidins-and an elapid, Pseudonaja textilis. We identified six novel, genetically encoded peptides: four from pit vipers, collectively named vipericidins, and two from the elapid. These new venom-derived cathelicidins exhibited potent killing activity against a number of bacterial strains (S. pyogenes, A. baumannii, E. faecalis, S. aureus, E. coli, K. pneumoniae, and P. aeruginosa), mostly with relatively less potent hemolysis, indicating their possible usefulness as lead structures for the development of new anti-infective agents. It is worth noting that these South American snake venom peptides are comparable in cytotoxicity (e.g., hemolysis) to human cathelicidin LL-37, and much lower than other membrane-active peptides such as mastoparan 7 and melittin from bee venom. Overall, the excellent bactericidal profile of vipericidins suggests they are a promising template for the development of broad-spectrum peptide antibiotics. PMID:25100358

  16. Batroxase, a new metalloproteinase from B. atrox snake venom with strong fibrinolytic activity.

    Science.gov (United States)

    Cintra, A C O; De Toni, L G B; Sartim, M A; Franco, J J; Caetano, R C; Murakami, M T; Sampaio, S V

    2012-07-01

    The structures and functional activities of metalloproteinases from snake venoms have been widely studied because of the importance of these molecules in envenomation. Batroxase, which is a metalloproteinase isolated from Bothrops atrox (Pará) snake venom, was obtained by gel filtration and anion exchange chromatography. The enzyme is a single protein chain composed of 202 amino acid residues with a molecular mass of 22.9 kDa, as determined by mass spectrometry analysis, showing an isoelectric point of 7.5. The primary sequence analysis indicates that the proteinase contains a zinc ligand motif (HELGHNLGISH) and a sequence C₁₆₄ I₁₆₅M₁₆₆ motif that is associated with a "Met-turn" structure. The protein lacks N-glycosylation sites and contains seven half cystine residues, six of which are conserved as pairs to form disulfide bridges. The three-dimensional structure of Batroxase was modeled based on the crystal structure of BmooMPα-I from Bothrops moojeni. The model revealed that the zinc binding site has a high structural similarity to the binding site of other metalloproteinases. Batroxase presented weak hemorrhagic activity, with a MHD of 10 μg, and was able to hydrolyze extracellular matrix components, such as type IV collagen and fibronectin. The toxin cleaves both α and β-chains of the fibrinogen molecule, and it can be inhibited by EDTA, EGTA and β-mercaptoethanol. Batroxase was able to dissolve fibrin clots independently of plasminogen activation. These results demonstrate that Batroxase is a zinc-dependent hemorrhagic metalloproteinase with fibrin(ogen)olytic and thrombolytic activity. PMID:22483847

  17. Pentastomídeos de répteis do Brasil: revisão dos cephalobaenidae

    Directory of Open Access Journals (Sweden)

    A. Arandas Rêgo

    1983-12-01

    Full Text Available Foram estudados os Cephalobaenidae (Pentastomida, depositados na coleção helmintológica do Instituto Oswaldo Cruz e na coleção de parasitologia do Instituto Butantan. São redescritas e discutidas as espécies, Cephalobaena tetrapoda, C. freitasi, C. giglioli, Raillietiella furcocerca e Mahafaliella venteli. Esses parasitas foram coletados dos répteis: Lachesis sp., Drymarchon c. corais, Xenodon merremii, Crotatus terrificus, Amphisbaena sp., Tropidurus torquatus, Bothrops atrox, Mabuya punctata e de Bufo paracnemis (anfíbio.In this work the author studies Cephalobaenidae parasites using specimens from the helminthological collection of the Oswaldo Cruz and Butantan Institutes. This material was collected from Lachesis sp., Drymarchon corais, Xenodon merremii, Crotalus terrificus, Amphisbaena sp., Tropidurus torquatus, Bothrops atrox, Mabuya punctata (Reptilia and Bufo paracnemis (Amphibia. The species studied are Cephalobaena tetrapoda Heymons, 1922, Cephalobaena giglioli (Hett, 1924 comb. n., Cephalobaena freitasi (Motta & Gomes, 1968 comb. n., Raillietiella furcocerca (Diesing, 1836 and Mahafaliella venteli (Motta, 1965. C. recurvocauda becomes a synonym of C. tetrapoda and as do the specimens that Motta called erronously R. furcocerca. Raillietiella giglioli is changed to Cephalobaena giglioli (Hett, 1924 comb. n. The author describes here the male of C. giglioli for the first time. Travassostulida freitasi and T. acutiacanthus enter in synonymy with C. freitasi, and T. acutiacanthus is considered to be a subspecies of C. freitasi. Raillietiella gomesi becomes a synonym of R. furcocerca. The author discusses Mahafaliella venteli, and also questions the validity of the genus Gretillaria proposed by Motta for some species of Raillietiella. The latter is considered a synonym of Raillietiella.

  18. A multifaceted analysis of viperid snake venoms by two-dimensional gel electrophoresis: an approach to understanding venom proteomics.

    Science.gov (United States)

    Serrano, Solange M T; Shannon, John D; Wang, Deyu; Camargo, Antonio C M; Fox, Jay W

    2005-02-01

    The complexity of Viperid venoms has long been appreciated by investigators in the fields of toxinology and medicine. However, it is only recently that the depth of that complexity has become somewhat quantitatively and qualitatively appreciated. With the resurgence of two-dimensional gel electrophoresis (2-DE) and the advances in mass spectrometry virtually all venom components can be visualized and identified given sufficient effort and resources. Here we present the use of 2-DE for examining venom complexity as well as demonstrating interesting approaches to selectively delineate subpopulations of venom proteins based on particular characteristics of the proteins such as antibody cross-reactivity or enzymatic activities. 2-DE comparisons between venoms from different species of the same genus (Bothrops) of snake clearly demonstrated both the similarity as well as the apparent diversity among these venoms. Using liquid chromatography/tandem mass spectrometry we were able to identify regions of the two-dimensional gels from each venom in which certain classes of proteins were found. 2-DE was also used to compare venoms from Crotalus atrox and Bothrops jararaca. For these venoms a variety of staining/detection protocols was utilized to compare and contrast the venoms. Specifically, we used various stains to visualize subpopulations of the venom proteomes of these snakes, including Coomassie, Silver, Sypro Ruby and Pro-Q-Emerald. Using specific antibodies in Western blot analyses of 2-DE of the venoms we have examined subpopulations of proteins in these venoms including the serine proteinase proteome, the metalloproteinase proteome, and the phospholipases A2 proteome. A functional assessment of the gelatinolytic activity of these venoms was also performed by zymography. These approaches have given rise to a more thorough understanding of venom complexity and the toxins comprising these venoms and provide insights to investigators who wish to focus on these venom

  19. Vipericidins: a novel family of cathelicidin-related peptides from the venom gland of South American pit vipers.

    Science.gov (United States)

    Falcao, C B; de La Torre, B G; Pérez-Peinado, C; Barron, A E; Andreu, D; Rádis-Baptista, G

    2014-11-01

    Cathelicidins are phylogenetically ancient, pleiotropic host defense peptides-also called antimicrobial peptides (AMPs)-expressed in numerous life forms for innate immunity. Since even the jawless hagfish expresses cathelicidins, these genetically encoded host defense peptides are at least 400 million years old. More recently, cathelicidins with varying antipathogenic activities and cytotoxicities were discovered in the venoms of poisonous snakes; for these creatures, cathelicidins may also serve as weapons against prey and predators, as well as for innate immunity. We report herein the expression of orthologous cathelicidin genes in the venoms of four different South American pit vipers (Bothrops atrox, Bothrops lutzi, Crotalus durissus terrificus, and Lachesis muta rhombeata)-distant relatives of Asian cobras and kraits, previously shown to express cathelicidins-and an elapid, Pseudonaja textilis. We identified six novel, genetically encoded peptides: four from pit vipers, collectively named vipericidins, and two from the elapid. These new venom-derived cathelicidins exhibited potent killing activity against a number of bacterial strains (S. pyogenes, A. baumannii, E. faecalis, S. aureus, E. coli, K. pneumoniae, and P. aeruginosa), mostly with relatively less potent hemolysis, indicating their possible usefulness as lead structures for the development of new anti-infective agents. It is worth noting that these South American snake venom peptides are comparable in cytotoxicity (e.g., hemolysis) to human cathelicidin LL-37, and much lower than other membrane-active peptides such as mastoparan 7 and melittin from bee venom. Overall, the excellent bactericidal profile of vipericidins suggests they are a promising template for the development of broad-spectrum peptide antibiotics.

  20. 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

  1. 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.

  2. Influenza em animais heterotérmicos Influenza in heterothermics

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    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

  3. [Epidemiology of accidents due to bites of poisonous snakes: a study of cases attended in 1988].

    Science.gov (United States)

    Ribeiro, L A; Jorge, M T; Iversson, L B

    1995-10-01

    The attendance given to patients by ta specialized Hospital, in S. Paulo, Brazil, during 1988 is studied. The study is based on the medical records of 322 patients and on questionnaires filled out by author during interviews with 209 patients or their companions. The 322 snake-bites occurred mainly between October and April, in the diurnal period, mainly in the afternoon. Most of patients were adult males, mainly between 10 and 20 years of age. The parts of the body most frequently affected were the feet, hands and legs. The snakes of the genera Bothrops, Crotalus and Micrurus were responsible, respectively, for 306 (95.0%) 14 (4.4%) and 2 (0.6%) of the accidents under study. Among the 160 snakes that were classified at the Herpetological Section of the IB, 152 were Bothrops; 142 B. Jararaca, mostly young reptiles, and 8 were of the genus Crotalus. Of the patients, 90.4% recovered completely, 2.2% presented sequelae, 7.5% were transferred and thus it was impossible to follow them up. Of the 209 persons interviewed, the occupational group most prone to snake bites was agricultural workers, followed by studentes; nearly 60% of the accidents ocurred during work; most of the patients had their inferior extremities unprotected at the moment of the bite. On hundred and sixty patients (76.6%) submitted to some from of treatment before coming to the HVB-IB, the more common being the use of a tourniquet (50.2%), local squeezing in an attempt to remove part of the venom (33.5), application of substances on the site of the snake bite (36.8%) and the ingestion of others (12.9%). Slightly over a quarter of the patients underwent some kind of medical treatment before coming to the HVB-IB, the most common being antissepsis (8.2%), administration of antivenom (6.2%), antihistamines (5.7%) and analgesics (5.3%). The snake was seen before it struch by 187 (89.5%) of the 209 persons interviewed and in most cases it adopted the strike posture just before the first bite. PMID:8731278

  4. 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

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    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

  5. Las serpientes venenosas de importancia en la salud publica del perú – The poisonous snakes of public health importance of peru

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    Navarrete Zamora Miluska B

    2010-07-01

    Full Text Available ResumenEn el Perú existen aproximadamente 33 especies de serpientesvenenosas distribuidas en 2 familias: Viperidae y Elapidae. Ladistribución geográfica es en los diferentes hábitats de la Costa, Sierra y Selva. El manejo de estos animales requiere de equipos especiales y de personal calificado para evitar los accidentes. El veneno producido por las serpientes es una modificación de las glándulas salivales que va a facilitar la digestión de sus presas. El ofidismo es la inoculación de éste veneno, y en el Perú se atribuye a Bothrops atrox como el animal agresor con mayor reporte de accidentes en humanos. El mecanismo de acción de estos venenos puede presentarse como una acción proteolítica, neurotóxica, coagulante, vasculotóxica, hemolítica, miotóxica, necrotóxica y hepatotóxica, que van a caracterizar a los diferentes accidentes ofídicos. El objetivo de ésta revisión fue proporcionar conocimientos acerca de las diferentes serpientes venenosas que hay en el Perú, su ubicación en el territorio nacional, los accidentes ofídicos por la inoculación del veneno y su tratamiento, asimismo una revisión epidemiológica de los casos presentados y de los últimos avances del uso del veneno en medicina.SummaryThere are 33 species of venomous snakes in Perú and these aredistributed in two families: Viperidae and Elapidae. They aregeographically distributed in different habitats in Costa, Sierra and Selva of Perú. Handling these animals requires special equipment andmust be done by qualified staff in order to avoid accidents. Venomproduced by snakes is a modification of salivary glands that facilitates digestion of its preys. Inoculation of the snake venom is called “ofidismo” (snakebite, in Perú Bothrops atrox is the aggressor animal with more ophidian accident reports. Mechanisms of these venoms can be present as proteolytic, neurotoxic, coagulate, vascular toxicity, hemolytic, necrotic toxicity and liver toxicity actions

  6. Involvement of Nitric Oxide on Bothropoides insularis Venom Biological Effects on Murine Macrophages In Vitro.

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    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

  7. Role of collagens and perlecan in microvascular stability: exploring the mechanism of capillary vessel damage by snake venom metalloproteinases.

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    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

  8. Low-Level Laser Therapy (904 nm) Counteracts Motor Deficit of Mice Hind Limb following Skeletal Muscle Injury Caused by Snakebite-Mimicking Intramuscular Venom Injection.

    Science.gov (United States)

    Vieira, Willians Fernando; Kenzo-Kagawa, Bruno; Cogo, José Carlos; Baranauskas, Vitor; Cruz-Höfling, Maria Alice da

    2016-01-01

    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. PMID:27392016

  9. Low-Level Laser Therapy (904 nm) Counteracts Motor Deficit of Mice Hind Limb following Skeletal Muscle Injury Caused by Snakebite-Mimicking Intramuscular Venom Injection

    Science.gov (United States)

    Vieira, Willians Fernando; Kenzo-Kagawa, Bruno; Cogo, José Carlos; da Cruz-Höfling, Maria Alice

    2016-01-01

    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. PMID:27392016

  10. Mutagenicity induced by the hydroalcoholic extract of the medicinal plant Plathymenia reticulata Benth

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    A Della Torre

    2011-01-01

    Full Text Available Plathymenia reticulata Benth has an anti-inflammatory effect and is capable of neutralizing the neuromuscular blockade induced by Bothrops jararacussu or Crotalus durissus terrificus venoms, probably by precipitating venom proteins (an effect caused by plant tannins. The present study aimed to evaluate the mutagenic activity of P. reticulata by using the Salmonella mutagenicity assay (Ames test and the micronucleus test in CHO-K1 cells. P. reticulata extract concentrations of 2.84, 5.68, 11.37, and 19.90 mg/plate were assayed by the Ames test using TA97a, TA98, TA100 and TA102 bacterial strains, with (+S9 and without (-S9 metabolic activation. Concentrations of 5, 1.6 and 0.5 μg/mL of P. reticulata extract were used for the micronucleus test. P. reticulata extract was mutagenic to TA98 (-S9 and showed signs of mutagenic activity in TA97a and TA102 (both -S9 strains. Micronucleus test CBPI values showed that the endogenous metabolic system increased the number of viable cells when compared to the non-activated samples and the micronucleus frequency increased when the cells were treated in the absence of S9. We concluded that P. reticulata extract may present direct mutagenic properties.

  11. 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.

  12. The use of ecological niche modeling to infer potential risk areas of snakebite in the Mexican state of Veracruz.

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    Carlos Yañez-Arenas

    Full Text Available Many authors have claimed that snakebite risk is associated with human population density, human activities, and snake behavior. Here we analyzed whether environmental suitability of vipers can be used as an indicator of snakebite risk. We tested several hypotheses to explain snakebite incidence, through the construction of models incorporating both environmental suitability and socioeconomic variables in Veracruz, Mexico.Ecological niche modeling (ENM was used to estimate potential geographic and ecological distributions of nine viper species' in Veracruz. We calculated the distance to the species' niche centroid (DNC; this distance may be associated with a prediction of abundance. We found significant inverse relationships between snakebites and DNCs of common vipers (Crotalus simus and Bothrops asper, explaining respectively 15% and almost 35% of variation in snakebite incidence. Additionally, DNCs for these two vipers, in combination with marginalization of human populations, accounted for 76% of variation in incidence.Our results suggest that niche modeling and niche-centroid distance approaches can be used to mapping distributions of environmental suitability for venomous snakes; combining this ecological information with socioeconomic factors may help with inferring potential risk areas for snakebites, since hospital data are often biased (especially when incidences are low.

  13. Comparative Analysis of Viperidae Venoms Antibacterial Profile: a Short Communication for Proteomics

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    Bruno L. Ferreira

    2011-01-01

    Full Text Available Bacterial infections involving multidrug-resistant strains are one of the ten leading causes of death and an important health problem in need for new antibacterial sources and agents. Herein, we tested and compared four snake venoms (Agkistrodon rhodostoma, Bothrops jararaca, B. atrox and Lachesis muta against 10 Gram-positive and Gram-negative drug-resistant clinical bacteria strains to identify them as new sources of potential antibacterial molecules. Our data revealed that, as efficient as some antibiotics currently on the market (minimal inhibitory concentration (MIC = 1–32 μg mL−1, A. rhodostoma and B. atrox venoms were active against Staphylococcus epidermidis and Enterococcus faecalis (MIC = 4.5 μg mL−1, while B. jararaca inhibited S. aureus growth (MIC = 13 μg ml−1. As genomic and proteomic technologies are improving and developing rapidly, our results suggested that A. rhodostoma, B. atrox and B. jararaca venoms and glands are feasible sources for searching antimicrobial prototypes for future design new antibiotics against drug-resistant clinical bacteria. They also point to an additional perspective to fully identify the pharmacological potential of these venoms by using different techniques.

  14. Epidemiologia dos acidentes ofídicos nos últimos 100 anos no Brasil: uma revisão

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    Bochner Rosany

    2003-01-01

    Full Text Available Neste trabalho foram analisados 22 artigos, 4 livros, 3 relatórios e 1 manual publicados no período de 1901 a 2000, que tratam de acidentes ofídicos ocorridos no Brasil. Concluiu-se que as análises epidemiológicas realizadas nos últimos 100 anos são baseadas nas mesmas variáveis já apontadas por Vital Brazil em seu Boletim para Observação de Accidente Ophidico, ou seja, variáveis referentes ao indivíduo, ao evento e ao atendimento. O perfil epidemiológico desses acidentes também se manteve inalterado ao longo dos anos, isto é, são mais comuns em pessoas do sexo masculino, em trabalhadores rurais, na faixa etária de 15 a 49 anos, atingem principalmente os membros inferiores e a maioria desses acidentes é atribuída às serpentes do gênero Bothrops.

  15. Freeze-dried snake antivenoms formulated with sorbitol, sucrose or mannitol: comparison of their stability in an accelerated test.

    Science.gov (United States)

    Herrera, María; Tattini, Virgilio; Pitombo, Ronaldo N M; Gutiérrez, José María; Borgognoni, Camila; Vega-Baudrit, José; Solera, Federico; Cerdas, Maykel; Segura, Alvaro; Villalta, Mauren; Vargas, Mariángela; León, Guillermo

    2014-11-01

    Freeze-drying is used to improve the long term stability of pharmaceutical proteins. Sugars and polyols have been successfully used in the stabilization of proteins. However, their use in the development of freeze-dried antivenoms has not been documented. In this work, whole IgG snake antivenom, purified from equine plasma, was formulated with different concentrations of sorbitol, sucrose or mannitol. The glass transition temperatures of frozen formulations, determined by Differential Scanning Calorimetry (DSC), ranged between -13.5 °C and -41 °C. In order to evaluate the effectiveness of the different stabilizers, the freeze-dried samples were subjected to an accelerated stability test at 40 ± 2 °C and 75 ± 5% relative humidity. After six months of storage at 40 °C, all the formulations presented the same residual humidity, but significant differences were observed in turbidity, reconstitution time and electrophoretic pattern. Moreover, all formulations, except antivenoms freeze-dried with mannitol, exhibited the same potency for the neutralization of lethal effect of Bothrops asper venom. The 5% (w:v) sucrose formulation exhibited the best stability among the samples tested, while mannitol and sorbitol formulations turned brown. These results suggest that sucrose is a better stabilizer than mannitol and sorbitol in the formulation of freeze-dried antivenoms under the studied conditions.

  16. Oral microbiota of Brazilian captive snakes

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    MG Fonseca

    2009-01-01

    Full Text Available The present work aimed to determine the oral microbiotic composition of snakes from São José do Rio Preto city, São Paulo State, Brazil. Ten snake species, comprising the families Boidae, Colubridae, Elapidae and Viperidae, were submitted to microbiological examination of their oral cavity, which indicated positivity for all buccal samples. Gram-negative bacilli, gram-negative cocci bacilli, gram-positive bacilli and gram-positive cocci were isolated from the snakes. Among isolated bacterium species, the occurrence of coagulase-negative staphylococci in the buccal cavity of Crotalus durissus (Viperiade, Eunectes murinus (Boidae, Mastigodryas bifossatus (Colubridae and Bacillus subtilis, common to oral cavity of Bothrops alternatus (Viperidae and Phalotris mertensi (Colubridae, was detected. It was observed higher diversity of isolated bacteria from the oral cavity of Micrurus frontalis (Elapidae and Philodryas nattereri (Colubridae, as well as the prevalence of gram-positive baccillus and gram-positive cocci. The composition of the oral microbiota of the studied snakes, with or without inoculating fangs, is diverse and also related to the formation of abscesses at the bite site in the victims of the ophidian accidents, and to pathogenic processes in the snakes that host these microorganisms.

  17. 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.

  18. Estudo histológico e histoquímico da glândula de Duvernoy de Clelia plumbea (Wied (Serpentes, Colubridae, Xenodontinae Histological and histochemical study of Duvernoy's gland from Clelia plumbea (Wied (Serpentes, Colubridae, Xenodontinae

    Directory of Open Access Journals (Sweden)

    Márcia Ferret Renner

    2000-09-01

    Full Text Available There are report two human envenenomations for species from Clelia Fitzinger, 1826 (opisthoglyph snake. The patients exhibited symptoms similar from bothropic accident. Then this work have with objective, the hystological and histochemical study of Duvernoy's gland from Clelia plumbea. For the hystological observation of the Duvernoy's gland were using, with paraffin's cutting edge, two methods: Hematoxylin + Eosin and Toluidine's Blue. With historesine's cutting edge were using three methods: Metilen's Blue, Toluidine's Blue + Floxine and Toluidine's Blue. For the histochemical detection were using six techniques, with paraffin's cutting edge: PAS, PAS + Alcian Blue pH 2,5, Alcian Blue pH 2,5, PAS + Salivar Amilase, Method of Lars Grimelius and Method of Lillie. It was evidenced the predominance of serous cells on Duvernoy's gland. In relation to the histochemical constituion of the duvernoy's gland, was verified the presence of glucoconjugates neuters, enzymes and cells with reduetive activity. By fact from this species to kill his preys by constriction, the predominance of serous cells in the duvernoy's gland, don' t must be direct related with the subjugation of his preys, but with the digestive processes. Histological and histochemical analysis showed cells of Duvernoy's gland are constituted for two populations: serous cells (prodution of enzymes and mucous cells (prodution of glucoconjugates.

  19. PO41, a snake venom metalloproteinase inhibitor isolated from Philander opossum serum.

    Science.gov (United States)

    Jurgilas, Patrícia B; Neves-Ferreira, Ana G C; Domont, Gilberto B; Perales, Jonas

    2003-11-01

    PO41 was isolated from Philander opossum serum by DEAE-Sephacel, Phenyl Superose and Superdex 200 chromatographies and showed a molecular mass of 41,330 Da by MALDI-TOF MS. Molecular masses of 81.5 and 84.5 kDa were obtained by size exclusion chromatography and dynamic laser light scattering, respectively, suggesting that PO41 is dimeric. Its isoelectric point was estimated to be lower than 3.5. PO41 presented similar amino terminal sequence to those of DM40 and DM43, two antihaemorrhagins previously isolated from Didelphis marsupialis serum and was recognized by polyclonal antibodies raised against D. marsupialis antibothropic fraction. To study the inhibitory properties of this protein, the metalloproteinases bothrolysin and jararhagin were isolated from Bothrops jararaca venom by chromatographies on Superdex 200 and Phenyl Superose. Jararhagin was further submitted to a Mono Q column. The proteolytic and haemorrhagic effects of these haemorrhagins were neutralized by PO41. Both snake venom metalloproteinases formed stable complexes with PO41. The stoichiometry of the complex PO41-jararhagin was one inhibitor subunit to one molecule of the enzyme. These results show that PO41 has physicochemical, structural, immunoreactive and biological properties similar to other metalloproteinase inhibitors belonging to the supergene family of immunoglobulins. PMID:14602117

  20. Isolation and characterization of DM40 and DM43, two snake venom metalloproteinase inhibitors from Didelphis marsupialis serum.

    Science.gov (United States)

    Neves-Ferreira, A G; Cardinale, N; Rocha, S L; Perales, J; Domont, G B

    2000-05-01

    From Didelphis marsupialis serum, two antihemorrhagic proteins were isolated by DEAE-Sephacel, Phenyl-Sepharose and Superdex 200 and characterized. Their masses by mass spectrometry were 40318 AMU for DM40 and 42373 and 43010 AMU for DM43, indicating the presence of isoforms for the last. Molecular masses of 44.8 and 47.3 were obtained by SDS-PAGE, respectively for DM40 and DM43. Both inhibitors showed isoelectric points lower than 3.5 and glycosylation percentages varying from 20.5 to 29.0%, as estimated by chemical deglycosylation and amino acid analysis. N-terminal sequences of the first 17 residues of DM40 and DM43 were identical except for the exchange of R9 for P9. Both were homologous to oprin, a similar inhibitor from Didelphis virginiana serum. No evidence of complex formation between DM40 and DM43 was observed either by native PAGE or gel filtration chromatography. In addition to the antihemorrhagic activity, DM40 and DM43 inhibited the hydrolysis of casein, fibrinogen and fibronectin by Bothrops jararaca venom. DM43 also showed antilethal, antiedematogenic and antihyperalgesic activities. None of the inhibitors showed enzymatic activity on casein. Both proteins formed stable complexes with jararhagin and inhibited its hemorrhagic effect as well as the enzymatic activity of this toxin on fluorogenic substrate. PMID:10779682

  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. PMID:12875875

  2. Use of snake venom fractions in the coagulation laboratory.

    Science.gov (United States)

    Marsh, N A

    1998-07-01

    Snake venom toxins are now regularly used in the coagulation laboratory for assaying haemostatic parameters and as coagulation reagents. Snake venom thrombin-like enzymes (SVTLE) are used for fibrinogen and fibrinogen breakdown product assay as well as detecting dysfibrinogenaemias. Significantly, because SVTLE are not inhibited by heparin, they can be used for defibrinating samples that contain the anticoagulant before assay of haemostatic variables. Prothrombin activators are found in many snake venoms and are used in prothrombin assays, for studying dysprothrombinaemias and preparing meizothrombin and non-enzymic prothrombin. Russell's viper (Daboia russelli) venom (RVV) contains a number of compounds useful in the assay of factors V, VII, X, platelet factor 3 and lupus anticoagulants. Activators from the taipan, Australian brown snake and saw-scaled viper have been used to assay lupus anticoagulants. Protein C and activated protein C resistance can be measured by means of RVV and Protac, a fast acting inhibitor from Southern copperhead snake venom and von Willebrand factor can be studied with Botrocetin from Bothrops jararaca venom. Finally, phospholipase A2 enzymes and the disintegrins, a family of Arg-Gly-Asp (RGD)-containing proteins found in snake venoms, show great potential for the study of haemostasis including, notably, platelet glycoprotein receptors GPIIb/IIIa and Ib. PMID:9712287

  3. Diagnostic uses of snake venom.

    Science.gov (United States)

    Marsh, N A

    2001-01-01

    Snake venom toxins are invaluable for the assay of coagulation factors and for the study of haemostasis generally. Thrombin-like enzymes (SVTLE) are used for fibrinogen and fibrinogen breakdown product assays as well as detecting dysfibrinogenaemias. Since SVTLE are not inhibited by heparin, they can be used for assaying antithrombin III in samples containing heparin. Snake venom prothrombin activators are utilised in prothrombin assays, whilst Russell's viper venom (RVV) can be used to assay clotting factors V, VII, X and lupus anticoagulants (LA). Activators from the taipan, Australian brown snake and saw-scaled viper have also been used to assay LA. Protein C (PC) and activated PC (APC) resistance can be measured by means of RVV, Protac (from Southern copperhead snake venom) and STA-Staclot (from Crotalus viridis helleri) whilst von Willebrand factor can be studied with Botrocetin (Bothrops jararaca). Finally, snake venom C-type lectins and metalloproteinase disintegrins are being used to study platelet glycoprotein receptors and show great potential for use in the routine coagulation laboratory. PMID:11910187

  4. The effective control of a bleeding injury using a medical adhesive containing batroxobin

    International Nuclear Information System (INIS)

    Many types of hemostatic agents have been studied for the effective control of bleeding. In this study, a powdery medical adhesive composed of aldehyded dextran and ε-poly (L-lysine) was used with the recombinant batroxobin. Batroxobin is a venomous component from the snake Bothrops atrox moojeni and catalyzes fibrinogen conversion to form soluble fibrin clots. This research aims to examine the performance of the batroxobin-containing adhesive for hemostasis, and evaluate its potential as a novel hemostatic adhesive. The fibrinogen conversion ability of batroxobin was evaluated by a fibrinogen clotting assay and a whole blood clotting assay. Both experiments demonstrated the effectiveness of the batroxobin-containing adhesive for blood clot formation. Animal experiments were also conducted. After a pricking wound was made in an ICR (imprinting control region) mouse liver, the adhesive and various concentrations of batroxobin were applied. The total amount of blood loss was reduced with increasing concentrations of batroxobin. For excessive bleeding conditions, the femoral artery wound model of SD (Sprague-Dawley) rats was adopted. With higher concentrations of batroxobin, hemostasis was more rapidly achieved. Histological analysis of the liver model also supports the hemostatic effects through fibrin clot formation. In conclusion, batroxobin and medical adhesive effectively facilitate blood coagulation, and could be developed for clinical use. (paper)

  5. The conserved structure of snake venom toxins confers extensive immunological cross-reactivity to toxin-specific antibody.

    Science.gov (United States)

    Harrison, R A; Wüster, W; Theakston, R D G

    2003-03-01

    We have demonstrated previously that antisera from mice immunised with DNA encoding the carboxy-terminal domain (JD9) of a potent haemorrhagic metalloproteinase, jararhagin, neutralised over 70% of the haemorrhagic activity of the whole Bothrops jararaca venom. Here, we demonstrate that the JD9-specific antibody possesses extensive immunological reactivity to venom components in snakes of distinct species and genera. The polyspecific immunological reactivity of the antibody showed a correlation with amino acid sequence identity and with predicted antigenic domains of JD9-analogues in venoms of snakes with closest phylogenetic links to B. jararaca. This study further promotes the potential of DNA immunisation to generate toxin-specific antibodies with polyspecific cover. An analysis of the reactivity of the JD9-specific antisera to B. atrox complex venoms that exhibited intraspecific variation in the venom proteome revealed, however, that the toxin-specific approach to antivenom development requires a more in-depth knowledge of the target molecules than is required for conventional antivenoms. PMID:12657313

  6. Characterization and cDNA cloning of aminopeptidase A from the venom of Gloydius blomhoffi brevicaudus.

    Science.gov (United States)

    Ogawa, Yuko; Murayama, Nobuhiro; Fujita, Yoshiaki; Yanoshita, Ryohei

    2007-06-15

    The aminopeptidase activities of snake venoms from Gloydius blomhoffi brevicaudus, Gloydius halys blomhoffii, Trimeresurus flavoviridis, Bothrops jararaca and Crotalus atrox were investigated. Aminopeptidase A (APA), aminopeptidase B and aminopeptidase N activities were present in all snake venoms. The strongest APA activity was found in venom from G. blomhoffi brevicaudus. The susceptibility to metallopeptidase inhibitors and the pH optimum of the partially purified enzyme from G. blomhoffi brevicaudus venom were similar to those of known APAs from mammals. A G. blomhoffi brevicaudus venom gland cDNA library was screened to isolate cDNA clones using probes based on highly conserved amino acid sequences in known APAs. Molecular cloning of APA from G. blomhoffi brevicaudus venom predicted that it was a type II integral membrane protein containing 958 amino acid residues with 17 potential N-linked glycosylation sites. It possessed a His-Glu-Xaa-Xaa-His-(Xaa)(18)-Glu zinc binding motif that allowed the classification of this protein as a member of the M1 family of zinc-metallopeptidases, or gluzincins. The deduced amino acid sequence shows approximately 60% sequence identity to mammalian APA sequences. This is the first study to report the primary structure of APA from a reptile. PMID:17383704

  7. Expression of mRNAs coding for VAP1/crotastatin-like metalloproteases in the venom glands of three South American pit vipers assessed by quantitative real-time PCR.

    Science.gov (United States)

    Tavares, N A C; Correia, J M; Guarnieri, M C; Lima-Filho, J L; Prieto-da-Silva, A R B; Rádis-Baptista, G

    2008-12-15

    Snake venom metalloproteases encompass a large family of toxins, with approximately 200 members already catalogued, which exhibit a diversity of structures and biological functions. From this relatively large number, only a dozen examples of apoptosis-inducing metalloproteases, like VAP1 and 2 from the venom of Crotalus atrox, are known. Since most VAP1-like toxins ever characterized were purified from the venom of Viperidae species inhabiting diverse places on earth, we investigate the expression of VAP-like metalloproteases in the venom gland of three representative pit vipers of the Brazilian territory. By molecular cloning and quantitative real-time polymerase chain reaction, using as calibrator gene the Crotalus durissus terrificus homolog of VAP1, named crotastatin, it is reported here that VAP1/crotastatin-like homologues in the venom gland of Bothrops atrox, C. d. cascavella and Lachesis m. rhombeata are expressed at different levels. Hence, batroxstatins, the crotastatin-like precursors from B. atrox, are expressed 87 times more than crotastatin-1, from C. d. cascavella, and 7.5-fold that lachestatins, from L. m. rhombeata. Moreover, in silico structural analysis of amino acid sequences indicates that batroxstatin-2, crotastatins and lachestatin-1 and -2 which share the archetypal motifs and metal- binding sites of VAP1, are subgrouped in a branch that comprises some apoptosis-inducing toxins. PMID:18926840

  8. Inhibition of mutalysin II, a metalloproteinase from bushmaster snake venom by human alpha2-macroglobulin and rabbit immunoglobulin.

    Science.gov (United States)

    Souza, C T; Moura, M B; Magalhaes, A; Heneine, L G; Olortegui, C C; Diniz, C R; Sanchez, E F

    2001-09-01

    Mutalysin II is a 22.5-kDa zinc endopeptidase isolated from Lachesis muta muta snake venom. In order to determine whether the inhibitors human alpha2-macroglobulin (alpha2-M) and rabbit antibody to mutalysin II share a common mechanism, we have investigated the inhibition of mutalysin II by these two different glycoproteins. The proteolytic activity of mutalysin II with dimethylcasein as substrate was completely inhibited by human alpha2-M and by a purified rabbit antibody to mutalysin II. The protection of fibrin(ogen) digestion by alpha2-M was slightly better than the protection offered by the antibody. In addition, the purified antibody reacted only with the metalloproteinase in bushmaster venom, as demonstrated by immunodiffusion. SDS-PAGE analysis of reduced samples showed that the interaction of mutalysin II with alpha2-M resulted in the formation of high molecular complex ( approximately 180000) and M(r) 90000 fragments generated by the venom enzyme. Also, fragments at 85 and 23 kDa were detected under non-reducing conditions after incubation of rabbit immunoglobulin with enzyme. Proteolysis of dimethylcasein as substrate revealed that the stoichiometry of inhibition was 1.0 mol of human alpha2-M and 1.5 mol of rabbit IgG antimutalysin II per mole of enzyme. Furthermore, dimethylcasein hydrolysis indicated that several viperid snake venoms, including Bothrops atrox, B. alternatus and Trimeresurus flavoviridis cross-reacted with the specific rabbit antibody to varying degrees. PMID:11544086

  9. Comparative analysis of viperidae venoms antibacterial profile: a short communication for proteomics.

    Science.gov (United States)

    Ferreira, Bruno L; Santos, Dilvani O; Dos Santos, André Luis; Rodrigues, Carlos R; de Freitas, Cícero C; Cabral, Lúcio M; Castro, Helena C

    2011-01-01

    Bacterial infections involving multidrug-resistant strains are one of the ten leading causes of death and an important health problem in need for new antibacterial sources and agents. Herein, we tested and compared four snake venoms (Agkistrodon rhodostoma, Bothrops jararaca, B. atrox and Lachesis muta) against 10 Gram-positive and Gram-negative drug-resistant clinical bacteria strains to identify them as new sources of potential antibacterial molecules. Our data revealed that, as efficient as some antibiotics currently on the market (minimal inhibitory concentration (MIC) = 1-32 μg mL(-1)), A. rhodostoma and B. atrox venoms were active against Staphylococcus epidermidis and Enterococcus faecalis (MIC = 4.5 μg mL(-1)), while B. jararaca inhibited S. aureus growth (MIC = 13 μg ml(-1)). As genomic and proteomic technologies are improving and developing rapidly, our results suggested that A. rhodostoma, B. atrox and B. jararaca venoms and glands are feasible sources for searching antimicrobial prototypes for future design new antibiotics against drug-resistant clinical bacteria. They also point to an additional perspective to fully identify the pharmacological potential of these venoms by using different techniques. PMID:18955360

  10. Factor V activation and inactivation by venom proteases.

    Science.gov (United States)

    Rosing, J; Govers-Riemslag, J W; Yukelson, L; Tans, G

    2001-01-01

    Blood coagulation factor V is a single-chain glycoprotein with M(r) = 330,000 which plays an important role in the procoagulant and anticoagulant pathways. Thrombin activates factor V into factor Va, a two-chain molecule which is composed of a heavy (M(r) = 105,000) and a light chain (M(r) = 71,000/74,000). Factor Va accelerates factor Xa-catalysed prothrombin activation more than 1,000-fold and under physiological conditions the cofactor activity of factor Va in prothrombin activation is down-regulated by activated protein C. Factor V can also be activated by a wide variety of snake venoms (e.g. from Vipera species, Naja naja oxiana, Bothrops atrox) and by proteases present in the bristles of a South American caterpillar (Lonomia achelous). Some venoms, notably of Vipera lebetina turanica and Lonomia achelous, contain proteases that are able to inactivate factor V or factor Va. Venom factor V activators are excellent tools in studying the structure-function relationship of factor V(a) and they are also used in diagnostic tests for quantification of plasma factor V levels and for the screening of defects in the protein C pathway. In this review, the structural and functional properties of animal venom factor V activators and inactivators is described. PMID:11910191

  11. 三种蛇毒类凝血酶对纤维蛋白原的作用方式%Action pattern of different snake venom thrombin-like enzyme on fibrinogen

    Institute of Scientific and Technical Information of China (English)

    沈文彧; 王宏英; 薛雁; 苏珊; 赵怀清

    2010-01-01

    目的 以纤维蛋白原为底物,考察巴西矛头蝮蛇(Bothrops atrox)、尖吻蝮蛇(Agkistrodon acutus)、长白白眉蝮蛇(Agkistrodon halys pullas)3种蛇毒类凝血酶对纤维蛋白原的作用方式.方法 采用SDS-PAGE及RP-HPLC法对作用结果进行检测.结果 3种蛇毒类凝血酶对纤维蛋白原的作用方式不完全相同,巴西矛头蝮蛇、尖吻蝮蛇2种蛇毒类凝血酶只作用于纤维蛋白原的α链,对β、γ链则元作用,长白白眉蝮蛇毒类凝血酶起初作用于纤维蛋白原的β链,对α链作用较弱,随着时间的延长对α链作用增强,对γ链无作用.结论 巴西矛头蝮蛇、尖吻蝮蛇2种蛇毒类凝血酶属于SVTLE-A型,而长白白眉蝮蛇毒类凝血酶则属于SVTLE-AB型.

  12. The effective control of a bleeding injury using a medical adhesive containing batroxobin.

    Science.gov (United States)

    You, Kyung Eun; Koo, Min-Ah; Lee, Dae-Hyung; Kwon, Byeong-Ju; Lee, Mi Hee; Hyon, Suong-Hyu; Seomun, Young; Kim, Jong-Tak; Park, Jong-Chul

    2014-04-01

    Many types of hemostatic agents have been studied for the effective control of bleeding. In this study, a powdery medical adhesive composed of aldehyded dextran and ε-poly (L-lysine) was used with the recombinant batroxobin. Batroxobin is a venomous component from the snake Bothrops atrox moojeni and catalyzes fibrinogen conversion to form soluble fibrin clots. This research aims to examine the performance of the batroxobin-containing adhesive for hemostasis, and evaluate its potential as a novel hemostatic adhesive. The fibrinogen conversion ability of batroxobin was evaluated by a fibrinogen clotting assay and a whole blood clotting assay. Both experiments demonstrated the effectiveness of the batroxobin-containing adhesive for blood clot formation. Animal experiments were also conducted. After a pricking wound was made in an ICR (imprinting control region) mouse liver, the adhesive and various concentrations of batroxobin were applied. The total amount of blood loss was reduced with increasing concentrations of batroxobin. For excessive bleeding conditions, the femoral artery wound model of SD (Sprague-Dawley) rats was adopted. With higher concentrations of batroxobin, hemostasis was more rapidly achieved. Histological analysis of the liver model also supports the hemostatic effects through fibrin clot formation. In conclusion, batroxobin and medical adhesive effectively facilitate blood coagulation, and could be developed for clinical use. PMID:24487019

  13. Characterising the enzymatic profile of crude tentacle extracts from the South Atlantic jellyfish Olindias sambaquiensis (Cnidaria: Hydrozoa).

    Science.gov (United States)

    Knittel, Paloma S; Long, Paul F; Brammall, Lucas; Marques, Antonio C; Almeida, Michelle T; Padilla, Gabriel; Moura-da-Silva, Ana M

    2016-09-01

    Jellyfish venoms are of medical and biotechnological importance, with toxins displaying antimicrobial, analgesic and anti-tumor activities. Although proteolytic enzymes have also been described, detailed characterisation of these proteins is scant in Olindias spp. High throughput mass spectrometry profiling of cnidarian venoms has become increasingly popular since the first description of the proteomic profile of putative toxins isolated from nematocysts of the hydrozoan jellyfish Olindias sambaquiensis describing the presence of orthologous enzymes as presented in venoms of advanced species as snakes. Rigorous bioinformatics analyses can aid functional annotation, but biochemical assays are prerequisite to unambiguously assign toxic function to a peptide or protein. Here we present results that experimentally confirm previously predicted proteomic analysis that crude venom extracts from tentacles of O. sambaquiensis are composed of polypeptides with metalloproteinase, serine proteinase and phospholipases A2 activities. Surprisingly, levels of serine proteinase and phospholipase A2 activities were comparable to those observed in venoms of Bothrops snakes which were used as positive controls in this study. Hence, these data offer new opportunities to explore serine proteinase and phospholipase A2 activities in the clinical sequelae following O. sambaquiensis envenomation, with future possible biopharmaceutical applications. PMID:27169682

  14. Taxocenose de serpentes no Planalto Médio do Rio Grande do Sul, Brasil Taxocenosis of snakes in the middle plateau region of Rio Grande do Sul, Brazil

    Directory of Open Access Journals (Sweden)

    Noeli Zanella

    2006-03-01

    Full Text Available Este trabalho foi desenvolvido na Fazenda da Brigada (28º14'39"S, 52º14'42"W, Passo Fundo, Planalto Médio do Rio Grande do Sul, e teve como objetivo contribuir com informações sobre riqueza, abundância de serpentes e comparação entre os métodos de amostragem. O estudo foi desenvolvido em duas áreas, floresta e campo, no período de janeiro de 2001 a março de 2003. Para a coleta de dados foram utilizados os seguintes métodos: procura limitada por tempo (PLT, encontros ocasionais por terceiros (EOT, encontros ocasionais pela equipe (EOE e armadilhas de interceptação e queda (AQ. Com a utilização dos quatro métodos foi possível registrar 19 espécies de serpentes, distribuídas em três famílias (Colubridae, Elapidae e Viperidae e coletar 284 indivíduos, sendo 128 por EOT, 36 por PLT, 112 por AQ e 8 por EOE. Utilizando todos os métodos, as três espécies mais abundantes foram: Thamnodynastes strigatus (Günther, 1858 (16,9%, Echinanthera cyanopleura (Cope, 1885 (15,2% e Bothrops alternatus Duméril, Bibron & Duméril, 1854 (13,4%. O estudo dessa taxocenose revela importantes características da ecologia de serpentes e poderá subsidiar futuros estudos de conservação para a área.The present study was conducted on the Brigada Farm (28º14'39"S, 52º14'42"W, Passo Fundo, middle plateau region of Rio Grande do Sul. The objective of the study was to provide data regarding the richness and abundance of snakes and to compare sampling methods. The study was carried out in two areas, i.e., forest and field, between January 2001 and March 2003. Data were collected using the following methods: time-constrained search (TCS, occasional encounters by local collectors (OEC, occasional encounters by the team (OET, and by pitfall traps (PT. Using the four methods, 19 snake species belonging to three families (Colubridae, Elapidae and Viperidae were recorded, and 284 individuals were collected, including 128 by OEC, 36 by TCS, 112 by PT, and

  15. Influência dos fatores abióticos e da disponibilidade de presas sobre comunidade de serpentes do Planalto Médio do Rio Grande do Sul Influence of abiotic factors and availability of prey on the occurrence of snakes in the middle plateau region of Rio Grande do Sul, Brazil

    Directory of Open Access Journals (Sweden)

    Noeli Zanella

    2009-03-01

    Full Text Available A influência dos fatores abióticos sobre a disponibilidade de presas e a dieta das espécies de serpentes mais abundantes do Planalto Médio do Rio Grande do Sul, foi estudada em duas áreas: floresta e campo. O trabalho foi desenvolvido utilizando serpentes coletadas com os métodos: procura limitada por tempo, encontros ocasionais, armadilhas de interceptação e queda, e serpentes depositadas na coleção de répteis da Universidade de Passo Fundo. Foram registradas as guildas alimentares das seis espécies mais abundantes: anurófagas (n = 2: Echinanthera cyanopleura (Cope, 1885 e Thamnodynates cf. strigatus (Günther, 1858; rodentívoras (n = 1: Bothrops alternatus Duméril, Bibron & Duméril, 1854; moluscófagas (n = 1: Tomodon dorsatus Duméril, Bibron & Duméril, 1854 e generalistas (n = 2: Liophis poecilogyrus (Wied-Neuwied, 1825 e Philodryas patagoniensis (Girard, 1858. Dos fatores abióticos analisados, a abundância de serpentes foi mais relacionada à temperatura máxima (R² = 0,66 e não apresentou relação significativa com a pluviosidade. A abundância de anfíbios apresentou relação positiva com a pluviosidade (R² = 0,54 e não foi significativa com a temperatura mínima. A abundância de serpentes não foi correlacionada com a abundância de anfíbios e roedores.The influence of abiotic factors on the availability of prey and on the diet of the most abundant snake species in the middle plateau region of Rio Grande do Sul was studied in two areas: forest and field. Snakes captured using time-constrained search, occasional encounters and pitfall traps and snakes deposited in the reptile collection of the University of Passo Fundo were studied. The feeding guilds of the six most abundant species were recorded: anurophagous (n = 2: Echinanthera cyanopleura (Cope, 1885 and Thamnodynates cf. strigatus (Günther, 1858; rodent-eating (n = 1: Bothrops alternatus Duméril, Bibron & Duméril, 1854; molluscophagous (n = 1: Tomodon

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

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    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

  17. Distribution and Ecology of the Western Ecuador Frog Leptodactylus labrosus (Amphibia: Anura: Leptodactylidae)%厄瓜多尔西部新热带蛙(两栖纲: 无尾目:细趾蟾科)的分布特征及生态习性

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Leptodactylus labrosus is a terrestrial sit-and-wait predator; its diet includes ground-level, fossorial, and flying insects, and ants numerically predominate. Bothrops asper is recorded as a predator of L. labrosus. L. labrosus lives mainly in deciduous and semi-deciduous forests, where it is restricted to wet microhabitats, and occasionally in evergreen forests. L. labrosus inhabits northern, central, and southern regions of western coastal Ecuador and northern and central western coastal Peru up to 700 m, and into the dry interandean valleys of southern Ecuador and northern Peru up to 1 300 m. Its distribution encompasses moistly seasonally dry forest in coastal Ecuador and Peru. It also occupies moister areas towards the slopes of the Andes where it is sympatric with three other congeneric species, but at sites of sympatry the species show habitat segregation. The distribution pattern of L. labrosus is shared by several other range-restricted amphibians corresponding to the Tumbesian region, which should be recognized as an endemic Amphibian area. The zone between the Choco and Tumbesian regions, where L. labrosus gets in sympatry with other Leptodactylus species, possess ecological and climatic characteristics that have shaped a unique fauna, including several endemic taxa; and it should be recognized as the west Ecuadorian endemic region.%Leptodactylus labrosus属于陆地坐-等型的捕食者, 其食物包括地表的、掘地的及飞行昆虫, 其中蚂蚁数量最多. 曾有报道Bothrops asper是其天敌. L. labrosus主要栖息于落叶及半落叶林潮湿的环境中, 偶尔也栖息于常绿林中. L. labrosus分布于厄瓜多尔西海岸的北部、中部、南部; 秘鲁西海岸的北部、中部, 分布地海拔可达600 m; 及位于南厄瓜多尔和北秘鲁海拔高达1 300 m的干燥的安第斯山谷. 其分布地区包括了厄瓜多尔和秘鲁海岸的潮湿的、季节性干燥的森林. 在安第斯

  18. Perfil eletroforético das proteínas séricas de serpentes Crotalus durissus terrificus (cascavel criadas em cativeiro Serum protein electrophoresis profile of the rattlesnake Crotalus durissus terrificus kept in captivity

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    Joandes Henrique Fonteque

    2009-06-01

    Full Text Available As serpentes peçonhentas dos gêneros Bothrops e Crotalus têm sido mantidas em cativeiro visando à extração de venenos para a produção de imunobiológicos. O conhecimento da fisiologia desses animais e as alterações na concentração de proteínas e suas frações séricas são importantes para a identificação precoce de importantes enfermidades que cursam com estados de hipoproteinemia e hiperproteinemia. O objetivo do trabalho foi determinar a concentração de proteína total e o perfil eletroforético das proteínas séricas de serpentes Crotalus durissus terrificus (cascavel criadas em cativeiro. Foram colhidas amostras de sangue da veia coccígea ventral de 21 serpentes adultas e sadias, divididas em dois grupos: Grupo 1 de 12 machos com peso médio de 588,89±193,55g, e Grupo 2 de nove fêmeas com peso médio de 708,33±194,04g. A proteína total sérica foi determinada pelo método de refratometria e a eletroforese em gel de agarose. Obtiveram-se valores da proteína total sérica (g/dL de 4,51±0,50 para machos e de 4,82±0,72 para fêmeas, e para machos e fêmeas de 4,64±0,61. Foram identificadas pela eletroforese quatro frações protéicas (g/dL: albumina, a, b, g-globulinas e calculada a relação albumina:globulina. As serpentes fêmeas apresentaram maiores valores para as variáveis, albumina e para a relação albumina/globulina (AG diferindo significativamente (PThe poisonous snakes of the genera Crotalus and Bothrops have been kept in captivity with the purpose of extracting poison for the production of immunobiological. Knowledge of the physiology of these animals and serum proteins concentration changes are important for early identification of major diseases which lead to states of hypoproteinemia and hyperproteinemia. The objective was to determine the concentration of total protein and serum protein electrophoresis profile of Crotalus durissus terrificus (rattlesnake in captivity. Blood samples were taken from

  19. Correlation of the inhibitory activity of phospholipase A2 snake venom and the antioxidant activity of Colombian plant extracts

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    Jaime A. Pereañez

    2010-12-01

    Full Text Available Snakebite continues to be a significant health problem in many countries of Latin America. Even though, there has been an improvement in the antivenom therapy, the local effects caused by myotoxic phospholipases A2 (PLA2 present in the venoms, still persist. In search for alternatives to antagonize the PLA2 activity of Bothrops asper's venom, 36 extracts belonging to seventeen families of vascular plants and bryophytes were screened. A significant inhibition of the enzymatic activity of PLA2 present in B. asper's whole venom was seen in eleven of these extracts. In addition, the antioxidant activity of all the extracts was evaluated. The results evidenced a significant statistical correlation between extracts with an inhibitory effect against PLA2 and those with an antioxidant activity. Moreover, the amount of phenols was quantified finding a relationship between the bioactivity and the presence of these compounds. Nine extracts were screened against a fraction of the venom rich in basic PLA2 (Fx-V B. asper, exhibiting an inhibitory effect on PLA2 activity of this fraction in a range from 30-80%. This activity was supported by the inhibition that these extracts presented on the cytotoxicity caused by Fx-V B. asper on murine skeletal muscle C2C12 myoblasts. The results obtained, could point to minimize efforts in the search of PLA2 inhibitors by focusing in samples with known antioxidant properties.Veneno de cobra continua a ser um problema importante de saúde em muitos países da América Latina. Apesar dos avanços na terapia antiveneno, os efeitos locais causados por fosfolipases A2 miotóxica (PLA2 presentes no veneno, ainda persistem. Em busca de alternativas para antagonizar a atividade da PLA2 do veneno de Bothrops asper, foram selecionados 36 extratos pertencentes a dezessete famílias de plantas vasculares e briófitas. Uma inibição significativa da atividade enzimática de PLA2 presente no veneno de B. asper foi observada em onze

  20. 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

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    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

  1. Non-venomous snake bite and snake bite without envenoming in a brazilian teaching hospital: analysis of 91 cases Acidentes por serpentes não-peçonhentas e acidentes por serpentes peçonhentas sem envenenamento em hospital universitário brasileiro: análise de 91 casos

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    Paulo Vitor Portella Silveira

    1992-12-01

    Full Text Available A retrospective survey of 473 cases of snake bite admitted to a Brazilian teaching hospital from 1984 to 1990 revealed 91 cases of bite without envenoming and/or caused by non-venomous snakes. In 17 of these cases the snake was identified, and one patient was bitten by a snake-like reptile (Amphisbaena mertensii. In 43 cases diagnosis was made on clinical grounds (fang marks in the absence of signs of envenoming. The other 30 cases were of patients who complained of being bitten but who did not show any sign of envenoming or fang mark. Most cases occurred in men (66;73%, in the 10-19 years age group (26;29%, in the lower limbs (51/74;69%, between 6 A. M. and 2 P.M. (49;61% and in the month of April (16; 18%. One patient bitten by Philodryas olfersii developed severe local pain, swelling and redness at the site of the bite, with normal clotting time. The patient bitten by Drymarcon corais was misdiagnosed as being bitten by a snake of the genus Bothrops, was given the specific antivenom, and developed anaphylaxis. One patient bitten by Sibynomorphus mikanii presented prolonged clotting time, and was also given antivenom as a case of Bothrops bite. Correct identification of venomous snakes by physicians is necessary to provide correct treatment to victims of snake bite, avoiding unnecessary distress to the patient, and overprescription of antivenom, which may eventually cause severe untoward effects.Um levantamento retrospectivo de 473 casos de acidentes ofídicos admitidos em um hospital-escola brasileiro de 1984 a 1990 revelou 91 casos sem envenenamento e/ou causados por serpentes não-peçonhentas. Em 17 casos a serpente foi identificada e um paciente foi mordido por um réptil que se assemelha às serpentes (Amphisbaena mertensii. Em 43 casos o diagnóstico foi clínico (sinal das presas na ausência de sinais de envenenamento. Os demais 30 casos foram de pacientes que se queixavam de terem sido mordidos mas que não apresentavam nem sinal de

  2. Venoms, toxins and derivatives from the Brazilian fauna: valuable sources for drug discovery.

    Science.gov (United States)

    De Marco Almeida, Flávia; de Castro Pimenta, Adriano Monteiro; Oliveira, Mônica Cristina; De Lima, Maria Elena

    2015-06-25

    Animal venoms have been widely investigated throughout the world. The great number of biotechnological articles as well as patent applications in the field of drug discovery based on these compounds indicates how important the source is. This review presents a list of the most studied Brazilian venomous animal species and shows the most recent patent applications filed from 2000 to 2013, which comprise Brazilian venoms, toxins and derivatives. We analyze the data according to the species, the type of products claimed and the nationality of the inventors. Fifty-five patent applications were found, involving 8 genera. Crotalus, Lachesis, Bothrops and Loxosceles represented 78% of the patent applications. The other 22% were represented by Phoneutria, Tityus, Acanthoscurria and Phyllomedusa. Most of the inventions (42%) involved anticancer, immunomodulator or antimicrobial drugs, while 13% involved anti-venoms and vaccines, 11% involved hypotensive compositions, 9% involved antinociceptive and/or anti-inflammatory compositions, and the other 25% involved methods, kits or compositions for various purposes. Brazilian inventors filed 49% of the patent applications, but other countries, mainly the United States of America, Germany, Russia and France, also filed patent applications claiming products comprising venoms, toxins and/or derivatives from the Brazilian fauna. Brazil holds an important number of patent applications which mostly belong to universities and research institutes, but the pharmaceutical industry in this field is still weak in Brazil. Although, Brazilian venomous animal species have been reported in drug discovery throughout the world, many species remain to be explored as valuable and promising tools for drug discovery and development. PMID:26109299

  3. 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

  4. Evaluation of myotoxic activity of Bothropstoxin-1 irradiated with 60Co gamma rays

    International Nuclear Information System (INIS)

    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 their immunogenic properties. Snake venoms and their isolated toxins showed structural modifications after gamma radiation process, in aqueous solution, mainly by water radiolysis sub products. Free radical scavengers, such as NaNO3 and t-butanol, present selective effects on specific radical from water radiolysis. The NaNO3 has affinity by aqueous electron, while the t-butanol has affinity by hydroxyl radical. At the present work, we have investigated the myotoxic activity of Bothropstoxin-1 (BTHX-1), a K49 phospholipase, present in Bothrops jararacussu crude venom, before and after irradiation process, with or without scavenger substances presence. BTHX-1 was irradiated with 2 kGy of 60Co gamma rays, in aqueous solution and in the presence of oxygen. BALB/c mice were inoculated with either native or irradiated toxin, with or without scavenger substances. After 3 hours, blood samples were collected and the myotoxic activity was evaluated by LDH (lactate dehydrogenase) release. The muscular tissue damage was directly related to the LDH amounts released. Irradiated Bothropstoxin-1, with or without NaNO3 substance, caused less damage than their native counterpart. But irradiated toxin, in the presence of t-butanol, was so myotoxic as the native BTHX-1. These results indicate that irradiation of toxic proteins can promote significant modifications on their structures, but still retaining many of the original biological properties of their native counterparts. Additionally, some scavengers substances can change these gamma radiation effects. (author)

  5. Argininosuccinate Synthetase Is a Functional Target for a Snake Venom Anti-hypertensive Peptide

    Science.gov (United States)

    Guerreiro, Juliano R.; Lameu, Claudiana; Oliveira, Eduardo F.; Klitzke, Clécio F.; Melo, Robson L.; Linares, Edlaine; Augusto, Ohara; Fox, Jay W.; Lebrun, Ivo; Serrano, Solange M. T.; Camargo, Antonio C. M.

    2009-01-01

    Bj-BPP-10c is a bioactive proline-rich decapeptide, part of the C-type natriuretic peptide precursor, expressed in the brain and in the venom gland of Bothrops jararaca. We recently showed that Bj-BPP-10c displays a strong, sustained anti-hypertensive effect in spontaneous hypertensive rats (SHR), without causing any effect in normotensive rats, by a pharmacological effect independent of angiotensin-converting enzyme inhibition. Therefore, we hypothesized that another mechanism should be involved in the peptide activity. Here we used affinity chromatography to search for kidney cytosolic proteins with affinity for Bj-BPP-10c and demonstrate that argininosuccinate synthetase (AsS) is the major protein binding to the peptide. More importantly, this interaction activates the catalytic activity of AsS in a dose-de pend ent manner. AsS is recognized as an important player of the citrulline-NO cycle that represents a potential limiting step in NO synthesis. Accordingly, the functional interaction of Bj-BPP-10c and AsS was evidenced by the following effects promoted by the peptide: (i) increase of NO metabolite production in human umbilical vein endothelial cell culture and of arginine in human embryonic kidney cells and (ii) increase of arginine plasma concentration in SHR. Moreover, α-methyl-dl-aspartic acid, a specific AsS inhibitor, significantly reduced the anti-hypertensive activity of Bj-BPP-10c in SHR. Taken together, these results suggest that AsS plays a role in the anti-hypertensive action of Bj-BPP-10c. Therefore, we propose the activation of AsS as a new mechanism for the anti-hypertensive effect of Bj-BPP-10c in SHR and AsS as a novel target for the therapy of hypertension-related diseases. PMID:19491403

  6. A rational design for the nanoencapsulation of poisonous animal venoms in liposomes prepared with natural phospholipids.

    Science.gov (United States)

    da Costa, Maria Helena Bueno; Sant'Anna, Osvaldo A; Quintilio, Wagner; Schwendener, Reto Albert; de Araujo, Pedro Soares

    2012-11-01

    Liposomes have been used since the 1970's to encapsulate drugs envisaging enhancement in efficacy and therapeutic index, avoidance of side effects and increase in the encapsulated agent stability. The major problem when encapsulating snake venoms is the liposomal membrane instability caused by venom phospholipases. Here the results obtained encapsulating Crotalus durissimus terrificus and a pool of Bothropic venoms within liposomes (LC and LB, respectively) used to produce anti-venom sera are presented. The strategy was to modify the immunization protocol to enhance antibody production and to minimize toxic effects by encapsulating inactivated venoms within stabilized liposomes. Chemically modified venoms were solubilized in a buffer containing an inhibitor and a chelating agent. The structures of the venoms were analyzed by UV, CD spectroscopy and ELISA. In spite of the differences in the helical content between natural and modified venoms, they were recognized by horse anti-sera. To maintain long-term stability, mannitol was used as a cryoprotectant. The encapsulation efficiencies were 59 % (LB) and 99 % (LC), as followed by filtration on Sephacryl S1000. Light scattering measurements led us to conclude that both, LB (119 ±47 nm) and LC (147±56 nm) were stable for 22 days at 4 °C, even after lyophilization. Genetically selected mice and mixed breed horses were immunized with these formulations. The animals did not show clinical symptoms of venom toxicity. Both, LB and LC enhanced by at least 30 % the antibody titers 25 days after injection and total IgG titers remained high 91 days after immunization. The liposomal formulation clearly exhibited adjuvant properties.

  7. Bothropstoxin-I reduces evoked acetylcholine release from rat motor nerve terminals: radiochemical and real-time video-microscopy studies.

    Science.gov (United States)

    Correia-de-Sá, Paulo; Noronha-Matos, José B; Timóteo, Maria A; Ferreirinha, Fátima; Marques, Patrícia; Soares, Andreimar M; Carvalho, Cicilia; Cavalcante, Walter L G; Gallacci, Márcia

    2013-01-01

    Understanding the biological activity profile of the snake venom components is fundamental for improving the treatment of snakebite envenomings and may also contribute for the development of new potential therapeutic agents. In this work, we tested the effects of BthTX-I, a Lys49 PLA(2) homologue from the Bothrops jararacussu snake venom. While this toxin induces conspicuous myonecrosis by a catalytically independent mechanism, a series of in vitro studies support the hypothesis that BthTX-I might also exert a neuromuscular blocking activity due to its ability to alter the integrity of muscle cell membranes. To gain insight into the mechanisms of this inhibitory neuromuscular effect, for the first time, the influence of BthTX-I on nerve-evoked ACh release was directly quantified by radiochemical and real-time video-microscopy methods. Our results show that the neuromuscular blockade produced by in vitro exposure to BthTX-I (1 μM) results from the summation of both pre- and postsynaptic effects. Modifications affecting the presynaptic apparatus were revealed by the significant reduction of nerve-evoked [(3)H]-ACh release; real-time measurements of transmitter exocytosis using the FM4-64 fluorescent dye fully supported radiochemical data. The postsynaptic effect of BthTX-I was characterized by typical histological alterations in the architecture of skeletal muscle fibers, increase in the outflow of the intracellular lactate dehydrogenase enzyme and progressive depolarization of the muscle resting membrane potential. In conclusion, these findings suggest that the neuromuscular blockade produced by BthTX-I results from transient depolarization of skeletal muscle fibers, consequent to its general membrane-destabilizing effect, and subsequent decrease of evoked ACh release from motor nerve terminals. PMID:23142504

  8. Synergism between basic Asp49 and Lys49 phospholipase A2 myotoxins of viperid snake venom in vitro and in vivo.

    Directory of Open Access Journals (Sweden)

    Diana Mora-Obando

    Full Text Available Two subtypes of phospholipases A2 (PLA2s with the ability to induce myonecrosis, 'Asp49' and 'Lys49' myotoxins, often coexist in viperid snake venoms. Since the latter lack catalytic activity, two different mechanisms are involved in their myotoxicity. A synergism between Asp49 and Lys49 myotoxins from Bothrops asper was previously observed in vitro, enhancing Ca2+ entry and cell death when acting together upon C2C12 myotubes. These observations are extended for the first time in vivo, by demonstrating a clear enhancement of myonecrosis by the combined action of these two toxins in mice. In addition, novel aspects of their synergism were revealed using myotubes. Proportions of Asp49 myotoxin as low as 0.1% of the Lys49 myotoxin are sufficient to enhance cytotoxicity of the latter, but not the opposite. Sublytic amounts of Asp49 myotoxin also enhanced cytotoxicity of a synthetic peptide encompassing the toxic region of Lys49 myotoxin. Asp49 myotoxin rendered myotubes more susceptible to osmotic lysis, whereas Lys49 myotoxin did not. In contrast to myotoxic Asp49 PLA2, an acidic non-toxic PLA2 from the same venom did not markedly synergize with Lys49 myotoxin, revealing a functional difference between basic and acidic PLA2 enzymes. It is suggested that Asp49 myotoxins synergize with Lys49 myotoxins by virtue of their PLA2 activity. In addition to the membrane-destabilizing effect of this activity, Asp49 myotoxins may generate anionic patches of hydrolytic reaction products, facilitating electrostatic interactions with Lys49 myotoxins. These data provide new evidence for the evolutionary adaptive value of the two subtypes of PLA2 myotoxins acting synergistically in viperid venoms.

  9. Using geographical information systems to identify populations in need of improved accessibility to antivenom treatment for snakebite envenoming in Costa Rica.

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    Erik Hansson

    Full Text Available INTRODUCTION: Snakebite accidents are an important health problem in rural areas of tropical countries worldwide, including Costa Rica, where most bites are caused by the pit-viper Bothrops asper. The treatment of these potentially fatal accidents is based on the timely administration of specific antivenom. In many regions of the world, insufficient health care systems and lack of antivenom in remote and poor areas where snakebites are common, means that efficient treatment is unavailable for many snakebite victims, leading to unnecessary mortality and morbidity. In this study, geographical information systems (GIS were used to identify populations in Costa Rica with a need of improved access to antivenom treatment: those living in areas with a high risk of snakebites and long time to reach antivenom treatment. METHOD/PRINCIPAL FINDINGS: Populations living in areas with high risk of snakebites were identified using two approaches: one based on the district-level reported incidence, and another based on mapping environmental factors favoring B. asper presence. Time to reach treatment using ambulance was estimated using cost surface analysis, thereby enabling adjustment of transportation speed by road availability and quality, topography and land use. By mapping populations in high risk of snakebites and the estimated time to treatment, populations with need of improved treatment access were identified. CONCLUSION/SIGNIFICANCE: This study demonstrates the usefulness of GIS for improving treatment of snakebites. By mapping reported incidence, risk factors, location of existing treatment resources, and the time estimated to reach these for at-risk populations, rational allocation of treatment resources is facilitated.

  10. Obtained and evaluation of antisera raised against irradiated crotalic whole venom or crotoxin in 60 Co source

    International Nuclear Information System (INIS)

    anticrotalic antisera. So they could be used in immuno assays to distinguish crotalic and bothropic accidents. (author)

  11. DOCLASP - Docking ligands to target proteins using spatial and electrostatic congruence extracted from a known holoenzyme and applying simple geometrical transformations

    Science.gov (United States)

    Chakraborty, Sandeep

    2016-01-01

    The ability to accurately and effectively predict the interaction between proteins and small drug-like compounds has long intrigued researchers for pedagogic, humanitarian and economic reasons. Protein docking methods (AutoDock, GOLD, DOCK, FlexX and Glide to name a few) rank a large number of possible conformations of protein-ligand complexes using fast algorithms. Previously, it has been shown that structural congruence leading to the same enzymatic function necessitates the congruence of electrostatic properties (CLASP). The current work presents a methodology for docking a ligand into a target protein, provided that there is at least one known holoenzyme with ligand bound - DOCLASP (Docking using CLASP). The contact points of the ligand in the holoenzyme defines a motif, which is used to query the target enzyme using CLASP. If there are significant matches, the holoenzyme and the target protein are superimposed based on congruent atoms. The same linear and rotational transformations are also applied to the ligand, thus creating a unified coordinate framework having the holoenzyme, the ligand and the target enzyme. In the current work, the dipeptidyl peptidase-IV inhibitor vildagliptin was docked to the PI-PLC structure complexed with myo-inositol using DOCLASP. Also, corroboration of the docking of phenylthiourea to the modelled structure of polyphenol oxidase (JrPPO1) from walnut is provided based on the subsequently solved structure of JrPPO1 (PDBid:5CE9). Analysis of the binding of the antitrypanosomial drug suramin to nine non-homologous proteins in the PDB database shows a diverse set of binding motifs, and multiple binding sites in the phospholipase A2-likeproteins from the Bothrops genus of pitvipers. The conformational changes in the suramin molecule on binding highlights the challenges in docking flexible ligands into an already ’plastic’ binding site. Thus, DOCLASP presents a method for ’soft docking’ ligands to proteins with low

  12. Epidemiologia do acidente por serpentes peçonhentas: estudo de casos atendidos em 1988

    Directory of Open Access Journals (Sweden)

    Lindioneza Adriano Ribeiro

    1995-10-01

    Full Text Available Foram avaliados aspectos epidemiológicos de acidentes por serpentes peçonhentas no Estado de São Paulo, Brasil, com base em prontuários de 322 pacientes e em entrevistas feitas com 209 deles e/ou seus acompanhantes. Os acidentes ocorreram principalmente com pessoas de 10 a 20 anos de idade, do sexo masculino, nos meses de outubro a abril e no período diurno. As regiões anatômicas mais freqüentemente picadas foram os pés, as mãos e as pernas. Bothrops, Crotalus e Micrurus foram responsáveis por, respectivamente, 95,0%, 4,4% e 0,6% dos casos. Não ocorreram óbitos, mas 2,2% dos pacientes apresentaram seqüelas. Dentre os 209 entrevistados, a ocupação de lavrador foi a mais freqüentemente relacionada ao acidente que, em aproximadamente 60% das vezes, ocorreu durante o trabalho. O total de 160 pacientes (76,6% submeteram-se a alguma forma de tratamento antes de chegarem a um serviço de saúde: foram mais comuns o uso de torniquete (50,2%, a expressão local na tentativa de retirar parte do veneno (33,5%, a colocação das mais diversas substâncias sobre o local da picada (36,8% e a ingestão de outras (12,9%; pouco mais de um quarto dos pacientes submeteram-se a alguma forma de tratamento médico antes de chegar ao HVB sendo mais comum a antissepsia (8,2%, a administração do antiveneno (6,2%, de anti-histamínicos (5,7% e de analgésicos (5,3%.

  13. Epidemiologia do acidente por serpentes peçonhentas: estudo de casos atendidos em 1988

    Directory of Open Access Journals (Sweden)

    Ribeiro Lindioneza Adriano

    1995-01-01

    Full Text Available Foram avaliados aspectos epidemiológicos de acidentes por serpentes peçonhentas no Estado de São Paulo, Brasil, com base em prontuários de 322 pacientes e em entrevistas feitas com 209 deles e/ou seus acompanhantes. Os acidentes ocorreram principalmente com pessoas de 10 a 20 anos de idade, do sexo masculino, nos meses de outubro a abril e no período diurno. As regiões anatômicas mais freqüentemente picadas foram os pés, as mãos e as pernas. Bothrops, Crotalus e Micrurus foram responsáveis por, respectivamente, 95,0%, 4,4% e 0,6% dos casos. Não ocorreram óbitos, mas 2,2% dos pacientes apresentaram seqüelas. Dentre os 209 entrevistados, a ocupação de lavrador foi a mais freqüentemente relacionada ao acidente que, em aproximadamente 60% das vezes, ocorreu durante o trabalho. O total de 160 pacientes (76,6% submeteram-se a alguma forma de tratamento antes de chegarem a um serviço de saúde: foram mais comuns o uso de torniquete (50,2%, a expressão local na tentativa de retirar parte do veneno (33,5%, a colocação das mais diversas substâncias sobre o local da picada (36,8% e a ingestão de outras (12,9%; pouco mais de um quarto dos pacientes submeteram-se a alguma forma de tratamento médico antes de chegar ao HVB sendo mais comum a antissepsia (8,2%, a administração do antiveneno (6,2%, de anti-histamínicos (5,7% e de analgésicos (5,3%.

  14. Practical applications of snake venom toxins in haemostasis.

    Science.gov (United States)

    Marsh, Neville; Williams, Vaughan

    2005-06-15

    Snake venom toxins affecting haemostasis have facilitated extensively the routine assays of haemostatic parameters in the coagulation laboratory. Snake venom thrombin-like enzymes (SVTLE) are used for fibrinogen/fibrinogen breakdown product assay and for the detection of fibrinogen dysfunction. SVTLE are not inhibited by heparin and can thus can be used for assaying antithrombin III and other haemostatic variables in heparin-containing samples. Snake venoms are a rich source of prothrombin activators and these are utilised in prothrombin assays, for studying dysprothrombinaemias and for preparing meizothrombin and non-enzymic forms of prothrombin. Russell's viper (Daboia russelli) venom (RVV) contains toxins which have been used to assay blood clotting factors V, VII, X, platelet factor 3 and, importantly, lupus anticoagulants (LA). Other prothrombin activators (from the taipan, Australian brown snake and saw-scaled viper) have now been used to assay LA. Protein C and activated protein C resistance can be measured by means of RVV and Protac, a fast acting inhibitor from Southern copperhead snake venom and von Willebrand factor can be studied with botrocetin from Bothrops jararaca venom. The disintegrins, a large family of Arg-Gly-Asp (RGD)-containing snake venom proteins, show potential for studying platelet glycoprotein receptors, notably, GPIIb/IIIa and Ib. Snake venom toxins affecting haemostasis are also used in the therapeutic setting: Ancrod (from the Malayan pit viper, Calloselasma rhodostoma), in particular, has been used as an anticoagulant to achieve 'therapeutic defibrination'. Other snake venom proteins show promise in the treatment of a range of haemostatic disorders. PMID:15922782

  15. [Snake bite accidents in children in Costa Rica: epidemiology and determination of risk factors in the development of abscess and necrosis].

    Science.gov (United States)

    Saborío, P; González, M; Cambronero, M

    1998-02-01

    A retrospective epidemiological analysis is presented of 80 snakebites in children admitted to the Pediatrics Service of Tony Facio Hospital in Limón, Costa Rica, between 1985 and 1995. An overall incidence rate of 20 snakebite accidents per 100,000 inhabitants per year was calculated for the region in that period. However, a higher incidence was described in some counties (36 and 30 cases per 100,000 inhabitants in Talamanca and Siquirres, respectively). Most of the cases occurred in February and November, between 16:00 and 19:00 hr. The mean age of the affected children was 8.67 +/- 2.66 years. No significant difference was found between genders. Thirty-three children affected (41.2%) were members of the local indigenous groups and 31 (38.8%) were residents of Talamanca County. The species of snake responsible were identified in 58.4% of cases, with Bothrops asper being the most important. The lower extremities were the most important primary site affected (86.3%). The most common clinical complications described in the study group were abscess formation (11.25%), necrosis (10%), renal failure (3.75%), compartmental syndrome (2.5%) and anaphylactic shock (1.25%). The overall mortality was 2.5%. Statistical analysis of the risk factors relating to the two most common complications showed that the condition of coming from Talamanca County (P = 0.02), damage in the proximal extremities (P = 0.02), a prothrombin time serum fibrinogen levels serum fibrinogen levels < 100 g/dl were associated with development of necrosis. All of the patients with necrosis and abscesses also experienced two or more of the risk factors that correlated with a sensitivity of 100% and a specificity of 67% for the development of abscess, and 87% sensitivity and 88% specificity for necrotic complications. According to these data, snakebite complications are a relevant health problem in Costa Rica. PMID:9620583

  16. Philodryas patagoniensis bite and local envenoming Picada por Philodryas patagoniensis e envenenamento local

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    Sérgio de Andrade Nishioka

    1994-06-01

    Full Text Available 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.Um menino de 5 anos de idade foi picado por um espécime de Philodryas patagomensis, uma serpente colubrídea tida como não peçonhenta, tendo apresentado sinais de envenenamento local caracterizados por edema e calor. Esta é a primeira vez que se descreve que Philodryas patagoniensis pode causar envenenamento local. Com base nos achados clínicos e na identificação errônea da serpente, o paciente foi tratado como vítima de acidente botrópico, tendo recebido antiveneno específico. São necessários esforços para proporcionar a médicos e outros profissionais da área de saúde informações para o reconhecimento de serpentes peçonhentas, o que evitaria o uso inapropriado de antiveneno e seus potenciais efeitos adversos. Exame do local da picada pode ser útil para o diagnóstico diferencial entre serpentes da subfamilia Crotalinae (peçonhentas e da familia Colubridae.

  17. Purification and characterization of a metalloproteinase, Porthidin-1, from the venom of Lansberg's hog-nosed pitvipers (Porthidium lansbergii hutmanni).

    Science.gov (United States)

    Girón, María E; Estrella, Amalid; Sánchez, Elda E; Galán, Jacob; Tao, W Andy; Guerrero, Belsy; Salazar, Ana M; Rodríguez-Acosta, Alexis

    2011-03-15

    Porthidium lansbergii hutmanni is a small pit viper found on Margarita Island, Venezuela. Local tissue damage is one of the most obvious characteristics of P. l. hutmanni envenomation, which can lead to diverse pathological effects, such as hemorrhage, edema, blistering, necrosis, lymphatic vessel damage and degradation of extracellular matrix. Metalloproteinases are one of the major components in venoms responsible for these effects. To date, very little is known or has been reported on P. l. hutmanni venom. Crude P. l. hutmanni venom had a LD(50) of 2.5 mg/kg and was considered very hemorrhagic (minimal hemorrhagic dose [MHD]: 0.98 μg) when compared to other hemorrhagic (Bothrops) venoms in Venezuela. Crude P. l. hutmanni venom also inhibited ADP-induced platelet aggregation. A metalloproteinase, Porthidin-1, from this venom was isolated by three chromatography steps (Sephadex G100, Superose 12 HR10/30 and Bioscale Q2). Porthidin-1 falls in the SVMP P-I class having a molecular weight of 23 kDa, verified by both SDS-PAGE and mass spectrometry. High-resolution mass spectrometry and a database search identified a peptide from Porthidin-1 (YNGDLDK) belonging to the SVMP family of proteins. Porthidin-1 contained hemorrhagic, fibrino(geno)lytic, caseinolytic and gelatinolytic activities, and these activities were capable of being neutralized by metalloproteinase inhibitors but not serine proteinase inhibitors. The peptide YNGDLDK shared similarities with five venom proteins with a BLAST e-value of <1. This work details the biochemical and pathophysiological effects that can result from envenomations, and highlights the importance and significance for characterizing unknown or poorly documented venoms from different geographical regions.

  18. Venoms, toxins and derivatives from the Brazilian fauna: valuable sources for drug discovery.

    Science.gov (United States)

    De Marco Almeida, Flávia; de Castro Pimenta, Adriano Monteiro; Oliveira, Mônica Cristina; De Lima, Maria Elena

    2015-06-25

    Animal venoms have been widely investigated throughout the world. The great number of biotechnological articles as well as patent applications in the field of drug discovery based on these compounds indicates how important the source is. This review presents a list of the most studied Brazilian venomous animal species and shows the most recent patent applications filed from 2000 to 2013, which comprise Brazilian venoms, toxins and derivatives. We analyze the data according to the species, the type of products claimed and the nationality of the inventors. Fifty-five patent applications were found, involving 8 genera. Crotalus, Lachesis, Bothrops and Loxosceles represented 78% of the patent applications. The other 22% were represented by Phoneutria, Tityus, Acanthoscurria and Phyllomedusa. Most of the inventions (42%) involved anticancer, immunomodulator or antimicrobial drugs, while 13% involved anti-venoms and vaccines, 11% involved hypotensive compositions, 9% involved antinociceptive and/or anti-inflammatory compositions, and the other 25% involved methods, kits or compositions for various purposes. Brazilian inventors filed 49% of the patent applications, but other countries, mainly the United States of America, Germany, Russia and France, also filed patent applications claiming products comprising venoms, toxins and/or derivatives from the Brazilian fauna. Brazil holds an important number of patent applications which mostly belong to universities and research institutes, but the pharmaceutical industry in this field is still weak in Brazil. Although, Brazilian venomous animal species have been reported in drug discovery throughout the world, many species remain to be explored as valuable and promising tools for drug discovery and development.

  19. Inhibition of hemorrhagic and edematogenic activities of snake venoms by a broad-spectrum protease inhibitor, murinoglobulin; the effect on venoms from five different genera in Viperidae family.

    Science.gov (United States)

    Ribeiro Filho, Wilker; Sugiki, Masahiko; Yoshida, Etsuo; Maruyama, Masugi

    2003-08-01

    In order to obtain basic data on the effect of broad-spectrum protease inhibitor against local symptoms of Viperidae snake envenomation, inhibitory capacity of rat murinoglobulin on local hemorrhagic and edematogenic activities of venoms from Crotalus atrox, Bothrops jararaca, Lachesis muta muta, Trimeresurus flavoviridis and Echis carinatus sochureki were examined. Murinoglobulin, pre-incubated with the crude venoms at 37 degrees C for 15 min, inhibited hemorrhagic activity of all five venoms to various extents. The activity of C. atrox was almost completely inhibited at the murinoglobulin/venom ratio (w/w) of 20. The activity of B. jararaca, Lachesis muta muta and T. flavoviridis venoms was considerably inhibited at the ratio of 20 (77.2, 80.0 and 86.2% inhibition, respectively), however some of the activity still remained even at the ratio of 40 (84.2, 79.8 and 86.2% inhibition, respectively). Among the five venoms, E. c. sochureki venom is quite resistant to murinoglobulin treatment and statistically significant inhibition was only found at the ratio of 40 (64.1% inhibition). Fibrinolytic and gelatinase activities were more susceptible to murinoglobulin inhibition. The treatment at the ratios of 10 and 20 almost completely inhibited respectively the fibrinolytic and the gelatinase activities of all the venoms. Murinoglobulin treatment also significantly inhibited the edematogenic activity of L. muta muta, T. flavoviridis and Echis carinatus sochureki. The treatment of murinoglobulin at the ratio of 40 considerably suppressed the swelling up to 60 min after subcutaneous injection of L. muta muta and E. c. sochureki venoms, and up to 30 min after T. flavoviridis venom injection. Murinoglobulin is a potent inhibitor against local effects of multiple snake venoms in Viperidae family. PMID:12906888

  20. Comparison of indirect and direct approaches using ion-trap and Fourier transform ion cyclotron resonance mass spectrometry for exploring viperid venom proteomes.

    Science.gov (United States)

    Fox, Jay W; Ma, Li; Nelson, Kristina; Sherman, Nicholas E; Serrano, Solange M T

    2006-05-01

    In a sense, the field of snake venom proteomics has been under investigation since the very earliest biochemical studies where it was soon recognized that venoms are comprised of complex mixtures of bioactive molecules, most of which are proteins. Only with the re-emergence of 2D polyacrylamide gel electrophoresis (2D PAGE) and the recent developments in mass spectrometry for the identification/characterization of proteins coupled with venom gland transcriptomes has the field of snake venom proteomics began to flourish and provide exciting insights into the protein composition of venoms and subsequently their pathological activities. In this manuscript we will briefly discuss the state of snake venom proteomics followed by the presentation of several straightforward experiments designed to explore approaches to investigating venom proteomics. The first set of experiments used 1D gel electrophoresis (1D PAGE) of Crotalus atrox venom followed by slice-by-slice analysis of the proteins using liquid chromatography/mass spectrometry/mass spectrometry (LC/MS/MS). In the second set of experiments, C. atrox and Bothrops jararaca venoms were subjected to in-solution digestion followed by Fourier transform ion cyclotron resonance (FTICR) LC/MS/MS. The peptide ion-maps of these venoms were compared along with the proteins identified. In addition, the results were compared to the results observed from the 1D PAGE approach. From these studies it is clear that sample de-complexation/fractionation before mass spectrometry is still the best approach for maximum proteome coverage. Furthermore, comparison of venom proteomes based on tryptic peptide identities between the proteomes is not particularly effective since there does not appear to be a sufficient number of such identical peptides, derived from related proteins, present in venoms. Finally, as has previously been recognized without either better databases of venom protein sequences or facile and rapid de novo sequencing

  1. Effects of batroxobin with continuous transcranial Doppler monitoring in patients with acute cerebral stroke: a randomized controlled trial.

    Science.gov (United States)

    Yitao, He; Kefu, Ma; Bingshan, Tang; Xuejun, Fu; Ying, Zhan; Zhili, Cai; Xin, Jiang; Guo, Yi

    2014-11-01

    Our objective was to determine whether continuous transcranial Doppler (TCD) monitoring could safely enhance the efficacy of batroxobin, a thrombin-like enzyme extracted from Bothrops atrox moojeni venom, in the treatment for acute cerebral stroke beyond the thrombolytic time window. Ninety patients suffering an acute cerebral stroke were recruited into the study within 12 hours after the onset of symptoms. Patients were randomized to receive batroxobin with (target group) or without 1 hour of continuous TCD monitoring (control group). Clinical evaluation of stroke was based on the National Institutes of Health Stroke Scale (NIHSS) score, Barthel index (BI), Thrombolysis in Brain Ischemia score (TIBI), the incidence of advancing stroke, and the recurrence of cerebral infarction. The patients receiving continuous TCD monitoring showed significant improvement in NIHSS score at 57 days post treatment compared with the control. Similarly, patients receiving continuous TCD monitoring also showed significant improvement in BI at 3 months compared with the controls. Consistently, both the incidence of advancing stroke after 1 week and the incidence of stroke recurrence after 3 months were significantly lower in TCD monitored group than control group. Moreover, the safety of the employment of TCD monitoring in the treatment of these patients was confirmed as there was no significant difference of the incidence of intracranial hemorrhage at 1 week after the treatment between the target and control groups. Taken together, our study showed that batroxobin, in combination with continuous TCD monitoring at the middle cerebral artery, reduced the incidence of advancing stroke and stroke recurrence after treatment without adverse effects in terms of poststroke intracranial hemorrhage. PMID:24684297

  2. Batroxobin binds fibrin with higher affinity and promotes clot expansion to a greater extent than thrombin.

    Science.gov (United States)

    Vu, Trang T; Stafford, Alan R; Leslie, Beverly A; Kim, Paul Y; Fredenburgh, James C; Weitz, Jeffrey I

    2013-06-01

    Batroxobin is a thrombin-like serine protease from the venom of Bothrops atrox moojeni that clots fibrinogen. In contrast to thrombin, which releases fibrinopeptide A and B from the NH2-terminal domains of the Aα- and Bβ-chains of fibrinogen, respectively, batroxobin only releases fibrinopeptide A. Because the mechanism responsible for these differences is unknown, we compared the interactions of batroxobin and thrombin with the predominant γA/γA isoform of fibrin(ogen) and the γA/γ' variant with an extended γ-chain. Thrombin binds to the γ'-chain and forms a higher affinity interaction with γA/γ'-fibrin(ogen) than γA/γA-fibrin(ogen). In contrast, batroxobin binds both fibrin(ogen) isoforms with similar high affinity (Kd values of about 0.5 μM) even though it does not interact with the γ'-chain. The batroxobin-binding sites on fibrin(ogen) only partially overlap with those of thrombin because thrombin attenuates, but does not abrogate, the interaction of γA/γA-fibrinogen with batroxobin. Furthermore, although both thrombin and batroxobin bind to the central E-region of fibrinogen with a Kd value of 2-5 μM, the α(17-51) and Bβ(1-42) regions bind thrombin but not batroxobin. Once bound to fibrin, the capacity of batroxobin to promote fibrin accretion is 18-fold greater than that of thrombin, a finding that may explain the microvascular thrombosis that complicates envenomation by B. atrox moojeni. Therefore, batroxobin binds fibrin(ogen) in a manner distinct from thrombin, which may contribute to its higher affinity interaction, selective fibrinopeptide A release, and prothrombotic properties. PMID:23612970

  3. 皖南尖吻蝮蛇毒中类去整合蛋白/富含半胱氨酸两结构域cDNA的克隆和表达%Cloning,Expression and Characteristics of Disintegrin-like/cysteine-rich Domains cDNA from Agkistrodon acutus Venom

    Institute of Scientific and Technical Information of China (English)

    刘庆都; 张家琳; 承新; 刘爱平; 刘兢

    2000-01-01

    为了探讨出血毒金属蛋白酶结构功能关系,通过RT-PCR方法,从皖南尖吻蝮蛇(Agkistrodon acutus)毒腺总RNA中扩增得到编码P-Ⅲ型出血毒金属蛋白酶的完整类去整合蛋白和富含半胱氨酸两个结构域cDNA(AA/DC).它全长964 bp的cDNA,开放阅读框架编码216个氨基酸残基,序列比较分析表明它同来自Bothrops jararaca的jararhagin-C、来自Crotalus atrox的catrocollastatin-C有很高的同源性.在类去整合蛋白结构域中,Ser-Glu-Cys-Asp(SECD)代替了去整合蛋白中相应部位的Arg-Gly-Asp(RGD)三肽序列.将编码区基因克隆入pGEX-2T栽体中,转化大肠杆菌TG-1,用IPTG诱导表达,表达产物具有抑制胶原诱导的血小板凝集活性,但不抑制ADP诱导的血小板凝集.该研究为进一步阐述蛇毒金属蛋白酶结构功能关系和药物开发奠定了基础.

  4. Epidemiologia do acidente por serpentes peçonhentas: estudo de casos atendidos em 1988 Epidemiology of poisonons snake-bites: a study of cases assisted in 1988

    Directory of Open Access Journals (Sweden)

    Lindioneza Adriano Ribeiro

    1995-10-01

    Full Text Available Foram avaliados aspectos epidemiológicos de acidentes por serpentes peçonhentas no Estado de São Paulo, Brasil, com base em prontuários de 322 pacientes e em entrevistas feitas com 209 deles e/ou seus acompanhantes. Os acidentes ocorreram principalmente com pessoas de 10 a 20 anos de idade, do sexo masculino, nos meses de outubro a abril e no período diurno. As regiões anatômicas mais freqüentemente picadas foram os pés, as mãos e as pernas. Bothrops, Crotalus e Micrurus foram responsáveis por, respectivamente, 95,0%, 4,4% e 0,6% dos casos. Não ocorreram óbitos, mas 2,2% dos pacientes apresentaram seqüelas. Dentre os 209 entrevistados, a ocupação de lavrador foi a mais freqüentemente relacionada ao acidente que, em aproximadamente 60% das vezes, ocorreu durante o trabalho. O total de 160 pacientes (76,6% submeteram-se a alguma forma de tratamento antes de chegarem a um serviço de saúde: foram mais comuns o uso de torniquete (50,2%, a expressão local na tentativa de retirar parte do veneno (33,5%, a colocação das mais diversas substâncias sobre o local da picada (36,8% e a ingestão de outras (12,9%; pouco mais de um quarto dos pacientes submeteram-se a alguma forma de tratamento médico antes de chegar ao HVB sendo mais comum a antissepsia (8,2%, a administração do antiveneno (6,2%, de anti-histamínicos (5,7% e de analgésicos (5,3%.The attendance given to patients by ta specialized Hospital, in S.Paulo, Brazil, during 1988 is studied. The study is based on the medical records of 322 patients and on questionnaires filled out by author during interviews with 209 patients or their companions. The 322 snake-bites occurred mainly between October and April, in the diurnal period, mainly in the afternoon. Most of patients were adult males, mainly between 10 and 20 years of age. The parts of the body most fequently affected were the feet, hands and legs. The snakes of the genera Bothrops, Crotalus and Micrurus were responsible

  5. Bites by the colubrid snake Philodryas patagoniensis: a clinical and epidemiological study of 297 cases.

    Science.gov (United States)

    de Medeiros, Carlos R; Hess, Priscila L; Nicoleti, Alessandra F; Sueiro, Leticia R; Duarte, Marcelo R; de Almeida-Santos, Selma M; França, Francisco O S

    2010-11-01

    We retrospectively analyzed 297 proven cases of Philodryas patagoniensis bites admitted to Hospital Vital Brazil (HVB), Butantan Institute, São Paulo, Brazil, between 1959 and 2008. Only cases in which the causative animal was brought and identified were included. Part of the snakes brought by the patients was still preserved in the collection maintained by the Laboratory of Herpetology. Of the 297 cases, in 199 it was possible to describe the gender of the snake, and seventy three (61.3%) of them were female. The length of snakes (snout-vent length) ranged from 160 to 1080 mm. In 117 snakes their state of preservation enabled the dissection and examination of their stomach contents. The stomach was empty in 106 snakes (89.1%). Most bites occurred in the seasons of spring and summer (n = 196, 66.0%) and during warmer periods of the day. The mean age of the victims was 24.1 +/- 15.1 years old and 206 (69.4%) patients were men. Around 92% of the patients sought medical care within 6 h after the bite. Both lower (n = 188, 63.3%) and upper limbs (n = 102, 34.3%) were most frequently bitten, especially the feet and hands (n = 205, 69.0%). The local clinical manifestations were pain (n = 151, 50.8%), transitory bleeding (n = 106, 35.7%), erythema (n = 47, 15.8%) and edema (n = 39, 13.1%). Ecchymosis was not observed. Only 7 (2.4%) patients reported systemic symptoms characterized by mild dizziness and 88 patients (29.6%) showed no evidence of envenoming. The whole blood clotting time was performed in 76 (25.6%) patients on admission and all of them had coagulable blood. Supportive treatment was offered to only 13.4% of patients, namely administration of antihistamines (n = 19, 6.4%) and analgesics (n = 12, 4.1%). Eight patients (2.7%) were mistreated with Bothrops antivenom before their admission to HVB. No sequels or relevant complications were observed in patients, and the prognostic was benign. Therefore, although P. patagoniensis accidents can cause mild local

  6. EPIDEMIOLOGIA DOS ACIDENTES COM ANIMAIS PEÇONHENTOS REGISTRADOS EM NOVA XAVANTINA – MT

    Directory of Open Access Journals (Sweden)

    Vivian Tallita Pinheiro Santana

    2015-11-01

    Full Text Available Justificativa e objetivo: No Brasil os acidentes com animais peçonhentos constituem um problema de Saúde Pública, principalmente em municípios do interior do país. Informações regionais atualizadas são importantes para o desenvolvimento de ações de vigilância epidemiológica. Através desta pesquisa determinaram-se os aspectos epidemiológicos dos acidentes com animais peçonhentos registrados no município de Nova Xavantina, Mato Grosso, entre janeiro de 2007 e maio de 2013. Método: Foi realizado um estudo epidemiológico transversal, não probabilístico. Resultados: No período estudado foram registrados 68 acidentes, destes 44 ocasionados por serpentes, 19 por escorpiões e quatro por aranhas. A maioria dos acidentes com serpentes foram provocadas pelos gêneros Bothrops (79,5%, Crotalus (11,5% e Micrurus (2,3%. Quanto às vítimas, prevaleceram indivíduos do sexo masculino (76,5%, na faixa etária entre 30 a 49 anos (41,2%, de raça parda (72,1%, com ensino fundamental (55,9% envolvidos em atividades do campo como agricultura e pecuária (41,2%. Predominaram os acidentes de gravidade moderada (48,5% e todos os casos graves foram provocados por serpentes. Na maioria dos agravos (75,0%, não foi registrada relação entre o acidente e a atividade ocupacional. As regiões corpóreas mais atingidas foram pé (39,7%, perna (20,6% e mão (11,8%. As manifestações locais predominantes foram dor, edema e equimose. As manifestações sistêmicas de maior ocorrência foram hemorrágicas e vagais. Conclusão: Portanto, o período avaliado foi caracterizado pelo predomínio de acidentes com serpentes, no sexo masculino, em atividades do campo e de gravidade moderada. Assim, campanhas educativas e a adoção de medidas de prevenção são fundamentais para diminuir o registro desses agravos neste município. DESCRITORES: Envenenamento; Acidente ofídico; Picadas de escorpião.

  7. Acidente ofídico no estado de Goiás

    Directory of Open Access Journals (Sweden)

    Pinho Fábia Maria Oliveira

    2004-01-01

    Full Text Available OBJETIVO: Avaliar aspectos epidemiológicos dos acidentes por serpe