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Sample records for crotalus durissus collilineatus

  1. A novel peptide from the ACEI/BPP-CNP precursor in the venom of Crotalus durissus collilineatus.

    Science.gov (United States)

    Higuchi, Shigesada; Murayama, Nobuhiro; Saguchi, Ken-ichi; Ohi, Hiroaki; Fujita, Yoshiaki; da Silva, Nelson Jorge; de Siqueira, Rodrigo José Bezerra; Lahlou, Saad; Aird, Steven D

    2006-10-01

    In crotaline venoms, angiotensin-converting enzyme inhibitors [ACEIs, also known as bradykinin potentiating peptides (BPPs)], are products of a gene coding for an ACEI/BPP-C-type natriuretic peptide (CNP) precursor. In the genes from Bothrops jararaca and Gloydius blomhoffii, ACEI/BPP sequences are repeated. Sequencing of a cDNA clone from venom glands of Crotalus durissus collilineatus showed that two ACEIs/BPPs are located together at the N-terminus, but without repeats. An additional sequence for CNP was unexpectedly found at the C-terminus. Homologous genes for the ACEI/BPP-CNP precursor suggest that most crotaline venoms contain both ACEIs/BPPs and CNP. The sequence of ACEIs/BPPs is separated from the CNP sequence by a long spacer sequence. Previously, there was no evidence that this spacer actually coded any expressed peptides. Aird and Kaiser (1986, unpublished) previously isolated and sequenced a peptide of 11 residues (TPPAGPDVGPR) from Crotalus viridis viridis venom. In the present study, analysis of the cDNA clone from C. d. collilineatus revealed a nearly identical sequence in the ACEI/BPP-CNP spacer. Fractionation of the crude venom by reverse phase HPLC (C(18)), and analysis of the fractions by mass spectrometry (MS) indicated a component of 1020.5 Da. Amino acid sequencing by MS/MS confirmed that C. d. collilineatus venom contains the peptide TPPAGPDGGPR. Its high proline content and paired proline residues are typical of venom hypotensive peptides, although it lacks the usual N-terminal pyroglutamate. It has no demonstrable hypotensive activity when injected intravenously in rats; however, its occurrence in the venoms of dissimilar species suggests that its presence is not accidental. Evidence suggests that these novel toxins probably activate anaphylatoxin C3a receptors.

  2. cDNA and deduced primary structure of basic phospholipase A2 with neurotoxic activity from the venom secretion of the Crotalus durissus collilineatus rattlesnake

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    F.H.R. Fagundes

    2010-03-01

    Full Text Available To illustrate the construction of precursor complementary DNAs, we isolated mRNAs from whole venom samples. After reverse transcription polymerase chain reaction (RT-PCR, we amplified the cDNA coding for a neurotoxic protein, phospholipase A2 D49 (PLA2 D49, from the venom of Crotalus durissus collilineatus (Cdc PLA2. The cDNA encoding Cdc PLA2 from whole venom was sequenced. The deduced amino acid sequence of this cDNA has high overall sequence identity with the group II PLA2 protein family. Cdc PLA2 has 14 cysteine residues capable of forming seven disulfide bonds that characterize this group of PLA2 enzymes. Cdc PLA2 was isolated using conventional Sephadex G75 column chromatography and reverse-phase high performance liquid chromatography (RP-HPLC. The molecular mass was estimated using matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF mass spectrometry. We tested the neuromuscular blocking activities on chick biventer cervicis neuromuscular tissue. Phylogenetic analysis of Cdc PLA2 showed the existence of two lines of N6-PLA2, denominated F24 and S24. Apparently, the sequences of the New World’s N6-F24-PLA2 are similar to those of the agkistrodotoxin from the Asian genus Gloydius. The sequences of N6-S24-PLA2 are similar to the sequence of trimucrotoxin from the genus Protobothrops, found in the Old World.

  3. Afibrinogenemia following snake bite (Crotalus durissus terrificus

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    C. F. S. Amaral

    1988-08-01

    Full Text Available This paper reports two cases of afibrinogenemia with normal platelet count following Crotalus durissus terrificus, snake bite Both patients presented high output acute renal failure and case two also had increased blood levels of CPK and LDH compatible with the diagnosis of rhabdomyolysis. Case one was given an unknown amount of antivenom and was treated with epsilonaminocaproic acid and a fresh whole blood transfusion and showed recovery of the coagulation disturbance 40 hours following these measures. Case two was given an adequate amount of crotalide antivenom and the coagulation tests performed 12 hours later showed a normal partial thromboplastin time and fibrinogen 86 mg/100ml. Case one presented no haemorrhagic disturbances. Case two presented persistent bleeding following venopuncture and after removal of impetigo crust in the legs. Acute renal failure was treated conservatively and both patients were discharged from the hospital with recovery of the renal function.

  4. Toxins from Crotalus durissus terrificus venom

    International Nuclear Information System (INIS)

    Rogero, J.R.

    1979-01-01

    Phospholipase A, crotapotin and crotamine were purified from the venom of crotalus durissus terrificus. Molecular weigths were respectively estimated in 13400, 8300 and 4880. By sephadex gel filtration, stable complex is formed by interaction (1:1 molar ratio) of Phospholipase A (LD=0,55mg/Kg, in mice) and crotapotin, obtaining an increase in the toxicity of Phospholipase A in 10 fold to LD 50 =0,05mg/Kg. These facts indicate crotoxin as a product of 1:1 molar ratio interaction of crotapotin and Phospholipase A. By sephadex gel filtration an unstable complex is formed by interaction of crotapotin and crotamine (LD 50 =0,8mg/Kg, in mice), increasing the toxicity of crotamine approximately 4 fold to LD 50 =0,2mg/Kg. In tritium-hydrogen exchange experiments, the back exchange kinetic of these tritium labelled proteins were measured in gel filtration columns of sephadex G 25 'coarse' showing for Phospholipase A two clearly distinguishable kinetic classes of exchangeable hydrogens. From the exchangeable hydrogens only 68% were rapidly exchanged and the occurrence of hydrogens envolved in alpha-helix was practically absent. Crotapotin has no hydrogens of alpha-helix and 83% of exchangeable hydrogens were rapidly exchanged with solvent. Crotamine, after a initial heating in tritiated water showed that 31% of exchangeable hydrogens were slowly exchanged with solvent. After 18 hours of heating, that number diminished for 11%. Crotoxin showed three exponential classes of exchangeable hydrogens and about 26 protons have alpha-helix characteristic exchange rate. A possible conformational change after Phospholipase A - crotapotin interaction is suggested. (Author) [pt

  5. Effects of gamma radiation on Crotalus durissus terrificus venom

    International Nuclear Information System (INIS)

    Murata, Y.

    1988-01-01

    A poll of crotomine positive Crotalus durissus terrificus venom was dissolved in 0.15 M NaCl and the supernatant irradiated using 60 CO. Doses of 100,250,750,1000,1500 and 2000 Cy were used at a dose rate of 1.190Gy/h. The presence of free SH, casein hydrolytic degradation, SDS-PAGE and molecular exclusion chromatography analysis together with LD 50 determination in mice were studied. The antigenic properties of samples were investigated by immunodiffision and immunoprecipitation. (M.A.C.) [pt

  6. Is the population of Crotalus durissus (Serpentes, Viperidae) expanding in Brazil?

    OpenAIRE

    Duarte, Marcelo Ribeiro; Menezes, Frederico Alcântara

    2013-01-01

    Crotalus durissus are found from Mexico to northern Argentina in a highly disjunct distribution. According to some studies, this species is prone to occupy areas disturbed by human activities and floods comprise a plausible method of dispersal as inferred for some North American rattlesnakes. Based on the literature, it seems plausible that Crotalus durissus expanded their natural distribution in Brazil due to floods, but only in a few municipalities in Rio de Janeiro State. Data entries of B...

  7. DSC analysis of irradiated proteins from Crotalus durissus terrificus

    International Nuclear Information System (INIS)

    Oliveira, Karina Corleto de; Silva, Monica Nascimento da; Goncalves, Karina de Oliveira; Spencer, Patrick Jack; Nascimento, Nanci do

    2011-01-01

    Full text: Snake bites are a serious public health problem, especially in subtropical countries. In Brazil, the serum, the only effective treatment in case of snake bites, is produced in horses which, despite the large size, have a reduced lifespan due to the high toxicity of the antigen. It is known that ionizing radiation effects - direct and indirect - can modify the molecular structure, affecting the biological properties of proteins. Ionizing radiation has been employed to attenuate the toxicity of snake venoms, aiming to generate an improved antigen with low toxicity. Two proteins, purified from Crotalus durissus terrificus (Cdt) venom were tested in this work: crotoxin and crotamine. Crotoxin, the main toxic compound of Cdt venom, is a heterodimeric protein composed of two subunits: crotapotin and phospholipase A2. Crotamine is a highly basic polypeptide (pI - 10.3), with myotoxic activity and molecular weight of 4882 Da. It is composed of 42 amino acids residues and reticulated by three disulfide bonds. This study aimed to investigate the effects of radiation on crotoxin and crotamine using Differential Scanning Calorimetry (DSC). After isolation of the toxins by chromatographic techniques, they were irradiated with 2.0 kGy from 60 Co source. The thermodynamics analysis, carried out in a METTLER TOLEDO, DSC 822e calorimeter, showed that irradiation promoted changes of the calorimetric profile. These changes suggest that, although radiation induced structural modifications of the protein, denaturation was only partial, since transition states could still be detected, suggesting that some structural elements were still present after irradiation. Taken together, our data suggest that following irradiation, the molecules underwent conformational changes, and that the remaining structural elements displayed a lower enthalpy, clearly indicating that the previously described loss of toxicity of irradiated toxins can be mostly ascribed to structural changes

  8. DSC analysis of irradiated proteins from Crotalus durissus terrificus

    Energy Technology Data Exchange (ETDEWEB)

    Oliveira, Karina Corleto de; Silva, Monica Nascimento da; Goncalves, Karina de Oliveira; Spencer, Patrick Jack; Nascimento, Nanci do [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)

    2011-07-01

    Full text: Snake bites are a serious public health problem, especially in subtropical countries. In Brazil, the serum, the only effective treatment in case of snake bites, is produced in horses which, despite the large size, have a reduced lifespan due to the high toxicity of the antigen. It is known that ionizing radiation effects - direct and indirect - can modify the molecular structure, affecting the biological properties of proteins. Ionizing radiation has been employed to attenuate the toxicity of snake venoms, aiming to generate an improved antigen with low toxicity. Two proteins, purified from Crotalus durissus terrificus (Cdt) venom were tested in this work: crotoxin and crotamine. Crotoxin, the main toxic compound of Cdt venom, is a heterodimeric protein composed of two subunits: crotapotin and phospholipase A2. Crotamine is a highly basic polypeptide (pI - 10.3), with myotoxic activity and molecular weight of 4882 Da. It is composed of 42 amino acids residues and reticulated by three disulfide bonds. This study aimed to investigate the effects of radiation on crotoxin and crotamine using Differential Scanning Calorimetry (DSC). After isolation of the toxins by chromatographic techniques, they were irradiated with 2.0 kGy from {sup 60}Co source. The thermodynamics analysis, carried out in a METTLER TOLEDO, DSC 822e calorimeter, showed that irradiation promoted changes of the calorimetric profile. These changes suggest that, although radiation induced structural modifications of the protein, denaturation was only partial, since transition states could still be detected, suggesting that some structural elements were still present after irradiation. Taken together, our data suggest that following irradiation, the molecules underwent conformational changes, and that the remaining structural elements displayed a lower enthalpy, clearly indicating that the previously described loss of toxicity of irradiated toxins can be mostly ascribed to structural changes

  9. The Evolutionary Implications of Hemipenial Morphology of Rattlesnake Crotalus durissus terrificus (Laurent, 1768 (Serpentes: Viperidae: Crotalinae.

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    Marcovan Porto

    Full Text Available Most amniotes vertebrates have an intromittent organ to deliver semen. The reptile Sphenodon and most birds lost the ancestral penis and developed a cloaca-cloaca mating. Known as hemipenises, the copulatory organ of Squamata shows unique features between the amniotes intromittent organ. They are the only paired intromittent organs across amniotes and are fully inverted and encapsulated in the tail when not in use. The histology and ultrastructure of the hemipenes of Crotalus durissus rattlesnake is described as the evolutionary implications of the main features discussed. The organization of hemipenis of Crotalus durissus terrificus in two concentric corpora cavernosa is similar to other Squamata but differ markedly from the organization of the penis found in crocodilians, testudinata, birds and mammals. Based on the available data, the penis of the ancestral amniotes was made of connective tissue and the incorporation of smooth muscle in the framework of the sinusoids occurred independently in mammals and Crotalus durissus. The propulsor action of the muscle retractor penis basalis was confirmed and therefore the named should be changed to musculus hemipenis propulsor.The retractor penis magnus found in Squamata has no homology to the retractor penis of mammals, although both are responsible for the retraction of the copulatory organ.

  10. Partial characterization of the venom of the Peruvian rattlesnake Crotalus durissus terrificus

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    César Remuzgo

    2014-06-01

    Full Text Available The venom of the rattlesnake Crotalus durissus terrificus from the region of Sandia, Puno, has been investigated for its protein content and some enzymatic activities, using for it the whole venom as well as the fractions obtained by gel filtration chromatography in Sephadex G-100. The protein percentage calculated by the method of Lowry was of 68,6% for the whole venom; 3 peaks were obtained during the fractionation; the first showed proteolytic activity, the second, amidolytic, clotting and phospholipase A2 activities, while the third, another proteolytic activity. Acetylcholinesterase activity was not found while L-aminoacid oxidase activity was found only in the whole venom.

  11. The adrenergic retulation of the cardiovascular system in the South American rattlesnake, Crotalus durissus

    DEFF Research Database (Denmark)

    Galli, G.L.J.; Jensen, Nini Skovgaard; Abe, A.S.

    2007-01-01

    The present study investigates adrenergic regulation of the systemic and pulmonary circulations of the anaesthetised South American rattlesnake, Crotalus durissus. Haemodynamic measurements were made following bolus injections of adrenaline and adrenergic antagonists administered through a systemic...... arterial catheter. Adrenaline caused a marked systemic vasoconstriction that was abolished by phentolamine, indicating this response was mediated through α-adrenergic receptors. Injection of phentolamine gave rise to a pronounced vasodilatation (systemic conductance (Gsys) more than doubled), while...... injection of propranolol caused a systemic vasoconstriction, pointing to a potent α-adrenergic, and a weaker β-adrenergic tone in the systemic vasculature of Crotalus. Overall, the pulmonary vasculature was far less responsive to adrenergic stimulation than the systemic circulation. Adrenaline caused...

  12. Studies of agregates produced during venom irradiation of Crotalus durissus terrificus with gamma ray

    International Nuclear Information System (INIS)

    Clissa, Patricia B.; Nascimento, Nanci do; Rogero, Jose R.

    1997-01-01

    Literature data show that 2.0 kGy dose of gamma radiation, generated by 60 Co source, reduces the toxic activity of Crotalus durissus terrificus venon, without altering its immunogenic capacity. When crotoxin, main toxin from crotalic venom, was irradiated with the same dose, toxicity was laos reduced and the immunogenicity was maintained. This fact was attributed to aggregates(compounds with high molecular weight generated during irradiation), that showed no toxicity but were able to induce the antibodies formation against native venom. Crotalus durissus terrificus venom was irradied with 2.0, 3.0, 5.0 and 10.0 kGy doses and submitted to molecular exclusion chromatography, in order to find an efficient dose that produces large amounts of non toxic but still immunogeneic aggregates. After being isolated, the products of irradiation were evaluated for the amount produced, molecular ateration, and toxic and immunogenic activities. The results from different doses irradiated venom were compared with native one, and 2.0 kGg dose was confirmed to be most efficient in the association of toxicity attenuation with maintenance of immunogenicity of the crotalic venom, while other doses, in spite of being efficient in the toxicity attenuation, they were not able to keep the immunogenicity property. So, the dose of 2.0 kGy could be used to immunize animals in order to improve anticrotalic sera production. (author). 14 refs., 6 figs., 4 tabs

  13. Gamma radiation effects on crotoxin (toxin of crotalus durissus terrificus venom)

    International Nuclear Information System (INIS)

    Souza Filho, J.N.; Rogero, J.R.

    1988-01-01

    The crotoxin is a great neurotoxin found on Crotalus durissus terrificus venom. This protein was isolated using molecular exclusion cromatography with Sephadex G-75 and irradiated in a source of 60 Co GAMMA-CELL in the concentration of 2 mg/ml 0.85% NaCl with dose rate of 1.19 x 10 3 Gy/hr. The doses used were 250, 500, 1000 and 2000 Gy. It was determinated for this samples, the proteic concentration, the diffusion immunoassay using crotalic antiserum and eletrophoresis (SDS-PAGE). The results showed some changes on the irradiated toxin. Preliminary results with doses of radiation of 100, 250, 500 and 1000 Gy showed that the letal dose 50% (LD50) in mice increase greatly with the increase of radiation's dose. (author) [pt

  14. Purification and radiodination of gyroxin, toxin like trombin, of crotalus durissus terrificus venom

    International Nuclear Information System (INIS)

    Camillo, Maria A.P.; Paes, Paulo C.A.; Ribela, Maria T.C.P.; Rogero, Jose R.

    1997-01-01

    Snake's venoms have attracted special interest in research because they are rich in bioactive compounds, which are scientific tools to study many physiological processes and are also important source of new therapeutic agents. Some thrombin-like enzymes are used as a reagent in laboratorial assays as well as in medical drugs. However, a neurotoxic syndrome called gyroxin syndrome or barrel rotation seems to be related to those enzymes. Pharmacokinetics and biodistribution studies of these toxins can contribute to clarifly this mechanisms and consequently, it will help to elaborate safer clinical prescriptions giving more information about possible nocive effects. This paper describes the first step of these studies with the thrombin-like enzyme (or gyroxin) from Crotalus durissus terrificus's venom. The purification and radioiodination methods as well as electrophoretic analysis and biological assay are presented. (author). 9 refs., 5 figs., 1 tab

  15. Effects of gamma radiation on crotoxin (toxin of Crotalus durissus terrificus venom)

    International Nuclear Information System (INIS)

    Souza Filho, J.N.; Rogero, J.R.

    1988-07-01

    The crotoxin in a great neurotoxin found on Crotalus durissus terrificus venom. This protein was isolated using molecular exclusion cromatrography with Sephadex G-75 and irradiated in a source of 60 Co GAMMACELL in the concentration of 2 mg/ml 0.85% NaCl with dose rate of 1.19 x 10 3 Gy/hr. The doses used were 250, 500, 1000 and 2000 Gy. It was determinated for this samples, the proteic concentration, the diffusion immunoassay using crotalic antiserum and eletrophoresis (SDS-PAGE). The results showed some changes on the irradiated toxin. Preliminary results with doses of radiation of 100, 250, 500 and 1000 Gy showed that the letal dose 50% (LD50) in mice increase greatly with the increase of radiation's dose. (author) [pt

  16. Daily and seasonal activity patterns of free range South-American rattlesnake (Crotalus durissus

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    ALEXANDRO M. TOZETTI

    2013-09-01

    Full Text Available This study aimed at describing daily and seasonal variation in the activity of a population of South-American rattlesnakes (Crotalus durissus in a savanna like habitat (Cerrado in Southeastern Brazil. Seasonal and daily activities of snakes were evaluated by the number of captures of snakes during road surveys, accidental encounters, and relocations by radio-tracking. Our results show that climatic variables such as air temperature and rainfall have little influence on the activity pattern of rattlesnakes. Our findings indicate that rattlesnakes spend most of the day resting and most of the night in ambush posture. The South-American rattlesnake is active throughout the year with a discrete peak in activity of males during the matting season. The possibility of maintaining activity levels even during the coldest and driest season can facilitate the colonization of several habitats in South America. This possibility currently facilitates the colonization of deforested areas by rattlesnakes.

  17. Daily and seasonal activity patterns of free range South-American rattlesnake (Crotalus durissus).

    Science.gov (United States)

    Tozetti, Alexandro M; Martins, Marcio

    2013-09-01

    This study aimed at describing daily and seasonal variation in the activity of a population of South-American rattlesnakes (Crotalus durissus) in a savanna like habitat (Cerrado) in Southeastern Brazil. Seasonal and daily activities of snakes were evaluated by the number of captures of snakes during road surveys, accidental encounters, and relocations by radio-tracking. Our results show that climatic variables such as air temperature and rainfall have little influence on the activity pattern of rattlesnakes. Our findings indicate that rattlesnakes spend most of the day resting and most of the night in ambush posture. The South-American rattlesnake is active throughout the year with a discrete peak in activity of males during the matting season. The possibility of maintaining activity levels even during the coldest and driest season can facilitate the colonization of several habitats in South America. This possibility currently facilitates the colonization of deforested areas by rattlesnakes.

  18. Effects of radiolysis products on crotamine (toxin from Crotalus durissus terrificus venom)

    International Nuclear Information System (INIS)

    Costa, T.A. da; Rogero, J.R.

    1988-07-01

    Ionizing Radiations changes the molecular structure due to chemical bond destruction. These chemical alterations are able to change the biological properties of the biomolecules. Crotamine was obtained from Crotalus durissus terrificus venom by molecular exclusion cromatography and irradiated in a concentration of 1 mg/ml in aqueous solution with gamma radiation produced by a 60 Co source. We used doses of 100 Gy, 2000 Gy and 5000 Gy (dose rate = 1,14 x 10 3 Gy/h). We performed the following experiments: proteic concentration, SDS-PAGE and immunodiffusion. Preliminary results showed a reduction of 55% in proteic concentration; with dose of 5000 Gy; an increase of the number of bands in SDS-PAGE suggesting the appearance of protein aggregates that was proportional as the dose increases. The immunodifusion data showed a reduction of the immunochemical activity against the Butantan antisera. (author) [pt

  19. Interaction study of water radiolysis products with Crotalus durissus terrificus miotoxin

    International Nuclear Information System (INIS)

    Silva, Murilo Casare da

    2008-01-01

    Ionizing radiation has been satisfactorily employed for venoms detoxification. In this report, the radiation was employed to verify the effects caused by the radiolysis products of water on the Crotamine, toxin purified from Crotalus durissus terrificus venom. These effects were analyzed using some substances called 'scavengers', those substances competes for specific reactive species hindering them to act on the toxins molecules. In order to study the possible structural damages caused on the toxins, circular dichroism, fluorescence, nuclear magnetic resonance, amino acids analysis and intravital microscopy were employed. Our results indicate that ionizing radiation caused structure alterations, mainly, in secondary and tertiary structure of crotamine. In the irradiated crotamine, was not possible to determine tridimensional structure. And the crotamine toxic effect was removed by ionizing radiation. (author)

  20. Geographic distribution and apparent decline of Crotalus durissus terrificus (Laurenti, 1768; Serpentes, Viperidae) in Uruguay

    OpenAIRE

    Prigioni, Carlos M.; Borteiro, Claudio; Kolenc, Francisco; Colina, Marcelo; González, Enrique M.

    2013-01-01

    The rattlesnake Crotalus durissus terrificus (Laurenti, 1768; Serpentes, Viperidae) is a rare species in Uruguay, where it reaches the southern boundaries of its distribution. The most recent distribution map available for this species in Uruguay is based on a few specimens that were collected during the 1950s and 1960s. Herein, we make a comprehensive account of rattlesnake records in this country obtained from herpetological literature and other bibliographic sources, specimens accessioned ...

  1. Isolation and characterization of delta toxin from the venom of Crotalus durissus terrificus; Isolamento e caracterizacao da delta toxina do veneno de Crotalus durissus terrificus

    Energy Technology Data Exchange (ETDEWEB)

    Campos, Lucelia de Almeida

    2006-07-01

    The Crotalus durissus terrificus venom has been so far described as being of low complexity, with four major components described: convulxin, gyroxin, crotoxin and crotamine. In recent studies, other components of this venom were characterized as, for example, an analgesic factor. In 1980, Vital Brazil predicted the existence of a toxin which could be involved in platelet aggregation, and named it delta toxin. However, this toxin has never been isolated or characterized. The aim of the present work was to purify and characterize this toxin. After FPLC size exclusion chromatography followed by reverse phase HPLC, an homogeneous fraction was obtained, with a molecular weight of 14,074.92 Da. When analyzed by SOS-PAGE, this toxin presented an anomalous behavior, with a molecular weight of 14 kDa, while in 2D gels, spots around 40 kDa and with an isoelectrical point between 4 and 5 were observed suggesting isoforms with glicosilation microheterogeneity. After trypsin digestion, the fragments were submitted to the swissprot databank showing high homology (43% coverage, 15 matching peptides) with trocarin, a prothrombin activator from Tropidechis carinatus. These data were further confirmed by aminoacid analysis. The toxin was tested for its ability to activate factor II and X using synthetic substrates. Our data indicate a direct activation of factor X. The same toxin also behaved as a potent direct platelet aggregation activator on washed platelets. Assays with specific inhibitors indicate that neither metalloproteinase, nor serinoproteinase or t lectin domains are involved in the aggregating activity, since EDTA, benzamidin and D-galactose did not inhibit the toxin. In the present work, we were able to identify, purify and characterize a new toxin from the brazilian rattlesnake. It behaved as predicted by Vital-Brazil and displayed direct factor X activating properties, also inducing platelet aggregation, even at low concentrations. Our data also indicate that it is

  2. Biochemical and biological evaluation of gyroxin isolated from Crotalus durissus terrificus venom

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    LC Barros

    2011-01-01

    Full Text Available Gyroxin, a thrombin-like enzyme isolated from Crotalus durissus terrificus venom and capable of converting fibrinogen into fibrin, presents coagulant and neurotoxic activities. The aim of the present study was to evaluate such coagulant and toxic properties. Gyroxin was isolated using only two chromatographic steps - namely gel filtration (Sephadex G-75 and affinity (Benzamidine Sepharose 6B - resulting in a sample of high purity, as evaluated by RP-HPLC C2/C18 and electrophoretic analysis that showed a molecular mass of 30 kDa. Gyroxin hydrolyzed specific chromogenic substrates, which caused it to be classified as a serine proteinase and thrombin-like enzyme. It was stable from pH 5.5 to 8.5 and inhibited by Mn²+, Cu²+, PMSF and benzamidine. Human plasma coagulation was more efficient at pH 6.0. An in vivo toxicity test showed that only behavioral alterations occurred, with no barrel rotation. Gyroxin was not able to block neuromuscular contraction in vitro, which suggests that its action, at the studied concentrations, has no effect on the peripheral nervous system.

  3. Occurrence of Cryptosporidium (Apicomplexa, Cryptosporidiidae in Crotalus durissus terrificus (Serpentes, Viperidae in Brazil

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    Karasawa Andréa Satie Matsubara

    2002-01-01

    Full Text Available The objective of the present study was to investigate the prevalence of Cryptosporidium (Apicomplexa, Cryptosporidiidae in the snake Crotalus durissus terrificus (Serpentes, Viperidae. Fifty animals were evaluated for the presence of oocysts of Cryptosporidium sp. at the time of arrival and 30 and 60 days later. Intestinal washings with saline solution (1% body weight, fecal samples, and organ scrapings were collected during the study. Oocysts were concentrated by an ether-phosphate-buffered saline sedimentation technique and then separated by a density gradient centrifugation technique. Smears were made with the sediment and submitted to modified acid-fast and auramine-rhodamine staining. Cryptosporidium-positive smears were used as controls for the experimental findings. The overall prevalence of Cryptosporidium sp. oocysts was 14%. Among the positive snakes, oocysts were detected only in the intestinal washing in two specimens, only in the feces in four specimens, and in both materials at least once in one specimen. The positive snakes were predominantly from Santa Maria da Serra city State of São Paulo (57.1%. We also observed that all of the examinations that presented positive results were obtained at least 27 days after the capture of the animals.

  4. Pulmonary mechanic and lung histology induced by Crotalus durissus cascavella snake venom.

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    Oliveira Neto, Joselito de; Silveira, João Alison de Moraes; Serra, Daniel Silveira; Viana, Daniel de Araújo; Borges-Nojosa, Diva Maria; Sampaio, Célia Maria Souza; Monteiro, Helena Serra Azul; Cavalcante, Francisco Sales Ávila; Evangelista, Janaina Serra Azul Monteiro

    2017-10-01

    This study have analyzed the pulmonary function in an experimental model of acute lung injury, induced by the Crotalus durissus cascavella venom (C. d. cascavella) (3.0 μg/kg - i.p), in pulmonary mechanic and histology at 1 h, 3 h, 6 h, 12 h and 24 h after inoculation. The C. d. cascavella venom led to an increase in Newtonian Resistance (R N ), Tissue Resistance (G) and Tissue Elastance (H) in all groups when compared to the control, particularly at 12 h and 24 h. The Histeresivity (η) increased 6 h, 12 h and 24 h after inoculation. There was a decrease in Static Compliance (C ST ) at 6 h, 12 h and 24 h and inspiratory capacity (IC) at 3 h, 6 h, 12 h and 24 h. C. d. cascavella venom showed significant morphological changes such as atelectasis, emphysema, hemorrhage, polymorphonuclear inflammatory infiltrate, edema and congestion. After a challenge with methacholine (MCh), R N demonstrated significant changes at 6, 12 and 24 h. This venom caused mechanical and histopathological changes in the lung tissue; however, its mechanisms of action need further studies in order to better elucidate the morphofunctional lesions. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Gross and histologic features of gastritis due to Ophidascaris arndti in tropical rattlesnakes ( Crotalus durissus

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    É.M. Mello

    Full Text Available ABSTRACT The tropical rattlesnake (Crotalus durissus is a snake of great importance for biomedical industry since its poison is used for the production of antiophidic serum and researches. Several conditions related to animal health, such as diseases and parasites, which can promote the reduction of poison production by these snakes should be investigated. Accordingly, the aim of this study was to characterize the gross and microscopic lesions related to the presence of Ophidascaris arndti in stomachs of tropical rattlesnakes. The gastrointestinal tract of thirty-five South American rattlesnakes captured in Southeastern region of Brazil were analyzed and nineteen animals showed infestation by the parasites, found in the small intestine and, especially, in the stomach of the hosts. Grossly, lesions were characterized by mucosal ulcers occasionally associated with hemorrhage. Microscopic alterations included histiocytic granulomas, fibrosis, necrosis, and hemorrhage. Based on these findings, the diagnosis of a parasitic granulomatous disease was made. The lesions may be related to the cause of death in captivity snakes, since the lesions can promote secondary infections by opportunistic bacteria. Moreover, the intense inflammatory response accompanied by fibrosis may be related to poor functioning of the gastric snakes, which it may exhibit frequent regurgitation of the food.

  6. Pro- and Anti-Inflammatory Cytokines Release in Mice Injected with Crotalus durissus terrificus Venom

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    A. Hernández Cruz

    2008-01-01

    Full Text Available The effects of Crotalus durissus terrificus venom (Cdt were analyzed with respect to the susceptibility and the inflammatory mediators in an experimental model of severe envenomation. BALB/c female mice injected intraperitoneally presented sensibility to Cdt, with changes in specific signs, blood biochemical and inflammatory mediators. The venom induced reduction of glucose and urea levels and an increment of creatinine levels in serum from mice. Significant differences were observed in the time-course of mediator levels in sera from mice injected with Cdt. The maximum levels of IL-6, NO, IL-5, TNF, IL-4 and IL-10 were observed 15 min, 30 min, 1, 2 and 4 hours post-injection, respectively. No difference was observed for levels of IFN-γ. Taken together, these data indicate that the envenomation by Cdt is regulated both pro- and anti-inflammatory cytokine responses at time-dependent manner. In serum from mice injected with Cdt at the two first hours revealed of pro-inflammatory dominance. However, with an increment of time an increase of anti-inflammatory cytokines was observed and the balance toward to anti-inflammatory dominance. In conclusion, the observation that Cdt affects the production of pro- and anti-inflammatory cytokines provides further evidence for the role played by Cdt in modulating pro/anti-inflammatory cytokine balance.

  7. Comprometimento respiratório secundário a acidente ofídico crotálico (Crotalus durissus Respiratory abnormalities following Crotalus durissus snakebite

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    Carlos Faria Santos Amaral

    1991-08-01

    Full Text Available São analisados três pacientes que apresentaram comprometimento da função respiratória após acidente por Crotalus durissus. As manifestações respiratórias surgiram nas primeiras 48 horas após a picada do ofídio e consistiram de dispnéia, taquipnéia, uso da musculatura acessória da respiração (casos 1 e 2 e batimento das aletas nasais (caso 2. Dois pacientes (casos 1 e 2 apresentaram insuficiência respiratória aguda. O diagnóstico desta complicação no caso 1 foi clínico pois o paciente apresentou apnéia. O paciente do caso 2, 24 horas após o acidente ofídico apresentou dificuldade respiratória intensa e períodos de apnéia sendo intubado, permanecendo em respiração espontânea. Houve agravamento dos sinais clínicos de insuficiência respiratória e a determinação de pH e gases do sangue arterial mostrou em relação ao exame inicial elevação da pressão parcial de gás carbônico (40 mmHg para 50,3 mm Hg caracterizando insuficiência ventilatória aguda. Ambos foram tratados com emprego de ventilação artificial mecânica, tendo o paciente do caso 1 permanecido no ventilador durante 33 dias e o do caso 2 durante 15 dias. Ambos desenvolveram insuficiência renal aguda, necessitaram de diálise peritoneal e recuperaram a função renal. A paciente do caso 3, apesar dos sintomas e sinais de comprometimento respiratório não apresentou alterações do pH e gases arteriais. Espirometria realizada 58 horas após o acidente mostrou capacidade vital forçada (CVF e volume espirado no primeiro segundo (VEF1 abaixo do previsto (60 e 67% respectivamente. As espirometrias realizadas nos dias subseqüentes evidenciaram melhora progressiva destes parâmetros. No 10º dia após o acidente constatou-se aumento de 20% da CVF e de 17% do VEF1 comparativamente ao exame inicial. A relação entre VEF1 e a CVF manteve-se praticamente inalterada e em valores próximos ao previsto, caracterizando distúrbio ventilatório do tipo

  8. Isolation and characterization of delta toxin from the venom of Crotalus durissus terrificus

    International Nuclear Information System (INIS)

    Campos, Lucelia de Almeida

    2006-01-01

    The Crotalus durissus terrificus venom has been so far described as being of low complexity, with four major components described: convulxin, gyroxin, crotoxin and crotamine. In recent studies, other components of this venom were characterized as, for example, an analgesic factor. In 1980, Vital Brazil predicted the existence of a toxin which could be involved in platelet aggregation, and named it delta toxin. However, this toxin has never been isolated or characterized. The aim of the present work was to purify and characterize this toxin. After FPLC size exclusion chromatography followed by reverse phase HPLC, an homogeneous fraction was obtained, with a molecular weight of 14,074.92 Da. When analyzed by SOS-PAGE, this toxin presented an anomalous behavior, with a molecular weight of 14 kDa, while in 2D gels, spots around 40 kDa and with an isoelectrical point between 4 and 5 were observed suggesting isoforms with glicosilation microheterogeneity. After trypsin digestion, the fragments were submitted to the swissprot databank showing high homology (43% coverage, 15 matching peptides) with trocarin, a prothrombin activator from Tropidechis carinatus. These data were further confirmed by aminoacid analysis. The toxin was tested for its ability to activate factor II and X using synthetic substrates. Our data indicate a direct activation of factor X. The same toxin also behaved as a potent direct platelet aggregation activator on washed platelets. Assays with specific inhibitors indicate that neither metalloproteinase, nor serinoproteinase or t lectin domains are involved in the aggregating activity, since EDTA, benzamidin and D-galactose did not inhibit the toxin. In the present work, we were able to identify, purify and characterize a new toxin from the brazilian rattlesnake. It behaved as predicted by Vital-Brazil and displayed direct factor X activating properties, also inducing platelet aggregation, even at low concentrations. Our data also indicate that it is

  9. Interaction study of water radiolysis products with Crotalus durissus terrificus miotoxin; Estudo das interacoes dos produtos de radiolise da agua com a miotoxina do veneno de Crotalus durissus terrificus

    Energy Technology Data Exchange (ETDEWEB)

    Silva, Murilo Casare da

    2008-07-01

    Ionizing radiation has been satisfactorily employed for venoms detoxification. In this report, the radiation was employed to verify the effects caused by the radiolysis products of water on the Crotamine, toxin purified from Crotalus durissus terrificus venom. These effects were analyzed using some substances called 'scavengers', those substances competes for specific reactive species hindering them to act on the toxins molecules. In order to study the possible structural damages caused on the toxins, circular dichroism, fluorescence, nuclear magnetic resonance, amino acids analysis and intravital microscopy were employed. Our results indicate that ionizing radiation caused structure alterations, mainly, in secondary and tertiary structure of crotamine. In the irradiated crotamine, was not possible to determine tridimensional structure. And the crotamine toxic effect was removed by ionizing radiation. (author)

  10. Experimental envenomation with Crotalus durissus terrificus venom in dogs treated with antiophidic serum - part II: laboratory aspects, electrocardiogram and histopathology

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    R. M. B. Nogueira

    2007-01-01

    Full Text Available The present work shows laboratory aspects, electrocardiogram and histopathology results during experimental envenomation by Crotalus durissus terrificus in dogs treated with antiophidic serum. Twenty-one dogs were divided into three groups of seven animals each. Group I received 1mg/kg venom (sc; Group II received 1mg/kg venom (sc, 50mg antiophidic serum (iv and fluid therapy including 0.9% NaCl solution (iv; and Group III received 1mg/kg venom (sc, 50mg antiophidic serum (iv and fluid therapy including 0.9% NaCl solution containing sodium bicarbonate diluted to the dose of 4mEq/kg. Urinalysis showed brown urine, proteinuria, occult blood and myoglobinuria. Respiratory acidosis and hypotension were also observed. At the venom inoculation site, there was discreet edema, popliteal lymph node response, musculature presenting whitish areas and necrotic myositis with myoregenerative activity. There was not evidence of electrocardiographical and biochemical alterations.

  11. Clinical and hematological alterations in dogs during experimental envenomation with Crotalus durissus terrificus venom and treated with antiophidic serum

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    R. M. B. Nogueira

    2006-04-01

    Full Text Available The present work aimed to evaluate the clinical and hematological aspects during experimental envenomation by Crotalus durissus terrificus in dogs treated with different antiophidic serum doses. Sixteen dogs were divided into two groups of eight animals each. Group I received 1mg/kg venom subcutaneously and 30mg antiophidic serum intravenously; Group II received 1mg/kg venom subcutaneously and 60mg antiophidic serum intravenously. In the clinical evaluation, we observed ataxia, moderate sedation, dilated pupils, sialorrhea, flaccid paralysis of mandibular muscles, and discreet edema at the site of venom inoculation. Evaluating red and white blood cells, we observed a decrease of hemoglobins, globular volume and erythrocytes, and an increase of plasmatic proteins, leukocytes, neutrophils, monocytes and lymphocytes. Clotting time increased and there was blood incoagulability with return to normal clotting time six hours after antiophidic serum administration. Animals treated with six antiophidic serum flasks had a faster recovery than the animals that received three serum flasks.

  12. Transferência do Hepatozoon tupinambis, parasita do lagarto Tupinambis teguixin, para a serpente cascavel (Crotalus durissus terrificus, por intermédio do mosquito Culex fatigans

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    Samuel B. Pessôa

    1974-01-01

    Full Text Available Os autores obtiveram a transferência do Hepatozoon tupinambis (Laveran e Salibeni, 1909 parasita do lagarto Teiidae, Tupinambis teguixin, L., para a serpente cascavel, Crotalus durissus terrificus (Laur., alimentando-a com mosquitos experimentalmente infectados. o parasita mantém os seus caracteres morfológicos no animal receptor, nos limites do tempo observado (cerca de 100 dias. O ofídio receptor apresentou cistos esquizogônicos do fígado.The authors achieved a transfer of the Hepatozoon tupinambis (Laveran and Salibeni, 1909, a parasite of the Sauria: Tupinambis teguixin, L., to a rattlesnake of the species Crotalus durissus terrificus, feeding the latter with experimentally infected mosquitoes. The parasite maintained its morphological characteristics in the recepting animal throughout the whole observation period (about 100 days. Schizogonic cysts were found in the liver of the rattlesnake.

  13. Comparison of wildlife and captivity rattlesnakes (Crotalus durissus terrificus microbiota Comparação da microbiota de cascavéis (Crotalus durissus terrificus de vida-livre e cativeiro

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    Rui S. Ferreira Junior

    2009-12-01

    Full Text Available The study evaluated and compared the aerobic microbiota from the oral cavity, cloaca and venom of Crotalus durissus terrificus snakes, recently caught from the wild and kept under quarantine (WQ, individual captivity (IC and collective captivity (CC. Antimicrobial drug effectiveness on isolated agents also was assayed. From group I, II and III were isolated, respectively, 29 (63.04%, 38 (90.48% and 21 (42.86% microorganisms from the cloaca; 15 (32.61%, 3 (7.14% and 25 (51.02% microorganisms from the oral cavity; and, 2 (4.35%, 1 (2.38% and 3 (6.12% microorganisms from venom. The most frequent bacteria were Pseudomonas aeruginosa, Proteus vulgaris and Morganella morganii, with sensitivity to amikacin, gentamicin, norfloxacin, sulfazotrin and tobramycin. Snakes kept in semi-open captivity exhibited the fewest microorganisms in oral cavities, perhaps due to the environment in captivity, with different temperature gradients, running water, absence of daily handling, circulating air, possibility of moving around, daily cleaning, and sunlight access.Este estudo avaliou e comparou a microflora aeróbica da cavidade oral, cloaca e veneno de serpentes Crotalus durissus terrificus recém-capturadas da natureza e mantidas sob quarentena (WQ, mantidas em cativeiro coletivo (CC e em cativeiro individual (IC. A eficácia de drogas antimicrobianas de agentes isolados foi também avaliada. Foram isolados microorganismos dos grupos I, II e III respectivamente: 29 (63.04%, 38 (90.48% e 21 (42.86% da cloaca; 15 (32.61%, 3 (7.14% e 25 (51.02% da cavidade oral, e finalmente 2 (4.35%, 1 (2.38% e 3 (6.12% do veneno. As bactérias mais frequentes foram Pseudomonas aeruginosa, Proteus vulgaris e Morganella morganii, com sensibilidade para amikacina, gentamicina, norfloxacina, sulfazotrina e tobramicina. Serpentes mantidas no cativeiro semi-aberto mostraram menor número de agentes infecciosos em cavidade oral, talvez devido ao ambiente de cativeiro com diferentes

  14. 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

  15. Crotoxin, the major toxin from the rattlesnake Crotalus durissus terrificus, inhibits ³H-choline uptake in guinea pig ileum

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    L.S. Kattah

    2000-09-01

    Full Text Available We examined the effect of crotoxin, the neurotoxic complex from the venom of the South American rattlesnake Crotalus durissus terrificus, on the uptake of ³H-choline in minces of smooth muscle myenteric plexus from guinea pig ileum. In the concentration range used (0.03-1 µM and up to 10 min of treatment, crotoxin decreased ³H-choline uptake by 50-75% compared to control. This inhibition was time dependent and did not seem to be associated with the disruption of the neuronal membrane, because at least for the first 20 min of tissue exposure to the toxin (up to 1 µM the levels of lactate dehydrogenase (LDH released into the supernatant were similar to those of controls. Higher concentrations of crotoxin or more extensive incubation times with this toxin resulted in elevation of LDH activity detected in the assay supernatant. The inhibitory effect of crotoxin on ³H-choline uptake seems to be associated with its phospholipase activity since the equimolar substitution of Sr2+ for Ca2+ in the incubation medium or the modification of the toxin with p-bromophenacyl bromide substantially decreased this effect. Our results show that crotoxin inhibits ³H-choline uptake with high affinity (EC25 = 10 ± 5 nM. We suggest that this inhibition could explain, at least in part, the blocking effect of crotoxin on neurotransmission.

  16. Comparison of the effect of Crotalus simus and Crotalus durissus ruruima venoms on the equine antibody response towards Bothrops asper venom: implications for the production of polyspecific snake antivenoms.

    Science.gov (United States)

    Dos-Santos, Maria Cristina; Arroyo, Cynthia; Solano, Sergio; Herrera, María; Villalta, Mauren; Segura, Alvaro; Estrada, Ricardo; Gutiérrez, José María; León, Guillermo

    2011-02-01

    Antivenoms are preparations of immunoglobulins purified from the plasma of animals immunized with snake venoms. Depending on the number of venoms used during the immunization, antivenoms can be monospecific (if venom from a single species is used) or polyspecific (if venoms from several species are used). In turn, polyspecific antivenoms can be prepared by purifying antibodies from the plasma of animals immunized with a mixture of venoms, or by mixing antibodies purified from the plasma of animals immunized separately with single venom. The suitability of these strategies to produce polyspecific antibothropic-crotalic antivenoms was assessed using as models the venoms of Bothrops asper, Crotalus simus and Crotalus durissus ruruima. It was demonstrated that, when used as co-immunogen, C. simus and C. durissus ruruima venoms exert a deleterious effect on the antibody response towards different components of B. asper venom and in the neutralization of hemorrhagic and coagulant effect of this venom when compared with a monospecific B. asper antivenom. Polyspecific antivenoms produced by purifying immunoglobulins from the plasma of animals immunized with venom mixtures showed higher antibody titers and neutralizing capacity than those produced by mixing antibodies purified from the plasma of animals immunized separately with single venom. Thus, despite the deleterious effect of Crotalus sp venoms on the immune response against B. asper venom, the use of venom mixtures is more effective than the immunization with separate venoms for the preparation of polyspecific bothropic-crotalic antivenoms. Copyright © 2010 Elsevier Ltd. All rights reserved.

  17. Influência da temperatura corporal de cascavéis (Crotalus durissus submetidas à anestesia com cetamina Influence of body temperature on rattlesnakes (Crotalus durissus anesthetized with ketamine

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    Adriano B. Carregaro

    2009-12-01

    Full Text Available O estudo objetivou verificar a influência da temperatura corporal nos parâmetros fisiológicos e nos períodos de indução e recuperação anestésicos de cascavéis (Crotalus durissus anestesiadas com cetamina. Os animais foram previamente submetidos à hipotermia (HIPO (The aim of the study was to verify the influence of the body temperature under physiological values and latency and recovery times on rattlesnakes anesthetized with ketamine. The animals were previously submitted to hypothermia (HYPO (<22°C and normothermia (30°C (NORMO and then, anesthetized with 80 mg/kg IM of ketamine. Latency and recovery times were evaluated by head tonus, muscular tonus and righting reflex. Heart rate (HR, time of apnea and body temperature were measured before and 5, 10, 15, 30, 60, 90 and 120 minutes after ketamine administration. Blood gases parameters were measured before, 30 and 60 minutes. It was not observed difference on latency time in both groups. The recovery time was higher on HYPO (5,5 hours compared to NORMO (3,5 hours. HR was higher on NORMO compared to HYPO. Time of apnea was the same pattern on both groups. Compared to basal levels, time of apnea was shorter between 5 to 30 min on both groups. Respiratory acidosis was observed only at 0 min in NORMO. SvO2 was higher after 30 min, the same as with PvO2 in both groups. PvCO2 reduced after 30 min in both groups. It was evident that body temperature exerts intense influence on the recovery time on rattlesnakes anesthetized with ketamine.

  18. Aspectos morfológicos e ultraestruturais de células sanguíneas de Crotalus durissus terrificus

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    Lívia M. Kindlovits

    Full Text Available RESUMO: A avaliação hematológica, de importância comprovada como um meio auxiliar de diagnóstico ao clínico de pequenos animais domésticos, vem se tornando comum em animais selvagens não apenas para a clínica, mas para a avaliação do manejo e como estudo auxiliar para a fisiologia das várias espécies. Tendo em vista o aumento da demanda para a produção de várias drogas de importância farmacêutica, a criação de serpentes peçonhentas vem se tornando comum a ponto destes animais já serem reconhecidos como sendo de produção. O conhecimento do manejo e da clínica destes animais ainda é escasso e a mortalidade é elevada nos criatórios, tornando urgente a ampliação destes. Embora alguns estudos hematológicos já tenham sido realizados em cascavéis (Crotalus durissus os dados analisados ainda são insipientes, notadamente em relação à caracterização das células do sangue e poucos estudos em microscopia eletrônica foram realizados em serpentes. Com o objetivo de caracterizar as células sanguíneas morfologicamente, sob microscopia óptica e ultraestrutural, foram coletadas amostras de sangue de 52 de indivíduos da subespécie Crotalus durissus terrificus para a realização de esfregaços sanguíneos e avaliação ultraestrutural. Concluiu-se que a coloração hematológica de Giemsa permite a avaliação morfológica e a diferenciação das células sanguíneas em serpentes assim como a visualização de hemoparasitos. A avaliação ultraestrutural permite evidenciar as organelas celulares e a diferenciação entre as células, inclusive entre os tipos leucocitários, porém ainda são necessários outros estudos para que seja elucidada a hipótese da existência dos eosinófilos na espécie estudada assim como é necessária melhor caracterização dos grânulos dos azurófilos para que se confirme uma possível diferença entre os monócitos típicos e os azurófilos.

  19. Hematological changes in sheep inoculated with natural and Cobalt60-irradiated Crotalus durissus terrificus venom (Laurenti, 1768

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    D. P. Netto

    2004-01-01

    Full Text Available Natural (NV and Cobalto60-irradiated (IrV Crotalus durissus terrificus venom were used to evaluate serum production capacity of sheep and possible hematological and biochemical effects. Freeze-dried venom aliquots were diluted in acidified saline solution (NaCl 150 mM, pH 3.0 and irradiated by a Cobalt 60 source at a dose of 5.54 x 102 Gy/h and a concentration of 2.000 Gy. Twelve sheep were divided into two groups of six animals. One group received irradiated venom (IrV and the other natural venom (NV. Three antigen doses (venom were administered at monthly intervals. Blood samples were collected weekly for analysis of serum neutralization potency and capacity, complete blood count (CBC, total plasma protein, fibrinogen, albumin, and globulin. At the end of the experiment, the animals were challenged with a LD50 for sheep and showed no signs of envenoming. The two groups did not present clinical alterations. Results of the total leukocyte count did not present interaction or time factor effect for both groups, but there was a different action between them, with the NV group presenting more cells than the IrV group. The leukocyte increase to 13,000/ml indicates that slight leukocytosis occurred in the week after the first inoculation in the NV group. There was no statistically significant difference between groups in the absolute count of segmented neutrophils, eosinophils, and lymphocytes but there were statistically significant oscillations in values at the different collecting times. The NV group presented an increase in the absolute neutrophil count after the first inoculation that persisted for 5 weeks. In the IrV group, the increase in neutrophils occurred only in the first week returning to normal in the following weeks. The alterations in the neutrophil count are indicative of systemic inflammatory response related to cytokine release; response was more marked in the NV group, showing its greater toxicity.

  20. Hematological changes in sheep inoculated with natural and Cobalt60-irradiated Crotalus durissus terrificus venom (Laurenti, 1768)

    International Nuclear Information System (INIS)

    Netto, D.P.; Alfieri, A.A.; Balarim, M.R.S.; Chiacchio, S.B.; Bicudo, P.L.; Nascimento, N.

    2004-01-01

    Natural (NV) and Cobalt 60 -irradiated (IrV) Crotalus durissus terrificus venom were used to evaluate serum production capacity of sheep and possible hematological and biochemical effects. Freeze-dried venom aliquots were diluted in acidified saline solution (NaCl 150 m M, p H 3.0) and irradiated by a Cobalt 60 source at a dose of 5.54 x 102 Gy/h and a concentration of 2.000 Gy. Twelve sheep were divided into two groups of six animals. One group received irradiated venom (IrV) and the other natural venom (NV). Three antigen doses (venom) were administered at monthly intervals. Blood samples were collected weekly for analysis of serum neutralization potency and capacity, complete blood count, total plasma protein, fibrinogen, albumin, and globulin. At the end of the experiment, the animals were challenged with a LD 50 for sheep and showed no signs of envenoming. The two groups did not present clinical alterations. Results of the total leukocyte count did not present interaction or time factor effect for both groups, but there was a different action between them, with the NV group presenting more cells than the IrV group. The leukocyte increase to 13,000/μl indicates that slight leucocytosis occurred in the week after the first inoculation in the NV group. There was no statistically significant difference between groups in the absolute count of segmented neutrophils, eosinophils, and lymphocytes but there were statistically significant oscillations in values at the different collecting times. The NV group presented an increase in the absolute neutrophil count after the first inoculation that persisted for 5 weeks. In the IrV group, the increase in neutrophils occurred only in the first week returning to normal in the following weeks. The alterations in the neutrophil count are indicative of systemic inflammatory response related to cytokine release; response was more marked in the N V group, showing its greater toxicity. (author)

  1. EFECTO DEL AZUL DE METILENO EN LA HIPOTENSIÓN Y BRADICARDIA ASOCIADAS A LA ADMINISTRACIÓN DE UNA DOSIS LETAL DEL VENENO TOTAL DE CASCABEL VENEZOLANA (Crotalus durissus cumanensis) EN RATAS

    DEFF Research Database (Denmark)

    Briceno, Elena; Zerpa, Hector; Ascanio, Elias

    2010-01-01

    The effects of pre treatment with methylene blue (MB) on the cardiovascular effect caused by the administration of total venom (TV) of Crotalus durissus cumanensis was studied in adult rats, allocated into two groups: group I, six rats treated with TV (single dose: 1.5 mg/kg, IV) and group II, si...

  2. A new structurally atypical bradykinin-potentiating peptide isolated from Crotalus durissus cascavella venom (South American rattlesnake).

    Science.gov (United States)

    Lopes, Denise M; Junior, Norberto E G; Costa, Paula P C; Martins, Patrícia L; Santos, Cláudia F; Carvalho, Ellaine D F; Carvalho, Maria D F; Pimenta, Daniel C; Cardi, Bruno A; Fonteles, Manassés C; Nascimento, Nilberto R F; Carvalho, Krishnamurti M

    2014-11-01

    Venom glands of some snakes synthesize bradykinin-potentiating peptides (BPP's) which increase bradykinin-induced hypotensive effect and decrease angiotensin I vasopressor effect by angiotensin-converting enzyme (ACE) inhibition. The present study shows a new BPP (BPP-Cdc) isolated from Crotalus durissus cascavella venom: Pro-Asn-Leu-Pro-Asn-Tyr-Leu-Gly-Ile-Pro-Pro. Although BPP-Cdc presents the classical sequence IPP in the C-terminus, it has a completely atypical N-terminal sequence, which shows very low homology with all other BPPs isolated to date. The pharmacological effects of BPP-Cdc were compared to BBP9a from Bothrops jararaca and captopril. BPP-Cdc (1 μM) significantly increased BK-induced contractions (BK; 1 μM) on the guinea pig ileum by 267.8% and decreased angiotensin I-induced contractions (AngI; 10 nM) by 62.4% and these effects were not significantly different from those of BPP9a (1 μM) or captopril (200 nM). Experiments with 4-week hypertensive 2K-1C rats show that the vasopressor effect of AngI (10 ng) was decreased by 50 μg BPP-Cdc (69.7%), and this result was similar to that obtained with 50 μg BPP9a (69.8%). However, the action duration of BPP-Cdc (60 min) was 2 times greater than that of BPP-9a (30 min). On the other hand, the hypotensive effect of BK (250 ng) was significantly increased by 176.6% after BPP-Cdc (50 μg) administration, value 2.5 times greater than that obtained with BPP9a administered at the same doses (71.4%). In addition, the duration of the action of BPP-Cdc (120 min) was also at least 4 times greater than that of BPP-9a (30 min). Taken together, these results suggest that BPP-Cdc presents more selective action on arterial blood system than BPP9a. Besides the inhibition of ACE, it may present other mechanisms of action yet to be elucidated. Copyright © 2014 Elsevier Ltd. All rights reserved.

  3. Crotoxin from Crotalus durissus terrificus is able to down-modulate the acute intestinal inflammation in mice.

    Directory of Open Access Journals (Sweden)

    Caroline de Souza Almeida

    Full Text Available Inflammatory bowel diseases (IBD is the result of dysregulation of mucosal innate and adaptive immune responses. Factors such as genetic, microbial and environmental are involved in the development of these disorders. Accordingly, animal models that mimic human diseases are tools for the understanding the immunological processes of the IBD as well as to evaluate new therapeutic strategies. Crotoxin (CTX is the main component of Crotalus durissus terrificus snake venom and has an immunomodulatory effect. Thus, we aimed to evaluate the modulatory effect of CTX in a murine model of colitis induced by 2,4,6- trinitrobenzene sulfonic acid (TNBS. The CTX was administered intraperitoneally 18 hours after the TNBS intrarectal instillation in BALB/c mice. The CTX administration resulted in decreased weight loss, disease activity index (DAI, macroscopic tissue damage, histopathological score and myeloperoxidase (MPO activity analyzed after 4 days of acute TNBS colitis. Furthermore, the levels of TNF-α, IL-1β and IL-6 were lower in colon tissue homogenates of TNBS-mice that received the CTX when compared with untreated TNBS mice. The analysis of distinct cell populations obtained from the intestinal lamina propria showed that CTX reduced the number of group 3 innate lymphoid cells (ILC3 and Th17 population; CTX decreased IL-17 secretion but did not alter the frequency of CD4+Tbet+ T cells induced by TNBS instillation in mice. In contrast, increased CD4+FoxP3+ cell population as well as secretion of TGF-β, prostaglandin E2 (PGE2 and lipoxin A4 (LXA4 was observed in TNBS-colitis mice treated with CTX compared with untreated TNBS-colitis mice. In conclusion, the CTX is able to modulate the intestinal acute inflammatory response induced by TNBS, resulting in the improvement of clinical status of the mice. This effect of CTX is complex and involves the suppression of the pro-inflammatory environment elicited by intrarectal instillation of TNBS due to the

  4. Hematological changes in sheep inoculated with natural and Cobalt{sub 60}-irradiated Crotalus durissus terrificus venom (Laurenti, 1768)

    Energy Technology Data Exchange (ETDEWEB)

    Netto, D.P.; Alfieri, A.A.; Balarim, M.R.S. [Universidade Estadual de Londrina, PR (Brazil). Faculdade de Medicina Veterinaria e Zootecnia. Dept. de Medicina Veterinaria Preventiva]. E-mail: rnetto@uel.br; Chiacchio, S.B.; Bicudo, P.L. [UNESP, Botucatu, SP (Brazil). Faculdade de Medicina Veterinaria e Zootecnia; Nascimento, N. [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil). Supervisao de Radiobiologia

    2004-07-01

    Natural (NV) and Cobalt{sub 60}-irradiated (IrV) Crotalus durissus terrificus venom were used to evaluate serum production capacity of sheep and possible hematological and biochemical effects. Freeze-dried venom aliquots were diluted in acidified saline solution (NaCl 150 m M, p H 3.0) and irradiated by a Cobalt 60 source at a dose of 5.54 x 102 Gy/h and a concentration of 2.000 Gy. Twelve sheep were divided into two groups of six animals. One group received irradiated venom (IrV) and the other natural venom (NV). Three antigen doses (venom) were administered at monthly intervals. Blood samples were collected weekly for analysis of serum neutralization potency and capacity, complete blood count, total plasma protein, fibrinogen, albumin, and globulin. At the end of the experiment, the animals were challenged with a LD{sub 50} for sheep and showed no signs of envenoming. The two groups did not present clinical alterations. Results of the total leukocyte count did not present interaction or time factor effect for both groups, but there was a different action between them, with the NV group presenting more cells than the IrV group. The leukocyte increase to 13,000/{mu}l indicates that slight leucocytosis occurred in the week after the first inoculation in the NV group. There was no statistically significant difference between groups in the absolute count of segmented neutrophils, eosinophils, and lymphocytes but there were statistically significant oscillations in values at the different collecting times. The NV group presented an increase in the absolute neutrophil count after the first inoculation that persisted for 5 weeks. In the IrV group, the increase in neutrophils occurred only in the first week returning to normal in the following weeks. The alterations in the neutrophil count are indicative of systemic inflammatory response related to cytokine release; response was more marked in the N V group, showing its greater toxicity. (author)

  5. Avaliação da resposta inflamatória hematológica em cascavéis (Crotalus durissus Linnaeus, 1758 inoculadas com BCG Assessment of blood inflammatory response in BCG stimulated rattlesnakes (Crotalus durissus Linnaeus, 1758

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    Wellington Bandeira da Silva

    2009-12-01

    Full Text Available A criação de serpentes peçonhentas em cativeiro para produção de soros antipeçonhas possui crescente importância para a saúde pública devido ao aumento do número de notificações de acidentes ofídicos a cada ano no Brasil. Iniciado no século XX, ainda hoje essa atividade apresenta alguns desafios como a instalação de doenças no plantel. O hemograma é um exame de triagem clínica que auxilia no diagnóstico de diversas moléstias que acometem diferentes espécies de animais, no entanto ainda pouco estudado em serpentes. A caracterização das alterações hematológicas em cascavéis inoculadas experimentalmente com BCG pode servir de base na utilização deste exame no auxílio ao diagnóstico de infecções bacterianas na espécie. Dessa forma, foram realizados exames hematológicos em 10 serpentes da espécie Crotalus durissus pertencentes ao plantel da Divisão de Herpetologia do Instituto Vital Brazil. Os animais foram divididos em dois grupos (Grupos 1 e 2, homogêneos entre si em relação ao peso e proporção sexual. Os dois grupos foram inoculados com BCG e submetidos à coleta de sangue antes da inoculação e em três momentos pós-inoculação (3º, 5º, e 7º dias para o Grupo 1 e 11º, 17º e 21º dias para o Grupo 2. O hemograma foi realizado por método semidireto pela utilização de líquido de Natt e Herrick e as lâminas foram coradas pelo Giemsa. Observou-se anemia discreta, com redução dos valores de concentração de hemoglobina corpuscular média e da hemoglobina globular média no Grupo 1 que foi relacionada à doença inflamatória. A trombocitopenia observada no Grupo 2 sugeriu a atuação deste tipo celular em processos inflamatórios. Um único animal do Grupo 1 apresentou granulocitose e alguns animais apresentaram discreta azurofilia. Observaram-se alterações morfológicas nos leucócitos. Os granulócitos apresentaram granulações grosseiras e os azurófilos apresentaram aumento de tamanho e

  6. Cytoskeleton, endoplasmic reticulum and nucleus alterations in CHO-K1 cell line after Crotalus durissus terrificus (South American rattlesnake venom treatment

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    B. P. Tamieti

    2007-01-01

    Full Text Available Snake venoms are toxic to a variety of cell types. However, the intracellular damages and the cell death fate induced by venom are unclear. In the present work, the action of the South American rattlesnake Crotalus durissus terrificus venom on CHO-K1 cell line was analyzed. The cells CHO-K1 were incubated with C. d. terrificus venom (10, 50 and 100g/ml for 1 and 24 hours, and structural alterations of actin filaments, endoplasmic reticulum and nucleus were assessed using specific fluorescent probes and agarose gel electrophoresis for DNA fragmentation. Significant structural changes were observed in all analyzed structures. DNA fragmentation was detected suggesting that, at the concentrations used, the venom induced apoptosis.

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

    Science.gov (United States)

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

    2016-01-01

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

  8. The comparison between the humoral response and the neutralizing capacity of sheep serum inoculated with natural venom and Co60 irradiated venom from Crotalus durissus terrificus (Laurenti, 1768)

    International Nuclear Information System (INIS)

    Netto, D.P.

    2000-01-01

    Crotalus durissus terrificus venom was irradiated with Co 60 to investigate the effects of antigen-irradiation on antivenom production in sheep. Twelve sheep were divided in two groups of 6. One group received irradiated, while the other received natural venom. Three doses of antigen were given at monthly intervals. The toxic activity of the venom was assessed by LD 50 in mice. Weekly blood samples were obtained to evaluate anti-crotalic serum titers by indirect ELISA, neutralization capacity, and serum potency. A complete blood count, plasma protein and fibrinogen concentration, and serum albumin and globulin were also determined. At end of the experiment, the animals were challenged with ovine LD 50 , without clinical abnormalities. (author)

  9. Internal dosimetry of radiopharmaceuticals derived of antitumor polypeptide isolated from venoms: Crotalus durissus terrifucus and Scorpaena plumieri;Dosimetria interna de radiofarmacos derivados de polipeptideos antitumorais isolados dos venenos de: Crotalus durissus terrificus e Scorpaena plumieri

    Energy Technology Data Exchange (ETDEWEB)

    Andrade, Henrique Martins de

    2009-07-01

    The identification of new diagnostic and therapeutic agents capable of inhibiting tumor growth is essential for improving the prognosis of patients suffering from malignant tumors (glioma, breast and others). In this context, natural products (plants and animals) are a rich source of substances with potential antitumor. Despite knowledge of the etiology and pathology of tumors little progress has been observed in the area of diagnosis. Molecules of snake venoms have been shown to play an important role not only in the survival and proliferation of tumor cells but also in the process of tumor cell adhesion, migration and angiogenesis. Polypeptides isolated from the venom of the snake, Crotalus durissus terrificus, Crtx, and Scorpaena plumieri fish, SPGP, have antitumor activity against malignant tumors. It was shown that similar radio iodines Crtx and SPGP, {sup 125}I-Crtx and {sup 125}I-SPGP, can interact specifically with malignant tumors and induce cell death. Prototype-based radiopharmaceuticals Crtx and SPGP containing radioiodine 1311 were able to produce diagnostic images to accumulate specifically in the tumor site. The present study aimed at evaluating the potential radiological safety and diagnostic/therapeutic efficacy of {sup 131}I-Crtx {sup l31}I-SPGP and (evaluated from the biokinetic data in mice bearing Ehrlich tumor) were treated by the MIRD formalism to carry out internal dosimetry studies. Absorbed doses due to the uptake of {sup 131}I-Crtx and {sup 131}I-SPGP were determined in various organs of mice and implanted into the tumor. The results obtained for the animal model were extrapolated to humans by assuming a similar concentration ratio among the various tissues between mice and humans. In extrapolation, we used the masses of human organs of the phantom of Cristy/Eckerman. Both radiation penetrating and non penetrating of {sup 131}I on the tissue were considered in dose calculations. The absorbed dose in the bone marrow due to the

  10. Description of three new species of Hepatozoon (Apicomplexa, Hepatozoidae) from Rattlesnakes (Crotalus durissus terrificus) based on molecular, morphometric and morphologic characters.

    Science.gov (United States)

    O'Dwyer, Lucia Helena; Moço, Tatiana Cristina; Paduan, Karina dos Santos; Spenassatto, Carine; da Silva, Reinaldo José; Ribolla, Paulo Eduardo Martins

    2013-10-01

    Hepatozoon spp. are commonly found infecting snakes. Since the latter are parasitized by diverse forms and data in the literature show divergence, we studied Hepatozoon spp. diversity on Crotalus durissus terrificus snakes using both molecular and morphological approaches. Naturally infected animals were employed. Blood was collected, blood smears were prepared and an aliquot was stored at -20°C for DNA extraction. Five specimens of C. durissus terrificus were selected, each of them infected with one gamont type. Morphological and morphometric analyses of the found gamonts led to their grouping into three populations. For molecular characterization, seven oligonucleotide pairs that amplify distinct regions of rDNA gene were tested by adopting the PCR technique. Only the oligonucleotide pairs HepF300/Hep900 and HEMO1/HEMO2 were efficient in amplifying and distinguishing different isolates of Hepatozoon spp. from snakes. The better results were obtained when both oligonucleotide pairs were used in association. Based on the molecular and morphologic differences, three new species were proposed: Hepatozoon cuestensis sp. nov.; Hepatozoon cevapii sp. nov. and Hepatozoon massardii sp. nov. This is the first description of new Hepatozoon species from snakes, based on molecular characterization and morphological data, in South America. Copyright © 2013 Elsevier Inc. All rights reserved.

  11. Internal dosimetry of radiopharmaceuticals derived of antitumor polypeptide isolated from venoms: Crotalus durissus terrifucus and Scorpaena plumieri

    International Nuclear Information System (INIS)

    Andrade, Henrique Martins de

    2009-01-01

    The identification of new diagnostic and therapeutic agents capable of inhibiting tumor growth is essential for improving the prognosis of patients suffering from malignant tumors (glioma, breast and others). In this context, natural products (plants and animals) are a rich source of substances with potential antitumor. Despite knowledge of the etiology and pathology of tumors little progress has been observed in the area of diagnosis. Molecules of snake venoms have been shown to play an important role not only in the survival and proliferation of tumor cells but also in the process of tumor cell adhesion, migration and angiogenesis. Polypeptides isolated from the venom of the snake, Crotalus durissus terrificus, Crtx, and Scorpaena plumieri fish, SPGP, have antitumor activity against malignant tumors. It was shown that similar radio iodines Crtx and SPGP, 125 I-Crtx and 125 I-SPGP, can interact specifically with malignant tumors and induce cell death. Prototype-based radiopharmaceuticals Crtx and SPGP containing radioiodine 1311 were able to produce diagnostic images to accumulate specifically in the tumor site. The present study aimed at evaluating the potential radiological safety and diagnostic/therapeutic efficacy of 131 I-Crtx l31 I-SPGP and (evaluated from the biokinetic data in mice bearing Ehrlich tumor) were treated by the MIRD formalism to carry out internal dosimetry studies. Absorbed doses due to the uptake of 131 I-Crtx and 131 I-SPGP were determined in various organs of mice and implanted into the tumor. The results obtained for the animal model were extrapolated to humans by assuming a similar concentration ratio among the various tissues between mice and humans. In extrapolation, we used the masses of human organs of the phantom of Cristy/Eckerman. Both radiation penetrating and non penetrating of 131 I on the tissue were considered in dose calculations. The absorbed dose in the bone marrow due to the administration of 131 I-Crtx was 0.01 mGy/370

  12. Avaliação da interação de compostos ativos hidratantes com modelo de biomembrana de Crotalus durissus por meio de calorimetria exploratória diferencial e espectroscopia RAMAN

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    Telma Kaneko

    2010-06-01

    Full Text Available O objetivo foi verificar as alterações no estrato córneo em modelos alternativos de membrana após a aplicação de ativos hidratantes envolvendo métodos biofísicos. O modelo de biomembrana utilizado foi a muda de pele de Crotalus durissus e os ativos hidratantes foram: uréia, silício orgânico, extrato vegetal de Imperata cylindrica, reação de xilitol e glicose e componentes de NMF. Os resultados da avaliação das alterações do modelo por meio de Espectroscopia Raman com Transformada de Fourier sugerem que os ativos hidratantes confirmam segurança necessária, pois não alteraram de forma acentuada a estrutura do estrato córneo. Utilizando-se Calorimetria Exploratória Diferencial pode-se indicar que a solução de silício orgânico e o gel hidrofílico com uréia apresentaram melhor poder hidratante. Palavras-chave: Estrato córneo. Hidratação. Métodos biofísicos. Crotalus durissus. FT-Raman, DSC. ABSTRACT Study of the interaction of moisturizers with a Crotalus durissus biomembrane by differential scanning calorimetry and RAMAN spectroscopy The objective of this research was to use biophysical techniques to investigate the alterations induced in a biomembrane model of the stratum corneum by the application of moisturizers. The biomembrane was obtained from the skin shed by the rattlesnake Crotalus durissus and the active moisturizing compounds were: urea, dimethylsilanol hyaluronate, Imperata cylindrical plant extract, carbohydrates and natural moisturizing factors (NMF components. Results from FT-Raman spectroscopy suggested that the moisturizers were safe, since they did not promote modifications in the structure of the stratum corneum. Differential scanning calorimetry results indicated that the solution containing the organic silicon compound and the gel with urea showed the best hydrating effects on the stratum corneum. Keywords: Stratum corneum. Hydration. Biophysical techniques. Crotalus durissus. FT-Raman, DSC.

  13. Rabdomióilise secundária a acidente ofídico crotálico (Crotalus durissus terrificus

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    Renato Almeida Magalhães

    1986-08-01

    Full Text Available Relatam-se dois casos de rabdomiólise secundária a envenenamento produzido por Crotalus durissus terrificus. O diagnóstico da rabdomiólise baseou-se na mialgia intensa e generalizada apresentada pelos pacientes e na constatação de níveis séricos elevados de CPK, TGO e DHL. A confirmação do diagnóstico foi obtida no caso n.° 2 pela detecção de mioglobina no soro através de imunoeletroforese contra soro antimioglobina humana e por biópsia muscular. Esse paciente desenvolveu também, como complicação do envenenamento ofídico, quadro clínico e laboratorial de insuficiência renal aguda. Essa complicação foi atribuída à ação nefrotóxica e hemolítica do veneno crotálico e à hipotensão arterial apresentada pelo paciente, não se afastando a possibilidade de que a rabdomiólise tenha sido um fator contribuinte para a sua instalação. Foram constatadas hipocalcemla, hiperuricemia e hiperfosfatemia grave na fase oligúria da insuficiência renal aguda, alterações peculiares quando essa condição está associada à rabdomiólise.

  14. Direct organogenesis of Mandevilla illustris (Vell) Woodson and effects of its aqueous extract on the enzymatic and toxic activities of Crotalus durissus terrificus snake venom.

    Science.gov (United States)

    Biondo, R; Soares, A M; Bertoni, B W; França, S C; Pereira, A M S

    2004-03-01

    In order to produce explants of Mandevilla illustris (Vell) Woodson for the "Cerrado in vitro", the Germplasm Bank of UNAERP, we carried out a micropropagation protocol using MS or MS/3 medium supplemented with different concentrations of 6-benzyladeninepurine (BA), Zeatin or 2-isopentenyladenine for nodal segment growth, and alpha-naphthaleneacetic acid, indole-3-butyric acid (IBA) or 1,4 dithiothreitol for rooting. For nodal segments, all the cytokinins tested yielded similar results. However, 2.22 micro M BA is more economical to use. MS/3 medium supplemented with 0.49 micro M IBA was the most appropriate medium for rooting, resulting in 29% rooted explants. The crude aqueous extract from the subterranean system (SS) of M. illustris was assayed for its inhibitory action on the enzymatic activity of Crotalus durissus terrificus snake venom, isolated basic phospholipase A2 (CB) and crotoxin. It totally inhibited the phospholipase activity of crude Cdt venom and CB toxin and inhibited the phospholipase activity of crotoxin by 49%. The toxic action of both the crude venom and crotoxin was partially inhibited-there was a prolonged survival time and a 40.0% decrease in lethality.

  15. Phospholipase A2 isolated from the venom of Crotalus durissus terrificus inactivates dengue virus and other enveloped viruses by disrupting the viral envelope.

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    Vanessa Danielle Muller

    Full Text Available The Flaviviridae family includes several virus pathogens associated with human diseases worldwide. Within this family, Dengue virus is the most serious threat to public health, especially in tropical and sub-tropical regions of the world. Currently, there are no vaccines or specific antiviral drugs against Dengue virus or against most of the viruses of this family. Therefore, the development of vaccines and the discovery of therapeutic compounds against the medically most important flaviviruses remain a global public health priority. We previously showed that phospholipase A2 isolated from the venom of Crotalus durissus terrificus was able to inhibit Dengue virus and Yellow fever virus infection in Vero cells. Here, we present evidence that phospholipase A2 has a direct effect on Dengue virus particles, inducing a partial exposure of genomic RNA, which strongly suggests inhibition via the cleavage of glycerophospholipids at the virus lipid bilayer envelope. This cleavage might induce a disruption of the lipid bilayer that causes a destabilization of the E proteins on the virus surface, resulting in inactivation. We show by computational analysis that phospholipase A2 might gain access to the Dengue virus lipid bilayer through the pores found on each of the twenty 3-fold vertices of the E protein shell on the virus surface. In addition, phospholipase A2 is able to inactivate other enveloped viruses, highlighting its potential as a natural product lead for developing broad-spectrum antiviral drugs.

  16. Humoral response and neutralization capacity of sheep serum inoculated with natural and Cobalt 60-irradiated Crotalus durissus terrificus venom (Laurenti, 1768)

    International Nuclear Information System (INIS)

    Netto, D.P.; Alfieri, A.A.; Chiacchio, S.B.; Bicudo, P.L.; Nascimento, N.

    2002-01-01

    The aim of this work was to investigate antigen irradiation on crotalic antivenom and the capacity of sheep as serum producers. Twelve sheep in two groups of six were inoculated with Crotalus durissus terrificus venom. One group was inoculated with natural venom (N V) and the other with Cobalt 60 gamma-irradiated venom (Ir V). Three antigen doses were given to the animals at monthly intervals for immunization. The toxic activity of the venom was assessed by LD 50 determination in mice. Blood samples were collected weekly analyses of serum neutralization capacity and potency. At the end of the experiment, the animals were challenged with a LD 50 for sheep showed no signs of envenoming. These results showed that toxicity of the irradiated venom was 4.4 times less than the natural venom. The sera from the irradiated group neutralized LD 50 14.6 times, and the sera from the natural group 4.4 times. Sera from the irradiated group were five times more potent. The two groups did not present clinical alterations. The results of this study show the potential for using sheep in crotalic antivenom production. The use of irradiated venom in sheep immunization induces a powerful and lasting humoral immune response shown by both the in vitro neutralization and potency tests and by the indirect ELISA antibody level detection technique. (author)

  17. Effect of Mikania glomerata (Asteraceae leaf extract combined with anti-venom serum on experimental Crotalus durissus (Squamata: Viperidae envenomation in rats

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    Rafael Stuani Floriano

    2009-12-01

    Full Text Available Crotalic envenomation represents the highest number of deaths when compared to other snakebite envenomations of medical interest. Crotalic venom has important characteristics such as neurotoxicity, myotoxicity, nephrotoxicity, and clotting and hemolytic action. We evaluated the clinical and laboratory aspects of Crotalus durissus terrificus experimental envenomation in Wistar rats treated with antivenom and the aqueous extract of the plant Mikania glomerata. The animals were divided into three groups: Group C (control; Group VS-venom and antivenom; Group VSM-venom, antivenom and aqueous extract of M. glomerata. Crotalic poison caused clinical and laboratory alterations in Wistar mice. Significant linical alterations were: temperature decrease, edema in the venom inoculated member, sedation and a locomotion decrease in groups VS and VSM when compared with group C. A faster recovery from sedation was observed only for animals of group VSM when compared to VS. There was an increase in the number of leukocytes, neutrophils and creatine kinase in the VS and VSM groups, compared to group C. Wistar rats showed a high resistance to crotalic venom. Additional studies with different doses, time of treatment, different administration methods and histopathological and immunological studies are necessary to understand the action of M. glomerata in crotalic accidents. Rev. Biol. Trop. 57 (4: 929-937. Epub 2009 December 01.El envenamiento crotálico representa el número más alto de muertes cuando es comparado con envenenamientos por mordeduras de otras serpientes de interés médico. El veneno crotálico tiene importantes características de acción neurotóxica, miotoxicidad, nefrotoxicidad, coagulación y acción hemolítica. Este trabajo evaluó los aspectos clínicos y de laboratorio del envenenamiento experimental con el veneno de la serpiente Crotalus durissus terrificus en las ratas Wistar tratadas con suero antiofídico y extracto acuoso de M

  18. Snakebites by Crotalus durissus ssp in children in Campinas, São Paulo, Brazil Acidentes por serpentes Crotalus durisssus ssp em crianças em Campinas, São Paulo, Brasil

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    Fábio BUCARETCHI

    2002-01-01

    Full Text Available From January, 1984 to March, 1999, 31 children under 15 y old (ages 1-14 y, median 8 y were admitted after being bitten by rattlesnakes (Crotalus durissus ssp. One patient was classified as "dry-bite", 3 as mild envenoming, 9 as moderate envenoming and 18 as severe envenoming. Most patients had neuromuscular manifestations, such as palpebral ptosis (27/31, myalgia (23/31 and weakness (20/31. Laboratory tests suggesting rhabdomyolysis included an increase in total blood creatine kinase (CK, 28/29 and lactate dehydrogenase (LDH, 25/25 levels and myoglobinuria (14/15. The main local signs and symptoms were slight edema (20/31 and erythema (19/31. Before antivenom (AV administration, blood coagulation disorders were observed in 20/25 children that received AV only at our hospital (incoagulable blood in 17/25. AV early reactions were observed in 20 of these 25 cases (9/9 patients not pretreated and 11/16 patients pretreated with hydrocortisone and histamine H1 and H2 antagonists. There were no significant differences in the frequency of patients with AV early reactions between the groups that were and were not pretreated (Fisher's exact test, p = 0.12. Patients admitted less than and more than 6 h after the bite showed the same risk of developing severe envenoming (Fisher's exact test, p = 1. No children of the first group (De janeiro de 1984 a março de 1999, 31 crianças com menos de 15 anos de idade (1 a 14 anos, mediana = 8 anos foram admitidas após terem sido picadas por Crotalus durissus ssp. Uma criança não apresentou manifestações clínicas de envenenamento, enquanto 3 foram classificadas como acidente leve, 9 como moderado e 18 como grave. A maioria das crianças apresentou envolvimento neuromuscular, tais como ptose palpebral (27/31, mialgia (23/31 e fraqueza (20/31. Alterações laboratoriais sugerindo rabdomiólise também foram observadas, como aumento das enzimas séricas CK (28/29 e LDH (25/25 e mioglobinúria (14/15. As

  19. Rabbit IgG antibodies against Phospholipase A2 from Crotalus durissus terrificus neutralize the lethal activity of the venom Los anticuerpos IgG de conejos anti-fosfolipasa A2 de Crotalus durissus terrificus neutralizan la actividad letal del veneno

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    Juan P. Rodríguez

    2006-12-01

    Full Text Available Crotalus durissus terrificus (C.d.t. (South American rattlesnake venom possesses myotoxic and neurotoxic activities, both of which are also expressed by crotoxin, the principal toxin of this venom. Crotoxin contains a basic phospholipase A2 (PLA2 and a non toxic acidic protein, crotapotin. We have produced and investigated the ability of IgG antibodies raised in rabbits against PLA2 to neutralize the lethality of the whole venom. PLA2 was isolated by gel filtration chromatography (Sephadex G-75. Specific antibodies were obtained by subcutaneous and intramuscular inoculation of PLA2 (700 µg with Freund adjuvant. Groups of six mice (20 + 2 g were inoculated with 0.5 ml i.p. of C. d. t. venom (4 µg or a mixture of venom that had been preincubated with the desired volume of IgG antibodies. Mortality, recorded 24 and 48 h after inoculation, showed that IgG anti-PLA2 were more effective than anticrotalic serum in neutralizing the lethal activity. These results demonstrate that it could be possible to obtain an anti-venom made by specific antibodies with a high level of protection against the lethal component of C.d.t. venom, and/or the inclusion of these antibodies as a supplement in heterologous anti-venoms.El veneno de Crotalus durissus terrificus (C.d.t. (Cascabel de Sud América posee actividad miotóxica y neurotóxica, actividades que también exhibe el complejo crotoxina, principal componente tóxico de este veneno. El complejo crotoxina está constituido por una fosfolipasa A2 básica (PLA2 y una proteína acídica no tóxica, el crotapotín. En este trabajo se estudió la capacidad neutralizante de anticuerpos IgG anti-PLA2 sobre la letalidad inducida por el veneno entero. El antígeno PLA2, fue aislado por cromatografía de filtración en gel (Sephadex G-75. Se inocularon conejos machos por vía subcutánea e intramuscular, con 700 µg de PLA2 y adyuvante para la obtención de anticuerpos específicos. La capacidad neutralizante del

  20. The comparison between the humoral response and the neutralizing capacity of sheep serum inoculated with natural venom and Co{sup 60} irradiated venom from Crotalus durissus terrificus (Laurenti, 1768)

    Energy Technology Data Exchange (ETDEWEB)

    Netto, D.P. [Universidade Estadual de Londrina, PR (Brazil). Centro de Ciencias Agrarias. Dept. de Medicina Veterinaria Preventiva]. E-mail: mnetto@uel.br

    2000-07-01

    Crotalus durissus terrificus venom was irradiated with Co{sup 60} to investigate the effects of antigen-irradiation on antivenom production in sheep. Twelve sheep were divided in two groups of 6. One group received irradiated, while the other received natural venom. Three doses of antigen were given at monthly intervals. The toxic activity of the venom was assessed by LD{sub 50} in mice. Weekly blood samples were obtained to evaluate anti-crotalic serum titers by indirect ELISA, neutralization capacity, and serum potency. A complete blood count, plasma protein and fibrinogen concentration, and serum albumin and globulin were also determined. At end of the experiment, the animals were challenged with ovine LD{sub 50}, without clinical abnormalities. (author)

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

    Directory of Open Access Journals (Sweden)

    Carlos Hubinger Tokarnia

    2014-04-01

    Full Text Available Foi realizada uma revisão dos quadros clínico-patológicos causados pelos venenos de Crotalus durissus terrificus e Bothrops spp. em bovinos, búfalos, ovinos equinos e suínos. Foram compilados os dados obtidos pela experimentação em animais de produção encontrados na literatura e os obtidos através de experimentação realizada por nossa equipe. Também foram revisados os casos naturais de envenenamento ofídico comunicados. Em dois Quadros foram lançados os mais importantes dados dessas revisões, que revelou diversos aspectos interessantes: 1 em nossos experimentos, o veneno de Crotalus durissus terrificus, quando injetado por via subcutânea em cavalos, causou um edema acentuado no local da aplicação, ao contrário do que tem sido observado em todas as outras espécies animais, aspecto não relatado na literatura; 2 em nossos experimentos, o veneno de diversas espécies de Bothrops, quando injetado por via subcutânea em bovinos, ovinos e equinos, não causou edema como em geral é relatado na literatura, e sim hemorragias subcutâneas acentuadas no local da aplicação. Nos casos não fatais este sangue era reabsorvido em poucos dias sem deixar sequelas. Exceção foi a reação ao veneno de Bothrops jararacussu, que causou edema nos ovinos experimentais, e tumefação acentuada que resultou em fístula com eliminação de líquido seroso nos equinos experimentais. O objetivo do presente estudo visa contribuir para o aperfeiçoamento do diagnóstico de acidentes ofídicos em animais de produção.

  2. Estabilidad de la actividad neutralizante del antiveneno ofídico conservado a 4°C y a 16°C contra el veneno de Cascabel (Crotalus durissus cumanensis de Colombia

    Directory of Open Access Journals (Sweden)

    Jaime Ramírez

    1995-12-01

    Full Text Available Se determinó la actividad neutralizante de la letalidad de varios lotes de antiveneno polivalente (no liofilizado, producidos por el Instituto Nacional de Salud, los cuales fueron retados contra una mezcla de venenos de cascabel (Crotalus durissus cumanensis de Colombia. Se compara la actividad de los diferentes lotes almacenados en oscuridad a las temperaturas de 4°C y 16°C. Se estableció el porcentaje remanente de la potencia de cada lote para cada temperatura en los diferentes años a lo largo del período del estudio. Se concluye que el antiveneno almacenando en las condiciones recomendadas, mantiene casi la totalidad de la capacidad de neutralización (97.9% luego de tres años de almacenamiento, lo quecorresponde a lafechade expiración del producto. En cambio, el antiveneno almacenado a 16°C y oscuridad, luego de dos años de almacenamiento, mantiene el 87.9% de su capacidad protectora contra veneno de cascabel, protección similara la encontrada bajo las mismas condiciones de almacenamiento, paraveneno de Taya X, pasados dos años de almacenamiento (l, sustentando la recomendación de descartar el producto, transcurridos dos años de almacenamiento en estas condiciones.

  3. Neuromuscular paralysis by the basic phospholipase A2 subunit of crotoxin from Crotalus durissus terrificus snake venom needs its acid chaperone to concurrently inhibit acetylcholine release and produce muscle blockage.

    Science.gov (United States)

    Cavalcante, Walter L G; Noronha-Matos, José B; Timóteo, Maria A; Fontes, Marcos R M; Gallacci, Márcia; Correia-de-Sá, Paulo

    2017-11-01

    Crotoxin (CTX), a heterodimeric phospholipase A 2 (PLA 2 ) neurotoxin from Crotalus durissus terrificus snake venom, promotes irreversible blockade of neuromuscular transmission. Indirect electrophysiological evidence suggests that CTX exerts a primary inhibitory action on transmitter exocytosis, yet contribution of a postsynaptic action of the toxin resulting from nicotinic receptor desensitization cannot be excluded. Here, we examined the blocking effect of CTX on nerve-evoked transmitter release measured directly using radioisotope neurochemistry and video microscopy with the FM4-64 fluorescent dye. Experiments were conducted using mice phrenic-diaphragm preparations. Real-time fluorescence video microscopy and liquid scintillation spectrometry techniques were used to detect transmitter exocytosis and nerve-evoked [ 3 H]-acetylcholine ([ 3 H]ACh) release, respectively. Nerve-evoked myographic recordings were also carried out for comparison purposes. Both CTX (5μg/mL) and its basic PLA 2 subunit (CB, 20μg/mL) had biphasic effects on nerve-evoked transmitter exocytosis characterized by a transient initial facilitation followed by a sustained decay. CTX and CB reduced nerve-evoked [ 3 H]ACh release by 60% and 69%, respectively, but only the heterodimer, CTX, decreased the amplitude of nerve-evoked muscle twitches. Data show that CTX exerts a presynaptic inhibitory action on ACh release that is highly dependent on its intrinsic PLA 2 activity. Given the high safety margin of the neuromuscular transmission, one may argue that the presynaptic block caused by the toxin is not enough to produce muscle paralysis unless a concurrent postsynaptic inhibitory action is also exerted by the CTX heterodimer. Copyright © 2017. Published by Elsevier Inc.

  4. Comparison of humoral immune response, neutralization capacity of anticrotalic serum in young ovines, clinical and weight evaluation between animals inoculated with Crotalus durissus terrificus venom, natural or Cobalt-60-irradiated

    International Nuclear Information System (INIS)

    Ferreira Junior, R.S.

    2005-01-01

    The Elisa technique was used to evaluate and compare the humoral immune response of young ovine to anticrotalic serum production. During serum production, the clinical and weight evaluation of the animals was performed. The parameters utilized were complete blood count, and dosage of urea, creatinine, aspartate aminotransferase, total proteins, albumin and globulin. The animals weight was verified fortnightly during the experiment. The neutralization capacity of the serum produced from the snake Crotalus durissus terrificus natural (NV) and Cobalt-60-irradiated venom (IrV) was evaluated by in vitro challenges. One group of six animals received natural venom, the second group received irradiated venom, and the third was the control group. The animals received six immunizations during 84 days with an interval of 14 days. There was a significant difference (p<5%) in the ELISA test for the profile of the antibodies produced by the experimental groups (NV< IrV). There was no significant difference (p<5%) for biochemical tests, complete blood count, and animals weight between the three groups tested. The group immunized with irradiated venom showed antibodies profile higher than the group immunized with natural venom. The neutralization capacity of the serum produced from the IrV was fivefold higher when compared to the serum produced with NV. The clinical and weight evaluation showed that the o vines in post-weaning phase did not have their physiological profiles altered, and showed an excellent increase in weight during the experimental period. These results indicate a new perspective for the utilization of o vines, aiming the commercial production of anticrotalic serum, which may be applied in the treatment of human and animal envenomation. The cost for its production may be reduced by the posterior utilization of hyperimmunized ovine in human feeding. (author)

  5. Comparison of humoral immune response, neutralization capacity of anticrotalic serum in young ovines, clinical and weight evaluation between animals inoculated with Crotalus durissus terrificus venom, natural or Cobalt-60-irradiated

    Energy Technology Data Exchange (ETDEWEB)

    Ferreira Junior, R.S. E-mail: rseabra@cevap.org.br

    2005-07-01

    The Elisa technique was used to evaluate and compare the humoral immune response of young ovine to anticrotalic serum production. During serum production, the clinical and weight evaluation of the animals was performed. The parameters utilized were complete blood count, and dosage of urea, creatinine, aspartate aminotransferase, total proteins, albumin and globulin. The animals weight was verified fortnightly during the experiment. The neutralization capacity of the serum produced from the snake Crotalus durissus terrificus natural (NV) and Cobalt-60-irradiated venom (IrV) was evaluated by in vitro challenges. One group of six animals received natural venom, the second group received irradiated venom, and the third was the control group. The animals received six immunizations during 84 days with an interval of 14 days. There was a significant difference (p<5%) in the ELISA test for the profile of the antibodies produced by the experimental groups (NV

  6. Effects of ionizing radiation on crotoxin (toxin of Crotalus durissus terrificus venom): molecular studies

    International Nuclear Information System (INIS)

    Souza Filho, J.N. de.

    1988-01-01

    It is know that the ionizing radiation is able to change significantly the biological and antigenic response of a toxin depending of the dose and irradiation's conditions, probable by structural alterations caused by radiation. In this work, the crotoxin, principal neurotoxin of the South American rattlesnake venom, was isolated using molecular exclusion chromatography with Sephadex G-75 and follwed by precipitation on the isoelectric point. Fractions in the concentration of 2 mg of protein/m1 0.85% NaCl were irradiated in a source of sup(60)Co GAMMACELL with dose rate of 1100 Gy/h using doses of 250, 500, 1000, 1500 and 2000 Gy. It was determinated for these samples, the proteic concentration (Lowry's method), the content sulphydryl (Ellman's method), the profile electrophoretic (SDS-PAGE), the toxicity by lethal dose 50% in mice and the antigenic response using crotalic antiserum by the diffusion imunoassay (Ouchterlony's method). The results showed the formation of aggregates and loss of protein in solution by precipitation. In the dose of 1000 Gy and higher it was possible to observe the presence of sulphydryl groups indicating the breakage of S-S bridges. The lethal dose 50% increased 2 times for the dose of 1000 Gy and 3.5 times for 1500 Gy shoding a detoxication. By the other hand, the antigenic response seems to be still intact at doses up to 1000 Gy. (author)

  7. Effects of ionizing radiation in crotamine from Crotalus durissus terrificus venous

    International Nuclear Information System (INIS)

    Costa, T.A. da.

    1988-01-01

    Ionizing radiations are sufficiently energetic and able to cleave any chemical bond. Molecules of substances present in solution have been shown either by direct or indirect action of ionizing radiations loss of biological activity. Crotamine obtained from C. d. terrificus venom by molecular exclusion cromatography was irradiated in aqueous solution of 1 mg/ml by gamma rays produced on a 60 Co source. Doses of 100 Gy, 2000 Gy and 5000 Gy (Dose rate = 1,14 x 10 3 Gy/h) in presence of O 2 were used for the comparison of properties of irradiated and non irradiated samples; the following assays were performed: determination of protein concentration, presence of free SH groups, SDS-Page and Ouchterlony's immunodiffusion. The Ellman's reaction for free SH groups was positive in the irradiated protein and negative one. The protein concentration was not altered up to the dose of 2000 Gy in relation to the native protein: at the dose of 5000 Gy was observed a loss of 55% of the proteic material. The SDS-PAGE analysis showed the formation of protein aggregates in the dose of 2000 Gy and 5000 Gy. A loss of the antigenic activity was observed both in the dose of 2000 Gy and 5000 Gy againt sera anticrotalic. Further studies was perfomed with crotamine irradiated at the dose of 2000 Gy in aqueous solution, O 2 absence and in presence of different concentrations of radioprotectors such as: cystein, sodium nitrate and tert-butanol. The level of the radioprotection to the toxin was estimated through the simple radical immunodiffusion technique. According to the technique the best radioprotective agent was cysteine. Since the antigenic determinants were preserved and in the SDS-PAGE analysis the crotamine structure was maintained entire like the non irradiated toxin. (author) [pt

  8. Geographic distribution and apparent decline of Crotalus durissus terrificus (Laurenti 1768; Serpentes, Viperidae) in Uruguay

    OpenAIRE

    Prigioni, Carlos M.; Borteiro, Claudio; Kolenc, Francisco; González, Enrique M.; Colina, Marcelo

    2013-01-01

    Herein, we make a comprehensive account of rattlesnake records in this country obtained from herpetological literature and other bibliographic sources, specimens accessioned in herpetological and non-specialized local collections, and new information gathered during field surveys. Asociación Herpetológica Argentina

  9. Effects of gamma radiation on isolated toxin of Crotalus durissus terrificus

    International Nuclear Information System (INIS)

    Souza Filho, J.N.; Rogero, J.R.

    1988-07-01

    It is know that the ionizing radiation is able to change significantly the biological and antigenic behaviour of a toxin depending of the dose and radiation's conditions, probably by structural alterations caused by radiation. In this paper, the crotoxin, principal neurotoxin of the south american rattlesnake venom, was isolated using molecular exclusion cromatography with Sephadex G-75 and followed by precipitation of the isoeletric point. (pI 4.7.). Fractions in the concentration of 2 mg toxin/ml 0.85 % NaCl were irradiated in a source of 60 Co GAMMACELL with dose rate of 1100 Gy/hr using doses of 250,500, 1000, 1500 and 2000 Gy. It was determinated for this samples, the proteic concentration (Lowry's method), the antigenic capability using crotalic antiserum by the diffusion imunoassay (Ouch - terlony's method), the DL50 in mice and eletrophoresis (SDS-PAGE). The results showed that the antigenic capability seems to be intact until dose of 1000 Gy. By the other hand, the LD50 in the same radiation dose increases more than two times when compared with the native sample. As the radiation dose increases, the solutions became turbid showing loss of protein probably by the presence of aggregates as can be seen by the determination of the proteic concentration and the eletrophoretic control. Other important changs were observed with the irrdiated samples in comparison with the native crotoxin that can given us additional information about dose-effect event. (author) [pt

  10. Microhabitat use by species of the genera Bothrops and Crotalus (Viperidae in semi-extensive captivity

    Directory of Open Access Journals (Sweden)

    CA Gomes

    2012-01-01

    Full Text Available Many factors influence microhabitat including climate and the occurrence of predators, prey and suitable shelters. The influence of predators in a semi-extensive breeding system is minimized due to frequent monitoring of the area. This situation enables the independent analysis of such other variables as refuges and temperature. Some specimens of the Viperidae family are kept in a semi-extensive breeding system at the Butantan Institute for display and study. These animals are widely distributed in the Atlantic Forest and Cerrado, two biomes with distinct climatic characteristics. We compared the daily activity pattern and microhabitat use of the species Bothrops jararaca and Crotalus durissus terrificus. Our main questions were whether rattlesnakes and lancehead snakes respond to habitat selection differently in similar climatic conditions and if they choose similar microhabitats. Species of the genus Bothrops were frequently found under shelters regardless of the time of day. On the other hand, snakes of the genus Crotalus were frequently found sheltered during the early morning, then migrated to sunnier areas and returned to shelters in late afternoon.

  11. Microscopic agglutination test on captive rattlesnakes : Data on serovars and titers

    Directory of Open Access Journals (Sweden)

    T.C.S. Rodrigues

    2016-06-01

    Full Text Available The microscopic agglutination test (MAT is considered the “golden standard” leptospirosis serodiagnostic test, but there is little information about it as it pertains to snakes. To fill this information gap, we provide data on serovars and titers of fifty-six Crotalus durissus collilineatus sera samples that tested positive by MAT (10.1016/j.actatropica.2016.02.006 (Rodrigues et al., 2016 [5]. These data are presented in a table, along with a description of the methodology used for sample collection and serologic testing.

  12. 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.

  13. Intraspecific venom variation in the medically significant Southern Pacific Rattlesnake (Crotalus oreganus helleri): biodiscovery, clinical and evolutionary implications.

    Science.gov (United States)

    Sunagar, Kartik; Undheim, Eivind A B; Scheib, Holger; Gren, Eric C K; Cochran, Chip; Person, Carl E; Koludarov, Ivan; Kelln, Wayne; Hayes, William K; King, Glenn F; Antunes, Agosthino; Fry, Bryan Grieg

    2014-03-17

    Due to the extreme variation of venom, which consequently results in drastically variable degrees of neutralization by CroFab antivenom, the management and treatment of envenoming by Crotalus oreganus helleri (the Southern Pacific Rattlesnake), one of the most medically significant snake species in all of North America, has been a clinician's nightmare. This snake has also been the subject of sensational news stories regarding supposed rapid (within the last few decades) evolution of its venom. This research demonstrates for the first time that variable evolutionary selection pressures sculpt the intraspecific molecular diversity of venom components in C. o. helleri. We show that myotoxic β-defensin peptides (aka: crotamines/small basic myotoxic peptides) are secreted in large amounts by all populations. However, the mature toxin-encoding nucleotide regions evolve under the constraints of negative selection, likely as a result of their non-specific mode of action which doesn't enforce them to follow the regime of the classic predator-prey chemical arms race. The hemorrhagic and tissue destroying snake venom metalloproteinases (SVMPs) were secreted in larger amounts by the Catalina Island and Phelan rattlesnake populations, in moderate amounts in the Loma Linda population and in only trace levels by the Idyllwild population. Only the Idyllwild population in the San Jacinto Mountains contained potent presynaptic neurotoxic phospholipase A2 complex characteristic of Mohave Rattlesnake (Crotalus scutulatus) and Neotropical Rattlesnake (Crotalus durissus terrificus). The derived heterodimeric lectin toxins characteristic of viper venoms, which exhibit a diversity of biological activities, including anticoagulation, agonism/antagonism of platelet activation, or procoagulation, appear to have evolved under extremely variable selection pressures. While most lectin α- and β-chains evolved rapidly under the influence of positive Darwinian selection, the β-chain lectin of

  14. The unequal influences of the left and right vagi on the control of the heart and pulmonary artery in the rattlesnake, Crotalus durissus

    DEFF Research Database (Denmark)

    Taylor, E.W.; Andrade, Denis V.; Abe, Augusto A.

    2009-01-01

    Autonomic control of the cardiovascular system in reptiles includes sympathetic components but heart rate (fH), pulmonary blood flow ( pul) and cardiac shunt patterns are primarily controlled by the parasympathetic nervous system. The vagus innervates both the heart and a sphincter on the pulmona....... This paves the way for an investigation of the role of the cardiac shunt in regulating metabolic rate, as chronic left vagotomy will cause a pronounced left–right shunt in recovered animals, whilst leaving intact control of the heart, via the right vagus....... artery. The present study reveals that whereas both the left and right vagi influence fH, it is only the left vagus that influences pulmonary vascular resistance. This is associated with the fact that rattlesnakes, in common with some other species of snakes, have a single functional lung, as the other...... lung regresses during development. Stimulation of the left cervical vagus in anaesthetised snakes slowed the heart and markedly reduced blood flow in the pulmonary artery whereas stimulation of the right cervical vagus slowed the heart and caused a small increase in stroke volume (VS) in both...

  15. Aspectos clínico-patológicos e laboratoriais do envenenamento crotálico experimental em bovinos Clinical and pathological aspects of experimental Crotalus poisoning in cattle

    Directory of Open Access Journals (Sweden)

    Flávio A.S. Graça

    2008-06-01

    Full Text Available Reproduziu-se experimentalmente o envenenamento crotálico, através da inoculação, por via subcutânea, do veneno de Crotalus durissus terrificus (cascavel sul-americana em dez bovinos mestiços. Dois animais foram utilizados como controle. O bovino que recebeu dose de 0,03mg/kg de peso corporal, morreu 7h40min após a inoculação. A dose de 0,015mg/kg causou a morte em quatro de sete bovinos inoculados, enquanto os dois animais que receberam 0,0075mg/kg adoeceram discretamente e se recuperaram. Os sintomas tiveram início entre 1h30min e 13h45min após a inoculação. A evolução oscilou entre 5h25min e 45h para os animais que morreram e entre 33h15min e 17 dias entre os animais que se recuperaram. Os principais sinais nervosos observados foram diminuição da resposta aos estímulos externos, reflexos hipotônicos, arrastar dos cascos no solo, aparente apatia, paralisia do globo ocular e da língua, decúbito esternal e lateral. Verificaram-se também adipsia e, por vezes, petéquias nas mucosas vaginal e conjuntival. Houve discreto a moderado aumento do tempo de sangramento e moderado aumento do tempo de tromboplastina parcial ativada. Houve moderada leucocitose com neutrofilia, linfopenia relativa, eosinopenia, monocitose e discreto aumento do número de bastões. Foi evidenciado significativo aumento dos níveis séricos de creatinaquinase, contudo, não foram observadas alterações significativas através da urinálise. À necropsia constataram-se edema quase imperceptível no local da inoculação, discretas petéquias e sufusões no epicárdio, omento, vesícula biliar e mucosa da bexiga em alguns dos animais envenenados experimentalmente. Os exames histopatológicos revelaram necrose (hialinização de grupos de miócitos ou em miócitos isolados em dez diferentes músculos esqueléticos examinados, próximos ou distantes do local de inoculação em todos os animais necropsiados. Concluí-se que o envenenamento por Crotalus Sul

  16. The impact of roads on the timber rattlesnake (Crotalus horridus) in eastern Texas

    Science.gov (United States)

    D. Craig Rudolph; Shirley J. Burgdorf; Richard N. Conner; James G. Dickson

    1998-01-01

    Roads and associated vehicular traffic have the potential to significantly impact vertebrate populations. In eastern Texas we compared the densities of paved and unpaved roads within 2 and 4 km radii of timber rattlesnake (Crotalus horridus) ocations and of random points. Road networks were significantly more dense at random points than at snake...

  17. Body size evolution in insular speckled rattlesnakes (Viperidae: Crotalus mitchellii.

    Directory of Open Access Journals (Sweden)

    Jesse M Meik

    2010-03-01

    Full Text Available Speckled rattlesnakes (Crotalus mitchellii inhabit multiple islands off the coast of Baja California, Mexico. Two of the 14 known insular populations have been recognized as subspecies based primarily on body size divergence from putative mainland ancestral populations; however, a survey of body size variation from other islands occupied by these snakes has not been previously reported. We examined body size variation between island and mainland speckled rattlesnakes, and the relationship between body size and various island physical variables among 12 island populations. We also examined relative head size among giant, dwarfed, and mainland speckled rattlesnakes to determine whether allometric differences conformed to predictions of gape size (and indirectly body size evolving in response to shifts in prey size.Insular speckled rattlesnakes show considerable variation in body size when compared to mainland source subspecies. In addition to previously known instances of gigantism on Angel de la Guarda and dwarfism on El Muerto, various degrees of body size decrease have occurred frequently in this taxon, with dwarfed rattlesnakes occurring mostly on small, recently isolated, land-bridge islands. Regression models using the Akaike information criterion (AIC showed that mean SVL of insular populations was most strongly correlated with island area, suggesting the influence of selection for different body size optima for islands of different size. Allometric differences in head size of giant and dwarf rattlesnakes revealed patterns consistent with shifts to larger and smaller prey, respectively.Our data provide the first example of a clear relationship between body size and island area in a squamate reptile species; among vertebrates this pattern has been previously documented in few insular mammals. This finding suggests that selection for body size is influenced by changes in community dynamics that are related to graded differences in area over

  18. The effects of hybridization on divergent venom phenotypes: Characterization of venom from Crotalus scutulatus scutulatus × Crotalus oreganus helleri hybrids.

    Science.gov (United States)

    Smith, Cara Francesca; Mackessy, Stephen P

    2016-09-15

    Hybridization between divergent species can be analyzed to elucidate expression patterns of distinct parental characteristics, as well as to provide information about the extent of reproductive isolation between species. A known hybrid cross between two rattlesnakes with highly divergent venom phenotypes provided the opportunity to examine occurrence of parental venom characteristics in the F1 hybrids as well as ontogenetic shifts in the expression of these characters as the hybrids aged. Although venom phenotypes of adult rattlesnake venoms are known for many species, the effect of hybridization on phenotype inheritance is not well understood, and effects of hybridization on venom ontogeny have not yet been investigated. The current study investigates both phenomena resulting from the hybridization of a male snake with type I degradative venom, Crotalus oreganus helleri (Southern Pacific Rattlesnake), and a female snake with type II highly toxic venom, Crotalus scutulatus scutulatus (Mojave Rattlesnake). SDS-PAGE, enzymology, Western blot and reversed phase HPLC (RP-HPLC) were used to characterize the venom of the C. o. helleri male, the C. s. scutulatus female and their two hybrid offspring as they aged. In general, Crotalus o. helleri × C. s. scutulatus hybrid venoms appeared to exhibit overlapping parental venom profiles, and several different enzyme activity patterns. Both hybrids expressed C. o. helleri father-specific myotoxins as well as C. s. scutulatus mother-specific Mojave toxin. Snake venom metalloprotease activity displayed apparent sex-influenced expression patterns, while hybrid serine protease activities were intermediate to parental activities. The C. s. scutulatus × C. o. helleri hybrid male's venom profile provided the strongest evidence that type I and type II venom characteristics are expressed simultaneously in hybrid venoms, as this snake contained distinctive characteristics of both parental species. However, the possibility of

  19. Characterization of ten microsatellite loci in midget faded rattlesnake (Crotalus oreganus concolor)

    Science.gov (United States)

    Oyler-McCance, Sara J.; Parker, Joshua M.

    2010-01-01

    Primers for 10 microsatellite loci were developed for midget faded rattlesnake (Crotalus oreganus concolor), a small bodied subspecies of the Western Rattlesnake, which is found in the Colorado Plateau of eastern Utah, western Colorado and southwestern Wyoming. In a screen of 23 individuals from the most northern portion of the subspecies range in southwestern Wyoming, the 10 loci were found to have levels of variability ranging from 4 to 11 alleles. No loci were found to be linked, although one locus revealed significant departures from Hardy–Weinberg equilibrium. These microsatellite loci will be applicable for population genetic analyses, which will ultimately aid in management efforts for this rare subspecies of rattlesnake.

  20. Vascular effects and electrolyte homeostasis of the natriuretic peptide isolated from Crotalus oreganus abyssus (North American Grand Canyon rattlesnake) venom

    NARCIS (Netherlands)

    Da Silva, S.L.; Dias-Junior, C.A.; Baldasso, P.A.; Damico, D.C.; Carvalho, B.M.; Garanto, A.; Acosta, G.; Oliveira, E.; Albericio, F.; Soares, A.M.; Marangoni, S.; Resende, R.R.

    2012-01-01

    Crotalus oreganus abyssus is a rattlesnake that is usually found in the Grand Canyon, United States of America. Knowledge regarding the composition of C. o. abyssus venom is scarce. New natriuretic peptides (NPs) have been isolated and characterized from the venoms of members of the Crotalinae

  1. Preparation of crotalus venom radiolabeled with technetium99m as a tool for biodistribution study

    International Nuclear Information System (INIS)

    Pujatti, Priscilla Brunelli; Santos, Raquel Gouvea dos; Simal, Carlos Jorge Rodrigues

    2005-01-01

    Technetium- 99m ( 99m Tc) has been the radionuclide of choice for nuclear medicine procedures and experimental research. Because of its optimal nuclear properties, 99m Tc is suitable for high efficiency detection with the advantage of reduced radiological waste. Crotalus venom (CV) has been shown to reduce tumors in clinical studies and tissue distribution studies are very important for clinical use. The goal of this work was to obtain CV labeled with 99m Tc which preserves its biological activity. After labeling, biological activity was assessed by hemolytic activity evaluation. Labeled and crude venom caused indirect hemolysis provided that the incubation medium contained an exogenous source of lecithin. High yield radiolabeled-CV was obtained and biological activity was preserved. The results suggest that 99m Tc-CV can be a useful tool for biodistribution studies. (author)

  2. Immunohistochemical changes in kidney glomerular and tubular proteins caused by rattlesnake (Crotalus vegrandis) venom Cambios inmunohistoquímicos en proteínas de túbulo y glomérulo renal causadas por el veneno de la serpiente de cascabel (Crotalus vegrandis)

    OpenAIRE

    María E. Girón; Irma Aguilar; Alexis Rodríguez-Acosta

    2003-01-01

    Renal damage is an important cause of death in patients who have survived the early effects of severe crotalid envenomation. Extracellular matrix of renal tissue is altered by Crotalus toxin activities. The aim of this study was to describe how cytoskeletal proteins and basal membrane components undergo substantial alterations under the action of Crotalus vegrandis crude venom and its hemorrhagic fraction (Uracoina-1) in mice. To detect the proteins in question, the immunoperoxidase method wi...

  3. Seasonal variation in hormonal responses of timber rattlesnakes (Crotalus horridus) to reproductive and environmental stressors.

    Science.gov (United States)

    Lutterschmidt, William I; Lutterschmidt, Deborah I; Mason, Robert T; Reinert, Howard K

    2009-08-01

    Data addressing adrenocortical modulation across taxonomic groups are limited, especially with regard to how female reproductive condition influences the sensitivity of the hypothalamus-pituitary-adrenal axis. We investigated seasonal and reproductive variation in basal and stress-induced hormone profiles in a population of free-ranging timber rattlesnakes (Crotalus horridus) in north-central Pennsylvania during spring (i.e., May), summer (i.e., July), and early fall (i.e., September). Baseline corticosterone concentrations varied seasonally and were significantly lower during the summer sampling period in July. We observed a significant negative relationship between baseline corticosterone and testosterone in male snakes, while baseline corticosterone and estradiol tended to be positively correlated in females. Treatment of snakes with 1 h of capture stress significantly increased corticosterone across all seasons. However, there was a significant interaction between corticosterone responses to capture stress and season, suggesting that adrenocortical function is modulated seasonally. Because elevated corticosterone may be associated with reproduction, we asked whether hormonal stress responses vary with female reproductive condition. Although sample sizes are low, reproductive snakes had significantly higher baseline and stress-induced corticosterone concentrations than non-reproductive or post-parturient females. Further, despite similar baseline corticosterone concentrations between non-reproductive and post-parturient rattlesnakes, post-parturient females responded to capture stress with a significantly higher increase in corticosterone. Collectively, these data suggest that the sensitivity of the hypothalamus-pituitary-adrenal axis varies both seasonally and with changing reproductive states.

  4. Biological and Proteolytic Variation in the Venom of Crotalus scutulatus scutulatus from Mexico

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    Miguel Borja

    2018-01-01

    Full Text Available Rattlesnake venoms may be classified according to the presence/absence and relative abundance of the neurotoxic phospholipases A 2 s (PLA 2 s, such as Mojave toxin, and snake venom metalloproteinases (SVMPs. In Mexico, studies to determine venom variation in Mojave Rattlesnakes (Crotalus scutulatus scutulatus are limited and little is known about the biological and proteolytic activities in this species. Tissue (34 and venom (29 samples were obtained from C. s. scutulatus from different locations within their distribution in Mexico. Mojave toxin detection was carried out at the genomic (by PCR and protein (by ELISA levels for all tissue and venom samples. Biological activity was tested on representative venoms by measuring LD 50 and hemorrhagic activity. To determine the approximate amount of SVMPs, 15 venoms were separated by RP-HPLC and variation in protein profile and proteolytic activity was evaluated by SDS-PAGE (n = 28 and Hide Powder Azure proteolytic analysis (n = 27. Three types of venom were identified in Mexico which is comparable to the intraspecific venom diversity observed in the Sonoran Desert of Arizona, USA: Venom Type A (∼Type II, with Mojave toxin, highly toxic, lacking hemorrhagic activity, and with scarce proteolytic activity; Type B (∼Type I, without Mojave toxin, less toxic than Type A, highly hemorrhagic and proteolytic; and Type A + B, containing Mojave toxin, as toxic as venom Type A, variable in hemorrhagic activity and with intermediate proteolytic activity. We also detected a positive correlation between SVMP abundance and hemorrhagic and proteolytic activities. Although more sampling is necessary, our results suggest that venoms containing Mojave toxin and venom lacking this toxin are distributed in the northwest and southeast portions of the distribution in Mexico, respectively, while an intergradation in the middle of both zones is present.

  5. Natural history of a northern population of twin-spotted rattlesnakes, Crotalus pricei

    Science.gov (United States)

    Prival, D.B.; Goode, M.J.; Swann, D.E.; Schwalbe, C.R.; Schroff, M.J.

    2002-01-01

    The twin-spotted rattlesnake (Crotalus pricei) is a small-bodied pitviper that has received little attention in the literature to date. The species reaches the northern limit of its range in southeastern Arizona, where it inhabits higher elevations than any of the state's 10 other rattlesnake species. During 1997-2000, we captured, measured, and marked 127 C. pricei in Arizona's Chiricahua Mountains between 2530 and 2900 m elevation. We also used radiotelemetry to track the movements of 16 C. pricei in the study area during 1997-1998. Mean (?? SE) snout-vent length of C. pricei was 387.8 ?? 8.3 mm (range = 168-572), and mean mass was 53.5 ?? 3.3 g (range = 3.6-188.5). Based on fecal analyses, lizards constituted the bulk of prey (74%), but the diet of C. pricei also included mammals, birds, and a conspecific. Mating was concentrated in August and early September and parturition took place during late July and August. Mean number of embryos was 3.94 ?? 0.34 (range = 1-6) and female reproduction appeared biennial or less frequent. Based on shed and growth rates, female C. pricei develop embryos at 4-5 years of age. Gravid females maintained warmer body temperatures relative to substrate temperature than nongravid females or males, presumably by spending more time basking than other snakes. Radiotelemetry revealed that movement patterns varied from year to year, as males moved over six times farther per week during the 1998 monsoon season (July to September) than during the 1997 monsoon season. Additionally, use of talus slopes by males decreased during 1998. During dry years, such as 1998, males may be forced off talus into cooler microclimates where resources are less concentrated than on talus.

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

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    Polianna L.M.M. Albuquerque

    2013-09-01

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

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

    Science.gov (United States)

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

    2013-01-01

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

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

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    Ezequiel Capeletti

    2016-09-01

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

  9. Snake venomics of Crotalus tigris: the minimalist toxin arsenal of the deadliest Nearctic rattlesnake venom. Evolutionary Clues for generating a pan-specific antivenom against crotalid type II venoms [corrected].

    Science.gov (United States)

    Calvete, Juan J; Pérez, Alicia; Lomonte, Bruno; Sánchez, Elda E; Sanz, Libia

    2012-02-03

    We report the proteomic and antivenomic characterization of Crotalus tigris venom. This venom exhibits the highest lethality for mice among rattlesnakes and the simplest toxin proteome reported to date. The venom proteome of C. tigris comprises 7-8 gene products from 6 toxin families; the presynaptic β-neurotoxic heterodimeric PLA(2), Mojave toxin, and two serine proteinases comprise, respectively, 66 and 27% of the C. tigris toxin arsenal, whereas a VEGF-like protein, a CRISP molecule, a medium-sized disintegrin, and 1-2 PIII-SVMPs each represent 0.1-5% of the total venom proteome. This toxin profile really explains the systemic neuro- and myotoxic effects observed in envenomated animals. In addition, we found that venom lethality of C. tigris and other North American rattlesnake type II venoms correlates with the concentration of Mojave toxin A-subunit, supporting the view that the neurotoxic venom phenotype of crotalid type II venoms may be described as a single-allele adaptation. Our data suggest that the evolutionary trend toward neurotoxicity, which has been also reported for the South American rattlesnakes, may have resulted by pedomorphism. The ability of an experimental antivenom to effectively immunodeplete proteins from the type II venoms of C. tigris, Crotalus horridus , Crotalus oreganus helleri, Crotalus scutulatus scutulatus, and Sistrurus catenatus catenatus indicated the feasibility of generating a pan-American anti-Crotalus type II antivenom, suggested by the identification of shared evolutionary trends among South and North American Crotalus species.

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

    OpenAIRE

    Ezequiel Capeletti; Michel Mendes; Diezza Biondo; Marcia Maria Dosciatti de Oliveira

    2016-01-01

    Snakes are reptiles that have large environmental adaptations, which favored it's distribution among the various ecosystems. In Brazil, there are found 392 species of snakes, while in Rio Grande do Sul (RS), this research field, 79 species have been described, of which there is the Viperidae family. The objective of this work is to verify the origin of snakes of genus Bothrops and Crotalus, according to the records of the last 15 years of the serpentarium the University of Caxias do Sul, repr...

  11. Morulustatin, A Disintegrin that Inhibits ADP-Induced Platelet Aggregation, Isolated from the Mexican Tamaulipan Rock Rattlesnake (Crotalus lepidus morulus)

    Science.gov (United States)

    Borja, Miguel; Galan, Jacob Anthony; Cantu, Esteban; Zugasti-Cruz, Alejandro; Rodríguez-Acosta, Alexis; Lazcano, David; Lucena, Sara; Suntravat, Montamas; Sánchez, y Elda Eliza

    2016-01-01

    The Tamaulipan rock rattlesnake (Crotalus lepidus morulus) is a montane snake that occurs in the humid pine-oak forest and the upper cloud forest of the Sierra Madre Oriental in southwestern Tamaulipas, central Nuevo Leon, and southeastern Coahuila in Mexico. Venom from this rattlesnake was fractionated by High-Performance Liquid Chromatography for the purpose of discovering disintegrin molecules. Disintegrins are non-enzymatic, small molecular weight peptides that interfere with cell-cell and cell-matrix interactions by binding to various cell receptors. Eleven fractions were collected by anion exchange chromatography and pooled into six groups (I, II, III, IV, V, and VI). Proteins of the six groups were analyzed by SDS-PAGE and western blot using antibodies raised against a disintegrin. The antibodies recognized different protein bands in five (II, III, IV, V, and VI) of six groups in a molecular mass range of 7 to 105 kDa. Western blot analysis revealed fewer protein bands in the higher molecular mass range and two bands in the disintegrin weight range in group II compared with the other four groups. Proteins in group II were further separated into nine fractions using reverse phase C18 chromatography. Fraction 4 inhibited platelet aggregation and was named morulustatin, which exhibited a single band with a molecular mass of approximately 7 kDa. Mass spectrometry analysis of fraction 4 revealed the identification of disintegrin peptides LRPGAQCADGLCCDQCR (MH+ 2035.84) and AGEECDCGSPANCCDAATCK (MH+ 2328.82). Morulustatin inhibited ADP-induced platelet aggregation in human whole blood and was concentration-dependent with an IC50 of 89.5 nM ± 12. PMID:28713196

  12. The relationship between plasma steroid hormone concentrations and the reproductive cycle in the Northern Pacific rattlesnake, Crotalus oreganus.

    Science.gov (United States)

    Lind, Craig M; Husak, Jerry F; Eikenaar, Cas; Moore, Ignacio T; Taylor, Emily N

    2010-05-01

    We describe the reproductive cycle of Northern Pacific rattlesnakes (Crotalus oreganus) by quantifying steroid hormone concentrations and observing reproductive behaviors in free-ranging individuals. Additionally, we examined reproductive tissues from museum specimens. Plasma steroid hormone concentrations were quantified for both male and female snakes throughout the active season (March-October). We measured testosterone (T), 5alpha-dihydrotestosterone (DHT), and corticosterone (B) concentrations in both sexes and 17beta-estradiol (E2) and progesterone (P) in females only. We observed reproductive behaviors (e.g., consortship, courtship, and copulation) in the field and measured testis and follicle size in male and female snakes from museum collections to relate steroid hormone concentrations to the timing of reproductive events. Our study revealed that C. oreganus in central California exhibits a bimodal pattern of breeding, with most mating behavior occurring in the spring and some incidences of mating behavior observed in late summer/fall. Each breeding period corresponded with elevated androgen (T or DHT) levels in males. Testes were regressed in the spring when the majority of reproductive behavior was observed in this population, and they reached peak volume in August and September during spermatogenesis. Although we did not detect seasonal variation in female hormone concentrations, some females had high E2 in the spring and fall, coincident with mating and with increased follicle size (indicating vitellogenesis) in museum specimens. Females with high E2 concentrations also had high T and DHT concentrations. Corticosterone concentrations in males and females were not related either to time of year or to concentrations of any other hormones quantified. Progesterone concentrations in females also did not vary seasonally, but this likely reflected sampling bias as females tended to be underground, and thus unobtainable, in summer months when P would be

  13. Effects of food supplementation on the physiological ecology of female Western diamond-backed rattlesnakes (Crotalus atrox).

    Science.gov (United States)

    Taylor, Emily N; Malawy, Michael A; Browning, Dawn M; Lemar, Shea V; DeNardo, Dale F

    2005-06-01

    Food availability is an important factor in the life histories of organisms because it is often limiting and thus can affect growth, mass change, reproduction, and behaviors such as thermoregulation, locomotion, and mating. Experimental studies in natural settings allow researchers to examine the effects of food on these parameters while animals are free to behave naturally. The wide variation among organisms in energy demands and among environmental food resources suggest that responses to changes in food availability may vary among organisms. Since most supplemental feeding field experiments have been conducted on species with high energy demands, we conducted a supplemental feeding study on free-ranging, female Western diamond-backed rattlesnakes (Crotalus atrox), a species with low energy demands and infrequent reproductive investment. Snakes were offered thawed rodents 1-4 times per week. Over two active seasons, we collected data on surface activity, home range size, growth, mass change, and reproduction of supplementally fed and control snakes. Fed and control snakes did not differ in surface activity levels (proportion of time encountered above versus below ground) or home range size. Fed snakes grew and gained mass faster, and had a dramatically higher occurrence of reproduction than control snakes. Also, fed snakes were in better body condition following reproduction than snakes that were not fed. However, litter characteristics such as offspring number and size were not increased by feeding, suggesting that these characteristics may be fixed. These data experimentally demonstrate that food availability can directly impact some life history traits (i.e., growth and reproduction for C. atrox), but not others (i.e., surface activity and home range size for C. atrox). The relationship between food availability and life history traits is affected in a complex way by ecological traits and physiological constraints, and thus interspecific variation in this

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

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    Alexis Rodríguez-Acosta,

    2016-10-01

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

  15. Intraspecies variation in the venom of the rattlesnake Crotalus simus from Mexico: different expression of crotoxin results in highly variable toxicity in the venoms of three subspecies.

    Science.gov (United States)

    Castro, Edgar Neri; Lomonte, Bruno; del Carmen Gutiérrez, María; Alagón, Alejandro; Gutiérrez, José María

    2013-07-11

    The composition and toxicological profile of the venom of the rattlesnake Crotalus simus in Mexico was analyzed at the subspecies and individual levels. Venoms of the subspecies C. s. simus, C. s. culminatus and C. s. tzabcan greatly differ in the expression of the heterodimeric neurotoxin complex 'crotoxin', with highest concentrations in C. s. simus, followed by C. s. tzabcan, whereas the venom of C. s. culminatus is almost devoid of this neurotoxic PLA2. This explains the large variation in lethality (highest in C. s. simus, which also exerts higher myotoxicity). Coagulant activity on plasma and fibrinogen occurs with the venoms of C. s. simus and C. s. tzabcan, being absent in C. s. culminatus which, in turn, presents higher crotamine-like activity. Proteomic analysis closely correlates with toxicological profiles, since the venom of C. s. simus has high amounts of crotoxin and of serine proteinases, whereas the venom of C. s. culminatus presents higher amounts of metalloproteinases and crotamine. This complex pattern of intraspecies venom variation provides valuable information for the diagnosis and clinical management of envenoming by this species in Mexico, as well as for the preparation of venom pools for the production and quality control of antivenoms. This study describes the variation in venom composition and activities of the three subspecies of Crotalus simus from Mexico. Results demonstrate that there is a notorious difference in these venoms, particularly regarding the content of the potent neurotoxic phospholipase A2 complex 'crotoxin'. In addition, other differences were observed regarding myotoxic and coagulant activities, and expression of the myotoxin 'crotamine'. These findings have implications in, at least, three levels: (a) the adaptive role of variations in venom composition; (b) the possible differences in the clinical manifestations of envenomings by these subspecies in Mexico; and (c) the design of venom mixtures for the preparation of

  16. Activity cycles and foraging behaviors of free-ranging sidewinder rattlesnakes (Crotalus cerastes): the ontogeny of hunting in a precocial vertebrate.

    Science.gov (United States)

    Clark, Rulon W; Dorr, Scott W; Whitford, Malachi D; Freymiller, Grace A; Putman, Breanna J

    2016-06-01

    Predators often employ a complex series of behaviors to overcome antipredator defenses and effectively capture prey. Although hunting behaviors can improve with age and experience, many precocial species are necessarily effective predators from birth. Additionally, many predators experience innate ontogenetic shifts in predatory strategies as they grow, allowing them to adapt to prey more appropriate for their increased size and energetic needs. Understanding how the relative roles of innate age-specific adaptation and learning have evolved requires information on how predation behavior develops in situ, in free-ranging predators. However, most of the research on the ontogeny of predation behavior is based on laboratory studies of captive animals, largely due to the difficulty of following newborn individuals in nature. Here, we take advantage of the unique tracks left by juveniles of a precocial viperid, the sidewinder rattlesnake (Crotalus cerastes), which we used to follow free-ranging snakes in the field. We recorded details of their ambush hunting behavior, and compared the behaviors of these juveniles to adult snakes that we monitored in the field via radio telemetry. Although juvenile and adult behaviors were similar in most respects, we did find that adults chose more effective ambush sites, which may be due to their increased experience. We also found that juveniles (but typically not adults) perform periodic tail undulations while in ambush, and that juveniles displayed slightly different activity cycles. Both of these latter differences are likely the result of age-specific adaptations for juveniles' greater reliance on lizards versus small mammals as prey. We also compared the general predatory behavior of sidewinders to that of other species in the genus Crotalus. These findings will provide important baseline field information for more detailed empirical research on the ontogeny of predation behavior in precocial vertebrates. Copyright © 2016

  17. Functional characterizations of venom phenotypes in the eastern diamondback rattlesnake (Crotalus adamanteus) and evidence for expression-driven divergence in toxic activities among populations

    Science.gov (United States)

    Margres, Mark J.; Walls, Robert; Suntravat, Montamas; Lucena, Sara; Sánchez, Elda E.; Rokyta, Darin R.

    2016-01-01

    Phenotypes frequently vary across and within species. The connection between specific phenotypic effects and function, however, is less understood despite being essential to our understanding of the adaptive process. Snake venoms are ideal for identifying functionally important phenotypic variation because venom variation is common, and venoms can be functionally characterized through simple assays and toxicity measurements. Previous work with the eastern diamondback rattlesnake (Crotalus adamanteus) used multivariate statistical approaches to identify six unique venom phenotypes. We functionally characterized hemolytic, gelatinase, fibrinogenolytic, and coagulant activity for all six phenotypes, as well as one additional venom, to determine if the statistically significant differences in toxin expression levels previously documented corresponded to differences in venom activity. In general, statistical differences in toxin expression predicted the identified functional differences, or lack thereof, in toxic activity, demonstrating that the statistical approach used to characterize C. adamanteus venoms was a fair representation of biologically meaningful differences. Minor differences in activity not accounted for by the statistical model may be the result of amino-acid differences and/or post-translational modifications, but overall we were able to link variation in protein expression levels to variation in function as predicted by multivariate statistical approaches. PMID:27179420

  18. Mating systems, reproductive success, and sexual selection in secretive species: a case study of the western diamond-backed rattlesnake, Crotalus atrox.

    Science.gov (United States)

    Clark, Rulon W; Schuett, Gordon W; Repp, Roger A; Amarello, Melissa; Smith, Charles F; Herrmann, Hans-Werner

    2014-01-01

    Long-term studies of individual animals in nature contribute disproportionately to our understanding of the principles of ecology and evolution. Such field studies can benefit greatly from integrating the methods of molecular genetics with traditional approaches. Even though molecular genetic tools are particularly valuable for species that are difficult to observe directly, they have not been widely adopted. Here, we used molecular genetic techniques in a 10-year radio-telemetric investigation of the western diamond-backed rattlesnake (Crotalus atrox) for an analysis of its mating system and to measure sexual selection. Specifically, we used microsatellite markers to genotype 299 individuals, including neonates from litters of focal females to ascertain parentage using full-pedigree likelihood methods. We detected high levels of multiple paternity within litters, yet found little concordance between paternity and observations of courtship and mating behavior. Larger males did not father significantly more offspring, but we found evidence for size-specific male-mating strategies, with larger males guarding females for longer periods in the mating seasons. Moreover, the spatial proximity of males to mothers was significantly associated with reproductive success. Overall, our field observations alone would have been insufficient to quantitatively measure the mating system of this population of C. atrox, and we thus urge more widespread adoption of molecular tools by field researchers studying the mating systems and sexual selection of snakes and other secretive taxa.

  19. Immunohistochemical changes in kidney glomerular and tubular proteins caused by rattlesnake (Crotalus vegrandis venom Cambios inmunohistoquímicos en proteínas de túbulo y glomérulo renal causadas por el veneno de la serpiente de cascabel (Crotalus vegrandis

    Directory of Open Access Journals (Sweden)

    María E. Girón

    2003-10-01

    Full Text Available Renal damage is an important cause of death in patients who have survived the early effects of severe crotalid envenomation. Extracellular matrix of renal tissue is altered by Crotalus toxin activities. The aim of this study was to describe how cytoskeletal proteins and basal membrane components undergo substantial alterations under the action of Crotalus vegrandis crude venom and its hemorrhagic fraction (Uracoina-1 in mice. To detect the proteins in question, the immunoperoxidase method with monoclonal and polyclonal antibodies was used. Cell types within renal lesions were characterized by phenotypic identification, by means of immunohistologic analysis of marker proteins using different primary antibodies against mesangial cells, endothelial cells, cytoskeletal proteins (intermediate filament, extracellular matrix and basal membranes. Samples for morphological study by standard procedures (biotin-streptavidin-peroxidase technique using light microscopy were processed. Positive and negative controls for each antigen tested in the staining assay were included. After crude venom and hemorrhagic fraction inoculation of mice, the disappearance of cytoskeletal vimentin and desmin and collagen proteins in the kidney was observed. In extracellular matrix and basal membranes, collagen type IV from envenomed animals tends to disappear from 24 h to 120 h after venom injection.El daño renal es una causa importante de muerte en pacientes que sobreviven a los efectos iniciales de los severos envenenamientos crotálicos. El objetivo de este estudio ha sido el describir como las proteínas del citoesqueleto y los componentes de membrana basal muestran alteraciones importantes en su manifestación, bajo la acción del veneno crudo de Crotalus vegrandis y una fracción hemorrágica (Uracoina-1 del mismo veneno ya que, la matriz extracelular del tejido renal es alterada por la actividad de estas toxinas. Para detectar las proteínas en cuestión se utiliz

  20. cDNA cloning of a snake venom metalloproteinase from the eastern diamondback rattlesnake (Crotalus adamanteus), and the expression of its disintegrin domain with anti-platelet effects

    Science.gov (United States)

    Suntravat, Montamas; Jia, Ying; Lucena, Sara E.; Sánchez, Elda E.; Pérez, John C.

    2013-01-01

    A 5′ truncated snake venom metalloproteinase was identified from a cDNA library constructed from venom glands of an eastern diamondback rattlesnake (Crotalus adamanteus). The 5′-rapid amplification of cDNA ends (RACE) was used to obtain the 1865 bp full-length cDNA sequence of a snake venom metalloproteinase (CamVMPII). CamVMPII encodes an open reading frame of 488 amino acids, which includes a signal peptide, a pro-domain, a metalloproteinase domain, a spacer, and an RGD-disintegrin domain. The predicted amino acid sequence of CamVMPII showed a 91%, 90%, 83%, and 82% sequence homology to the P-II class enzymes of C. adamanteus metalloproteinase 2, C. atrox CaVMP-II, Gloydius halys agkistin, and Protobothrops jerdonii jerdonitin, respectively. Disintegrins are potent inhibitors of both platelet aggregation and integrin-dependent cell adhesion. Therefore, the disintegrin domain (Cam-dis) of CamVMPII was amplified by PCR, cloned into a pET-43.1a vector, and expressed in Escherichia coli BL21. Affinity purified recombinantly modified Cam-dis (r-Cam-dis) with a yield of 8.5 mg/L culture medium was cleaved from the fusion tags by enterokinase cleavage. r-Cam-dis was further purified by two-step chromatography consisting of HiTrap™ Benzamidine FF column, followed by Talon Metal affinity column with a final yield of 1 mg/L culture. r-Cam-dis was able to inhibit all three processes of platelet thrombus formation including platelet adhesion with an estimated IC50 of 1 nM, collagen- and ADP-induced platelet aggregation with the estimated IC50s of 18 and 6 nM, respectively, and platelet function on clot retraction. It is a potent anti-platelet inhibitor, which should be further investigated for drug discovery to treat stroke patients or patients with thrombotic disorders. PMID:23313448

  1. Isolation and characterization of two disintegrins inhibiting ADP-induced human platelet aggregation from the venom of Crotalus scutulatus scutulatus (Mohave Rattlesnake)

    International Nuclear Information System (INIS)

    Sanchez, Elda E.; Galan, Jacob A.; Russell, William K.; Soto, Julio G.; Russell, David H.; Perez, John C.

    2006-01-01

    Disintegrins and disintegrin-like proteins are molecules found in the venom of four snake families (Atractaspididae, Elapidae, Viperidae, and Colubridae). The disintegrins are nonenzymatic proteins that inhibit cell-cell interactions, cell-matrix interactions, and signal transduction, and may have potential in the treatment of strokes, heart attacks, cancers, and osteoporosis. Prior to 1983, the venom of Crotalus scutulatus scutulatus (Mohave Rattlesnake) was known to be only neurotoxic; however, now there is evidence that these snakes can contain venom with: (1) neurotoxins; (2) hemorrhagins; and (3) both neurotoxins and hemorrhagins. In this study, two disintegrins, mojastin 1 and mojastin 2, from the venom of a Mohave rattlesnake collected in central Arizona (Pinal County), were isolated and characterized. The disintegrins in these venoms were identified by mass-analyzed laser desorption ionization/time-of-flight/time-of-flight (MALDI/TOF/TOF) mass spectrometry as having masses of 7.436 and 7.636 kDa. Their amino acid sequences are similar to crotratroxin, a disintegrin isolated from the venom of the western diamondback rattlesnake (C. atrox). The amino acid sequence of mojastin 1 was identical to the amino acid sequence of a disintegrin isolated from the venom of the Timber rattlesnake (C. horridus). The disintegrins from the Mohave rattlesnake venom were able to inhibit ADP-induced platelet aggregation in whole human blood both having IC 5 s of 13.8 nM, but were not effective in inhibiting the binding of human urinary bladder carcinoma cells (T24) to fibronectin

  2. Wound healing activity and mechanisms of action of an antibacterial protein from the venom of the eastern diamondback rattlesnake (Crotalus adamanteus.

    Directory of Open Access Journals (Sweden)

    Ramar Perumal Samy

    Full Text Available Basic phospholipase A2 was identified from the venom of the eastern diamondback rattlesnake. The Crotalus adamanteus toxin-II (CaTx-II induced bactericidal effects (7.8 µg/ml on Staphylococcus aureus, while on Burkholderia pseudomallei (KHW, and Enterobacter aerogenes were killed at 15.6 µg/ml. CaTx-II caused pore formation and membrane damaging effects on the bacterial cell wall. CaTx-II was not cytotoxic on lung (MRC-5, skin fibroblast (HEPK cells and in mice. CaTx-II-treated mice showed significant wound closure and complete healing by 16 days as compared to untreated controls (**P<0.01. Histological examination revealed enhanced collagen synthesis and neovascularization after treatment with CaTx-II versus 2% Fusidic Acid ointment (FAO treated controls. Measurement of tissue cytokines revealed that interleukin-1 beta (IL-1β expression in CaTx-II treated mice was significantly suppressed versus untreated controls. In contrast, cytokines involved in wound healing and cell migration i.e., monocyte chemotactic protein-1 (MCP-1, fibroblast growth factor-basic (FGF-b, chemokine (KC, granulocyte-macrophage colony-stimulating factor (GM-CSF were significantly enhanced in CaTx-II treated mice, but not in the controls. CaTx-II also modulated nuclear factor-kappa B (NF-κB activation during skin wound healing. The CaTx-II protein highlights distinct snake proteins as a potential source of novel antimicrobial agents with significant therapeutic application for bacterial skin infections.

  3. 78 FR 73877 - Endangered Species; Marine Mammals; Receipt of Applications for Permit

    Science.gov (United States)

    2013-12-09

    ..., Division of Management Authority, U.S. Fish and Wildlife Service, 4401 North Fairfax Drive, Room 212... (Tragopan blythii) Harpy eagle (Harpia harpyja) Bali mynah (Leucopsar rothschildi) Applicant: The Living...: Bali starling (Leucopsar rothschildi) Aruba Island rattlesnake (Crotalus durissus unicolor) Applicant...

  4. Comparative venomics of the Prairie Rattlesnake (Crotalus viridis viridis) from Colorado: Identification of a novel pattern of ontogenetic changes in venom composition and assessment of the immunoreactivity of the commercial antivenom CroFab®.

    Science.gov (United States)

    Saviola, Anthony J; Pla, Davinia; Sanz, Libia; Castoe, Todd A; Calvete, Juan J; Mackessy, Stephen P

    2015-05-21

    Here we describe and compare the venomic and antivenomic characteristics of both neonate and adult Prairie Rattlesnake (Crotalus viridis viridis) venoms. Although both neonate and adult venoms contain unique components, similarities among protein family content were seen. Both neonate and adult venoms consisted of myotoxin, bradykinin-potentiating peptide (BPP), phospholipase A2 (PLA2), Zn(2+)-dependent metalloproteinase (SVMP), serine proteinase, L-amino acid oxidase (LAAO), cysteine-rich secretory protein (CRISP) and disintegrin families. Quantitative differences, however, were observed, with venoms of adults containing significantly higher concentrations of the non-enzymatic toxic compounds and venoms of neonates containing higher concentrations of pre-digestive enzymatic proteins such as SVMPs. To assess the relevance of this venom variation in the context of snakebite and snakebite treatment, we tested the efficacy of the common antivenom CroFab® for recognition of both adult and neonate venoms in vitro. This comparison revealed that many of the major protein families (SVMPs, CRISP, PLA2, serine proteases, and LAAO) in both neonate and adult venoms were immunodepleted by the antivenom, whereas myotoxins, one of the major toxic components of C. v. viridis venom, in addition to many of the small peptides, were not efficiently depleted by CroFab®. These results therefore provide a comprehensive catalog of the venom compounds present in C. v. viridis venom and new molecular insight into the potential efficacy of CroFab® against human envenomations by one of the most widely distributed rattlesnake species in North America. Comparative proteomic analysis of venoms of neonate and adult Prairie Rattlesnake (Crotalus viridis viridis) from a discrete population in Colorado revealed a novel pattern of ontogenetic shifts in toxin composition for viperid snakes. The observed stage-dependent decrease of the relative content of disintegrins, catalytically active D49-PLA2s

  5. A low-cost method to test cytotoxic effects of Crotalus vegrandis (Serpentes: Viperidae venom on kidney cell cultures Un método de bajo costo para probar los efectos citotóxicos del veneno de Crotalus vegrandis (Serpentes: Viperidae en cultivos de células renales

    Directory of Open Access Journals (Sweden)

    María E. Girón

    2005-06-01

    Full Text Available The pathogenesis of the renal lesion upon envenomation by snakebite has been related to myolysis, hemolysis, hypotension and/or direct venom nephrotoxicity caused by the venom. Both primary and continuous cell culture systems provide an in vitro alternative for quantitative evaluation of the toxicity of snake venoms. Crude Crotalus vegrandis venom was fractionated by molecular exclusion chromatography. The toxicity of C. vegrandis crude venom, hemorrhagic, and neurotoxic fractions were evaluated on mouse primary renal cells and a continuous cell line of Vero cells maintained in vitro. Cells were isolated from murine renal cortex and were grown in 96 well plates with Dulbecco's Modified Essential Medium (DMEM and challenged with crude and venom fractions. The murine renal cortex cells exhibited epithelial morphology and the majority showed smooth muscle actin determined by immune-staining. The cytotoxicity was evaluated by the tetrazolium colorimetric method. Cell viability was less for crude venom, followed by the hemorrhagic and neurotoxic fractions with a CT50 of 4.93, 18.41 and 50.22 µg/mL, respectively. The Vero cell cultures seemed to be more sensitive with a CT50 of 2.9 and 1.4 µg/mL for crude venom and the hemorrhagic peak, respectively. The results of this study show the potential of using cell culture system to evaluate venom toxicity.La patogénesis de la lesion renal ha sido relacionada a la miolisis, hemólisis, hipotensión y/o el efecto directo del veneno. Tanto el cultivo primario o el cultivo celular continuo proveen una alternativa in vitro para la evaluación cuantitativa de la toxicidad de venenos de serpiente. El veneno crudo de Crotalus vegrandis fue fraccionado por una cromatografía de exclusión molecular. La toxicidad del veneno crudo de C. vegrandis, sus fracciones hemorrágicas y neurotóxicas fueron evaluadas en células renales primarias de ratón y una línea continua de células Vero mantenidas in vitro. Las c

  6. Nowhere to Go but Up: Impacts of Climate Change on Demographics of a Short-Range Endemic (Crotalus willardi obscurus) in the Sky-Islands of Southwestern North America.

    Science.gov (United States)

    Davis, Mark A; Douglas, Marlis R; Webb, Colleen T; Collyer, Michael L; Holycross, Andrew T; Painter, Charles W; Kamees, Larry K; Douglas, Michael E

    2015-01-01

    Biodiversity elements with narrow niches and restricted distributions (i.e., 'short range endemics,' SREs) are particularly vulnerable to climate change. The New Mexico Ridge-nosed Rattlesnake (Crotalus willardi obscurus, CWO), an SRE listed under the U.S. Endangered Species Act within three sky islands of southwestern North America, is constrained at low elevation by drought and at high elevation by wildfire. We combined long-term recapture and molecular data with demographic and niche modeling to gauge its climate-driven status, distribution, and projected longevity. The largest population (Animas) is numerically constricted (N = 151), with few breeding adults (Nb = 24) and an elevated inbreeding coefficient (ΔF = 0.77; 100 years). Mean home range (0.07 km2) is significantly smaller compared to other North American rattlesnakes, and movements are within, not among sky islands. Demographic values, when gauged against those displayed by other endangered/Red-Listed reptiles [e.g., Loggerhead Sea Turtle (Caretta caretta)], are either comparable or markedly lower. Survival rate differs significantly between genders (femalesky islands. CWO is a rare organism in a unique environment, with a conserved niche and a predisposition towards extinction. It is a bellwether for the eventual climate-driven collapse of the Madrean pine-oak ecosystem, one of Earth's three recognized megadiversity centers.

  7. Analysis of Brazilian snake venoms by neutron activation analysis

    International Nuclear Information System (INIS)

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

    1991-01-01

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

  8. Nowhere to Go but Up: Impacts of Climate Change on Demographics of a Short-Range Endemic (Crotalus willardi obscurus in the Sky-Islands of Southwestern North America.

    Directory of Open Access Journals (Sweden)

    Mark A Davis

    Full Text Available Biodiversity elements with narrow niches and restricted distributions (i.e., 'short range endemics,' SREs are particularly vulnerable to climate change. The New Mexico Ridge-nosed Rattlesnake (Crotalus willardi obscurus, CWO, an SRE listed under the U.S. Endangered Species Act within three sky islands of southwestern North America, is constrained at low elevation by drought and at high elevation by wildfire. We combined long-term recapture and molecular data with demographic and niche modeling to gauge its climate-driven status, distribution, and projected longevity. The largest population (Animas is numerically constricted (N = 151, with few breeding adults (Nb = 24 and an elevated inbreeding coefficient (ΔF = 0.77; 100 years. Mean home range (0.07 km2 is significantly smaller compared to other North American rattlesnakes, and movements are within, not among sky islands. Demographic values, when gauged against those displayed by other endangered/Red-Listed reptiles [e.g., Loggerhead Sea Turtle (Caretta caretta], are either comparable or markedly lower. Survival rate differs significantly between genders (female

  9. Discovery of human scFvs that cross-neutralize the toxic effects of B. jararacussu and C. d. terrificus venoms.

    Science.gov (United States)

    Silva, Luciano C; Pucca, Manuela B; Pessenda, Gabriela; Campos, Lucas B; Martinez, Edson Z; Cerni, Felipe A; Barbosa, José E

    2018-01-01

    Accidents involving venomous snakes are a public health problem worldwide, causing a large number of deaths per year. In Brazil, the majority of accidents are caused by the Bothrops and Crotalus genera, which are responsible for approximately 80% of severe envenoming cases. The cross-neutralization of snake venoms by antibodies is an important issue for development of more effective treatments. Our group has previously reported the construction of human monoclonal antibody fragments towards Bothrops jararacussu and Crotalus durissus terrificus' venoms. This study aimed to select human single-chain variable fragments (scFvs) that recognize both bothropic and crotalic crude venoms following venoms neutralizing capacity in vitro and in vivo. The cross-reactivity of Cro-Bothrumabs were demonstrated by ELISA and in vitro and in vivo experiments showed that a combination of scFvs neutralizes in vitro toxic activities (e.g. indirect hemolysis and plasma-clotting) of crotalic and bothropic venoms as well as prolonged survival time of envenomed animals. Our results may contribute to the development of the first human polyvalent antivenom against Bothrops jararacussu and Crotalus durissus terrificus venoms, overcoming some undesirable effects caused by conventional serotherapy. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Crotacetin, a novel snake venom C-type lectin, is homolog of convulxin

    Directory of Open Access Journals (Sweden)

    G. Rádis-Baptista

    2005-12-01

    Full Text Available Snake venom (sv C-type lectins encompass a group of hemorrhagic toxins, which are able to interfere with hemostasis. They share significant similarity in their primary structures with C-type lectins of other animals, and also present a conserved carbohydrate recognition domain (CRD. A very well studied sv C-type lectin is the heterodimeric toxin, convulxin (CVX, from the venoms of South American rattlesnakes, Crotalus durissus terrificus and C. d. cascavella. It consists of two subunits, alfa (CVXalpha , 13.9 kDa and beta (CVXbeta , 12.6 kDa, joined by inter and intra-chain disulfide bounds, and is arranged in a tetrameric alpha4beta4 conformation. Convulxin is able to activate platelet and induce their aggregation by acting via p62/GPVI collagen receptor. Several cDNA precursors, homolog of CVX subunits, were cloned by PCR homology screening. As determined by computational analysis, one of them, named crotacetin beta subunit, was predicted as a polypeptide with a tridimensional conformation very similar to other subunits of convulxin-like snake toxins. Crotacetin was purified from C. durissus venoms by gel permeation and reverse phase high performance liquid chromatography. The heterodimeric crotacetin is expressed in the venoms of several C. durissus subspecies, but it is prevalent in the venom of C. durissus cascavella. As inferred from homology modeling, crotacetin induces platelet aggregation but noticeably exhibits antimicrobial activity against Gram-positive and Gram-negative bacteria.

  11. Structural studies of the oligomeric formation crotoxin and their complexes; Estudos estruturais da formacao oligomerica da crotoxina e seus complexos

    Energy Technology Data Exchange (ETDEWEB)

    Morelli, Humberto A.S.; Fernandes, Carlos A.H.; Magro, Angelo J.; Fontes, Marcos R.M.; Fernandez, Roberto M., E-mail: rmorato@ibb.unesp.br [Universidade Estadual Paulista Julio de Mesquita Filho (UNESP), Botucatu, SP (Brazil). Inst. de Biociencias. Dept. de Fisica e Biofisica; Bicev, Renata N.; Oliveira, Cristiano L.P. de [Universidade de Sao Paulo (USP), Sao Paulo, SP (Brazil). Inst. de Fisica. Dept. de Fisica Experimental; Barioni, Marina B.; Ito, Amando S. [Universidade de Sao Paulo (USP), Ribeirao Preto, SP (Brazil). Fac. de Filosofia, Ciencias e Letras. Dept. de Fisica

    2013-08-15

    The crotoxin is a phospholipase A{sub 2} that is the mainly responsible for the neurotoxic effect of the snakes' venom of the species Crotalus durissus, popularly known as rattlesnakes. In the crotoxin there are two subunits: an acidic, non-toxic and devoid of enzymatic action, (called crotoxin A or CA); and a basic, toxic with enzymatic activity (called crotoxin B or CB). The main objective of this work is to study the CA, CB and the complex CA + CB using the Small-Angle X-ray Scattering (SAXS), Optical absorption spectrometry and static fluorescence techniques. (author)

  12. Prevalence of Hepatozoon spp. (Apicomplexa, Hepatozoidae among recently captured Brazilian snakes Prevalência de Hepatozoon spp. (Apicomplexa, Hepatozoidae em serpentes recém-capturadas no Brasil

    Directory of Open Access Journals (Sweden)

    L.H. O'Dwyer

    2003-06-01

    Full Text Available The goal of this study was to determine the prevalence of Hepatozoon spp. infection in recently captured snakes from Botucatu, São Paulo State, Brazil. Blood was collected from all snakes by ventral tail venipuncture. Blood smears were air dried, fixed with methanol, and stained with 10% Giemsa solution. The slides were microscopically examined for detection of hemoparasites by light microscopy at 250x magnification. A total of 238 snakes from 23 species were examined, of which 135 (56.7% were venomous and 103 (43.3% non-venomous snakes. The more numerous venomous species sampled were Crotalus durissus terrificus (n=108 and Bothrops jararaca (n=17 and non-venomous snakes were Oxyrhopus guibei (n=35, Boa constrictor amarali (n=18, and Waglerophis merremi (n=13. Hepatozoon spp. infection was detected in 39 (16.4% snakes. The prevalence in venomous and non-venomous snakes was 20.0% and 11.7%, respectively. The highest prevalences observed were 38.9% for Boa constrictor amarali, 35.3% for Bothrops jararaca, and 19.4% for Crotalus durissus terrificus.O presente estudo teve como objetivo determinar a prevalência da infecção por Hepatozoon spp. em serpentes recém-capturadas da região de Botucatu, São Paulo. O sangue foi coletado de todas as serpentes por punção da veia caudal. Os esfregaços foram secos ao ar, fixados com metanol e corados com solução de Giemsa a 10%. Examinaram-se 238 serpentes pertencentes a 23 espécies, das quais 135 (56,7% eram venenosas e 103 (43,3% não venenosas. As espécies venenosas mais representativas foram Crotalus durissus terrificus (n=108 e Bothrops jararaca (n=17 e as não venenosas foram Oxyrhopus guibei (n=35, Boa constrictor amarali (n=18 e Waglerophis merremi (n=13. A infecção por Hepatozoon spp. foi detectada em 39 (16,4% serpentes. As prevalências em serpentes venenosas e não venenosas foram 20,0% e 11,7%, respectivamente. As maiores prevalências foram 38,9% para Boa constrictor amarali, 35

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

    Directory of Open Access Journals (Sweden)

    Rejane Maria Lira-da-Silva

    1994-01-01

    Full Text Available Great part of lhe avaiable data about snakes reprodution refers to species coming from subtropical and temperate regions. In Brazil, the data is rather rare and can be found in various works where information is restricted. Results from studies developed with five viviparous snakes - Crotalus durissus cascavella (Wagler, 1824. Bothrops erythromelas(Amaral, 1923, B. leucurus (Wagler, 1824, Helicops leopardinus (Schlegel, 1873 and Thamnodynastes strigilis (Thiinberg, 1787 - which come from the Northeast of Brazil (Bahia are described. Data about pregnancy and birth, number, sex ratio, length and weight of neonates is given and discussed.

  14. Crotamine and crotoxin interact with tumor cells and trigger cell death

    International Nuclear Information System (INIS)

    Soares, Marcella Araugio; Pujatti, Priscilla Brunelli; Santos, Raquel Gouvea dos; Dias, Consuelo Latorre Fortes; Chavez Olortegui, Carlos Delfin; Santos, Wagner Gouvea dos

    2007-01-01

    Crotoxin (Crtx) and Crotamine (Crota) are polypeptides isolated from Crotalus durissus terrificus snake venom (CV). Previous reports have been shown therapeutic effects of Crotalus durissus terrificus venom and Crtx on skin, breast and lung tumours, although, the mechanisms of this antitumoral effect are still unknown. The aim of this work was to investigate the antitumoral effect of Crtx and Crota on brain tumours cells (GH3 and RT2) in vitro and their capacity of interaction with these tumour cells membranes. Cell survival after Crtx and Crota treatment was evaluated by MTT assay in different times post-treatment and apoptosis was evaluated by DAPI staining. In order to evaluate the specific interaction of Crtx and Crota, these polypeptides were radiolabelled, using 125 I as radiotracer and binding assays were performed. The results were compared with the binding in nontumoral brain tissue. Crtx and Crota induced apoptosis on both tumour cells lineages but, Crota was more powerful than Crtx 90% and 20% cell death for RT2 cells; 80% and 20% cell death for GH3 cells, respectively). Both 125 I-Crtx and 125 I-Crota bound specifically in glioblastoma membranes. Nonetheless, CV polypeptides recognised glioblastoma cells with higher specificity than normal brain tissue. These results suggest that the Crtx and Crota interactions with the plasmatic membrane of tumour cells may be the first step of the cascade of signalling that trigger their antitumoral effect. (author)

  15. Toxicity attenuation optimization of crotalic venom by gamma radiation and studies of its immunogenic properties

    International Nuclear Information System (INIS)

    Clissa, Patricia Bianca

    1997-01-01

    Literature data show that 2.0 kGy dose of gamma radiation, generated by 60 source, reduces the toxic activity of Crotalus durissus terrificus venom, without altering its immunogenic capacity. When crotoxin, main toxin from crotalic venom, was irradiated with the same dose, toxicity was also reduced and the immunogenicity was maintained. This fact was attributed to aggregates (compounds with high molecular weight generated during irradiation), that showed no toxicity but were able to induce the antibodies formation against native venom. Crotalus durissus terrificus venom was irradiated with 2.0, 3.0, 5.0 and 10.0 kGy doses and submitted to molecular exclusion chromatography, in order to find an efficient dose that produces large amounts of non toxic but still immunogenic aggregates. After being isolated, the products of irradiation were evaluated for the amount produced, molecular alteration, and toxic and immunogenic activities. These parameters were also analyzed for the whole venom irradiated. The results from different doses irradiated venom were compared with native one, and 2.0 kGy dose was confirmed to be the most efficient in the association of toxicity attenuation with maintenance of immunogenicity of the crotalic venom, while other doses, in spite of being efficient in the toxicity attenuation, they were not able to keep the immunogenicity property. So, the dose of 2.0 kGy could be used to immunize animals in order to improve anticrotalic sera production. (author)

  16. Research Article Genetic diversity and structure of Crotalus ...

    Indian Academy of Sciences (India)

    anonimo

    agriculture fields, urbanization, or other human land use activity (Hoekstra et al. ... are the urban expansion and the habitat loss and fragmentation (Campbell and ..... practices and 2) the local people kill the rattlesnakes for fear or beliefs that ...

  17. Gamma radiation damage in crotamine (venom of Brazilian rattlesnake)

    International Nuclear Information System (INIS)

    Costa, T.A.; Rogero, J.R.

    1988-01-01

    Ionizing Radiations changes the molecular structure due to chemical bond destruction. These chemical alterations is able to change the biological properties of the macro-molecules. Crotamine was obtained from Crotalus durissus terrificus venom by molecular exclusion cromatography and irradiated in concentration of 2 mg/ml of NaCl 0,85% with gamma radiation produced by a 60 Co source. We used doses of 100 Gy, 250 Gy, 500 Gy, 1000 Gy and 2000 Gy (dose rate = 1,19.10 3 Gy/h). We performed the following experiments: presence of free SH groups, proteic concentration,SDS-PAGE and immunodifusion. Preliminary results showed an increase of the number of bands in SDS-PAGE suggesting the appearence of protein aggregates that proportional to the dose increasing. The immunodiffusion data showed no modification of the immunochemical activity against theButantan anti - sera. (author) [pt

  18. Influence of the main reactive species formed during the detoxication process of toxins by ionizing radiation; Influencia das principais especies reativas formadas durante o processo de destoxicacao de toxinas por radiacao ionizante

    Energy Technology Data Exchange (ETDEWEB)

    Silva, Murilo Casare da

    2003-07-01

    Ionizing radiation has been satisfactorily employed for venoms detoxification. In this report, the radiation was employed to verify the effects caused by the radiolysis products of water on two toxins (Crotoxin and Crotamine) purified from Crotalus durissus terrificus venom. These effects were analyzed using some substances called 'scavengers', those substances competes for specific reactive species hindering them to act on the toxins molecules. In order to study the possible structural damages caused on the toxins, UV spectra, fluorescence, mass spectrometry, enzymatic activity were employed. In addition, biochemical techniques were employed to evaluate the decrease of toxicity and the immunogenicity of toxins before and after the irradiation. Our results indicate that the irradiation promotes structural damages, even at low doses. These modifications lead to a gradual decrease in toxicity, however, the immunogenic properties of the toxins are preserved. (author)

  19. Effects of gamma radiation on snake venoms

    International Nuclear Information System (INIS)

    Nascimento, N.; Spencer, P.J.; Andrade, H.F.; Guarnieri, M.C.; Rogero, J.R.

    1998-01-01

    Ionizing radiation is able to detoxify several venoms, including snake venoms, without affecting significantly their immunogenic properties. In order to elucidate this phenomena, we conceived a comparative pharmacological study between native and irradiated (2,000 Gy) crotoxin, the main toxin of the South American rattlesnake Crotalus durissus terrificus. Crotoxin was isolated and purified by molecular exclusion chromatography, pI precipitation and, subsequently submitted to irradiation. Gel filtration of the irradiated toxin resulted in some high molecular weight aggregates formation. Crotoxin toxicity decreased two folds after irradiation, as determined by LD 50 in mice. Native and irradiated crotoxin biodistribution ocurred in the same general manner, with renal elimination. However, in contrast to irradiated crotoxin, the native form was initially retained in kidneys. A later concentration (2-3 hr) appeared in phagocytic mononuclear cells rich organs (liver and spleen) and neural junction rich organs (muscle and brain)

  20. Influence of the main reactive species formed during the detoxication process of toxins by ionizing radiation

    International Nuclear Information System (INIS)

    Silva, Murilo Casare da

    2003-01-01

    Ionizing radiation has been satisfactorily employed for venoms detoxification. In this report, the radiation was employed to verify the effects caused by the radiolysis products of water on two toxins (Crotoxin and Crotamine) purified from Crotalus durissus terrificus venom. These effects were analyzed using some substances called 'scavengers', those substances competes for specific reactive species hindering them to act on the toxins molecules. In order to study the possible structural damages caused on the toxins, UV spectra, fluorescence, mass spectrometry, enzymatic activity were employed. In addition, biochemical techniques were employed to evaluate the decrease of toxicity and the immunogenicity of toxins before and after the irradiation. Our results indicate that the irradiation promotes structural damages, even at low doses. These modifications lead to a gradual decrease in toxicity, however, the immunogenic properties of the toxins are preserved. (author)

  1. Evaluation of snake venom phospholipase A{sub 2}: hydrolysis of non-natural esters

    Energy Technology Data Exchange (ETDEWEB)

    Pirolla, Renan A.S.; Baldasso, Paulo A.; Marangoni, Sergio; Moran, Paulo J.S.; Rodrigues, Jose Augusto R., E-mail: jaugusto@iqm.unicamp.b [University of Campinas (UNICAMP), SP (Brazil). Inst. of Chemistry. Dept. of Organic Chemistry

    2011-07-01

    Phospholipase A2 from the rattlesnake Crotalus durissus terrificus was employed for the first time to test its enantioselectivity on the hydrolysis of different non-natural esters. It was observed that the structure of this small enzyme is restrictive in the choice of its lipase action with non-natural substrates. Two forms of the enzyme were used; free and as its cross-linked enzyme aggregate (CLEA). With all substrates, the free enzyme showed activity similar to the CLEA preparation. The advantage of the CLEA phospholipase is the possibility to reuse it in several consecutive reactions without a decrease of activity and selectivity with good but higher yields and ee than with the free enzyme. (author)

  2. Commentary on: "Vascular distensibilities have minor effects on intracardiac shunt patterns in reptiles" by Filogonio et al. (2017).

    Science.gov (United States)

    Hillman, Stanley S; Hedrick, Michael S; Kohl, Zachary F

    2017-06-01

    The recent study by Filogonio et al. (2017) suggested that net cardiac shunt patterns in two species of reptiles (Trachemys scripta and Crotalus durissus) were not significantly influenced by the vascular distensibilities of the systemic and pulmonary vasculatures. This is in contrast to a previously published study (Hillman et al., 2014) in the toad (Rhinella marina) in which net cardiac shunts were predicted primarily by the physical properties of vascular distensibility rather than physiological control of resistance of the systemic and pulmonary vasculature. We analyze the data and conclusions reached by Filogonio et al. (2017) regarding the role of vascular distensibilities in determining net cardiac shunt patterns in reptiles in comparison with toads. In our view, the conclusions reached by Filogonio et al. (2017) are not supported by the data primarily because vascular distensibilities were not measured in the reptiles analyzed in their study. Copyright © 2017 Elsevier GmbH. All rights reserved.

  3. Minimal volume regulation after shrinkage of red blood cells from five species of reptiles

    DEFF Research Database (Denmark)

    Kristensen, Karina; Berenbrink, Michael; Koldkjær, Pia

    2008-01-01

    Red blood cells (RBCs) from most vertebrates restore volume upon hypertonic shrinkage and the mechanisms underlying this regulatory volume increase (RVI) have been studied extensively in these cells. Despite the phylogenetically interesting position of reptiles, very little is known about their red...... cell function. The present study demonstrates that oxygenated RBCs in all major groups of reptiles exhibit no or a very reduced RVI upon ~ 25% calculated hyperosmotic shrinkage. Thus, RBCs from the snakes Crotalus durissus and Python regius, the turtle Trachemys scripta and the alligator Alligator...... was not characterized. It seems, therefore, that the RVI response based on NHE activation was lost among the early sauropsids that gave rise to modern reptiles and birds, while it was retained in mammals. An RVI response has then reappeared in birds, but based on activation of the NKCC. Alternatively, the absence...

  4. Molecular exclusion chromatographic analysis on 60Co irradiated rattlesnake

    International Nuclear Information System (INIS)

    Murata, Y.; Rogero, J.R.

    1988-07-01

    Ionizing radiations are sufficiently energetic to be able of breaking chemical bonds and as result of that molecules of substances present in the irradiated will be chemically changed and their biological properties affected also. In this paper the effects of gamma radiation of Co-60 on the rattlesnake venom were studied. A pool of positive crotamina Crotalus durissus terrificus venom was dissolved in 0,15 M NaCl and irradiated with Co-60. Doses of 100, 250, 500, 750, 1500 and 2000 Gy were applied at the dose rate of 1190 Gy/h. With doses over 500 Gy the solutions became turbid, suggesting the presence of aggregates and structural changes of the proteins. The concentration of proteins from the filtered solution was measured by the Lowry method. One ml samples were measured at 230 nm. The partition coefficients and the areas of the three main fraction obtained were calculated. (author) [pt

  5. {sup 131}I-CRTX internal dosimetry: animal model and human extrapolation

    Energy Technology Data Exchange (ETDEWEB)

    Andrade, Henrique Martins de; Ferreira, Andrea Vidal; Soares, Marcella Araugio; Silveira, Marina Bicalho; Santos, Raquel Gouvea dos [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN-CNEN-MG), Belo Horizonte, MG (Brazil)], e-mail: hma@cdtn.br

    2009-07-01

    Snake venoms molecules have been shown to play a role not only in the survival and proliferation of tumor cells but also in the processes of tumor cell adhesion, migration and angiogenesis. {sup 125}I-Crtx, a radiolabeled version of a peptide derived from Crotalus durissus terrificus snake venom, specifically binds to tumor and triggers apoptotic signalling. At the present work, {sup 125}I-Crtx biokinetic data (evaluated in mice bearing Erlich tumor) were treated by MIRD formalism to perform Internal Dosimetry studies. Doses in several organs of mice were determinate, as well as in implanted tumor, for {sup 131}I-Crtx. Doses results obtained for animal model were extrapolated to humans assuming a similar concentration ratio among various tissues between mouse and human. In the extrapolation, it was used human organ masses from Cristy/Eckerman phantom. Both penetrating and non-penetrating radiation from {sup 131}I in the tissue were considered in dose calculations. (author)

  6. 131I-CRTX internal dosimetry: animal model and human extrapolation

    International Nuclear Information System (INIS)

    Andrade, Henrique Martins de; Ferreira, Andrea Vidal; Soares, Marcella Araugio; Silveira, Marina Bicalho; Santos, Raquel Gouvea dos

    2009-01-01

    Snake venoms molecules have been shown to play a role not only in the survival and proliferation of tumor cells but also in the processes of tumor cell adhesion, migration and angiogenesis. 125 I-Crtx, a radiolabeled version of a peptide derived from Crotalus durissus terrificus snake venom, specifically binds to tumor and triggers apoptotic signalling. At the present work, 125 I-Crtx biokinetic data (evaluated in mice bearing Erlich tumor) were treated by MIRD formalism to perform Internal Dosimetry studies. Doses in several organs of mice were determinate, as well as in implanted tumor, for 131 I-Crtx. Doses results obtained for animal model were extrapolated to humans assuming a similar concentration ratio among various tissues between mouse and human. In the extrapolation, it was used human organ masses from Cristy/Eckerman phantom. Both penetrating and non-penetrating radiation from 131 I in the tissue were considered in dose calculations. (author)

  7. Identification and functional analysis of a novel bradykinin inhibitory peptide in the venoms of New World Crotalinae pit vipers

    International Nuclear Information System (INIS)

    James Graham, Robert Leslie; Graham, Ciaren; McClean, Stephen; Chen, Tianbao; O'Rourke, Martin; Hirst, David; Theakston, David; Shaw, Chris

    2005-01-01

    A novel undecapeptide has been isolated and structurally characterized from the venoms of three species of New World pit vipers from the subfamily, Crotalinae. These include the Mexican moccasin (Agkistrodon bilineatus), the prairie rattlesnake (Crotalus viridis viridis), and the South American bushmaster (Lachesis muta). The peptide was purified from all three venoms using a combination of gel permeation chromatography and reverse-phase HPLC. Automated Edman degradation sequencing and MALDI-TOF mass spectrometry established its peptide primary structure as: Thr-Pro-Pro-Ala-Gly-Pro-Asp-Val-Gly-Pro-Arg-OH, with a non-protonated molecular mass of 1063.18 Da. A synthetic replicate of the peptide was found to be an antagonist of bradykinin action at the rat vascular B2 receptor. This is the first bradykinin inhibitory peptide isolated from snake venom. Database searching revealed the peptide to be highly structurally related (10/11 residues) with a domain residing between the bradykinin-potentiating peptide and C-type natriuretic peptide domains of a recently cloned precursor from tropical rattlesnake (Crotalus durissus terrificus) venom gland. BIP thus represents a novel biological entity from snake venom

  8. Structural aspects of crotalic venom proteins modified by ionizing radiation; Aspectos estruturais de proteinas do veneno crotalico modificadas por radiacao ionizante

    Energy Technology Data Exchange (ETDEWEB)

    Oliveira, Karina Corleto de

    2010-07-01

    Snake bites are a serious public health problem, especially in subtropical countries. In Brazil, the Ministry of Health notified around 26 000 accidents in 2008. The genus Crotalus (rattlesnakes) accounts for approximately 7% of the total, with a high mortality rate of 72% when untreated with the specific serum, the only effective treatment in case of snake bites. In Brazil, the serum is produced in horses which, despite the large size, have a reduced lifespan due to the high toxicity of the antigen. Ionizing radiation has proven to be an excellent tool for reducing the toxicity of venoms and isolated toxins, resulting in better immunogens for serum production, and contributing to the welfare of serum producing animals. Since the action of gamma radiation on venoms and toxins has not been yet fully clarified from the structural point of view, we proposed in this paper, to characterize two toxins of the species Crotalus durissus terrificus: crotoxin and crotamine. After isolation of the toxins of interest by chromatographic techniques, they were subjected to structural analysis with the application of the following methods: Fluorescence, Circular Dichroism, Differential Calorimetry and Infrared Spectroscopy. These tests showed that both crotamine as crotoxin when subjected to gamma radiation, showed changes in their structural conformation compared with the samples in the native state. Such changes probably occur in the secondary and tertiary structure and may explain the changes on the biological activity of these toxins. (author)

  9. Structural aspects of crotalic venom proteins modified by ionizing radiation

    International Nuclear Information System (INIS)

    Oliveira, Karina Corleto de

    2010-01-01

    Snake bites are a serious public health problem, especially in subtropical countries. In Brazil, the Ministry of Health notified around 26 000 accidents in 2008. The genus Crotalus (rattlesnakes) accounts for approximately 7% of the total, with a high mortality rate of 72% when untreated with the specific serum, the only effective treatment in case of snake bites. In Brazil, the serum is produced in horses which, despite the large size, have a reduced lifespan due to the high toxicity of the antigen. Ionizing radiation has proven to be an excellent tool for reducing the toxicity of venoms and isolated toxins, resulting in better immunogens for serum production, and contributing to the welfare of serum producing animals. Since the action of gamma radiation on venoms and toxins has not been yet fully clarified from the structural point of view, we proposed in this paper, to characterize two toxins of the species Crotalus durissus terrificus: crotoxin and crotamine. After isolation of the toxins of interest by chromatographic techniques, they were subjected to structural analysis with the application of the following methods: Fluorescence, Circular Dichroism, Differential Calorimetry and Infrared Spectroscopy. These tests showed that both crotamine as crotoxin when subjected to gamma radiation, showed changes in their structural conformation compared with the samples in the native state. Such changes probably occur in the secondary and tertiary structure and may explain the changes on the biological activity of these toxins. (author)

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

    Directory of Open Access Journals (Sweden)

    Gustavo A. Sandoval

    2012-02-01

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

  11. Growth rates and post-release survival of captive neonate timber rattlesnakes Crotalus horridus

    Science.gov (United States)

    Richard N. Conner; D. Craig Rudolph; Daniel Saenz; Richard R. Schaefer; Shirley J. Burgdorf

    2003-01-01

    The need for conservation and management of rare species is becoming increasingly important as wildlife species and their habitat continue to decline. Translocation of wild captured adults to augment and reintroduce populations has been successfully used for some endangered avian species (see Carrie et al. 1999; Rudolph et al. 1992). In general, success rates for...

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

    Directory of Open Access Journals (Sweden)

    de Roodt, Adolfo Rafael

    2010-06-01

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

  13. Novo método de dosagem de soros antipeçonhentos em camundongos lactentes: I - dosagem do soro anticrotálico New method for dosage of antivenoms sera in baby mice: I-Anticrotalic serum's dosage

    Directory of Open Access Journals (Sweden)

    Bruno Soerensen

    1984-08-01

    Full Text Available A atividade biológica dos soros antipeçonhentos pode ser determinada "in vivo" em pombos, coelhos, cobaios e camundongos adultos, não existindo, entretanto, nenhum método que possa ser recomendado internacionalmente. Estudou-se comparativamente aos métodos tradicionais de dosagem de soros em pombos e camundongos adultos, a validade do uso de camundongos lactentes de 6 a 7 dias, pesando 4 a 5g, inoculados pela via subcutânea. Inicialmente foi determinada a toxidez do veneno de Crotalus durissus terríficus através do estudo da sintomatologia do envenenamento e da atividade letal. O estudo comparativo dos três métodos forneceu maior concordância de resultados em DE100 entre as dosagens realizadas com camundongos lactentes e adultos. A DE100 e DL50 determinadas em camundongos lactentes forneceu resultados mais constantes que os dos métodos de camundongos adultos e pombos. O uso deste novo método permite eliminar a dificuldade de obtenção do atual animal de prova; a utilização de um maior número de animais por ponto de avaliação biológica, possibilitando uma maior precisão e conseguindo-se uma uniformidade nas características exigidas neste tipo de dosagem como peso, idade e linhagem, visando a reprodução sistemática dos resultados.The biological activity of antiserum against venoms can be assessed by "in vivo" tests using pigeons, rabbits, guinea-pigs and adult mice although there is no International standardized reaction for it. The traditional methods using pigeons and adult mice were comparatively studied with those using mice of 4-5 g (6-7 days of age, inoculated by subcutaneous injection. The toxicity of the Crotalus durissus terrificus venom was initially determined by studying the symptomatology after administration of venom and lethal activity. The comparative study of these three methods showed that the best concordance in the results of the ED100 was obtained using adult and baby mice. The most homogeneous ED100

  14. Estabilidade e estudo de penetração cutânea in vitro da rutina veiculada em uma emulsão cosmética através de um modelo de biomembrana alternativo Stability and in vitro penetration study of rutin incorporated in a cosmetic emulsion through an alternative model biomembrane

    Directory of Open Access Journals (Sweden)

    André Rolim Baby

    2008-06-01

    Full Text Available A rutina é empregada como antioxidante e na prevenção da fragilidade capilar. Estudos de penetração in vitro através da pele humana seria a situação ideal, entretanto, há dificuldades de sua obtenção e manutenção de sua viabilidade. Entre os demais modelos de membrana, a muda de pele de cobra se apresenta como estrato córneo puro, fornecendo barreira similar ao humano e é obtida sem a morte do animal. Os objetivos desta pesquisa foram desenvolver e avaliar a estabilidade de uma emulsão cosmética contendo rutina e, como promotor de penetração cutânea, o propilenoglicol; e avaliar a penetração e a retenção cutânea in vitro da referida substância ativa da formulação, empregando um modelo de biomembrana alternativo. A emulsão foi desenvolvida com rutina e propilenoglicol, ambos a 5,0% p/p. Quantificou-se a rutina das emulsões por espectrofotometria a 361,0 nm, método previamente validado. A penetração e retenção cutânea in vitro foram realizadas em células de difusão vertical com muda de pele de cobra de Crotalus durissus, como modelo de biomembrana alternativo, e água destilada e álcool etílico absoluto 99,5% (1:1, como fluido receptor. O experimento foi conduzido em um período de seis horas, a 37,0 ± 0,5 ºC e agitação constante de 300 rpm. Empregou-se o método espectrofotométrico validado a 410,0 nm para a quantificação da rutina após penetração e retenção cutânea. A emulsão não promoveu a penetração cutânea da rutina através da muda de pele de C. durissus, retendo 0,931 ± 0,0391 mg de rutina/mg de muda de pele de cobra. Nas condições de armazenamento a 25,0 ± 2,0 ºC; 5,0 ± 0,5 ºC e 45,0 ± 0,5 ºC, a emulsão apresentou-se quimicamente estável durante 30 dias. De acordo com os resultados, a emulsão não favoreceu a penetração cutânea da rutina, mas apenas sua retenção no estrato córneo de C. durissus, condição considerada estável no período de 30 dias.Rutin is

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

    International Nuclear Information System (INIS)

    Paula, Regina A. de.

    1995-01-01

    Snake bite is a great Public Health problem in our country. The accidents with snakes from Crotalus genus are the most severe. About 1% of the victims die without seratherapy. The antivenons are obtained from hyper immune horse plasma. During the production these animals present signs of envenoming that result in a decrease of organic resistance besides the horses maintenance is very expensive and the producers are fewer, so the sera production is restrict. Many techniques which could reduce the venoms toxicity and increase the sera production using chemical and physical agents have been studied. The gamma rays are excellent tool to detoxify venoms and toxins. It is able to modify protein structures that decrease lethally, toxic and enzymatic activities without modifying the immunogenicity. So, it is important evaluate the sera production in rabbits using gamma rays detoxified venom and crotoxin as immunogen and their power as reagents in immuno assays. In order to obtain the antisera, Crotalus durissus terrificus whole venom or isolated crotoxin was irradiated with 2.000 Gy in 60 Co source, in a 150 mM NaCl solution, and inoculated in rabbits. The sera production were screened by immunoprecipitation, immuno enzymatic (ELISA) and immunoradiometric (IRMA) assays. The specificity was studied by immuno-electrophoresis, ELISA and western blot techniques. The neutralizing power was evaluated by neutralization of phospholipase A 2 activity of toxin in vitro. The antisera were used as reagents in antigen capture assays ELISA and IRMA immuno assays to detect circulant antigens in sera of mice experimentally inoculated with crotalic venom or crotoxin. The results showed that both detoxified venom or crotoxin were good immunogens, and they were able to induce antibodies that could recognize non-irradiated venom or isolated crotoxin. The data suggest that those antibodies present more specificity and higher in vitro neutralizing power, when compared with commercial

  16. Biodistribution and SPECT imaging of 125/131I-crotoxin on mice bearing Ehrlich solid tumour

    International Nuclear Information System (INIS)

    Soares, Marcella Araugio; Santos, Raquel Gouvea dos; Silveira, Marina B.; Simal, Carlos

    2009-01-01

    The search of specific radiopharmaceuticals to be used in breast tumour diagnosis is relevant to complement the techniques applied in conventional medicine. Crotalus durissus terrificus venom (CV) and its main polypeptide, Crotoxin (Crtx), are natural source of several bioactive substances with therapeutical potential. The aim of this work was to evaluate the binding of Crtx with tumour targets in vivo, as well as, evaluate its applicability for breast tumours diagnosis. Crtx was labelled with 125/131 I using lactoperoxidase method and radiochemical analysis was performed by chromatography. 125 I-Crtx was used for biodistribution and pharmacokinetics studies on swiss mice bearing Ehrlich solid tumour, while 131 I-Crtx was used for single photon emission computed tomography (SPECT) imaging. Crtx presented specific binding sites on Ehrlich tumour cells and had a rapid blood clearance (T 1/2 = 201.1 min.). Intratumoral administration increased significantly the activity delivered into the tumour site (128-fold higher) and reduced the kidney burden (7.2-fold lower). 131 I-Crxt demonstrated to interact with tumour cells for until 72 hours allowing good quality images of tumour. Our results indicate the biotechnological potential of Crtx as template for radiopharmaceutical design for cancer diagnosis. (author)

  17. The complementarity-determining region sequences in IgY antivenom hypervariable regions

    Directory of Open Access Journals (Sweden)

    David Gitirana da Rocha

    2017-08-01

    Full Text Available The data presented in this article are related to the research article entitled "Development of IgY antibodies against anti-snake toxins endowed with highly lethal neutralizing activity" (da Rocha et al., 2017 [1]. Complementarity-determining region (CDR sequences are variable antibody (Ab sequences that respond with specificity, duration and strength to identify and bind to antigen (Ag epitopes. B lymphocytes isolated from hens immunized with Bitis arietans (Ba and anti-Crotalus durissus terrificus (Cdt venoms and expressing high specificity, affinity and toxicity neutralizing antibody titers were used as DNA sources. The VLF1, CDR1, CDR2, VLR1 and CDR3 sequences were validated by BLASTp, and values corresponding to IgY VL and VH anti-Ba or anti-Cdt venoms were identified, registered [Gallus gallus IgY Fv Light chain (GU815099/Gallus gallus IgY Fv Heavy chain (GU815098] and used for molecular modeling of IgY scFv anti-Ba. The resulting CDR1, CDR2 and CDR3 sequences were combined to construct the three - dimensional structure of the Ab paratope.

  18. Morphologic and morphometric analysis of Hepatozoon spp. (Apicomplexa, Hepatozoidae of snakes

    Directory of Open Access Journals (Sweden)

    Moço Tatiana Cristina

    2002-01-01

    Full Text Available Hepatozoon species are the most abundant hemoparasites of snakes. Its identification has been based mainly on the morphologic characterization of the gamonts in the peripheral blood of the vertebrate host and also of the cysts found in the internal organs of the vertebrate and invertebrate hosts. Using a computerized image analysis system, we studied five species of Hepatozoon from recently captured snakes in Botucatu, State of São Paulo, Brazil, to evaluate the importance of the morphology and morphometry of the gamonts for the characterization of Hepatozoon species and to analyze the morphologic changes induced in the erythrocytes by the parasite. The studied species were H. terzii of Boa constrictor amarali, Hepatozoon sp. of Crotalus durissusterrificus, H. philodryasi of Philodryas patagoniensis, and H. migonei and H. cyclagrasi of Hydrodynastes gigas. We observed three different groups, one of them including the species H. terzii, H. philodryasi and Hepatozoon sp. of C. durissus terrificus; and the other two consisting of H. migonei and H. cyclagrasi. Degree of alterations in the erythrocytes was variable and it may be useful for characterization of Hepatozoon species.

  19. Comparative study of native and irradiated crotoxin. Biochemical and pharmacological effects

    International Nuclear Information System (INIS)

    Nascimento, N. do.

    1991-01-01

    Ionizing radiation is able to detoxify several venoms, including snake venom, without affecting significantly their antigenic and immunogenic properties. In order to elucidate this phenomena, we conceived a comparative biochemical and pharmacological study between native and gamma irradiated (2000Gy) crotoxin, main toxin of south american rattlesnake Crotalus durissus terrificus. Crotoxin was isolated and purified from crude venom by molecular exclusion chromatography, pI precipitation and then irradiated. Immunodiffusion, electrophoresis and gel filtration showed that the molecular integrity was preserved after irradiation with some higher molecular weight aggregate formation and maintenance of its antigenic capacity. The antibodies induced by irradiated toxin had a similar titer to the antibodies induced by native crotoxin; however with higher protective effects in mice. Crotoxin toxicity became 15 times lower after irradiation, as determined by LD sub(50) in mice. Native and irradiated crotoxin biodistribution occurred with a similar general pattern, with renal elimination. In contrast to irradiated crotoxin, the native crotoxin is initially retained in kidneys. A later concentration (2-3hs) occurs in phagocyticmononuclear cells rich organs (liver and spleen) and neural junctions rich organs (muscle and brain). (author)

  20. Oral microbiota of Brazilian captive snakes

    Directory of Open Access Journals (Sweden)

    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.

  1. Substituted thiobenzoic acid S-benzyl esters as potential inhibitors of a snake venom phospholipase A2: Synthesis, spectroscopic and computational studies

    Science.gov (United States)

    Henao Castañeda, I. C.; Pereañez, J. A.; Jios, J. L.

    2012-11-01

    4-Chlorothiobenzoic acid S-benzyl ester (I), 3-nitrothiobenzoic acid S-benzyl ester (II), 4-nitrothiobenzoic acid S-benzyl ester (III) and 4-methylthiobenzoic acid S-benzyl ester (IV) were prepared and characterized by 1H and 13C NMR, Mass spectrometry and IR spectroscopy. Quantum chemical calculations were performed with Gaussian 09 to calculate the geometric parameters and vibrational spectra. Phospholipase A2 (PLA2) was purified from Crotalus durissus cumanensis venom by molecular exclusion chromatography, followed by reverse phase-high performance liquid chromatography. Two studies of the inhibition of phospholipase A2 activity were performed using phosphatidilcholine and 4-nitro-3-octanoyloxybenzoic acid as substrates, in both cases compound II showed the best inhibitory ability, with 74.89% and 69.91% of inhibition, respectively. Average percentage of inhibition was 52.49%. Molecular docking was carried out with Autodock Vina using as ligands the minimized structures of compounds (I-IV) and as protein PLA2 (PDB code 2QOG). The results suggest that compounds I-IV could interact with His48 at the active site of PLA2. In addition, all compounds showed Van der Waals interactions with residues from hydrophobic channel of the enzyme. This interaction would impede normal catalysis cycle of the PLA2.

  2. Structural aspects of crotoxin modified by ionizing radiation; Aspectos estruturais da crotoxina modificada pela radiacao ionizante

    Energy Technology Data Exchange (ETDEWEB)

    Oliveira, Karina Corleto de; Albero, Felipe Guimaraes; Zezell, Denise Maria; Spencer, Patrick Jack; Nascimento, Nanci do, E-mail: kcorleto@usp.b [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)

    2010-03-15

    Ionizing radiation has proven to be an excellent tool for reducing the toxicity of venoms and isolated toxins, resulting in better immunogens for serum production, the only effective treatment in case of snake bites, and contributing to the welfare of serum-producing animals. Since the action of gamma radiation of venoms and toxins has not been yet fully clarified from the structural point of view, we proposed in this paper, to characterize the crotoxin, a venom protein of the species Crotalus durissus terrificus by Circular Dichroism (CD) and Infrared Spectroscopy (FTIR) techniques. After chromatographic techniques, crotoxin was irradiated with 2.0 kGy ({sup 60}Co source). The CD spectra obtained of native and irradiated crotoxin solutions showed changes between the samples in characteristic regions of -sheet and-helix . The Infrared analyse showed expressive changes in the spectra of the native and irradiated crotoxin (amide I band region). These tests showed that crotoxin when subjected to gamma radiation, showed changes in their structural conformation compared with the samples in the native state. Such changes probably occur in the secondary structure and may explain its neurotoxic activity loss. (author)

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

    Directory of Open Access Journals (Sweden)

    Janaína Lima de Souza

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

  4. Protease activated receptors (PARS) mediation in gyroxin biological activity

    International Nuclear Information System (INIS)

    Silva, Jose Alberto Alves da

    2009-01-01

    Gyroxin is a serine protease enzyme from the South American rattlesnake (Crotalus durissus terrificus) venom; it is only partially characterized and has multiple activities. Gyroxin induces blood coagulation, blood pressure decrease and a neurotoxic behavior named barrel rotation. The mechanisms involved in this neurotoxic activity are not known. Whereas gyroxin is a member of enzymes with high potential to become a new drug with clinical applications such as thrombin, batroxobin, ancrod, tripsyn and kalicrein, it is important to find out how gyroxin works. The analysis on agarose gel electrophoresis and circular dichroism confirmed the molecules' integrity and purity. The gyroxin intravenous administration in mice proved its neurotoxicity (barrel rotation). In vivo studies employing intravital microscopy proved that gyroxin induces vasodilation with the participation of protease activated receptors (PARs), nitric oxide and Na+K+ATPase. The leukocytes' adherence and rolling counting indicated that gyroxin has no pro inflammatory activity. Gyroxin induced platelet aggregation, which was blocked by inhibitors of PAR1 and PAR4 receptors (SCH 79797 and tcY-NH 2 , respectively). Finally, it was proved that the gyroxin temporarily alter the permeability of the blood brain barrier (BBB). Our study has shown that both the protease-activated receptors and nitric oxide are mediators involved in the biological activities of gyroxin. (author)

  5. Traditional uses of medicinal animals in the semi-arid region of northeastern Brazil

    Directory of Open Access Journals (Sweden)

    Alves Rômulo Romeu Nóbrega

    2012-10-01

    Full Text Available Abstract The present work presents an inventory of the traditional medicinal uses of animals in the municipality of Bom Sucesso in Paraíba State (PB in the semiarid northeastern region of Brazil. Information was obtained through the use of semi-structured interviews with 50 people who use zootherapeutic products. A total of 25 animal species used for medicinal purposes were identified (18 vertebrates and seven invertebrates distributed among five taxonomic categories; the groups with the largest numbers of citations were: mammals (8 citations, insects (7, and reptiles (5. The most cited animal species were: Tubinambis merianae “teju” lizards (44 citations; Apis mellifera Italian honeybees (318 citations; Gallus gallus chickens (31 citations; Ovis aries sheep (31 citations; Crotalus durissus rattlesnakes (14 citations; Boa constrictor (12 citations; and Bos taurus cattle (12 citations. A significant number of illnesses and conditions treated with animal-based medicines were cited, and the category with the greatest number of citations was “problems affecting the respiratory system”. Our results suggest that the use of zootherapeutics in the region is persistent, and that knowledge about these curative practices is an integral part of the regional culture. As such, studies concerning the uses of zootherapeutics are important windows to understanding human/environmental/cultural interactions and a pathway to conciliating regional cultures with efforts to conserve the native fauna.

  6. 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.

  7. Reply to the commentary by Hillman et al. on: "Vascular distensibilities have minor effects on intracardiac shunt patterns in reptiles" by Filogonio et al. (2017).

    Science.gov (United States)

    Filogonio, Renato; Costa Leite, Cléo Alcantara; Wang, Tobias

    2017-06-01

    Our meta-analysis (Filogonio et al., 2017) on central vascular blood flows in a snake (Crotalus durissus) and a turtle (Trachemys scripta) was motivated by Hillman et al.'s (2014) analysis on amphibians to investigate whether cardiac shunt patterns depend on cardiac output and vascular distensibilities. In contrast to Hillman et al. (2014), we did not uncover a general trend that supports the notion that cardiac shunts in reptiles are dictated by vascular distensibilities. In addition to our response to the criticism raised by Hillman et al. (2017), we suggest that future experiments should consider (i) both compliance and distensibility of the major arteries; (ii) differences in volume of the systemic and pulmonary circuits to account for the accommodation of stroke volume; and (iii) an evaluation of the pulsatile pressures in both the ventricle and the major arteries to consider the timing of the ventricular ejection provided by opening of the ventricular valves. We hope these suggestions may help future clarification of the relative importance of passive arterial mechanical properties compared to autonomic regulation in determining intracardiac shunts in both amphibians and reptiles. Copyright © 2017 Elsevier GmbH. All rights reserved.

  8. Biodistribution and SPECT imaging of {sup 125/131}I-crotoxin on mice bearing Ehrlich solid tumour

    Energy Technology Data Exchange (ETDEWEB)

    Soares, Marcella Araugio; Santos, Raquel Gouvea dos [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil)], e-mail: marcellaaraugio@yahoo.com.br, e-mail: santosr@cdtn.br; Silveira, Marina B.; Simal, Carlos [Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, MG (Brazil). Faculdade de Medicina; Dias, Consuelo L. Fortes [Fundacao Ezequiel Dias (FUNED), Belo Horizonte, MG (Brazil)

    2009-07-01

    The search of specific radiopharmaceuticals to be used in breast tumour diagnosis is relevant to complement the techniques applied in conventional medicine. Crotalus durissus terrificus venom (CV) and its main polypeptide, Crotoxin (Crtx), are natural source of several bioactive substances with therapeutical potential. The aim of this work was to evaluate the binding of Crtx with tumour targets in vivo, as well as, evaluate its applicability for breast tumours diagnosis. Crtx was labelled with {sup 125/131}I using lactoperoxidase method and radiochemical analysis was performed by chromatography. {sup 125}I-Crtx was used for biodistribution and pharmacokinetics studies on swiss mice bearing Ehrlich solid tumour, while {sup 131}I-Crtx was used for single photon emission computed tomography (SPECT) imaging. Crtx presented specific binding sites on Ehrlich tumour cells and had a rapid blood clearance (T{sub 1/2}= 201.1 min.). Intratumoral administration increased significantly the activity delivered into the tumour site (128-fold higher) and reduced the kidney burden (7.2-fold lower). {sup 131}I-Crxt demonstrated to interact with tumour cells for until 72 hours allowing good quality images of tumour. Our results indicate the biotechnological potential of Crtx as template for radiopharmaceutical design for cancer diagnosis. (author)

  9. Protease activated receptors (PARS) mediation in gyroxin biological activity; Mediacao dos receptores ativados por proteases (PARs) em atividades biologicas da giroxina

    Energy Technology Data Exchange (ETDEWEB)

    Silva, Jose Alberto Alves da

    2009-07-01

    Gyroxin is a serine protease enzyme from the South American rattlesnake (Crotalus durissus terrificus) venom; it is only partially characterized and has multiple activities. Gyroxin induces blood coagulation, blood pressure decrease and a neurotoxic behavior named barrel rotation. The mechanisms involved in this neurotoxic activity are not known. Whereas gyroxin is a member of enzymes with high potential to become a new drug with clinical applications such as thrombin, batroxobin, ancrod, tripsyn and kalicrein, it is important to find out how gyroxin works. The analysis on agarose gel electrophoresis and circular dichroism confirmed the molecules' integrity and purity. The gyroxin intravenous administration in mice proved its neurotoxicity (barrel rotation). In vivo studies employing intravital microscopy proved that gyroxin induces vasodilation with the participation of protease activated receptors (PARs), nitric oxide and Na+K+ATPase. The leukocytes' adherence and rolling counting indicated that gyroxin has no pro inflammatory activity. Gyroxin induced platelet aggregation, which was blocked by inhibitors of PAR1 and PAR4 receptors (SCH 79797 and tcY-NH{sub 2}, respectively). Finally, it was proved that the gyroxin temporarily alter the permeability of the blood brain barrier (BBB). Our study has shown that both the protease-activated receptors and nitric oxide are mediators involved in the biological activities of gyroxin. (author)

  10. Effects of 60Co gamma radiation on crotamine

    Directory of Open Access Journals (Sweden)

    Boni-Mitake M.

    2001-01-01

    Full Text Available Ionizing radiation can change the molecular structure and affect the biological properties of biomolecules. This has been employed to attenuate animal toxins. Crotamine is a strongly basic polypeptide (pI 10.3 from Crotalus durissus terrificus venom composed of 42 amino acid residues. It induces skeletal muscle spasms leading to a spastic paralysis of hind limbs in mice. The objective of the present study was to carry out a biochemical study and a toxic activity assay on native and irradiated crotamine. Crotamine was purified from C.d. terrificus venom by Sephadex G-100 gel filtration followed by ion-exchange chromatography, and irradiated at 2 mg/ml in 0.15 M NaCl with 2.0 kGy gamma radiation emitted by a 60Co source. The native and irradiated toxins were evaluated in terms of structure and toxic activity (LD50. Irradiation did not change the protein concentration, the electrophoretic profile or the primary structure of the protein although differences were shown by spectroscopic techniques. Gamma radiation reduced crotamine toxicity by 48.3%, but did not eliminate it.

  11. Physico-chemical characterization of gamma rays irradiated crotamine

    Energy Technology Data Exchange (ETDEWEB)

    Oliveira, Karina Corleto de; Spencer, Patrick Jack; Nascimento, Nanci do [Instituto de Pesquisas Energeticas e Nucleares (IPEN-CNEN/SP), Sao Paulo, SP (Brazil)], e-mail: kcorleto@usp.br

    2009-07-01

    Ionizing radiation can change the molecular structure and affect the biological properties of biomolecules. It has been employed to attenuate animal toxins. Crotamine, a toxin from Crotalus durissus terrificus (Cdt), is a highly basic polypeptide (pI - 10.3), with myotoxic activity and molecular weight of 4882 Da. It is composed of 42 amino acids residues and reticulated by three disulfide bonds. This study aimed the characterization of irradiated crotamine using Circular Dichroism (CD), Fluorescence Spectroscopy and Differential Scanning Calorimetry (DSC) techniques. We used size exclusion and ion-exchange chromatography to purify it from Cdt crude venom. The pure crotamine was irradiated with 2.0 kGy from a {sup 60}Co source. Native and irradiated crotamine were analyzed in a fluorescence spectrophotometer (Hitachi F-4500), under excitation wavelength at 275 nm and the emission was scanned from 300 to 500 nm. The analysis of fluorescence quenching showed that the irradiated form displayed a lower quantum yield when compared to the native form. CD spectra, obtained from a Jasco, J-180 spectropolarimeter, of native and irradiated crotamine solutions, showed a discrete change between the samples, from apparently ordered conformation to a random coil. Finally, the thermodynamics analysis, realized in a calorimeter METTLER TOLEDO, DSC 822e, showed that irradiation promoted changes in the calorimetric profile. Our results indicate that irradiation leads to progressive changes in the structure of the toxin, which could explain the decrease in myotoxic activity. (author)

  12. Effects of {sup 60} Co gamma radiation on crotamine

    Energy Technology Data Exchange (ETDEWEB)

    Boni-Mitake, M. [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil). Dept. de Radioprotecao Ocupacional; Costa, H.; Spencer, P.J. [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil). Dept. de Bioengenharia; Rogero, J.R [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil). Diretoria de Materiais; Vassilieff, V.S. [UNESP, Botucatu, SP (Brazil). Instituto de Biociencias. Ceatox - Centro de Assistencia Toxicologica]. E-mail: mbmitake@net.ipen.br

    2001-12-01

    Ionizing radiation can change the molecular structure and affect the biological properties of biomolecules. This has been employed to attenuate animal toxins. Crotamine is a strongly basic polypeptide (p I 10.3) from Crotalus durissus terrificus venom composed of 42 amino acid residues. It induces skeletal muscle spasms leading to a spastic paralysis of hind limbs in mice. The objective of the present study was to carry out a biochemical study and a toxic activity assay on native and irradiated crotamine. Crotamine was purified from C.d. terrificus venom by Sephadex G-100 gel filtration followed by ion-exchange chromatography, and irradiated at 2 mg/ml in 0.15 M NaCl with 2.0 kGy gamma radiation emitted by a 60 Co source. The native and irradiated toxins were evaluated in terms of structure and toxic activity (Ld50). Irradiation did not change the protein concentration, the electrophoretic profile or the primary structure of the protein although differences were shown by spectroscopic techniques. Gamma radiation reduced crotamine toxicity by 48.3%, but did not eliminate it. (author)

  13. Immunochemical aspects of crotoxim and its subunits

    International Nuclear Information System (INIS)

    Nakazone, A.K.

    1979-01-01

    Crotamine and crotoxin with the subunits - phospholipase A and crotapotin - were obtained by purification from Crotalus durissus terrificus venom. Interaction studies of the subunits using crotalic antiserum, indicated that: crotoxin is formed of crotapotin and phospholipase A with the molar ratio of 1 to 1; using crotapotin 125 I the presence of a soluble complex was shown with the same antiserum. Immunological precipitation reactions demonstrated that crotapotin is antigenic: crotapotin and phospholipase A presented similar antigenic determinants; crotoxin antiserum reacted with each one of the submits; when the subunits are mixed to form synthetic crotoxin some antigenic determinants are masked in the process of interaction. Crotamine, interacted with crotapotin 1:1, without hidden antigenic determinants crotapotin antigenic site seems to be formed by, at least, one lysine. Enzimatical activity of phospholipase A apreared to be dependent on some reaction conditions when its arginine residues are blocked. Tyrosines of phospholipase A are more susceptible to labelling with 131 I than crotapotin. Gama irradiation of aqueous solutions of the subunits produced modifications in the ultraviolet spectra. A decrease of the enzymatic activity occured as a function of radiation dosis. Immunological activities of crotapotin and phospholipase A were not altered [pt

  14. Physico-chemical characterization of gamma rays irradiated crotamine

    International Nuclear Information System (INIS)

    Oliveira, Karina Corleto de; Spencer, Patrick Jack; Nascimento, Nanci do

    2009-01-01

    Ionizing radiation can change the molecular structure and affect the biological properties of biomolecules. It has been employed to attenuate animal toxins. Crotamine, a toxin from Crotalus durissus terrificus (Cdt), is a highly basic polypeptide (pI - 10.3), with myotoxic activity and molecular weight of 4882 Da. It is composed of 42 amino acids residues and reticulated by three disulfide bonds. This study aimed the characterization of irradiated crotamine using Circular Dichroism (CD), Fluorescence Spectroscopy and Differential Scanning Calorimetry (DSC) techniques. We used size exclusion and ion-exchange chromatography to purify it from Cdt crude venom. The pure crotamine was irradiated with 2.0 kGy from a 60 Co source. Native and irradiated crotamine were analyzed in a fluorescence spectrophotometer (Hitachi F-4500), under excitation wavelength at 275 nm and the emission was scanned from 300 to 500 nm. The analysis of fluorescence quenching showed that the irradiated form displayed a lower quantum yield when compared to the native form. CD spectra, obtained from a Jasco, J-180 spectropolarimeter, of native and irradiated crotamine solutions, showed a discrete change between the samples, from apparently ordered conformation to a random coil. Finally, the thermodynamics analysis, realized in a calorimeter METTLER TOLEDO, DSC 822e, showed that irradiation promoted changes in the calorimetric profile. Our results indicate that irradiation leads to progressive changes in the structure of the toxin, which could explain the decrease in myotoxic activity. (author)

  15. 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.

  16. Crystallization and preliminary X-ray crystallographic analysis of two vascular apoptosis-inducing proteins (VAPs) from Crotalus atrox venom

    International Nuclear Information System (INIS)

    Igarashi, Tomoko; Oishi, Yuko; Araki, Satohiko; Mori, Hidezo; Takeda, Soichi

    2006-01-01

    Vascular apoptosis-inducing protein 1 (VAP1) and VAP2 from C. atrox venom were crystallized in variety of different crystal forms. Diffraction data sets were obtained to 2.5 and 2.15 Å resolution for VAP1 and VAP2, respectively. VAPs are haemorrhagic snake-venom toxins belonging to the reprolysin family of zinc metalloproteinases. In vitro, VAPs induce apoptosis specifically in cultured vascular endothelial cells. VAPs have a modular structure that bears structural homology to mammalian ADAMs (a disintegrin and metalloproteinases). VAP1 is a homodimer with a MW of 110 kDa in which the monomers are connected by a single disulfide bridge. VAP2 is homologous to VAP1 and exists as a monomer with a MW of 55 kDa. In the current study, several crystal forms of VAP1 and VAP2 were obtained using the vapour-diffusion method and diffraction data sets were collected using SPring-8 beamlines. The best crystals of VAP1 and VAP2 generated data sets to 2.5 and 2.15 Å resolution, respectively

  17. CLONING, EXPRESSION, AND HEMOSTATIC ACTIVITIES OF A DISINTEGRIN, r-MOJASTIN 1, FROM THE MOHAVE RATTLESNAKE (Crotalus scutulatus scutulatus)

    Science.gov (United States)

    Sánchez, Elda E.; Lucena, Sara E.; Reyes, Steven; Soto, Julio G.; Cantu, Esteban; Lopez-Johnston, Juan Carlos; Guerrero, Belsy; Salazar, Ana Maria; Rodríguez-Acosta, Alexis; Galán, Jacob A.; Tao, W. Andy; Pérez, John C.

    2012-01-01

    Interactions with exposed subendothelial extracellular proteins and cellular integrins (endothelial cells, platelets and lymphocytes) can cause alterations in the hemostatic system associated with atherothrombotic processes. Many molecules found in snake venoms induce pathophysiological changes in humans, cause edema, hemorrhage, and necrosis. Disintegrins are low molecular weight, non-enzymatic proteins found in snake venom that mediate changes by binding to integrins of platelets or other cells and prevent binding of the natural ligands such as fibrinogen, fibronectin or vitronectin. Disintegrins are of great biomedical importance due to their binding affinities resulting in the inhibition of platelet aggregation, adhesion of cancer cells, and induction of signal transduction pathways. RT-PCR was used to obtain a 216 bp disintegrin cDNA from a C. s. scutulatus snake venom gland. The cloned recombinant disintegrin called r-mojastin 1 codes for 71 amino acids, including 12 cysteines, and an RGD binding motif. r-Mojastin 1 inhibited platelet adhesion to fibronectin with an IC50 of 58.3 nM and ADP-induced platelet aggregation in whole blood with an IC50 of 46 nM. r-Mojastin 1 was also tested for its ability to inhibit platelet ATP release using PRP resulting with an IC50 of 95.6 nM. MALDI-TOF mass spectrum analysis showed that r-mojastin has a mass of 7.9509 kDa. PMID:20598348

  18. Alterações hematológicas em bovinos submetidos ao envenenamento crotálico

    Directory of Open Access Journals (Sweden)

    P. M. Ferreira

    2005-03-01

    Full Text Available RESUMO: O quadro clínico observado ao exame do animal acidentado pela cascavél sul americana (Crotalus durissus terrificus , portadora de veneno contendo quantidades significativas da toxina crotamina, é de difícil diferenciação clínica com outras doenças. Com o objetivo de se estabelecer alterações hematológicas que auxiliem o diagnóstico desta condição mórbida, cinco fêmeas bovinas mestiças, com idade variando entre dois e três anos, foram inoculadas com veneno crotálico do tipo crotamina positivo, utilizando a dose de 0,03 mg por quilo de pêso vivo, na região glútea, por via intramuscular. Colheu-se amostras de sangue para realizar os exames laboratoriais antes e a cada duas horas após a inoculação, até a morte dos animais. Verificou-se que: o número de hemácias, o volume corpuscular médio ( VCM , a hemoglobina corpuscular média ( HCM , os números de leucócitos totais, de linfócitos, de neutrófilos e de monócitos, tiveram aumentos significativos e que os números de leucócitos totais apresentaram comportamento atípico, caracterizado por leucocitoses com valores atingindo até 60.000 celulas/mm,3 a cada seis horas, alternadas por quedas para em torno de 13.000 células/mm3. Pelo exposto, concluiu-se que a observação destas alterações poderá auxiliar o estabelecimento do diagnóstico desta afecção nos bovinos. PALAVRAS CHAVE: Envenenamento crotálico; hemograma; bovinos. SUMMARY: The clinical findings observed at examination of an animal attacked by South American rattle snake (Crotalus durissus terrificus , that has poison containing significative crotamine toxin quantites are difficult to distinguish from other diseases. With the aim of establishing haematological parameters of diagnostic value for this condition diagnosis, five crossbred female cattle, from two to three years old, were inoculated with

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

    Directory of Open Access Journals (Sweden)

    Julio C. Mendoza

    2012-03-01

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

  20. Crotoxin in humans: analysis of the effects on extraocular and facial muscles

    Directory of Open Access Journals (Sweden)

    Geraldo de Barros Ribeiro

    2012-12-01

    Full Text Available PURPOSE: Crotoxin is the main neurotoxin of South American rattlesnake Crotalus durissus terrificus. The neurotoxic action is characterized by a presynaptic blockade. The purpose of this research is to assess the ability of crotoxin to induce temporary paralysis of extraocular and facial muscles in humans. METHODS: Doses of crotoxin used ranged from 2 to 5 units (U, each unit corresponding to one LD50. We first applied 2U of crotoxin in one of the extraocular muscles of 3 amaurotic individuals to be submitted to ocular evisceration. In the second stage, we applied crotoxin in 12 extraocular muscles of 9 patients with strabismic amblyopia. In the last stage, crotoxin was used in the treatment of blepharospasm in another 3 patients. RESULTS: No patient showed any systemic side effect or change in vision or any eye structure problem after the procedure. The only local side effects observed were slight conjunctival hyperemia, which recovered spontaneously. In 2 patients there was no change in ocular deviation after 2U crotoxin application. Limitation of the muscle action was observed in 8 of the 12 applications. The change in ocular deviation after application of 2U of crotoxin (9 injections was in average 15.7 prism diopters (PD. When the dose was 4U (2 applications the change was in average 37.5 PD and a single application of 5U produced a change of 16 PD in ocular deviation. This effect lasted from 1 to 3 months. Two of the 3 patients with blepharospasm had the hemifacial spasm improved with crotoxin, which returned after 2 months. CONCLUSIONS: This study provides data suggesting that crotoxin may be a useful new therapeutic option for the treatment of strabismus and blepharospasm. We expect that with further studies crotoxin could be an option for many other medical areas.

  1. Structural analysis of irradiated crotoxin by spectroscopic techniques

    Energy Technology Data Exchange (ETDEWEB)

    Oliveira, Karina C. de; Fucase, Tamara M.; Silva, Ed Carlos S. e; Chagas, Bruno B.; Buchi, Alisson T.; Viala, Vincent L.; Spencer, Patrick J.; Nascimento, Nanci do, E-mail: kcorleto@usp.br [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil). Centro de Biotecnologia

    2013-07-01

    Snake bites are a serious public health problem, especially in subtropical countries. In Brazil, the serum, the only effective treatment in case of snake bites, is produced in horses which, despite of their large size, have a reduced lifespan due to the high toxicity of the antigen. Ionizing radiation has been successfully employed to attenuate the biological activity of animal toxins. Crotoxin, the main toxic compound from Crotalus durissus terrificus (Cdt), is a heterodimeric protein composed of two subunits: crotapotin and phospholipase A{sub 2}. Previous data indicated that this protein, following irradiation process, undergoes unfolding and/or aggregation, resulting in a much lower toxic antigen. The exact mechanisms and structural modifications involved in aggregation process are not clear yet. This work investigates the effects of ionizing radiation on crotoxin employing Infrared Spectroscopy, Circular Dichroism and Dynamic Light Scattering techniques. The infrared spectrum of lyophilized crotoxin showed peaks corresponding to the vibrational spectra of the secondary structure of crotoxin, including β-sheet, random coil, α-helix and β-turns. We calculated the area of these spectral regions after adjusting for baseline and normalization using the amide I band (1590-1700 cm{sup -1}), obtaining the variation of secondary structures of the toxin following irradiation. The Circular Dichroism spectra of native and irradiated crotoxin suggests a conformational change within the molecule after the irradiation process. This data indicates structural changes between the samples, apparently from ordered conformation towards a random coil. The analyses by light scattering indicated that the irradiated crotoxin formed multimers with an average molecular radius 100 folds higher than the native toxin. (author)

  2. Biochemical and pharmacological characterization of irradiated crotamine by gamma rays of 60Co

    International Nuclear Information System (INIS)

    Oliveira, Karina Corleto

    2014-01-01

    The serum production in Brazil, the only effective treatment in cases of snakebites, uses horses that although large size, have reduced l lifespan compared with horses not immunized. Ionizing radiation has been shown as an excellent tool in reducing the toxicity of venoms and toxins isolated, and promote the achievement of better immunogens for serum production, and contributing to the welfare of serum-producing animals. It is known, however, that the effects of ionizing radiation on protein are characterized by various chemical modifications, such as fragmentation, cross-linking due to aggregation and oxidation products generated by water radiolysis. However, the action of gamma radiation on toxins is not yet fully understood structurally and pharmacologically, a fact that prevents the application of this methodology in the serum production process. So we proposed in this paper the characterization of crotamine, an important protein from the venom of Crotalus durissus terrificus species, irradiated with 60 Co gamma rays. After isolating the toxin by chromatographic techniques and testing to prove the obtaining of pure crotamine, it was irradiated with gamma rays and subjected to structural analysis, Fluorescence and Circular Dichroism. Using high hydrostatic pressure tests were also conducted in order to verify that the conformational changes caused by radiation suffer modifications under high pressures. From the pharmacological point of view, muscle contraction tests were conducted with the objective of limiting the action of crotamine in smooth muscle as well as the change in the action of toxin caused structural changes to the front. Analysis of Circular Dichroism and Fluorescence showed changes in structural conformation of crotamine when subjected to gamma radiation and that such changes possibly occurring in the secondary and tertiary structure of the protein. The observed in pharmacological tests showed that the irradiated crotamine was less effective in

  3. A snake venom group IIA PLA2 with immunomodulatory activity induces formation of lipid droplets containing 15-d-PGJ2 in macrophages.

    Science.gov (United States)

    Giannotti, Karina Cristina; Leiguez, Elbio; Carvalho, Ana Eduarda Zulim de; Nascimento, Neide Galvão; Matsubara, Márcio Hideki; Fortes-Dias, Consuelo Latorre; Moreira, Vanessa; Teixeira, Catarina

    2017-06-22

    Crotoxin B (CB) is a catalytically active group IIA sPLA 2 from Crotalus durissus terrificus snake venom. In contrast to most GIIA sPLA 2 s, CB exhibits anti-inflammatory effects, including the ability to inhibit leukocyte functions. Lipid droplets (LDs) are lipid-rich organelles associated with inflammation and recognized as a site for the synthesis of inflammatory lipid mediators. Here, the ability of CB to induce formation of LDs and the mechanisms involved in this effect were investigated in isolated macrophages. The profile of CB-induced 15-d-PGJ 2 (15-Deoxy-Delta-12,14-prostaglandin J 2 ) production and involvement of LDs in 15-d-PGJ 2 biosynthesis were also investigated. Stimulation of murine macrophages with CB induced increased number of LDs and release of 15-d-PGJ 2 . LDs induced by CB were associated to PLIN2 recruitment and expression and required activation of PKC, PI3K, MEK1/2, JNK, iPLA 2 and PLD. Both 15-d-PGJ 2 and COX-1 were found in CB-induced LDs indicating that LDs contribute to the inhibitory effects of CB by acting as platform for synthesis of 15-d-PGJ 2 , a pro-resolving lipid mediator. Together, our data indicate that an immunomodulatory GIIA sPLA 2 can directly induce LD formation and production of a pro-resolving mediator in an inflammatory cell and afford new insights into the roles of LDs in resolution of inflammatory processes.

  4. Study of crotoxin on the induction of paralysis in extraocular muscle in animal model

    Directory of Open Access Journals (Sweden)

    Geraldo de Barros Ribeiro

    2012-10-01

    Full Text Available PURPOSE: Crotoxin is the major toxin of the venom of the South American rattlesnake Crotalus durissus terrificus, capable of causing a blockade of the neurotransmitters at the neuromuscular junction. The objective of this study was to appraise the action and effectiveness of the crotoxin induced paralysis of the extraocular muscle and to compare its effects with the botulinum toxin type A (BT-A. METHODS: The crotoxin, with LD50 of 1.5 µg, was injected into the superior rectus muscle in ten New Zealand rabbits. The concentration variance was 0.015 up to 150 µg. Two rabbits received 2 units of botulinum toxin type A for comparative analysis. The evaluation of the paralysis was performed using serial electromyography. After the functional recovery of the muscles, which occurred after two months, six rabbits were sacrificed for anatomopathology study. RESULTS: The animals did not show any evidence of systemic toxicity. Transitory ptosis was observed in almost every animal and remained up to fourteen days. These toxins caused immediate blockade of the electrical potentials. The recovery was gradual in the average of one month with regeneration signs evident on the electromyography. The paralysis effect of the crotoxin on the muscle was proportional to its concentration. The changes with 1.5 µg crotoxin were similar to those produced by the botulinum toxin type A. The histopathology findings were localized to the site of the injection. No signs of muscle fiber's necrosis were seen in any sample. The alterations induced by crotoxin were also proportional to the concentration and similar to botulinum toxin type A in concentration of 1.5 µg. CONCLUSION: Crotoxin was able to induce transitory paralysis of the superior rectus muscle. This effect was characterized by reduction of action potentials and non-specific signs of fibrillation. Crotoxin, in concentration of 1.5 µg was able to induce similar effects as botulinum toxin type A.

  5. Oxygenation properties and isoform diversity of snake hemoglobins.

    Science.gov (United States)

    Storz, Jay F; Natarajan, Chandrasekhar; Moriyama, Hideaki; Hoffmann, Federico G; Wang, Tobias; Fago, Angela; Malte, Hans; Overgaard, Johannes; Weber, Roy E

    2015-11-01

    Available data suggest that snake hemoglobins (Hbs) are characterized by a combination of unusual structural and functional properties relative to the Hbs of other amniote vertebrates, including oxygenation-linked tetramer-dimer dissociation. However, standardized comparative data are lacking for snake Hbs, and the Hb isoform composition of snake red blood cells has not been systematically characterized. Here we present the results of an integrated analysis of snake Hbs and the underlying α- and β-type globin genes to characterize 1) Hb isoform composition of definitive erythrocytes, and 2) the oxygenation properties of isolated isoforms as well as composite hemolysates. We used species from three families as subjects for experimental studies of Hb function: South American rattlesnake, Crotalus durissus (Viperidae); Indian python, Python molurus (Pythonidae); and yellow-bellied sea snake, Pelamis platura (Elapidae). We analyzed allosteric properties of snake Hbs in terms of the Monod-Wyman-Changeux model and Adair four-step thermodynamic model. Hbs from each of the three species exhibited high intrinsic O2 affinities, low cooperativities, small Bohr factors in the absence of phosphates, and high sensitivities to ATP. Oxygenation properties of the snake Hbs could be explained entirely by allosteric transitions in the quaternary structure of intact tetramers, suggesting that ligation-dependent dissociation of Hb tetramers into αβ-dimers is not a universal feature of snake Hbs. Surprisingly, the major Hb isoform of the South American rattlesnake is homologous to the minor HbD of other amniotes and, contrary to the pattern of Hb isoform differentiation in birds and turtles, exhibits a lower O2 affinity than the HbA isoform. Copyright © 2015 the American Physiological Society.

  6. Structural analysis of irradiated crotoxin by spectroscopic techniques

    International Nuclear Information System (INIS)

    Oliveira, Karina C. de; Fucase, Tamara M.; Silva, Ed Carlos S. e; Chagas, Bruno B.; Buchi, Alisson T.; Viala, Vincent L.; Spencer, Patrick J.; Nascimento, Nanci do

    2013-01-01

    Snake bites are a serious public health problem, especially in subtropical countries. In Brazil, the serum, the only effective treatment in case of snake bites, is produced in horses which, despite of their large size, have a reduced lifespan due to the high toxicity of the antigen. Ionizing radiation has been successfully employed to attenuate the biological activity of animal toxins. Crotoxin, the main toxic compound from Crotalus durissus terrificus (Cdt), is a heterodimeric protein composed of two subunits: crotapotin and phospholipase A 2 . Previous data indicated that this protein, following irradiation process, undergoes unfolding and/or aggregation, resulting in a much lower toxic antigen. The exact mechanisms and structural modifications involved in aggregation process are not clear yet. This work investigates the effects of ionizing radiation on crotoxin employing Infrared Spectroscopy, Circular Dichroism and Dynamic Light Scattering techniques. The infrared spectrum of lyophilized crotoxin showed peaks corresponding to the vibrational spectra of the secondary structure of crotoxin, including β-sheet, random coil, α-helix and β-turns. We calculated the area of these spectral regions after adjusting for baseline and normalization using the amide I band (1590-1700 cm -1 ), obtaining the variation of secondary structures of the toxin following irradiation. The Circular Dichroism spectra of native and irradiated crotoxin suggests a conformational change within the molecule after the irradiation process. This data indicates structural changes between the samples, apparently from ordered conformation towards a random coil. The analyses by light scattering indicated that the irradiated crotoxin formed multimers with an average molecular radius 100 folds higher than the native toxin. (author)

  7. Crotoxin stimulates an M1 activation profile in murine macrophages during Leishmania amazonensis infection.

    Science.gov (United States)

    Farias, L H S; Rodrigues, A P D; Coêlho, E C; Santos, M F; Sampaio, S C; Silva, E O

    2017-09-01

    American tegumentary leishmaniasis is caused by different species of Leishmania. This protozoan employs several mechanisms to subvert the microbicidal activity of macrophages and, given the limited efficacy of current therapies, the development of alternative treatments is essential. Animal venoms are known to exhibit a variety of pharmacological activities, including antiparasitic effects. Crotoxin (CTX) is the main component of Crotalus durissus terrificus venom, and it has several biological effects. Nevertheless, there is no report of CTX activity during macrophage - Leishmania interactions. Thus, the main objective of this study was to evaluate whether CTX has a role in macrophage M1 polarization during Leishmania infection murine macrophages, Leishmania amazonensis promastigotes and L. amazonensis-infected macrophages were challenged with CTX. MTT [3-(4,5dimethylthiazol-2-yl)-2,5-diphenyl tetrasodium bromide] toxicity assays were performed on murine macrophages, and no damage was observed in these cells. Promastigotes, however, were affected by treatment with CTX (IC50 = 22·86 µg mL-1) as were intracellular amastigotes. Macrophages treated with CTX also demonstrated increased reactive oxygen species production. After they were infected with Leishmania, macrophages exhibited an increase in nitric oxide production that converged into an M1 activation profile, as suggested by their elevated production of the cytokines interleukin-6 and tumour necrosis factor-α and changes in their morphology. CTX was able to reverse the L. amazonensis-mediated inhibition of macrophage immune responses and is capable of polarizing macrophages to the M1 profile, which is associated with a better prognosis for cutaneous leishmaniasis treatment.

  8. Biochemical, Pharmacological, and Structural Characterization of New Basic Bbil-TX from Bothriopsis bilineata Snake Venom

    Directory of Open Access Journals (Sweden)

    Victor Corasolla Carregari

    2013-01-01

    Full Text Available Bbil-TX, a PLA2, was purified from Bothriopsis bilineata snake venom after only one chromatographic step using RP-HPLC on μ-Bondapak C-18 column. A molecular mass of 14243.8 Da was confirmed by Q-Tof Ultima API ESI/MS (TOF MS mode mass spectrometry. The partial protein sequence obtained was then submitted to BLASTp, with the search restricted to PLA2 from snakes and shows high identity values when compared to other PLA2s. PLA2 activity was presented in the presence of a synthetic substrate and showed a minimum sigmoidal behavior, reaching its maximal activity at pH 8.0 and 25–37∘C. Maximum PLA2 activity required Ca2+ and in the presence of Cd2+, Zn2+, Mn2+, and Mg2+ it was reduced in the presence or absence of Ca2+. Crotapotin from Crotalus durissus cascavella rattlesnake venom and antihemorrhagic factor DA2-II from Didelphis albiventris opossum sera under optimal conditions significantly inhibit the enzymatic activity. Bbil-TX induces myonecrosis in mice. The fraction does not show a significant cytotoxic activity in myotubes and myoblasts (C2C12. The inflammatory events induced in the serum of mice by Bbil-TX isolated from Bothriopsis bilineata snake venom were investigated. An increase in vascular permeability and in the levels of TNF-a, IL-6, and IL-1 was was induced. Since Bbil-TX exerts a stronger proinflammatory effect, the phospholipid hydrolysis may be relevant for these phenomena.

  9. Biochemical and pharmacological studies of native and irradiated crotamine with gamma radiation of Co60

    International Nuclear Information System (INIS)

    Mitake, M.B.

    2000-01-01

    Ionizing radiation can change the molecular structure and affect the biological properties of biomolecules. This has been employed to attenuate animal toxins. Crotamine is a strongly basic polypeptide from South American rattlesnake venom, composed of 42 amino acid residues. It induces skeletal muscle spasms, leading to a spastic paralysis of hind limbs in mice. The objective was to carry out biochemical and pharmacological studies of native and irradiated crotamine with Co. Crotamine was purified from Crotalus durissus terrificus venom by Sephadex G-100 gel filtration followed by ion exchange chromatography, using a Fast performance Liquid Chromatography (FPLC) system. It was irradiated at 2 mg/ml in 0.15 m NaCl with 2.0 kGy gamma radiation emitted by a Co source. Native and irradiated crotamine were evaluated by biochemical characterization, toxic activity (LD50), and biodistribution. The native and irradiated crotamine were labeled with 29.6 MBq of I using chloramine T method and separated in a Sephadex G-50 column. Male Swiss mice (35 @ 5 g) were injected IP with 0.1 mL (2.4x10 cpm/mouse) of I native crotamine or with 0.4 mL (1.3 x 10 cpm/mouse) of I irradiated crotamine. The animals were sacrificed by ether inhalation at 0.08, 0.25, 0.5,1, 2, 3, 4, 8, 12, and 24 hours. Blood, spleen, liver, kidneys, brain, lungs, heart, and skeletal muscle were collected in order to determine radioactivity content. The results showed that gamma radiation did not change protein concentration, electrophoretic profile, or protein primary structure, although differences could be seen by spectroscopic techniques. Gamma radiation reduced crotamine toxicity, but did not eliminate bioactivity. Biodistribution studies showed that native and irradiated crotamine have hepatic metabolism and renal elimination. Native and irradiated crotamine have an affinity to skeletal muscle and did not cross the blood-brain barrier. (author)

  10. 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

  11. Binding studies of the antitumoral radiopharmaceutical 125I-Crotoxin to Ehrlich ascites tumor cells

    International Nuclear Information System (INIS)

    Silveira, Marina B.; Santos, Raquel G. dos; Dias, Consuelo L. Fortes; Cassali, Geovanni D.

    2009-01-01

    The development of tools for functional diagnostic imaging is mainly based on radiopharmaceuticals that specifically target membrane receptors. Crotoxin (Crtx), a polypeptide isolated from Crotalus durissus terrificus venom, has been shown to have an antitumoral activity and is a promising bioactive tracer for tumor detection. More specific radiopharmaceuticals are being studied to complement the techniques applied in the conventional medicine against breast cancer, the most frequent cause of death from malignant disease in women. Crtx's effect has been shown to be related with the overexpression of epidermal growth factor receptor (EGFR), present in high levels in 30 to 60% of breast tumor cells. Our objective was to evaluate Crtx as a tracer for cancer diagnosis, investigating its properties as an EGFR-targeting agent. Ehrlich ascites tumor cells (EAT cells) were used due to its origin and similar characteristics to breast tumor cells, specially the presence of EGFR. Crtx was labeled with 125I and binding experiments were performed. To evaluate the specific binding in vitro of Crtx, competition binding assay was carried out in the presence of increasing concentrations of non-labelled crotoxin and epidermal growth factor (EGF). Specific binding of 125I-Crtx to EAT cells was determined and the binding was considered saturable, with approximately 70% of specificity, high affinity (Kd = 19.7 nM) and IC50 = 1.6 x 10-11 M. Our results indicate that Crtx's interaction with EAT cells is partially related with EGFR and increases the biotechnological potential of Crtx as a template for radiopharmaceutical design for cancer diagnosis. (author)

  12. Ação neuro-muscular do veneno crotálico: dados preliminares Neuromuscular action of crotalid venom: preliminar data

    Directory of Open Access Journals (Sweden)

    Maria Dorvalina Silva

    1996-03-01

    Full Text Available Estudamos 6 pacientes, 2 cães e um coelho com intoxicação crotálica. Avaliamos a condução nervosa periférica sensitiva e motora, a transmissão neuromuscular e eletromiografias. As biópsias de músculo foram processadas por histoquímica. Os 6 pacientes apresentaram mononeuropatia sensitiva no nervo periférico adjacente ao local da inoculação do veneno e encontramos evidências histoquímicas de miopatia mitocondrial. Os defeitos da transmissão neuromuscular foram mínimos. A maioria dos autores admite que veneno crotálico determina síndrome miastênica. Nossos achados indicam que ptose palpebral, facies miastênico e fraqueza muscular observados após acidente crotálico, correspondem provavelmente a miopatia mitocondrial, muitas vezes transitória e reversível.We studied 6 patients and 2 dogs that have been bitten by South American rattlesnake Crotalus durissus terrificus and one rabbit inoculated with crotalid venom. We analized sensory and motor peripheral nerve conduction, repetitive stimulation for studying neuromuscular transmission and electromyographies. Muscle biopsies were processed by histochemistry. All patients had peripheral mononeuropathy of the closest sensitive nerve to the area of snakebite. The neuromuscular transmission alterations were minimal. Muscle histochemistry of 4 patients, 2 dogs and 1 rabbit showed findings of mitochondrial myopathy. The majority of authors admit that crotalid venom causes myastenic syndrome. Our findings suggest that palpebral ptosis, myastenic facies and muscular weakness observed after crotalid poisoning are, probably, due to transient and reversible mitochondrial myopathy. As far as we know, this is the first report on the ability of the venom of this rattlesnake to cause local sensitive mononeuropathy and the first muscle histochemistry showing mitochondrial myopathy in humans poisoned by crotalid venom.

  13. Hypoxic pulmonary vasoconstriction in reptiles: a comparative study of four species with different lung structures and pulmonary blood pressures.

    Science.gov (United States)

    Skovgaard, Nini; Abe, Augusto S; Andrade, Denis V; Wang, Tobias

    2005-11-01

    Low O2 levels in the lungs of birds and mammals cause constriction of the pulmonary vasculature that elevates resistance to pulmonary blood flow and increases pulmonary blood pressure. This hypoxic pulmonary vasoconstriction (HPV) diverts pulmonary blood flow from poorly ventilated and hypoxic areas of the lung to more well-ventilated parts and is considered important for the local matching of ventilation to blood perfusion. In the present study, the effects of acute hypoxia on pulmonary and systemic blood flows and pressures were measured in four species of anesthetized reptiles with diverse lung structures and heart morphologies: varanid lizards (Varanus exanthematicus), caimans (Caiman latirostris), rattlesnakes (Crotalus durissus), and tegu lizards (Tupinambis merianae). As previously shown in turtles, hypoxia causes a reversible constriction of the pulmonary vasculature in varanids and caimans, decreasing pulmonary vascular conductance by 37 and 31%, respectively. These three species possess complex multicameral lungs, and it is likely that HPV would aid to secure ventilation-perfusion homogeneity. There was no HPV in rattlesnakes, which have structurally simple lungs where local ventilation-perfusion inhomogeneities are less likely to occur. However, tegu lizards, which also have simple unicameral lungs, did exhibit HPV, decreasing pulmonary vascular conductance by 32%, albeit at a lower threshold than varanids and caimans (6.2 kPa oxygen in inspired air vs. 8.2 and 13.9 kPa, respectively). Although these observations suggest that HPV is more pronounced in species with complex lungs and functionally divided hearts, it is also clear that other components are involved.

  14. Binding studies of the antitumoral radiopharmaceutical 125I-Crotoxin to Ehrlich ascites tumor cells

    Energy Technology Data Exchange (ETDEWEB)

    Silveira, Marina B.; Santos, Raquel G. dos [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil); Dias, Consuelo L. Fortes [Fundacao Ezequiel Dias (FUNED), Belo Horizonte, MG (Brazil)], e-mail: consuelo@pq.cnpq.br; Cassali, Geovanni D. [Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, MG (Brazil). Lab. de Patologia Comparada], e-mail: cassalig@icb.ufmg.br

    2009-07-01

    The development of tools for functional diagnostic imaging is mainly based on radiopharmaceuticals that specifically target membrane receptors. Crotoxin (Crtx), a polypeptide isolated from Crotalus durissus terrificus venom, has been shown to have an antitumoral activity and is a promising bioactive tracer for tumor detection. More specific radiopharmaceuticals are being studied to complement the techniques applied in the conventional medicine against breast cancer, the most frequent cause of death from malignant disease in women. Crtx's effect has been shown to be related with the overexpression of epidermal growth factor receptor (EGFR), present in high levels in 30 to 60% of breast tumor cells. Our objective was to evaluate Crtx as a tracer for cancer diagnosis, investigating its properties as an EGFR-targeting agent. Ehrlich ascites tumor cells (EAT cells) were used due to its origin and similar characteristics to breast tumor cells, specially the presence of EGFR. Crtx was labeled with 125I and binding experiments were performed. To evaluate the specific binding in vitro of Crtx, competition binding assay was carried out in the presence of increasing concentrations of non-labelled crotoxin and epidermal growth factor (EGF). Specific binding of 125I-Crtx to EAT cells was determined and the binding was considered saturable, with approximately 70% of specificity, high affinity (Kd = 19.7 nM) and IC50 = 1.6 x 10-11 M. Our results indicate that Crtx's interaction with EAT cells is partially related with EGFR and increases the biotechnological potential of Crtx as a template for radiopharmaceutical design for cancer diagnosis. (author)

  15. Biochemical and pharmacological characterization of irradiated crotamine by gamma rays of {sup 60}Co; Caracterizacao bioquimica e farmacologica da crotamina irradiada por raios gama de {sup 60}Co

    Energy Technology Data Exchange (ETDEWEB)

    Oliveira, Karina Corleto

    2014-07-01

    The serum production in Brazil, the only effective treatment in cases of snakebites, uses horses that although large size, have reduced l lifespan compared with horses not immunized. Ionizing radiation has been shown as an excellent tool in reducing the toxicity of venoms and toxins isolated, and promote the achievement of better immunogens for serum production, and contributing to the welfare of serum-producing animals. It is known, however, that the effects of ionizing radiation on protein are characterized by various chemical modifications, such as fragmentation, cross-linking due to aggregation and oxidation products generated by water radiolysis. However, the action of gamma radiation on toxins is not yet fully understood structurally and pharmacologically, a fact that prevents the application of this methodology in the serum production process. So we proposed in this paper the characterization of crotamine, an important protein from the venom of Crotalus durissus terrificus species, irradiated with {sup 60}Co gamma rays. After isolating the toxin by chromatographic techniques and testing to prove the obtaining of pure crotamine, it was irradiated with gamma rays and subjected to structural analysis, Fluorescence and Circular Dichroism. Using high hydrostatic pressure tests were also conducted in order to verify that the conformational changes caused by radiation suffer modifications under high pressures. From the pharmacological point of view, muscle contraction tests were conducted with the objective of limiting the action of crotamine in smooth muscle as well as the change in the action of toxin caused structural changes to the front. Analysis of Circular Dichroism and Fluorescence showed changes in structural conformation of crotamine when subjected to gamma radiation and that such changes possibly occurring in the secondary and tertiary structure of the protein. The observed in pharmacological tests showed that the irradiated crotamine was less effective

  16. Crotoxin in humans: analysis of the effects on extraocular and facial muscles.

    Science.gov (United States)

    Ribeiro, Geraldo de Barros; Almeida, Henderson Celestino de; Velarde, David Toledo

    2012-01-01

    Crotoxin is the main neurotoxin of South American rattlesnake Crotalus durissus terrificus. The neurotoxic action is characterized by a presynaptic blockade. The purpose of this research is to assess the ability of crotoxin to induce temporary paralysis of extraocular and facial muscles in humans. Doses of crotoxin used ranged from 2 to 5 units (U), each unit corresponding to one LD50. We first applied 2U of crotoxin in one of the extraocular muscles of 3 amaurotic individuals to be submitted to ocular evisceration. In the second stage, we applied crotoxin in 12 extraocular muscles of 9 patients with strabismic amblyopia. In the last stage, crotoxin was used in the treatment of blepharospasm in another 3 patients. No patient showed any systemic side effect or change in vision or any eye structure problem after the procedure. The only local side effects observed were slight conjunctival hyperemia, which recovered spontaneously. In 2 patients there was no change in ocular deviation after 2U crotoxin application. Limitation of the muscle action was observed in 8 of the 12 applications. The change in ocular deviation after application of 2U of crotoxin (9 injections) was in average 15.7 prism diopters (PD). When the dose was 4U (2 applications) the change was in average 37.5 PD and a single application of 5U produced a change of 16 PD in ocular deviation. This effect lasted from 1 to 3 months. Two of the 3 patients with blepharospasm had the hemifacial spasm improved with crotoxin, which returned after 2 months. This study provides data suggesting that crotoxin may be a useful new therapeutic option for the treatment of strabismus and blepharospasm. We expect that with further studies crotoxin could be an option for many other medical areas.

  17. The effects of Western Diamondback Rattlesnake (Crotalus atrox) venom on the production of antihemorrhagins and/or antibodies in the Virginia opossum (Didelphis virginiana).

    Science.gov (United States)

    McKeller, Morgan R; Pérez, John C

    2002-04-01

    Opossums are animals that are naturally resistant to the proteolytic effects of Crotalid venoms. Opossums possess proteinase inhibitors in their sera that bind to and neutralize hemorrhagic and other proteolytic activity in many snake venoms. The proteinase inhibitors are not antibodies since they have different molecular weights (60kDa) and pI (4.2). The purpose of this study was to determine if opossums were capable of producing antibodies against venom and/or increasing the production of proteinase inhibitors (specifically antihemorrhagins). Five different venom immunization protocols were used to determine the effects of the venom in the opossums. The dosages ranged from 1mg of venom per immunization to 350mg/kg body weight of venom per immunization. The antihemorrhagic response was increased, but there is no evidence to suggest that an opossum can produce antibodies against venom. The lack of an antibody response is most likely due to the natural proteinase inhibitors clearing the venom from the opossum's body before an antibody response can occur.

  18. Male Snakes Allocate Time and Energy according to Individual Energetic Status: Body Condition, Steroid Hormones, and Reproductive Behavior in Timber Rattlesnakes, Crotalus horridus.

    Science.gov (United States)

    Lind, Craig M; Beaupre, Steven J

    2015-01-01

    Life-history theory predicts that organisms will hedge current reproductive investment against potential costs in terms of survivorship and future fecundity. However, little is known regarding the endocrine mechanisms underlying bet-hedging strategies in free-ranging male vertebrates. We examined the relationships among individual energetic status, steroid hormones, mate search, and reproductive behavior in free-ranging male timber rattlesnakes. Snakes were monitored over four active seasons in order to test two hypotheses: (1) males adjust the amount of time and energy allocated toward reproduction according to the level of individual energy stores, and (2) observed condition-dependent reproductive allocation is associated with circulating concentrations of steroid hormones (testosterone and corticosterone) thought to regulate reproductive behaviors in vertebrates. A positive relationship between body condition and testosterone was observed in both the field and the laboratory. Male mate search effort was positively correlated with both body condition and testosterone. Body condition and testosterone concentrations were negatively related to time allocated toward foraging during the breeding season. A strong effect of year was observed in the analysis of testosterone and search effort, suggesting that multiple environmental factors impact hormone production and reproductive investment. Corticosterone was not related to any measured variable. Therefore, our results did not indicate a clear role of corticosterone in mediating observed relationships between energetic status and behavior. Observed relationships are consistent with the hypothesis that males allocate time and energy toward reproduction according to individual energetic status and that testosterone plays a role in mediating the trade-off between current reproductive investment and residual reproductive value.

  19. Pancreatic and snake venom presynaptically active phospholipases A2 inhibit nicotinic acetylcholine receptors.

    Science.gov (United States)

    Vulfius, Catherine A; Kasheverov, Igor E; Kryukova, Elena V; Spirova, Ekaterina N; Shelukhina, Irina V; Starkov, Vladislav G; Andreeva, Tatyana V; Faure, Grazyna; Zouridakis, Marios; Tsetlin, Victor I; Utkin, Yuri N

    2017-01-01

    Phospholipases A2 (PLA2s) are enzymes found throughout the animal kingdom. They hydrolyze phospholipids in the sn-2 position producing lysophospholipids and unsaturated fatty acids, agents that can damage membranes. PLA2s from snake venoms have numerous toxic effects, not all of which can be explained by phospholipid hydrolysis, and each enzyme has a specific effect. We have earlier demonstrated the capability of several snake venom PLA2s with different enzymatic, cytotoxic, anticoagulant and antiproliferative properties, to decrease acetylcholine-induced currents in Lymnaea stagnalis neurons, and to compete with α-bungarotoxin for binding to nicotinic acetylcholine receptors (nAChRs) and acetylcholine binding protein. Since nAChRs are implicated in postsynaptic and presynaptic activities, in this work we probe those PLA2s known to have strong presynaptic effects, namely β-bungarotoxin from Bungarus multicinctus and crotoxin from Crotalus durissus terrificus. We also wished to explore whether mammalian PLA2s interact with nAChRs, and have examined non-toxic PLA2 from porcine pancreas. It was found that porcine pancreatic PLA2 and presynaptic β-bungarotoxin blocked currents mediated by nAChRs in Lymnaea neurons with IC50s of 2.5 and 4.8 μM, respectively. Crotoxin competed with radioactive α-bungarotoxin for binding to Torpedo and human α7 nAChRs and to the acetylcholine binding protein. Pancreatic PLA2 interacted similarly with these targets; moreover, it inhibited radioactive α-bungarotoxin binding to the water-soluble extracellular domain of human α9 nAChR, and blocked acetylcholine induced currents in human α9α10 nAChRs heterologously expressed in Xenopus oocytes. These and our earlier results show that all snake PLA2s, including presynaptically active crotoxin and β-bungarotoxin, as well as mammalian pancreatic PLA2, interact with nAChRs. The data obtained suggest that this interaction may be a general property of all PLA2s, which should be proved by

  20. Biochemical and pharmacological studies of native and irradiated crotamine with gamma radiation of 60Co

    International Nuclear Information System (INIS)

    Mitake, Malvina Boni

    2000-01-01

    Ionizing radiation can change the molecular structure and affect the biological properties of biomolecules. This has been employed to attenuate animal toxins. Crotamine is a strongly basic polypeptide from the South American rattlesnake venom, composed of 42 amino acid residues. It induces skeletal muscle spasms leading to a spastic paralysis of hind limbs in mice. The objective of this thesis was carry out biochemical and pharmacological studies of native and irradiated crotamine with 60 Co. Crotamine was purified from Crotalus durissus terrificus venom by Sephadex G-100 gel filtration followed by ion exchange chromatography, using a Fast Performance Liquid Chromatography (FPLC) system. It was irradiated at 2 mg/ml in 0.15 M Na Cl with 2.0 kGy gamma radiation emitted by a 60 Co source. The native and irradiated crotamine were evaluated by biochemical characterization, toxic activity (LD 50 and biodistribution. The native and irradiated crotamine were labelled with 29.6 MBq of 125 I using chloramine T method, and separated in a Sephadex G-50 column. Male Swiss mice (35± 5 g), were injected i.p. with o.1 mL (2.4 x 10 6 cpm/mouse) of 125 I native crotamine or with 0.4 mL (1.3 x 10 6 cpm/mouse) of 125 I irradiated crotamine. At 0.08; 0.25; 0.5; 1; 2; 4; 8; 12 and 24 hours the animal were killed by ether inhalation. Blood, spleen, liver, kidneys, brain, lungs, heart, and skeletal muscle were collected in order to determine radioactivity content. The results showed that gamma radiation did not change the protein concentration, the electroforetic profile or the primary structure of the protein, although differences were shown by spectroscopic techniques. The gamma radiation diminished the toxicity of crotamine, but it did not abolish bioactivity. Biodistribution studies showed that native and irradiated crotamine have hepatic metabolism and renal elimination. The native and irradiated crotamine have affinity by skeletal muscle and they did not pass the blood - brain

  1. Modulation of the pharmacological effects of enzymatically-active PLA2 by BTL-2, an isolectin isolated from the Bryothamnion triquetrum red alga

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    Nagano Celso S

    2008-06-01

    Full Text Available Abstract Background An interaction between lectins from marine algae and PLA2 from rattlesnake was suggested some years ago. We, herein, studied the effects elicited by a small isolectin (BTL-2, isolated from Bryothamnion triquetrum, on the pharmacological and biological activities of a PLA2 isolated from rattlesnake venom (Crotalus durissus cascavella, to better understand the enzymatic and pharmacological mechanisms of the PLA2 and its complex. Results This PLA2 consisted of 122 amino acids (approximate molecular mass of 14 kDa, its pI was estimated to be 8.3, and its amino acid sequence shared a high degree of similarity with that of other neurotoxic and enzymatically-active PLA2s. BTL-2 had a molecular mass estimated in approximately 9 kDa and was characterized as a basic protein. In addition, BTL-2 did not exhibit any enzymatic activity. The PLA2 and BTL-2 formed a stable heterodimer with a molecular mass of approximately 24–26 kDa, estimated by molecular exclusion HPLC. In the presence of BTL-2, we observed a significant increase in PLA2 activity, 23% higher than that of PLA2 alone. BTL-2 demonstrated an inhibition of 98% in the growth of the Gram-positive bacterial strain, Clavibacter michiganensis michiganensis (Cmm, but only 9.8% inhibition of the Gram-negative bacterial strain, Xanthomonas axonopodis pv passiflorae (Xap. PLA2 decreased bacterial growth by 27.3% and 98.5% for Xap and Cmm, respectively, while incubating these two proteins with PLA2-BTL-2 inhibited their growths by 36.2% for Xap and 98.5% for Cmm. PLA2 significantly induced platelet aggregation in washed platelets, whereas BTL-2 did not induce significant platelet aggregation in any assay. However, BTL-2 significantly inhibited platelet aggregation induced by PLA2. In addition, PLA2 exhibited strong oedematogenic activity, which was decreased in the presence of BTL-2. BTL-2 alone did not induce oedema and did not decrease or abolish the oedema induced by the 48

  2. Obtainment of crotoxin/phema-g-LDPE and crotoxin/PCL systems;Obtencao dos sistemas bioconjugados crotoxina/PEBD-g-phema e crotoxina/PCL

    Energy Technology Data Exchange (ETDEWEB)

    Lorenzetti, Solange Gubbelini

    2006-07-01

    The aim of the work was the obtainment of polymeric matrices immobilized with crotoxin purified from crude venom of rattle snake. A matrix was processed by gamma irradiation by the grafting of a hydrogel onto a polymeric film which resulted in a copolymer for the chemical immobilization of crotoxin. The second matrix was attained by the entrapment of crotoxin in microspheres of epsilon-polycaprolactone. After the purification, the crotoxin proceeding from the snake Crotalus durissus terrificus was evaluated biochemical and biologically. The lethal dose (LD50%) of the toxin was 0.09/kg animal. The test of cytotoxicity not revealed any significant difference between the tumoral cells and the respective normal control cells in culture. Grafting copolymers were used as scaffold for the chemical immobilization of the purified crotoxin. For this purpose the low density polyethylene (LDPE) and the hydrophilic monomer 2-hydroxy-ethyl-methacrylate (HEMA) were copolymerized in a {sup 60}Co source. The copolymers (LDPE-g-PHEMA) showed grafting levels in the range of 2 and 50 %. In the infrared spectroscopy analysis (FTIR-ATR) it was observed in the copolymer, carbonyl groups (C=O) and hydroxyl groups -OH due to the grafting of PHEMA. The MEV micrographs showed a smooth surface for the virgin LDPE and a rough surface for the LDPE-g-PHEMA, owing to the presence of grafted PHEMA. The hydrophilicity was observed by the determination of water content in the copolymer after immersion in water. By the diffusion coefficient it was noted that from 30 % grafting degree, the copolymers become less hydrophilic due to the crosslinking increase among the chains in PHEMA. The biocompatibility of the LDPE-g-PHEMA was proved by the cytotoxicity test. At the end, the immobilized copolymer, the entrapped crotoxin and the free crotoxin was tested 'in vivo'. During 20 days, C3H strain mice were observed in their weight, behavior and motor changes. The results demonstrated that the group

  3. The use of zootherapeutics in folk veterinary medicine in the district of Cubati, Paraíba State, Brazil

    Directory of Open Access Journals (Sweden)

    da S Mourão José

    2007-09-01

    Full Text Available Abstract Background The present work addresses the use of zootherapy in folk veterinary medicine (ethnoveterinary by the residents of the municipal district of Cubati, microregion of Seridó, Paraíba State, Brazil. It sought to identify the principal animals used as medicinal sources for zootherapeutics and to contribute to the preservation and sustainability of this traditional knowledge. Methods Field research was undertaken on a weekly or biweekly basis during the period November, 2006, to January, 2007. Free, semi-structured, and open interviews were made with local residents of the municipal district of Cubati (in both urban and rural settings as well as with venders in public markets. A total of 25 individuals of both sexes were interviewed (with ages varying from 26 to 78 years although only 16 were finally chosen as informants as these people demonstrated the greatest degree of knowledge concerning zootherapeutics. Graphs and percentages were generated using Microsoft© Excel 2007 software, and the species were identified by photographic registration and subsequent bibliographical surveys. Results Mammals constitute the main medicinal zootherapeutic source for folk veterinary medicines in the studied area, both in terms of the total number of species used and the frequency of their citation. Sheep (Ovis aries, pigs (Sus scrofa, cattle (Bos taurus, and foxes (Cerdocyon thous were mentioned by 62.5, 43.75, 37.5, and 31.25% of the informants, respectively, as being used in folk veterinary medicine. Additionally, chameleons (Iguana iguana, chickens (Gallus domesticus, and rattlesnakes (Crotalus durissus were mentioned by 75, 43.75, and 31.25% of the informants, respectively. Relatively simple animal illnesses, such as furuncles, or injuries resulting from embedded thorns or skin eruptions are responsible for the largest number of zootherapeutic treatment, while, diseases of greater complexity, such as rabies and brucellosis, were not even

  4. Use of I-131- CRTX for targeting malignant adenocarcinoma in mice: biodistribution and radiation dosimetry

    International Nuclear Information System (INIS)

    Santos, Raquel Gouvea dos; Soares, Marcella Araugio; Andrade, Henrique Martins de; Santos, Marcos Antonio da Cunha

    2008-01-01

    Snake venoms molecules have been shown to play a role not only in the survival and proliferation of tumour cells but also in the processes of tumour cell adhesion, migration and angiogenesis. We have shown that 125 I-Crtx, a radiolabeled version of a protein derived from Crotalus durissus terrificus snake venom (Cdt), specifically binds to tumor and triggers apoptotic death. This study reports the biokinetic profile of 99m Tc-Cdt and 125/131 I-Crtx in Swiss mice bearing Ehrlich solid tumor and MIRD formulation was applied to calculate the absorbed radiation doses for various organs and tumor site. Biokinetic evaluations were performed up to 24 h after intravenous (i.v) or intratumor (i.tu.) injection of 99m Tc-Cdt or 125/131 I-Crtx. Time-activity curves were generated for the main organs by fitting the organ specific mass mean counts. The radiation dose from 131 I-Crtx was calculated based on non penetrating radiation in the mouse model. Biokinetics data from 99m Tc-Cdt after i.v. injection in mice tumor model showed rapid blood clearance (T 1/2 = 36.1 ± 2.4 min.), slow tumor clearance (T 1/2 : 108.3 ± 19.5 min.) and indicated the kidneys as the main excretion pathway. Interaction studies in vitro demonstrated that 125 I-Crtx recognize specific sites on Erlich tumor cell membrane. Upon intravenous and intratumor administration of 131 I-Crtx in mice bearing Erlich tumor, it was observed high uptake in tumor site in vivo (Ã =72kBq x h/g) resulting in a high absorbed dose radiation to tumor site. Distributions of 125/131 I-Crtx i.v. were only significant in tumor, stomach, liver and kidneys, reflecting non-specific uptake of Crtx in normal excretion tissues in vivo. Intratumoral administration reduced significantly the radiation dose to the kidneys (42-fold lower) and increased the uptake by the tumor site (128- fold higher). 131 I control was run in a parallel experiment and showed no significant tumor uptake. In conclusion, 131 I-Crtx had a high concentration in

  5. DNA-interactive properties of crotamine, a cell-penetrating polypeptide and a potential drug carrier.

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    Pei-Chun Chen

    Full Text Available Crotamine, a 42-residue polypeptide derived from the venom of the South American rattlesnake Crotalus durissus terrificus, has been shown to be a cell-penetrating protein that targets chromosomes, carries plasmid DNA into cells, and shows specificity for actively proliferating cells. Given this potential role as a nucleic acid-delivery vector, we have studied in detail the binding of crotamine to single- and double-stranded DNAs of different lengths and base compositions over a range of ionic conditions. Agarose gel electrophoresis and ultraviolet spectrophotometry analysis indicate that complexes of crotamine with long-chain DNAs readily aggregate and precipitate at low ionic strength. This aggregation, which may be important for cellular uptake of DNA, becomes less likely with shorter chain length. 25-mer oligonucleotides do not show any evidence of such aggregation, permitting the determination of affinities and size via fluorescence quenching experiments. The polypeptide binds non-cooperatively to DNA, covering about 5 nucleotide residues when it binds to single (ss or (ds double stranded molecules. The affinities of the protein for ss- vs. ds-DNA are comparable, and inversely proportional to salt levels. Analysis of the dependence of affinity on [NaCl] indicates that there are a maximum of ∼3 ionic interactions between the protein and DNA, with some of the binding affinity attributable to non-ionic interactions. Inspection of the three-dimensional structure of the protein suggests that residues 31 to 35, Arg-Trp-Arg-Trp-Lys, could serve as a potential DNA-binding site. A hexapeptide containing this sequence displayed a lower DNA binding affinity and salt dependence as compared to the full-length protein, likely indicative of a more suitable 3D structure and the presence of accessory binding sites in the native crotamine. Taken together, the data presented here describing crotamine-DNA interactions may lend support to the design of more

  6. Obtainment of crotoxin/phema-g-LDPE and crotoxin/PCL systems

    International Nuclear Information System (INIS)

    Lorenzetti, Solange Gubbelini

    2006-01-01

    The aim of the work was the obtainment of polymeric matrices immobilized with crotoxin purified from crude venom of rattle snake. A matrix was processed by gamma irradiation by the grafting of a hydrogel onto a polymeric film which resulted in a copolymer for the chemical immobilization of crotoxin. The second matrix was attained by the entrapment of crotoxin in microspheres of epsilon-polycaprolactone. After the purification, the crotoxin proceeding from the snake Crotalus durissus terrificus was evaluated biochemical and biologically. The lethal dose (LD50%) of the toxin was 0.09/kg animal. The test of cytotoxicity not revealed any significant difference between the tumoral cells and the respective normal control cells in culture. Grafting copolymers were used as scaffold for the chemical immobilization of the purified crotoxin. For this purpose the low density polyethylene (LDPE) and the hydrophilic monomer 2-hydroxy-ethyl-methacrylate (HEMA) were copolymerized in a 60 Co source. The copolymers (LDPE-g-PHEMA) showed grafting levels in the range of 2 and 50 %. In the infrared spectroscopy analysis (FTIR-ATR) it was observed in the copolymer, carbonyl groups (C=O) and hydroxyl groups -OH due to the grafting of PHEMA. The MEV micrographs showed a smooth surface for the virgin LDPE and a rough surface for the LDPE-g-PHEMA, owing to the presence of grafted PHEMA. The hydrophilicity was observed by the determination of water content in the copolymer after immersion in water. By the diffusion coefficient it was noted that from 30 % grafting degree, the copolymers become less hydrophilic due to the crosslinking increase among the chains in PHEMA. The biocompatibility of the LDPE-g-PHEMA was proved by the cytotoxicity test. At the end, the immobilized copolymer, the entrapped crotoxin and the free crotoxin was tested 'in vivo'. During 20 days, C3H strain mice were observed in their weight, behavior and motor changes. The results demonstrated that the group injected with

  7. Biochemical and pharmacological studies of native and irradiated crotamine with gamma radiation of {sup 60}Co; Estudo bioquimico e farmacologico das crotaminas nativa e irradiada com radiacao gama de {sup 60}Co

    Energy Technology Data Exchange (ETDEWEB)

    Mitake, Malvina Boni

    2000-07-01

    Ionizing radiation can change the molecular structure and affect the biological properties of biomolecules. This has been employed to attenuate animal toxins. Crotamine is a strongly basic polypeptide from the South American rattlesnake venom, composed of 42 amino acid residues. It induces skeletal muscle spasms leading to a spastic paralysis of hind limbs in mice. The objective of this thesis was carry out biochemical and pharmacological studies of native and irradiated crotamine with {sup 60} Co. Crotamine was purified from Crotalus durissus terrificus venom by Sephadex G-100 gel filtration followed by ion exchange chromatography, using a Fast Performance Liquid Chromatography (FPLC) system. It was irradiated at 2 mg/ml in 0.15 M Na Cl with 2.0 kGy gamma radiation emitted by a {sup 60} Co source. The native and irradiated crotamine were evaluated by biochemical characterization, toxic activity (LD{sub 50} and biodistribution. The native and irradiated crotamine were labelled with 29.6 MBq of {sup 125} I using chloramine T method, and separated in a Sephadex G-50 column. Male Swiss mice (35{+-} 5 g), were injected i.p. with o.1 mL (2.4 x 10{sup 6} cpm/mouse) of {sup 125} I native crotamine or with 0.4 mL (1.3 x 10{sup 6} cpm/mouse) of {sup 125} I irradiated crotamine. At 0.08; 0.25; 0.5; 1; 2; 4; 8; 12 and 24 hours the animal were killed by ether inhalation. Blood, spleen, liver, kidneys, brain, lungs, heart, and skeletal muscle were collected in order to determine radioactivity content. The results showed that gamma radiation did not change the protein concentration, the electroforetic profile or the primary structure of the protein, although differences were shown by spectroscopic techniques. The gamma radiation diminished the toxicity of crotamine, but it did not abolish bioactivity. Biodistribution studies showed that native and irradiated crotamine have hepatic metabolism and renal elimination. The native and irradiated crotamine have affinity by skeletal

  8. Pancreatic and snake venom presynaptically active phospholipases A2 inhibit nicotinic acetylcholine receptors.

    Directory of Open Access Journals (Sweden)

    Catherine A Vulfius

    Full Text Available Phospholipases A2 (PLA2s are enzymes found throughout the animal kingdom. They hydrolyze phospholipids in the sn-2 position producing lysophospholipids and unsaturated fatty acids, agents that can damage membranes. PLA2s from snake venoms have numerous toxic effects, not all of which can be explained by phospholipid hydrolysis, and each enzyme has a specific effect. We have earlier demonstrated the capability of several snake venom PLA2s with different enzymatic, cytotoxic, anticoagulant and antiproliferative properties, to decrease acetylcholine-induced currents in Lymnaea stagnalis neurons, and to compete with α-bungarotoxin for binding to nicotinic acetylcholine receptors (nAChRs and acetylcholine binding protein. Since nAChRs are implicated in postsynaptic and presynaptic activities, in this work we probe those PLA2s known to have strong presynaptic effects, namely β-bungarotoxin from Bungarus multicinctus and crotoxin from Crotalus durissus terrificus. We also wished to explore whether mammalian PLA2s interact with nAChRs, and have examined non-toxic PLA2 from porcine pancreas. It was found that porcine pancreatic PLA2 and presynaptic β-bungarotoxin blocked currents mediated by nAChRs in Lymnaea neurons with IC50s of 2.5 and 4.8 μM, respectively. Crotoxin competed with radioactive α-bungarotoxin for binding to Torpedo and human α7 nAChRs and to the acetylcholine binding protein. Pancreatic PLA2 interacted similarly with these targets; moreover, it inhibited radioactive α-bungarotoxin binding to the water-soluble extracellular domain of human α9 nAChR, and blocked acetylcholine induced currents in human α9α10 nAChRs heterologously expressed in Xenopus oocytes. These and our earlier results show that all snake PLA2s, including presynaptically active crotoxin and β-bungarotoxin, as well as mammalian pancreatic PLA2, interact with nAChRs. The data obtained suggest that this interaction may be a general property of all PLA2s, which

  9. Study of crotoxin mechanism of action to mammary carcinomas and evaluation of its potential as a radiopharmaceutical

    International Nuclear Information System (INIS)

    Silveira, Marina Bicalho

    2010-01-01

    Crotoxin, the main component of Crotalus durissus terrificus snake venom, has been studied since 1938. It is a natural polypeptidic complex with pharmacological potential because of its antitumoral properties which has attracted great interest for diagnosis and therapy of oncological diseases. However, Crotoxin mechanism of action and sites of specific interaction on tumor cells are still misunderstood. Breast cancer is the second most frequent type in the world and the most common cancer in women. About 30 to 60% of mammary tumors overexpress epidermal growth factor receptor (EGFR), a transmembrane protein related to cell proliferation. Since literature has reported that Crotoxin antitumoral effect is more potent on cells with EGFR overexpression the objectives of this work were to evaluate Crotoxin cytotoxic effects on mammary tumor cells human breast carcinoma (MCF-7) and Ehrlich tumor cells (murine ascitics carcinoma), and to investigate the specific molecular interaction of Crotoxin on Ehrlich tumor cells. Initially, Crotoxin was radiolabelled with iodine-125 ( 125 I-Crotoxin) and iodine-131 ( 131 I-Crotoxin). Saturation and competition assay were carried out to characterize Crotoxin in vitro interaction; Crotoxin biodistribution studies and singlephoton emission computed tomography (SPECT) of mice bearing Ehrlich tumor have been evaluated to describe in vivo interaction. Our results showed that Crotoxin presented cytotoxic effect against Ehrlich with DL 50 in vitro (concentration of compound which is lethal for 50% of cells) of about one micromolar, but did not present significant effect against MCF-7. Morphological alterations characteristic of apoptosis suggests programmed cell death. 125 I-Crotoxin interaction with Ehrlich tumor cells was saturable with approximately 70% specificity, and presented K d =24.98 nmol/L and B max =16,570 sites/cell for low affinity binding sites and K d =0.06 nmol/L and B max =210 sites/cell high affinity binding sites

  10. Crotoxin in humans: analysis of the effects on extraocular and facial muscles Crotoxina em humanos: estudo da ação em músculos extraoculares e faciais

    Directory of Open Access Journals (Sweden)

    Geraldo de Barros Ribeiro

    2012-12-01

    Full Text Available PURPOSE: Crotoxin is the main neurotoxin of South American rattlesnake Crotalus durissus terrificus. The neurotoxic action is characterized by a presynaptic blockade. The purpose of this research is to assess the ability of crotoxin to induce temporary paralysis of extraocular and facial muscles in humans. METHODS: Doses of crotoxin used ranged from 2 to 5 units (U, each unit corresponding to one LD50. We first applied 2U of crotoxin in one of the extraocular muscles of 3 amaurotic individuals to be submitted to ocular evisceration. In the second stage, we applied crotoxin in 12 extraocular muscles of 9 patients with strabismic amblyopia. In the last stage, crotoxin was used in the treatment of blepharospasm in another 3 patients. RESULTS: No patient showed any systemic side effect or change in vision or any eye structure problem after the procedure. The only local side effects observed were slight conjunctival hyperemia, which recovered spontaneously. In 2 patients there was no change in ocular deviation after 2U crotoxin application. Limitation of the muscle action was observed in 8 of the 12 applications. The change in ocular deviation after application of 2U of crotoxin (9 injections was in average 15.7 prism diopters (PD. When the dose was 4U (2 applications the change was in average 37.5 PD and a single application of 5U produced a change of 16 PD in ocular deviation. This effect lasted from 1 to 3 months. Two of the 3 patients with blepharospasm had the hemifacial spasm improved with crotoxin, which returned after 2 months. CONCLUSIONS: This study provides data suggesting that crotoxin may be a useful new therapeutic option for the treatment of strabismus and blepharospasm. We expect that with further studies crotoxin could be an option for many other medical areas.OBJETIVO: A crotoxina é a principal neurotoxina da cascavel sul-americana Crotalus durissus terrificus e sua ação neurotóxica caracteriza-se por um bloqueio pr

  11. Quadro clínico-patológico do envenenamento crotálico experimental em bubalinos comparado com o de bovinos

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    José Diomedes Barbosa

    2011-11-01

    Full Text Available O estudo teve por objetivo verificar a sensibilidade dos bubalinos à peçonha de Crotalus durissus terriiicus, estudar o quadro clínico-patológico e laboratorial nessa espécie e estabelecer comparações com o verificado em bovinos. A inoculação do veneno liofilizado de Crotalus durissus terriiicus, diluído em 1ml de solução fisiológica, foi feita na região da articulação úmero-rádio-ulnar, por via subcutânea, em três bubalinos (doses de 0,015; 0,03; e 0,066mg/kg e em dois bovinos (doses de 0,03 e 0,066mg/kg. O bubalino que recebeu a dose de 0,03mg/ kg apresentou sinais clínicos graves recuperou-se seis dias após, e o bovino que recebeu a mesma dose morreu com evolução de 22h56min. A dose de 0,066mg/kg causou a morte tanto do bovino quanto do bubalino, com evolução clínica de 4h23min e 8h12min, respectivamente. O bubalino que recebeu a dose de 0,015mg/kg, recuperou-se com evolução de 48 horas. Os sinais clínicos tiveram início dentro de 3h58min no bubalino que morreu, e nos bubalinos que adoeceram mas se recuperaram, dentro de 17h25min e 24h00min após a inoculação do veneno. Nos dois bovinos que morreram (com doses de 0,03 e 0,066mg/kg, os primeiros sinais clínicos foram observados 6h10min e 6h31min após a inoculação do veneno. A inoculação do veneno produziu nos búfalos e bovinos um quadro nervoso de paralisia flácida. Os principais sinais observados tanto nos búfalos quanto nos bovinos, foram discreto aumento de volume no local da inoculação, dificuldade respiratória caracterizada por respiração predominantemente abdominal, apatia, sialorreia, dificuldade para se levantar quando estimulados, evolução para decúbito esternal permanente, seguido de decúbito lateral e movimentos de pedalagem, e diminuição dos reflexos relacionados aos pares de nervos cranianos. Nos bubalinos adicionalmente foi observado aumento da base de sustentação, arrastar das pinças dos membros posteriores, marcha lenta e

  12. Study of crotoxin on the induction of paralysis in extraocular muscle in animal model Estudo da crotoxina na indução de paralisia da musculatura extraocular em modelo animal

    Directory of Open Access Journals (Sweden)

    Geraldo de Barros Ribeiro

    2012-10-01

    Full Text Available PURPOSE: Crotoxin is the major toxin of the venom of the South American rattlesnake Crotalus durissus terrificus, capable of causing a blockade of the neurotransmitters at the neuromuscular junction. The objective of this study was to appraise the action and effectiveness of the crotoxin induced paralysis of the extraocular muscle and to compare its effects with the botulinum toxin type A (BT-A. METHODS: The crotoxin, with LD50 of 1.5 µg, was injected into the superior rectus muscle in ten New Zealand rabbits. The concentration variance was 0.015 up to 150 µg. Two rabbits received 2 units of botulinum toxin type A for comparative analysis. The evaluation of the paralysis was performed using serial electromyography. After the functional recovery of the muscles, which occurred after two months, six rabbits were sacrificed for anatomopathology study. RESULTS: The animals did not show any evidence of systemic toxicity. Transitory ptosis was observed in almost every animal and remained up to fourteen days. These toxins caused immediate blockade of the electrical potentials. The recovery was gradual in the average of one month with regeneration signs evident on the electromyography. The paralysis effect of the crotoxin on the muscle was proportional to its concentration. The changes with 1.5 µg crotoxin were similar to those produced by the botulinum toxin type A. The histopathology findings were localized to the site of the injection. No signs of muscle fiber's necrosis were seen in any sample. The alterations induced by crotoxin were also proportional to the concentration and similar to botulinum toxin type A in concentration of 1.5 µg. CONCLUSION: Crotoxin was able to induce transitory paralysis of the superior rectus muscle. This effect was characterized by reduction of action potentials and non-specific signs of fibrillation. Crotoxin, in concentration of 1.5 µg was able to induce similar effects as botulinum toxin type A.OBJETIVO: A

  13. Mice plasma fibrinogen consumption by thrombin-like enzyme present in rattlesnake venom from the North-East region of Argentina Consumo de fibrinógeno plasmático en ratones por acción de enzima con actividad trombínica presente en el veneno de cascabel del nordeste argentino

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    Silvana L. Maruñak

    2004-12-01

    Full Text Available Due to variability of venom components from the same species of snakes that inhabit different regions, particular properties of the venom of Crotalus durissus terrificus that inhabits the North-East of Argentina were studied. Gyroxin, a thrombin-like enzyme, was isolated from this venom by gel filtration and affinity chromatography, it was found to be homogeneous according to SDS-PAGE, with a molecular weight of 33 kDa. "Gyroxin syndrome" in mice was tested and it showed changes in the animal behavior, confirming that the isolated thrombin-like enzyme is gyroxin. Effects of this enzyme and the crude venom on mice plasmatic fibrinogen levels were determined. The mice plasma fibrinogen decreased rapidly until incoagulability during the first hour after thrombin-Iike enzyme injection, then reaching its normal level 10 hours after injection; whereas crude venom resulted in a 60% decrease of the mice plasma fibrinogen, reaching its normal level after the same period of time. After 1 hour of gyroxin inoculation, intravascular coagulation was observed in histological cuttings of lung, cardiac muscle and liver. The isolated enzyme showed strong hydrolyzing activity on fibrinogen and fibrin in vitro, whereas the crude venom exhibited weak hydrolyzing activity on both substrates. It is probable that this very low activity is due to the low percentage of the enzyme in the crude venom. Decreasing of plasmatic fibrinogen levels may be due to either the coagulant or hydrolyzing actions of the enzyme.Teniendo en cuenta la variabilidad de los componentes del veneno de serpientes de una misma especie que habitan regiones diferentes, se decidió estudiar las propiedades particulares del veneno de Crotalus durissus terrificus que habita el nordeste de Argentina, Giroxina, una enzima con actividad trombínica, fue aislada del veneno por cromatografía de filtración por gel y de afinidad; se comprobó su homogeneidad y se determinó su peso molecular, 33 kDa, por

  14. Environmental Impact Analysis Process. Environmental Impact Statement. Supersonic Flight Operations in the Reserve Military Operations Area, Holloman AFB, New Mexico

    Science.gov (United States)

    1983-09-01

    tailed Rattlesnake Crotalus molossus Western Rattlesnake Crotalus viridis CAUTION Most snake bites occur to people who are trying to kill or catch snakes...Strawberry Fragaria bracteata Yarrow Achillea lanulosa Yellow Sweetclover Melilotus indicus IX L-2 List of Preparers Jim Battis; AFGL/LWH, Ha’-com

  15. Study of crotoxin mechanism of action to mammary carcinomas and evaluation of its potential as a radiopharmaceutical; Estudo do mecanismo de acao da crotoxina em tumores mamarios e avaliacao do seu potenctial radiofarmaceutico

    Energy Technology Data Exchange (ETDEWEB)

    Silveira, Marina Bicalho

    2010-07-01

    Crotoxin, the main component of Crotalus durissus terrificus snake venom, has been studied since 1938. It is a natural polypeptidic complex with pharmacological potential because of its antitumoral properties which has attracted great interest for diagnosis and therapy of oncological diseases. However, Crotoxin mechanism of action and sites of specific interaction on tumor cells are still misunderstood. Breast cancer is the second most frequent type in the world and the most common cancer in women. About 30 to 60% of mammary tumors overexpress epidermal growth factor receptor (EGFR), a transmembrane protein related to cell proliferation. Since literature has reported that Crotoxin antitumoral effect is more potent on cells with EGFR overexpression the objectives of this work were to evaluate Crotoxin cytotoxic effects on mammary tumor cells human breast carcinoma (MCF-7) and Ehrlich tumor cells (murine ascitics carcinoma), and to investigate the specific molecular interaction of Crotoxin on Ehrlich tumor cells. Initially, Crotoxin was radiolabelled with iodine-125 ({sup 125}I-Crotoxin) and iodine-131 ({sup 131}I-Crotoxin). Saturation and competition assay were carried out to characterize Crotoxin in vitro interaction; Crotoxin biodistribution studies and singlephoton emission computed tomography (SPECT) of mice bearing Ehrlich tumor have been evaluated to describe in vivo interaction. Our results showed that Crotoxin presented cytotoxic effect against Ehrlich with DL{sub 50} in vitro (concentration of compound which is lethal for 50% of cells) of about one micromolar, but did not present significant effect against MCF-7. Morphological alterations characteristic of apoptosis suggests programmed cell death. {sup 125}I-Crotoxin interaction with Ehrlich tumor cells was saturable with approximately 70% specificity, and presented K{sub d}=24.98 nmol/L and B{sub max}=16,570 sites/cell for low affinity binding sites and K{sub d}=0.06 nmol/L and B{sub max}=210 sites

  16. Garrison Dam/Lake Sakakawea Master Plan with Integrated Programmatic Environmental Assessment, Missouri River, Montana: Update of Design Memorandum MGR-107D

    Science.gov (United States)

    2007-12-14

    Toxicodendron radicans), wild strawberry ( Fragaria virginiana), violets (Oxalis violacea), and wild parsley (Lomatium orientale) fill in the...prairie rattlesnakes (Crotalus viridis ), yellowbelly racers (Coluber constrictor), western hognose snake (Heterdon nasicus), and smooth green snakes

  17. Environmental Assessment for Atlantic White Cedar Restoration Project at Dare County Range, Seymour Johnson Air Force Base, North Carolina

    Science.gov (United States)

    2014-12-23

    rattlesnakes inhabit forested areas, and in the mountains, they will often hibernate together in large numbers (Davidson Herpetology , 2013). Timber...YorktownAquiferVol1-Missimer1992.pdf Davidson Herpetology . (2013). Timber Rattlesnake (Crotalus horridus). Retrieved November 19, 2013, from http

  18. Snake mortality associated with late season radio-transmitter implantation

    Science.gov (United States)

    D. Craig Rudolph; Shirley J. Burgdorf; Richard R. Schaefer; Richard N. Conner; Robert T. Zappalorth

    1998-01-01

    Radio-telemetry is an increasingly used procedure to obtain data on the biology of free-living snakes (Reinert 1992, 1994). In Texas and Louisiana we have been using the surgical technique of Weatherhead and Anderka (1984) to implant transmitters in timber rattlesnakes (Crotalus horridus) and Louisiana pine snakes (Pituophis melanoleucus...

  19. Spatial ecology of timber rattlesnakes on the hardwood ecosystem experiment: pre-treatment results

    Science.gov (United States)

    Brian J. MacGowan; Zachary J. Walker

    2013-01-01

    The timber rattlesnake (Crotalus horridus) is a species of conservation concern throughout much of its geographic range and may serve as a sentinel species in investigations of the effects of timber harvesting on forest reptiles. Our objective was to determine the effect of even-aged timber management regimes on timber rattlesnake home range and...

  20. Monterey Peninsula Water Supply Project. Supplemental Draft Environmental Impact Report/Statement II. Appendices

    Science.gov (United States)

    1993-02-01

    ROSACEAE Acaena californica: C-S, CPI Adenostoma fasciculctmm: CH-, CS, C-L Aichemilla arvensis: CP Fragaria vesca ssp. cabjfomica: NP, LO, M-LI Hetermeles...Thamnophis elegans * Western Aquatic Garter Snake T. couchi * Common Garter Snake T. sinalis * Western Rattlesnake Crotalus viridis TOTAL 22 8 3 3 BIRDS

  1. Responses of timber rattlesnakes to fire: Lessons from two prescribed burns

    Science.gov (United States)

    Steven J. Beaupre; Lara E. Douglas

    2012-01-01

    Timber rattlesnakes (Crotalus horridus) are excellent model organisms for understanding the effects of large scale habitat manipulations because of their low-energy lifestyle, rapid response to changes in resource environment, uniform diet (small mammals), and simple behaviors. We present two case studies that illustrate interactions between timber...

  2. Taste buds in the palatal mucosa of snakes | Berkhoudt | African ...

    African Journals Online (AJOL)

    An examination of the oral mucosa of Crotalus and several Scolecophidia revealed the presence of taste buds. The taste buds in these two divergent groups of snakes are similar in appearance, and correspond to previous descriptions of gustatory organs in other reptiles. Few taste buds were present in any specimen, and ...

  3. Novel Cytochrome P45OlBl as a Mammary Cancer Risk Factor.

    Science.gov (United States)

    1998-07-01

    3.1.4.1; from Crotalus atrox) were purchased from Sigma Chemical Co. (St Louis, MO). RNase Tl (EC 3.1.21.3; from Aspergillus oryzae ) and RNase (DNase...change under serum- c T c T free conditions, in which LEC express BEC characteristics in response to hormones present in the media (19-20). We have...fibroblasts from different tissue sources would show different CYP expression characteristics . Human BF [tumors and peripheral] and human skin

  4. Irreversible inactivation of snake venom l-amino acid oxidase by covalent modification during catalysis of l-propargylglycine☆

    Science.gov (United States)

    Mitra, Jyotirmoy; Bhattacharyya, Debasish

    2013-01-01

    Snake venom l-amino acid oxidase (SV-LAAO, a flavor-enzyme) has attracted considerable attention due to its multifunctional nature, which is manifest in diverse clinical and biological effects such as inhibition of platelet aggregation, induction of cell apoptosis and cytotoxicity against various cells. The majority of these effects are mediated by H2O2 generated during the catalytic conversion of l-amino acids. The substrate analog l-propargylglycine (LPG) irreversibly inhibited the enzyme from Crotalus adamanteus and Crotalus atrox in a dose- and time-dependent manner. Inactivation was irreversible which was significantly protected by the substrate l-phenylalanine. A Kitz–Wilson replot of the inhibition kinetics suggested formation of reversible enzyme–LPG complex, which occurred prior to modification and inactivation of the enzyme. UV–visible and fluorescence spectra of the enzyme and the cofactor strongly suggested formation of covalent adduct between LPG and an active site residue of the enzyme. A molecular modeling study revealed that the FAD-binding, substrate-binding and the helical domains are conserved in SV-LAAOs and both His223 and Arg322 are the important active site residues that are likely to get modified by LPG. Chymotrypsin digest of the LPG inactivated enzyme followed by RP-HPLC and MALDI mass analysis identified His223 as the site of modification. The findings reported here contribute towards complete inactivation of SV-LAAO as a part of snake envenomation management. PMID:23772385

  5. 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.

  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. Phospholipase a properties of several snake venom preparations.

    Science.gov (United States)

    Nutter, L J; Privett, O S

    1966-07-01

    The hydrolytic properties of the venoms of seven species of snakes,Crotalus adamanteus, Ancistrodon contortrix, Naja naja, Bothrops atrox, Ophiophagus hannah, Crotalus atrox andVipera russeli, were studied with purified lecithins and mixtures of lecithins of known fatty acid and class composition as substrates.The relative rates of hydrolysis of the fatty acids by the above venoms were studied by analysis of the products of the reaction at intervals during the course of the reaction. Of the seven venoms studied, that ofOphiophagus hannah was the only one which did not give some degree of preferential rate of hydrolysis of individual fatty acids.In general, saturated fatty acids were liberated faster than unsaturated fatty acids; differences in the rates of the hydrolysis of individual saturate and unsaturated fatty acids were also observed. Individual classes of lecithin were also hydrolyzed at different rates. For the determination of the distribution of the fatty acids between the alpha- and beta-position of lecithin, the reaction should be carried to completion. If the reaction requires a prolonged time to go to completion, it should be carried out under nitrogen to prevent autoxidation.

  8. /sup 239/Pu contamination in snakes inhabiting the Rocky Flats Plant site

    Energy Technology Data Exchange (ETDEWEB)

    Geiger, R.A.; Winsor, T.F.

    1975-01-01

    For approximately four years studies have been under way at the Rocky Flats plant to determine contamination patterns and concentrations of Pu in the biota. Contamination of the Rocky Flats environs has resulted from at least three incidents, a September 1957 fire, a May 1969 fire, and leaking barrels containing plutonium-laden cutting oil. The latter incident was considered by far the major source of the plutonium contamination. Results are reported from a study conducted to determine whether snake tissues of the area contained detectable amounts of /sup 239/Pu and, if so, at what concentrations. Eastern yellow-bellied racers (Coluber constrictor flaviventris, bullsnakes (Pituophis melanoleucus sayi, and prairie rattlesnakes (Crotalus viridis viridis, were collected for /sup 239/Pu bioassay of lung, liver, and bone tissues. Snakes were captured using drift fences terminating in funnel traps and by opportunistic sampling. Results led to the conclusion that snakes are not an important organism in the redistribution of /sup 239/Pu. (CH)

  9. 239Pu contamination in snakes inhabiting the Rocky Flats Plant site

    International Nuclear Information System (INIS)

    Geiger, R.A.; Winsor, T.F.

    1975-01-01

    For approximately four years studies have been under way at the Rocky Flats plant to determine contamination patterns and concentrations of Pu in the biota. Contamination of the Rocky Flats environs has resulted from at least three incidents, a September 1957 fire, a May 1969 fire, and leaking barrels containing plutonium-laden cutting oil. The latter incident was considered by far the major source of the plutonium contamination. Results are reported from a study conducted to determine whether snake tissues of the area contained detectable amounts of 239 Pu and, if so, at what concentrations. Eastern yellow-bellied racers (Coluber constrictor flaviventris, bullsnakes (Pituophis melanoleucus sayi, and prairie rattlesnakes (Crotalus viridis viridis, were collected for 239 Pu bioassay of lung, liver, and bone tissues. Snakes were captured using drift fences terminating in funnel traps and by opportunistic sampling. Results led to the conclusion that snakes are not an important organism in the redistribution of 239 Pu

  10. Professor José Roberto Giglio and toxinnology in Brazil: 48 years in research

    Directory of Open Access Journals (Sweden)

    A. M. Soares

    2007-01-01

    Full Text Available This work succinctly describes the professional and scientific life of Dr. José R. Giglio, one of the most outstanding Brazilian researchers in the field of Toxinology. During his long and successful career, he has made major contributions, especially in elucidating the function, structure, and mechanisms of action of animal venom proteins (from snakes, scorpions and spiders as well as the characterization of antibodies and several inhibitors of venoms and toxins. We present here a brief history of Dr. Giglio’s personal and professional life, also reporting some of his numerous published scientific articles on venoms from snakes (Bothrops, Crotalus, and other genera, scorpions (Tityus sp, spiders (Phoneutria sp, their isolated toxins and natural inhibitors. Thus, this work is a tribute to Dr. Giglio in his 73rd birthday, having devoted 48 years of his life studying animal venoms, an effort that has continued even after his formal retirement from university duties.

  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. Influenza em animais heterotérmicos Influenza in heterothermics

    Directory of Open Access Journals (Sweden)

    Dalva Assunção Portari Mancini

    2004-06-01

    Full Text Available O objetivo foi pesquisar Ortomyxovirus em animais heterotérmicos. Coletou-se sangue de serpentes dos gêneros Bothrops e Crotalus e de sapo e rãs dos gêneros Bufo e Rana, para a detecção dos receptores de hemácias e anticorpos específicos, ao vírus influenza, pelos testes de hemaglutinação e inibição da hemaglutinação, respectivamente. Pelo teste de hemaglutinação, verificou-se que serpentes e sapos em cativeiro apresentaram receptores em suas hemácias para o vírus influenza, humano e eqüino do tipo A e tipo B. O mesmo ocorreu com serpentes recém chegadas. Quanto ao teste de inibição da hemaglutinação dos soros dos répteis observou-se títulos protetores de anticorpos aos vírus influenza tipo A (origens humana e eqüina e tipo B. Com soro de sapo não se observou reação de inibição da hemaglutinação porém, 83,3% das rãs obtiveram médias de 40UIH para algumas cepas. Conclui-se que animais heterotérmicos podem oferecer condições de hospedeiros aos vírus influenza, assim como susceptibilidade à infecção.The objective was to study Orthomyxovirus in heterothermic animals. Blood samples from snakes (genus Bothrops and Crotalus and from toads and frogs (genus Bufo and Rana were collected to evaluate the red cell receptors and antibodies specific to influenza virus by the hemagglutination and hemagglutination inhibition tests, respectively. Both snakes and toads kept in captivity presented receptors in their red cells and antibodies specific to either influenza virus type A (human and equine origin or influenza type B. The same was observed with recently captured snakes. Concerning the influenza hemagglutination inhibition antibodies protective levels were observed in the reptiles' serum, against influenza type A and type B. Unlike the toads, 83.3% of the frogs presented mean levels of Ab 40HIU for some influenza strains. It was concluded that heterothermic animals could offer host conditions to the influenza

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

    Directory of Open Access Journals (Sweden)

    Orlando C. de O. Barreto

    1990-02-01

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

  14. Photorepair mutants of Arabidopsis

    International Nuclear Information System (INIS)

    Jiang, C.Z.; Yee, J.; Mitchell, D.L.; Britt, A.B.

    1997-01-01

    UV radiation induces two major DNA damage products, the cyclobutane pyrimidine dimer (CPD) and, at a lower frequency, the pyrimidine (6-4) pyrimidinone dimer (6-4 product). Although Escherichia coli and Saccharomyces cerevisiae produce a CPD-specific photolyase that eliminates only this class of dimer, Arabidopsis thaliana, Drosophila melanogaster, Crotalus atrox, and Xenopus laevis have recently been shown to photoreactivate both CPDs and 6-4 products. We describe the isolation and characterization of two new classes of mutants of Arabidopsis, termed uvr2 and uvr3, that are defective in the photoreactivation of CPDs and 6-4 products, respectively. We demonstrate that the CPD photolyase mutation is genetically linked to a DNA sequence encoding a type II (metazoan) CPD photolyase. In addition, we are able to generate plants in which only CPDs or 6-4 products are photoreactivated in the nuclear genome by exposing these mutants to UV light and then allowing them to repair one or the other class of dimers. This provides us with a unique opportunity to study the biological consequences of each of these two major UV-induced photoproducts in an intact living system

  15. Endogenous hepadnaviruses, bornaviruses and circoviruses in snakes.

    Science.gov (United States)

    Gilbert, C; Meik, J M; Dashevsky, D; Card, D C; Castoe, T A; Schaack, S

    2014-09-22

    We report the discovery of endogenous viral elements (EVEs) from Hepadnaviridae, Bornaviridae and Circoviridae in the speckled rattlesnake, Crotalus mitchellii, the first viperid snake for which a draft whole genome sequence assembly is available. Analysis of the draft assembly reveals genome fragments from the three virus families were inserted into the genome of this snake over the past 50 Myr. Cross-species PCR screening of orthologous loci and computational scanning of the python and king cobra genomes reveals that circoviruses integrated most recently (within the last approx. 10 Myr), whereas bornaviruses and hepadnaviruses integrated at least approximately 13 and approximately 50 Ma, respectively. This is, to our knowledge, the first report of circo-, borna- and hepadnaviruses in snakes and the first characterization of non-retroviral EVEs in non-avian reptiles. Our study provides a window into the historical dynamics of viruses in these host lineages and shows that their evolution involved multiple host-switches between mammals and reptiles. © 2014 The Author(s) Published by the Royal Society. All rights reserved.

  16. Prairie rattlesnake envenomation in 27 New World camelids.

    Science.gov (United States)

    Sonis, J M; Hackett, E S; Callan, R J; Holt, T N; Hackett, T B

    2013-01-01

    Morbidity and case fatality from rattlesnake envenomation is regionally specific because of variability in relative toxicity of the species of snake encountered. A previous report of rattlesnake envenomation in New World camelids (NWC) from the western coastal United States documented high case fatality rates and guarded prognosis for survival. To describe clinical findings, treatments, and outcome of NWC with prairie rattlesnake (Crotalus viridis viridis) envenomation in the Rocky Mountain region of the United States. Twenty-seven NWC admitted to the Colorado State University Veterinary Teaching Hospital for evaluation of acute rattlesnake envenomation between 1992 and 2012. Medical records of NWC evaluated for rattlesnake envenomation as coded by the attending clinician and identified by a database search were reviewed retrospectively. Month of admission, signalment, area of bite, clinical and clinicopathologic data, treatments, and outcome were recorded. Twenty-five llamas and 2 alpacas were admitted for envenomation. Llamas were overrepresented compared to hospital caseload. The face was the most common site of envenomation, observed in 96% of recorded cases. Presenting clinical signs included fever, tachypnea, tachycardia, and respiratory distress. Nine animals required a tracheotomy. Median hospitalization time was 3 days and overall survival rate was 69%. Case fatality rate for prairie rattlesnake envenomation in NWC was lower than that reported in the Western coastal region of the United States and similar to that reported for prairie rattlesnake envenomation in horses. Copyright © 2013 by the American College of Veterinary Internal Medicine.

  17. A STUDY OF THE COMPETITION OF LECITHIN AND ANTITOXIN FOR CL. WELCHII LECITHINASE

    Science.gov (United States)

    Zamecnik, Paul C.; Lipmann, Fritz

    1947-01-01

    Lecithin has been found to interfere with the combining reaction of Cl. welchii alpha toxin (lecithinase) and its antitoxin. If the lecithinase is first brought into contact with lecithin, and the antitoxin is then added, the antitoxin fails to stop the enzymatic reaction, but gradually decelerates it. If the lecithinase is brought into contact with both lecithin and antitoxin at the same instant, it appears to combine in part with each, and the enzymatic process takes place at a reduced rate, which gradually declines further. If the lecithinase is first brought into contact with antitoxin, before the lecithin is added, the enzymatic reaction is completely inhibited. This ability of lecithin to inhibit the antitoxin-toxin combination cannot be explained adequately as a non-specific coating of the toxin-enzyme by the lecithin. It is rather suggested that lecithin and antitoxin compete specifically for combination with the same regions on the enzyme molecule. Lecithin has similarly been found to interfere with the combination of Crotalus terrificus venom and its antiserum. The above findings provide a partial explanation for the lack of effectiveness of antitoxin when given late in the course of Cl. welchii infection. PMID:19871624

  18. Common Cutaneous Bacteria Isolated from Snakes Inhibit Growth of Ophidiomyces ophiodiicola.

    Science.gov (United States)

    Hill, Aubree J; Leys, Jacob E; Bryan, Danny; Erdman, Fantasia M; Malone, Katherine S; Russell, Gabrielle N; Applegate, Roger D; Fenton, Heather; Niedringhaus, Kevin; Miller, Andrew N; Allender, Matthew C; Walker, Donald M

    2018-03-01

    There is increasing concern regarding potential impacts of snake fungal disease (SFD), caused by Ophidiomyces ophiodiicola (Oo), on free-ranging snake populations in the eastern USA. The snake cutaneous microbiome likely serves as the first line of defense against Oo and other pathogens; however, little is known about microbial associations in snakes. The objective of this study was to better define the composition and immune function of the snake cutaneous microbiome. Eight timber rattlesnakes (Crotalus horridus) and four black racers (Coluber constrictor) were captured in Arkansas and Tennessee, with some snakes exhibiting signs of SFD. Oo was detected through real-time qPCR in five snakes. Additional histopathological techniques confirmed a diagnosis of SFD in one racer, the species' first confirmed case of SFD in Tennessee. Fifty-eight bacterial and five fungal strains were isolated from skin swabs and identified with Sanger sequencing. Non-metric multidimensional scaling and PERMANOVA analyses indicated that the culturable microbiome does not differ between snake species. Fifteen bacterial strains isolated from rattlesnakes and a single strain isolated from a racer inhibited growth of Oo in vitro. Results shed light on the culturable cutaneous microbiome of snakes and probiotic members that may play a role in fighting an emergent disease.

  19. Snake venomics and antivenomics of Bothrops atrox venoms from Colombia and the Amazon regions of Brazil, Perú and Ecuador suggest the occurrence of geographic variation of venom phenotype by a trend towards paedomorphism.

    Science.gov (United States)

    Núñez, Vitelbina; Cid, Pedro; Sanz, Libia; De La Torre, Pilar; Angulo, Yamileth; Lomonte, Bruno; Gutiérrez, José María; Calvete, Juan J

    2009-11-02

    The venom proteomes of Bothrops atrox from Colombia, Brazil, Ecuador, and Perú were characterized using venomic and antivenomic strategies. Our results evidence the existence of two geographically differentiated venom phenotypes. The venom from Colombia comprises at least 26 different proteins belonging to 9 different groups of toxins. PI-metalloproteinases and K49-PLA(2) molecules represent the most abundant toxins. On the other hand, the venoms from Brazilian, Ecuadorian, and Peruvian B. atrox contain predominantly PIII-metalloproteinases. These toxin profiles correlate with the venom phenotypes of adult and juvenile B. asper from Costa Rica, respectively, suggesting that paedomorphism represented a selective trend during the trans-Amazonian southward expansion of B. atrox through the Andean Corridor. The high degree of crossreactivity of a Costa Rican polyvalent (Bothrops asper, Lachesis stenophrys, Crotalus simus) antivenom against B. atrox venoms further evidenced the close evolutionary kinship between B. asper and B. atrox. This antivenom was more efficient immunodepleting proteins from the venoms of B. atrox from Brazil, Ecuador, and Perú than from Colombia. Such behaviour may be rationalized taking into account the lower content of poorly immunogenic toxins, such as PLA(2) molecules and PI-SVMPs in the paedomorphic venoms. The immunological profile of the Costa Rican antivenom strongly suggests the possibility of using this antivenom for the management of snakebites by B. atrox in Colombia and the Amazon regions of Ecuador, Perú and Brazil.

  20. Skin lipid structure controls water permeability in snake molts.

    Science.gov (United States)

    Torri, Cristian; Mangoni, Alfonso; Teta, Roberta; Fattorusso, Ernesto; Alibardi, Lorenzo; Fermani, Simona; Bonacini, Irene; Gazzano, Massimo; Burghammer, Manfred; Fabbri, Daniele; Falini, Giuseppe

    2014-01-01

    The role of lipids in controlling water exchange is fundamentally a matter of molecular organization. In the present study we have observed that in snake molt the water permeability drastically varies among species living in different climates and habitats. The analysis of molts from four snake species: tiger snake, Notechis scutatus, gabon viper, Bitis gabonica, rattle snake, Crotalus atrox, and grass snake, Natrix natrix, revealed correlations between the molecular composition and the structural organization of the lipid-rich mesos layer with control in water exchange as a function of temperature. It was discovered, merging data from micro-diffraction and micro-spectroscopy with those from thermal, NMR and chromatographic analyses, that this control is generated from a sophisticated structural organization that changes size and phase distribution of crystalline domains of specific lipid molecules as a function of temperature. Thus, the results of this research on four snake species suggest that in snake skins different structured lipid layers have evolved and adapted to different climates. Moreover, these lipid structures can protect, "safety", the snakes from water lost even at temperatures higher than those of their usual habitat. Copyright © 2013 Elsevier Inc. All rights reserved.

  1. Limitations of climatic data for inferring species boundaries: insights from speckled rattlesnakes.

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    Jesse M Meik

    Full Text Available Phenotypes, DNA, and measures of ecological differences are widely used in species delimitation. Although rarely defined in such studies, ecological divergence is almost always approximated using multivariate climatic data associated with sets of specimens (i.e., the "climatic niche"; the justification for this approach is that species-specific climatic envelopes act as surrogates for physiological tolerances. Using identical statistical procedures, we evaluated the usefulness and validity of the climate-as-proxy assumption by comparing performance of genetic (nDNA SNPs and mitochondrial DNA, phenotypic, and climatic data for objective species delimitation in the speckled rattlesnake (Crotalus mitchellii complex. Ordination and clustering patterns were largely congruent among intrinsic (heritable traits (nDNA, mtDNA, phenotype, and discordance is explained by biological processes (e.g., ontogeny, hybridization. In contrast, climatic data did not produce biologically meaningful clusters that were congruent with any intrinsic dataset, but rather corresponded to regional differences in atmospheric circulation and climate, indicating an absence of inherent taxonomic signal in these data. Surrogating climate for physiological tolerances adds artificial weight to evidence of species boundaries, as these data are irrelevant for that purpose. Based on the evidence from congruent clustering of intrinsic datasets, we recommend that three subspecies of C. mitchellii be recognized as species: C. angelensis, C. mitchellii, and C. Pyrrhus.

  2. Black Bear Reactions to Venomous and Non-venomous Snakes in Eastern North America

    Science.gov (United States)

    Rogers, Lynn L; Mansfield, Susan A; Hornby, Kathleen; Hornby, Stewart; Debruyn, Terry D; Mize, Malvin; Clark, Rulon; Burghardt, Gordon M

    2014-01-01

    Bears are often considered ecological equivalents of large primates, but the latter often respond with fear, avoidance, and alarm calls to snakes, both venomous and non-venomous, there is sparse information on how bears respond to snakes. We videotaped or directly observed natural encounters between black bears (Ursus americanus) and snakes. Inside the range of venomous snakes in Arkansas and West Virginia, adolescent and adult black bears reacted fearfully in seven of seven encounters upon becoming aware of venomous and non-venomous snakes; but in northern Michigan and Minnesota where venomous snakes have been absent for millennia, black bears showed little or no fear in four encounters with non-venomous snakes of three species. The possible roles of experience and evolution in bear reactions to snakes and vice versa are discussed. In all areas studied, black bears had difficulty to recognize non-moving snakes by smell or sight. Bears did not react until snakes moved in 11 of 12 encounters with non-moving timber rattlesnakes (Crotalus horridus) and four species of harmless snakes. However, in additional tests in this study, bears were repulsed by garter snakes that had excreted pungent anal exudates, which may help explain the absence of snakes, both venomous and harmless, in bear diets reported to date. PMID:25635152

  3. Molecular modeling of the inhibition of enzyme PLA2 from snake venom by dipyrone and 1-phenyl-3-methyl-5-pyrazolone

    Science.gov (United States)

    Silva, S. L. Da; Comar, M., Jr.; Oliveira, K. M. T.; Chaar, J. S.; Bezerra, E. R. M.; Calgarotto, A. K.; Baldasso, P. A.; Veber, C. L.; Villar, J. A. F. P.; Oliveira, A. R. M.; Marangoni, S.

    Phospholipases A2 (PLA2) are enzymes that trigger the degradation cascade of the arachidonic acid, leading to the formation of pro-inflammatory eicosanoids. The selective inhibition of PLA2s is crucial in the search for a more efficient anti-inflammatory drug with fewer side effects than the drugs currently used. Hence, we studied the influences caused by two pyrazolonic inhibitors: dipyrone (DIP) and 1-phenyl-3-methyl-5-pyrazolone (PMP) on the kinetic behavior of PLA2 from Crotalus adamanteus venom. Molecular modeling results, by DFT and MM approaches, showed that DIP is strongly associated to the active site of PLA2 through three hydrogen bonds, whereas PMP is associated to the enzyme just through hydrophobic interactions. In addition, only PMP presents an intramolecular hydrogen bond that make difficult the formation of more efficient interactions with PLA2. These results help in the understanding of the experimental observations. Experimentally, the results showed that PLA2 from C. adamanteus present a typical Michaelian behavior. In addition, the calculated kinetic parameters showed that, in the presence of DIP or PMP, the maximum enzymatic velocity (VMAX) value was kept constant, whereas the Michaelis constant (KM) values increased and the inhibition constant (KI) decreased, indicating competitive inhibition. These results show that the phenyl-pyrazolonic structures might help in the development and design of new drugs able to selectively inhibit PLA2.

  4. 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.

  5. Roads, interrupted dispersal, and genetic diversity in timber rattlesnakes.

    Science.gov (United States)

    Clark, Rulon W; Brown, William S; Stechert, Randy; Zamudio, Kelly R

    2010-08-01

    Anthropogenic habitat modification often creates barriers to animal movement, transforming formerly contiguous habitat into a patchwork of habitat islands with low connectivity. Roadways are a feature of most landscapes that can act as barriers or filters to migration among local populations. Even small and recently constructed roads can have a significant impact on population genetic structure of some species, but not others. We developed a research approach that combines fine-scale molecular genetics with behavioral and ecological data to understand the impacts of roads on population structure and connectivity. We used microsatellite markers to characterize genetic variation within and among populations of timber rattlesnakes (Crotalus horridus) occupying communal hibernacula (dens) in regions bisected by roadways. We examined the impact of roads on seasonal migration, genetic diversity, and gene flow among populations. Snakes in hibernacula isolated by roads had significantly lower genetic diversity and higher genetic differentiation than snakes in hibernacula in contiguous habitat. Genetic-assignment analyses revealed that interruption to seasonal migration was the mechanism underlying these patterns. Our results underscore the sizeable impact of roads on this species, despite their relatively recent construction at our study sites (7 to 10 generations of rattlesnakes), the utility of population genetics for studies of road ecology, and the need for mitigating effects of roads.

  6. 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.

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

    Science.gov (United States)

    Yañez-Arenas, Carlos; Peterson, A Townsend; Mokondoko, Pierre; Rojas-Soto, Octavio; Martínez-Meyer, Enrique

    2014-01-01

    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).

  8. Serpentes peçonhentas e ofidismo em Cruzeiro do Sul, Alto Juruá, Estado do Acre, Brasil Venomous snakes and ophidism in Cruzeiro do Sul, Alto Juruá, State of Acre, Brazil

    Directory of Open Access Journals (Sweden)

    Paulo Sérgio Bernarde

    2012-03-01

    Full Text Available Apesar da importância dos acidentes ofídicos na Saúde Pública, são relativamente poucas as pesquisas realizadas sobre esse tema no Brasil. Devido aos poucos estudos sobre ofidismo na Amazônia e especialmente no estado do Acre, trabalhos epidemiológicos são de grande relevância. Esse estudo apresenta a lista de serpentes peçonhentas e aspectos epidemiológicos dos acidentes ofídicos em Cruzeiro do Sul, região do Alto Juruá (Acre, verificando quais gêneros de serpentes são responsáveis pelos envenenamentos e aspectos que envolvem o acidente e o atendimento hospitalar. Os dados epidemiológicos foram coligidos a partir do SINAN (Sistema de Informações de Agravos de Notificação, no setor de vigilância epidemiológica do Hospital Regional do Juruá, no município de Cruzeiro do Sul. Sete espécies de serpentes peçonhentas foram registradas nesse município: três viperídeos (Bothrops atrox, Bothriopsis bilineata e Lachesis muta e quatro elapídeos (Micrurus hemprichii, M. lemniscatus, M. remotus and M. surinamensis. Durante o período de dois anos (agosto de 2007 a julho de 2009 foram registrados 195 casos de acidentes ofídicos. Cinquenta e um por cento dos acidentes foi classificado como laquético (Lachesis, seguido pelo botrópico (Bothrops e Bothriopsis com 38% e crotálico (Crotalus com 2%. Em 9% dos casos o gênero da serpente envolvida não foi informado. A maioria dos acidentes envolveu indivíduos adultos do gênero masculino em área rural, afetados principalmente nos membros inferiores. Os casos ocorreram mais frequentemente nos meses de novembro a abril, coincidindo com os maiores níveis pluviométricos. A maioria dos acidentes foi atribuída equivocadamente à serpente L. muta, tendo sido provavelmente causados por B. atrox.Despite the importance of accidents involving snakebites, research on this theme in Brazil is relatively rare, especially in the Amazon region. Due to the poor number of studies about ophidism

  9. Management of Tissue Loss After Agkistrodon Snakebite: Appropriate Use of Crotalidae-Fab Antivenin.

    Science.gov (United States)

    Larson, Kenneth W; Schaefer, Keith R; Austin, Cindy; Norton, Rhy; Finley, Phillip J

    2016-01-01

    Although initially created for the treatment of rattlesnake (genus: Crotalus) bites, Crotalidae-Fab antivenin is used to treat many different pit viper envenomations. However, the efficacy of Crotalidae-Fab in preventing tissue loss from copperhead (Agkistrodon contortrix) or cottonmouth (Agkistrodon piscivorus) snakebites remains unclear. Recent reports show that Agkistrodon-related bites rarely require treatment beyond simple observation and pain control. The purpose of this study was to examine the amount of tissue loss in patients who received Crotalidae-Fab compared with those who did not after an Agkistrodon bite. After institutional review board approval, a retrospective study was completed at a Level 1 trauma center. Between 2009 and 2013, a total of 57 snakebites were identified. Of the 57 bites, the snake species was documented in 36 cases including 31 copperheads, 1 cottonmouth, and 4 rattlesnakes. The other 21 bites were from unknown or nonvenomous species. Of the 32 Agkistrodon-related bites, 15 patients received Crotalidae-Fab (average of 3 vials administered) and 17 did not receive Crotalidae-Fab. None of the 32 patients, regardless of treatment option, had tissue loss or required surgical interventions. Only 1 patient received Crotalidae-Fab and debridement of a vesicle associated with the bite. No clinically significant differences were observed between the groups. These findings support previous literature that failed to show added benefit of Crotalidae-Fab treatment for Agkistrodon bites beyond patient comfort and pain control. Evaluation of current protocols for Agkistrodon envenomations is warranted. Snakebite wound education in trauma physicians and nurses may decrease unnecessary use of antivenom medication.

  10. BjussuSP-I: a new thrombin-like enzyme isolated from Bothrops jararacussu snake venom.

    Science.gov (United States)

    Sant' Ana, Carolina D; Ticli, Fabio K; Oliveira, Leandro L; Giglio, Jose R; Rechia, Carem G V; Fuly, André L; Selistre de Araújo, Heloisa S; Franco, João J; Stabeli, Rodrigo G; Soares, Andreimar M; Sampaio, Suely V

    2008-11-01

    A thrombin-like enzyme named BjussuSP-I, isolated from B. jararacussu snake venom, is an acidic single chain glycoprotein with approximately 6% sugar, Mr=61,000 under reducing conditions and pI approximately 3.8, representing 1.09% of the chromatographic A(280) recovery. BjussuSP-I is a glycosylated serine protease containing both N-linked carbohydrates and sialic acid in its structure. BjussuSP-I showed a high clotting activity upon human plasma, which was inhibited by PMSF, leupeptin, heparin and 1,10-phenantroline. This enzyme showed high stability regarding coagulant activity when analyzed at different temperatures (-70 to 37 degrees C), pHs (4.5 to 8.0), and presence of two divalent metal ions (Ca(2+) and Mg(2+)). It also displayed TAME esterase and proteolytic activities toward natural (fibrinogen and fibrin) and synthetic (BAPNA) substrates, respectively, being also inhibited by PMSF and leupeptin. BjussuSP-I can induce production of polyclonal antibodies able to inhibit its clotting activity, but unable to inhibit its proteolytic activity on fibrinogen. The enzyme also showed crossed immunoreactivity against 11 venom samples of Bothrops, 1 of Crotalus, and 1 of Calloselasma snakes, in addition of LAAO isolated from B. moojeni venom. It displayed neither hemorrhagic, myotoxic, edema-inducing profiles nor proteolytic activity on casein. BjussuSP-I showed an N-terminal sequence (VLGGDECDINEHPFLA FLYS) similar to other thrombin-like enzymes from snake venoms. Based on its biochemical, enzymatic and pharmacological characteristics, BjussuSP-I was identified as a new thrombin-like enzyme isoform from Bothrops jararacussu snake venom.

  11. Anthropogenic impacts drive niche and conservation metrics of a cryptic rattlesnake on the Colorado Plateau of western North America.

    Science.gov (United States)

    Douglas, M R; Davis, M A; Amarello, M; Smith, J J; Schuett, G W; Herrmann, H-W; Holycross, A T; Douglas, M E

    2016-04-01

    Ecosystems transition quickly in the Anthropocene, whereas biodiversity adapts more slowly. Here we simulated a shifting woodland ecosystem on the Colorado Plateau of western North America by using as its proxy over space and time the fundamental niche of the Arizona black rattlesnake (Crotalus cerberus). We found an expansive (= end-of-Pleistocene) range that contracted sharply (= present), but is blocked topographically by Grand Canyon/Colorado River as it shifts predictably northwestward under moderate climate change (= 2080). Vulnerability to contemporary wildfire was quantified from available records, with forested area reduced more than 27% over 13 years. Both 'ecosystem metrics' underscore how climate and wildfire are rapidly converting the Plateau ecosystem into novel habitat. To gauge potential effects on C. cerberus, we derived a series of relevant 'conservation metrics' (i.e. genetic variability, dispersal capacity, effective population size) by sequencing 118 individuals across 846 bp of mitochondrial (mt)DNA-ATPase8/6. We identified five significantly different clades (net sequence divergence = 2.2%) isolated by drainage/topography, with low dispersal (F ST = 0.82) and small sizes (2N ef = 5.2). Our compiled metrics (i.e. small-populations, topographic-isolation, low-dispersal versus conserved-niche, vulnerable-ecosystem, dispersal barriers) underscore the susceptibility of this woodland specialist to a climate and wildfire tandem. We offer adaptive management scenarios that may counterbalance these metrics and avoid the extirpation of this and other highly specialized, relictual woodland clades.

  12. Good vibrations: Assessing the stability of snake venom composition after researcher-induced disturbance in the laboratory.

    Science.gov (United States)

    Claunch, Natalie M; Holding, Matthew L; Escallón, Camilo; Vernasco, Ben; Moore, Ignacio T; Taylor, Emily N

    2017-07-01

    Phenotypic plasticity contributes to intraspecific variation in traits of many animal species. Venom is an integral trait to the success and survival of many snake species, and potential plasticity in venom composition is important to account for in the context of basic research as well as in human medicine for treating the various symptoms of snakebite and producing effective anti-venoms. Researchers may unknowingly induce changes in venom variation by subjecting snakes to novel disturbances and potential stressors. We explored phenotypic plasticity in snake venom composition over time in captive Pacific rattlesnakes (Crotalus oreganus) exposed to vibration treatment, compared to an undisturbed control group. Venom composition did not change significantly in response to vibration, nor was there a detectable effect of overall time in captivity, even though snakes re-synthesized venom stores while subjected to novel disturbance in the laboratory. This result indicates that venom composition is a highly repeatable phenotype over short time spans and that the composition of venom within adult individuals may be resistant to or unaffected by researcher-induced disturbance. On the other hand, the change in venom composition, measured as movement along the first principle component of venom phenotype space, was associated with baseline corticosterone (CORT) levels in the snakes. While differential forms of researcher-induced disturbance may not affect venom composition, significant changes in baseline CORT, or chronic stress, may affect the venom phenotype, and further investigations will be necessary to assess the nature of the relationship between CORT and venom protein expression. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Crovirin, a snake venom cysteine-rich secretory protein (CRISP with promising activity against Trypanosomes and Leishmania.

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    Camila M Adade

    2014-10-01

    Full Text Available The neglected human diseases caused by trypanosomatids are currently treated with toxic therapy with limited efficacy. In search for novel anti-trypanosomatid agents, we showed previously that the Crotalus viridis viridis (Cvv snake venom was active against infective forms of Trypanosoma cruzi. Here, we describe the purification of crovirin, a cysteine-rich secretory protein (CRISP from Cvv venom with promising activity against trypanosomes and Leishmania.Crude venom extract was loaded onto a reverse phase analytical (C8 column using a high performance liquid chromatographer. A linear gradient of water/acetonitrile with 0.1% trifluoroacetic acid was used. The peak containing the isolated protein (confirmed by SDS-PAGE and mass spectrometry was collected and its protein content was measured. T. cruzi trypomastigotes and amastigotes, L. amazonensis promastigotes and amastigotes and T. brucei rhodesiense procyclic and bloodstream trypomastigotes were challenged with crovirin, whose toxicity was tested against LLC-MK2 cells, peritoneal macrophages and isolated murine extensor digitorum longus muscle. We purified a single protein from Cvv venom corresponding, according to Nano-LC MS/MS sequencing, to a CRISP of 24,893.64 Da, henceforth referred to as crovirin. Human infective trypanosomatid forms, including intracellular amastigotes, were sensitive to crovirin, with low IC50 or LD50 values (1.10-2.38 µg/ml. A considerably higher concentration (20 µg/ml of crovirin was required to elicit only limited toxicity on mammalian cells.This is the first report of CRISP anti-protozoal activity, and suggests that other members of this family might have potential as drugs or drug leads for the development of novel agents against trypanosomatid-borne neglected diseases.

  14. Contrasting modes and tempos of venom expression evolution in two snake species.

    Science.gov (United States)

    Margres, Mark J; McGivern, James J; Seavy, Margaret; Wray, Kenneth P; Facente, Jack; Rokyta, Darin R

    2015-01-01

    Selection is predicted to drive diversification within species and lead to local adaptation, but understanding the mechanistic details underlying this process and thus the genetic basis of adaptive evolution requires the mapping of genotype to phenotype. Venom is complex and involves many genes, but the specialization of the venom gland toward toxin production allows specific transcripts to be correlated with specific toxic proteins, establishing a direct link from genotype to phenotype. To determine the extent of expression variation and identify the processes driving patterns of phenotypic diversity, we constructed genotype-phenotype maps and compared range-wide toxin-protein expression variation for two species of snake with nearly identical ranges: the eastern diamondback rattlesnake (Crotalus adamanteus) and the eastern coral snake (Micrurus fulvius). We detected significant expression variation in C. adamanteus, identified the specific loci associated with population differentiation, and found that loci expressed at all levels contributed to this divergence. Contrary to expectations, we found no expression variation in M. fulvius, suggesting that M. fulvius populations are not locally adapted. Our results not only linked expression variation at specific loci to divergence in a polygenic, complex trait but also have extensive conservation and biomedical implications. C. adamanteus is currently a candidate for federal listing under the Endangered Species Act, and the loss of any major population would result in the irrevocable loss of a unique venom phenotype. The lack of variation in M. fulvius has significant biomedical application because our data will assist in the development of effective antivenom for this species. Copyright © 2015 by the Genetics Society of America.

  15. Full-Length Venom Protein cDNA Sequences from Venom-Derived mRNA: Exploring Compositional Variation and Adaptive Multigene Evolution.

    Science.gov (United States)

    Modahl, Cassandra M; Mackessy, Stephen P

    2016-06-01

    Envenomation of humans by snakes is a complex and continuously evolving medical emergency, and treatment is made that much more difficult by the diverse biochemical composition of many venoms. Venomous snakes and their venoms also provide models for the study of molecular evolutionary processes leading to adaptation and genotype-phenotype relationships. To compare venom complexity and protein sequences, venom gland transcriptomes are assembled, which usually requires the sacrifice of snakes for tissue. However, toxin transcripts are also present in venoms, offering the possibility of obtaining cDNA sequences directly from venom. This study provides evidence that unknown full-length venom protein transcripts can be obtained from the venoms of multiple species from all major venomous snake families. These unknown venom protein cDNAs are obtained by the use of primers designed from conserved signal peptide sequences within each venom protein superfamily. This technique was used to assemble a partial venom gland transcriptome for the Middle American Rattlesnake (Crotalus simus tzabcan) by amplifying sequences for phospholipases A2, serine proteases, C-lectins, and metalloproteinases from within venom. Phospholipase A2 sequences were also recovered from the venoms of several rattlesnakes and an elapid snake (Pseudechis porphyriacus), and three-finger toxin sequences were recovered from multiple rear-fanged snake species, demonstrating that the three major clades of advanced snakes (Elapidae, Viperidae, Colubridae) have stable mRNA present in their venoms. These cDNA sequences from venom were then used to explore potential activities derived from protein sequence similarities and evolutionary histories within these large multigene superfamilies. Venom-derived sequences can also be used to aid in characterizing venoms that lack proteomic profiles and identify sequence characteristics indicating specific envenomation profiles. This approach, requiring only venom, provides

  16. Rapid screening and identification of ACE inhibitors in snake venoms using at-line nanofractionation LC-MS.

    Science.gov (United States)

    Mladic, Marija; de Waal, Tessa; Burggraaff, Lindsey; Slagboom, Julien; Somsen, Govert W; Niessen, Wilfried M A; Manjunatha Kini, R; Kool, Jeroen

    2017-10-01

    This study presents an analytical method for the screening of snake venoms for inhibitors of the angiotensin-converting enzyme (ACE) and a strategy for their rapid identification. The method is based on an at-line nanofractionation approach, which combines liquid chromatography (LC), mass spectrometry (MS), and pharmacology in one platform. After initial LC separation of a crude venom, a post-column flow split is introduced enabling parallel MS identification and high-resolution fractionation onto 384-well plates. The plates are subsequently freeze-dried and used in a fluorescence-based ACE activity assay to determine the ability of the nanofractions to inhibit ACE activity. Once the bioactive wells are identified, the parallel MS data reveals the masses corresponding to the activities found. Narrowing down of possible bioactive candidates is provided by comparison of bioactivity profiles after reversed-phase liquid chromatography (RPLC) and after hydrophilic interaction chromatography (HILIC) of a crude venom. Additional nanoLC-MS/MS analysis is performed on the content of the bioactive nanofractions to determine peptide sequences. The method described was optimized, evaluated, and successfully applied for screening of 30 snake venoms for the presence of ACE inhibitors. As a result, two new bioactive peptides were identified: pELWPRPHVPP in Crotalus viridis viridis venom with IC 50  = 1.1 μM and pEWPPWPPRPPIPP in Cerastes cerastes cerastes venom with IC 50  = 3.5 μM. The identified peptides possess a high sequence similarity to other bradykinin-potentiating peptides (BPPs), which are known ACE inhibitors found in snake venoms.

  17. Recombinant rubistatin (r-Rub), an MVD disintegrin, inhibits cell migration and proliferation, and is a strong apoptotic inducer of the human melanoma cell line SK-Mel-28.

    Science.gov (United States)

    Carey, Clayton M; Bueno, Raymund; Gutierrez, Daniel A; Petro, Christopher; Lucena, Sara E; Sanchez, Elda E; Soto, Julio G

    2012-02-01

    Disintegrins are low molecular weight peptides isolated from viper venom. These peptides bind to integrin receptors using a conserved binding motif sequence containing an RGD or similar motif. As a consequence, disintegrins can inhibit platelet aggregation and inhibit cell migration, proliferation, and initiate apoptosis in cancer cell lines. Rubistatin is a MVD disintegrin cloned from a Crotalus ruber ruber venom gland. The biological activity of MVD disintegrins is poorly understood. Recombinant rubistatin (r-Rub) was cloned into a pET32b plasmid and expressed in reductase-deficient Escherichia coli. Expression was induced with IPTG and the resulting fusion peptide was affinity purified, followed by thrombin cleavage, and removal of vector coded sequences. r-Rub peptide inhibited ADP-induced platelet aggregation by 54% ± 6.38 in whole blood. We assessed the ability of r-Rub to initiate apoptosis in three human cancer cell lines. Cultures of SK-Mel-28, HeLA, and T24 cells were grown for 24 h with 2.5 μM r-Rub followed by Hoechst staining. Chromatin fragmentation was observed in treated SK-Mel-28, but not in T24 or HeLA cells. A TUNEL assay revealed that 51.55% ± 5.28 of SK-Mel-28 cells were apoptotic after 18 h of treatment with 3.5 μM of r-Rub. Cell migration and proliferation assays were performed in order to further characterize the biological effects of r-Rub on SK-Mel-28 cells. At 3 μM, r-Rub inhibited cell migration by 44.4% ± 0.5, while at 3.5 μM it was able to inhibit cell proliferation by 83% ± 6.0. Copyright © 2011 Elsevier Ltd. All rights reserved.

  18. Acções phylacticas não especificas em relação aos venenos ophidicos: tratamento auxiliar dos accidentes ophidicos

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    J. Vellard

    1929-01-01

    Full Text Available Dans l'intoxication expérimentale du pigeon avec les venins de Crotalus terrificus, de Lachesis jararaca et de Naya tripudians, nous avons étudié l'action de 12 médicaments, possédant des affinités neurotropes plus ou moins accusées, arrivant aux conclusions suivantes: 1 - Quelques-uns des ces médicaments exercent une action protectrice spéciale contre ces venins, permettant, injectés une heure avant le vénin, d'eviter tout symptôme d'intoxication chez le pigeon. Certains n'agissent qu'injectés em même temps que le vénin. D'autres sont complètement inactifs. Aucun d'êntre eux ne possède de propriétés curatives. 2 - En plus de ce pouvoir protecteur, ou phylactique, un certain nombre d'entre eux possède sur les venins, une action neutralisante "in vitro", s'accentuant après une heure de contact. 3 - Les conditions dans lesquelles s'exercent ces deux actions, protectrice et neutralisante, sont très variables suivant les venins et les médicaments. Sur leur mécanisme très complexe, on ne peut actuellement formuler que des hypothèses. 4 - Au point de vue pratique, certains de ces médicaments, surtout la spartéine et l'adrénaline, peuvent être utilisés avec avantage comme complément du traitement sérotérapique spécifique dans les accidents déterminés par les animaux venimeux.

  19. Using multiscale spatial models to assess potential surrogate habitat for an imperiled reptile.

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    Jennifer M Fill

    Full Text Available In evaluating conservation and management options for species, practitioners might consider surrogate habitats at multiple scales when estimating available habitat or modeling species' potential distributions based on suitable habitats, especially when native environments are rare. Species' dependence on surrogates likely increases as optimal habitat is degraded and lost due to anthropogenic landscape change, and thus surrogate habitats may be vital for an imperiled species' survival in highly modified landscapes. We used spatial habitat models to examine a potential surrogate habitat for an imperiled ambush predator (eastern diamondback rattlesnake, Crotalus adamanteus; EDB at two scales. The EDB is an apex predator indigenous to imperiled longleaf pine ecosystems (Pinus palustris of the southeastern United States. Loss of native open-canopy pine savannas and woodlands has been suggested as the principal cause of the species' extensive decline. We examined EDB habitat selection in the Coastal Plain tidewater region to evaluate the role of marsh as a potential surrogate habitat and to further quantify the species' habitat requirements at two scales: home range (HR and within the home range (WHR. We studied EDBs using radiotelemetry and employed an information-theoretic approach and logistic regression to model habitat selection as use vs.We failed to detect a positive association with marsh as a surrogate habitat at the HR scale; rather, EDBs exhibited significantly negative associations with all landscape patches except pine savanna. Within home range selection was characterized by a negative association with forest and a positive association with ground cover, which suggests that EDBs may use surrogate habitats of similar structure, including marsh, within their home ranges. While our HR analysis did not support tidal marsh as a surrogate habitat, marsh may still provide resources for EDBs at smaller scales.

  20. Phospholipase A2-treated human high-density lipoprotein and cholesterol movements: exchange processes and lecithin: cholesterol acyltransferase reactivity.

    Science.gov (United States)

    Chollet, F; Perret, B P; Chap, H; Douste-Blazy, L

    1986-02-12

    Human HDL3 (d 1.125-1.21 g/ml) were treated by an exogenous phospholipase A2 from Crotalus adamenteus in the presence of albumin. Phosphatidylcholine hydrolysis ranged between 30 and 90% and the reisolated particle was essentially devoid of lipolysis products. (1) An exchange of free cholesterol was recorded between radiolabelled erythrocytes at 5-10% haematocrit and HDL3 (0.6 mM total cholesterol) from 0 to 12-15 h. Isotopic equilibration was reached. Kinetic analysis of the data indicated a constant rate of free cholesterol exchange of 13.0 microM/h with a half-time of equilibration around 3 h. Very similar values of cholesterol exchange, specific radioactivities and kinetic parameters were measured when phospholipase-treated HDL replaced control HDL. (2) The lecithin: cholesterol acyltransferase reactivity of HDL3, containing different amounts of phosphatidylcholine, as achieved by various degrees of phospholipase A2 treatment, was measured using a crude preparation of lecithin: cholesterol acyltransferase (the d 1.21-1.25 g/ml plasma fraction). The rate of esterification was determined between 0 and 12 h. Following a 15-30% lipolysis, the lecithin: cholesterol acyltransferase reactivity of HDL3 was reduced about 30-40%, and then continued to decrease, though more slowly, as the phospholipid content was further lowered in the particle. (3) The addition of the lecithin: cholesterol acyltransferase preparation into an incubation medium made of labelled erythrocytes and HDL3 promoted a movement of radioactive cholesterol out of cells, above the values of exchange, and an accumulation of cholesteryl esters in HDL. This reflected a mass consumption of free cholesterol, from both the cellular and the lipoprotein compartments upon the lecithin: cholesterol acyltransferase action. As a consequence of a decreased reactivity, phospholipase-treated HDL (with 2/3 of phosphatidylcholine hydrolyzed) proved much less effective in the lecithin: cholesterol acyltransferase

  1. The kinetics of the phospholipase A2-catalyzed hydrolysis of Egg phosphatidylcholine in unilamellar vesicles. Product inhibition and its relief by serum albumin.

    Science.gov (United States)

    Kupferberg, J P; Yokoyama, S; Kézdy, F J

    1981-06-25

    Only the lecithin in the outer leaflet (representing 70% of the total) of egg lecithin unilamellar vesicles is hydrolyzed by Crotalus atrox phospholipase A2. Hydrolyzed vesicles remain intact and impermeable to ionic solutes. The fatty acids produced in the hydrolysis remain on the vesicle and are only partially ionized at neutral pH due to electrostatic repulsions. About 40% of the lysolecithin product is desorbed from the vesicle. In the presence of a large excess of bovine serum albumin, the reaction is first order with respect to both the enzyme and the substrate. At 21 degrees C, pH 7.2, I = 0.16 M, and [Ca2+] = 7 mM, the second order rate constant is kex(2) = 1.5 X 10(6) M-1 s-1. In the absence of albumin, the reaction is inhibited competitively by both the monomeric (KIm = 4.5 X 10(-8) M) and micellar (nKIa = 3.7 X 10(-7) M) forms of lysolecithin ([critical micelle concentration] = 4.3 X 10(-6) M). Bovine serum albumin complexes two molecules of lysolecithin with a dissociation constant, Kb = 5 X 10(-8) M. With substoichiometric albumin, the reaction is biphasic, and, when the albumin is saturated with lysolecithin, the kinetics become similar to those observed in the absence of albumin. The action of phospholipase A2 shows that in unilamellar vesicles there is only one major lecithin conformation in the outer leaflet, or that all conformations are rapidly interconvertible.

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

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    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%.

  3. Perfil epidemiológico del accidente ofídico en las tierras bajas de Guatemala

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    Dennis Guerra-Centeno

    2017-03-01

    Full Text Available Se describe el perfil epidemiológico del accidente ofídico en las tierras bajas de Guatemala a partir del análisis de 305 expedientes de casos atendidos en los hospitales regionales de Escuintla y San Benito, Petén, entre 2008 y 2013. De los 305 casos, 169 correspondieron al Hospital Regional de Escuintla y 136 al Hospital Regional de San Benito Petén. Los casos del Hospital Regional de Escuintla correspondieron a pacientes provenientes de los departamentos de Escuintla, Santa Rosa, Chimaltenango, Retalhuleu, Suchitepéquez, Sacatepéquez, Sololá y Guatemala. Los casos del Hospital Regional de San Benito Petén correspondieron a pacientes provenientes del área centro y norte de Petén, y algunos referidos del área sur. La edad media de la víctima fue 25.2 años. La enfermedad afectó más a hombres (61.3% que a mujeres (38.7%. La mayoría de los afectados (28.3% eran agricultores. Las especies de serpiente que se consignaron en los expedientes con mayor frecuencia fueron Bothrops asper (serpiente barba amarilla en la región norte y Crotalus simus (serpiente cascabel en la región sur. La mayoría de las mordeduras ocurrieron en los miembros inferiores. Los eventos de mordedura ocurrieron durante el día y durante la noche en proporciones similares. El tiempo medio que tarda la víctima en llegar al hospital fue de 5.6 horas. Se describen también otros aspectos epidemiológicos y se discuten las implicaciones ecológicas y sociales de los datos generados.

  4. The detection of hemorrhagic proteins in snake venoms using monoclonal antibodies against Virginia opossum (Didelphis virginiana) serum.

    Science.gov (United States)

    Sánchez, E E; García, C; Pérez, J C; De La Zerda, S J

    1998-10-01

    Most snakes and a few warm-blooded animals have a resistance to snake venoms because of naturally occurring antihemorrhagins found in their sera. The antihemorrhagins in serum of Virginia opossum (Didelphis virginiana) neutralize hemorrhagic activity by binding to hemorrhagins in snake venoms. The binding characteristic of antihemorrhagins in D. virginiana serum was used to develop a five-step western blot. The detection of hemorrhagic proteins were measured indirectly with antihemorrhagins in Virginia opossum serum and with DV-2LD#2, a monoclonal antibody specific for Virginia opossum antihemorrhagins. Snake venoms were separated by native-PAGE, transferred to a Millipore Immobilon-P membrane and then incubated with crude Virginia opossum serum. The hemorrhagins in snake venom bind to antihemorrhagins in Virginia opossum serum which react with DV-2LD#2 a monoclonal antibody that is specific for Virginia opossum antihemorrhagins. DV-2LD#2 monoclonal antibody inhibits antihemorrhagic activity in Virginia opossum serum when mixed in equal amounts. The inhibition of antihemorrhagins by DV-2LD#2 monoclonal antibody suggests specificity. DV-2LD#2 monoclonal antibody does not recognize antihemorrhagins in gray woodrat (Neotoma micropus) serum. The five-step western blot reveals two well-defined bands which represent hemorrhagins found in Western diamondback rattlesnake (Crotalus atrox) venom. Venoms from 15 different snake species were examined to determine the usefulness of the five-step western blot. Other hemorrhagic venoms (Great Basin rattlesnake (C. viridis lutosus), Prairie rattlesnake (C. viridis viridis), Tancitaran dusky rattlesnake (C. pusillus), Northern Mojave rattlesnake (C. scutulatus scutulatus type B) and Northern Pacific rattlesnake (C. v. oreganus)) had one single band in the five-step western blot. DV-2LD#2 did not bind to the non-hemorrhagic venoms and reacted with 50% of the hemorrhagic venoms used in this study. The monoclonal antibody, CAH

  5. Human cytokine response to Texas crotaline envenomation before and after antivenom administration.

    Science.gov (United States)

    Crocker, Patrick; Zad, Omid; Milling, Truman; Maxson, Todd; King, Benjamin; Whorton, Elbert

    2010-10-01

    The aim of this study was to characterize the human cytokine response to Texas crotaline envenomation before and after antivenom administration. This study enrolled crotaline bite victims presenting to a regional trauma center and children's hospital from March to November 2007 and age-matched unbitten controls. Blood spot cards were obtained from bite victims at presentation and at 1 and 6 hours after antivenom administration. One control sample was drawn from each of the age-matched controls selected from urgent care patients presenting for minor complaints. Samples were delivered to a laboratory using a proprietary method for quantitative evaluation of a large number of biomarkers in parallel with bead-based multiplex immunoassays. After obtaining informed consent, 14 crotaline bite victims (age range, 5-85 years; median age, 45 years; 50% female) (Snakebite Severity Score, 2-7; median, 3) and 14 age-matched controls were enrolled. There were 7 copperhead (Agkistrodon contortrix) bites, 4 rattlesnake (probably Western Diamondback Crotalus atrox) bites, 2 cottonmouth (Agkistrodon piscivorus) bites, and 1 bite from a snake that was not identified by the victim. In t tests, the means in the presentation samples for apolipoprotein A-I (Apo A-I), Apo C3, interleukin 4 (IL-4), myeloperoxidase, plasminogen activator inhibitor 1 (PAI-1), epidermal growth factor, and regulated upon activation, normal t-cell expressed and secreted were significantly lower and Apo H was significantly higher in the bite patients than in the controls. In the 1-hour sample, α(1)-antitrypsin, Apo A-I, Apo C3, eotaxin, IL-4, myeloperoxidase, and PAI-1 levels were lower and prostatic acid phosphatase and cancer antigen 125 levels were higher in the bite patients than in the controls. And in the 6-hour sample, α(1)-antitrypsin, Apo A-I, Apo C3, endothelin-1, IL-4, macrophage inflammatory protein 1β, myeloperoxidase, and epidermal growth factor levels were lower and Apo H level was higher in

  6. Construction of concept maps as tool for Biochemistry learning

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    Silvia Lopes de Menezes

    2006-07-01

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

  7. Perfil bioquímico sorológico de bovinos inoculados experimentalmente com veneno crotálico iodado livre e iodado incorporado em liposomas Serological biochemical profile of bovines poisoned experimentally with free iodized crotalic poison and iodized incorporated in lipossomes

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    L.A. Lago

    2004-10-01

    Full Text Available Investigou-se o perfil sorológico de bovinos inoculados com veneno crotálico detoxificado pelo método de iodação e iodação com encapsulação em liposomas. Onze fêmeas com idade média de 18 meses e peso médio de160kg, foram inoculadas com 0,03mg/kg de peso vivo do veneno crotálico do tipo crotamina positivo. Cinco animais receberam o veneno iodado livre, cinco o iodado encapsulado em liposomas e um animal recebeu o mesmo veneno na forma natural, para controle da letalidade da amostra de veneno utilizada. Não houve alterações significativas na concentração de proteínas totais, uréia e creatinina em ambos os tratamentos. Foi observada diferença significativa na concentração de creatinafosfoquinase a partir de duas horas após a inoculação do veneno. A iodação do veneno crotálico, com ou sem incorporação em liposomas, suprimiu seus efeitos deletérios sobre o perfil bioquímico de bovinos.It was studied the activity of the Crotalus venom detoxified encapsulated in lipossomes or non-encapsulated by the method of controlled iodination on the biochemical profile of bovines. Eleven 18-months-old female bovines, weighting 160kg in average were used in this study. Groups of five animals were inoculated intramuscularly with a dose of 0,03mg/kg with either the detoxified non-encapsulated venom or the detoxified encapsulated venom. One control animal received the same dose of non detoxified venom by the same route. No significant differences were observed between the study groups for the parameters of total protein, urea and creatinine. Differences were observed for creatine phosfokinase at times T O and T12 for the group treated with the iodinated venom encapsulated in liposomes. The detoxification process abolished the activity of the venom in the encapsulated or nonencapsulated form on the biochemical parameters and renal activity of bovine.

  8. Profiling the venom gland transcriptomes of Costa Rican snakes by 454 pyrosequencing

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    Sanz Libia

    2011-05-01

    Full Text Available Abstract Background A long term research goal of venomics, of applied importance for improving current antivenom therapy, but also for drug discovery, is to understand the pharmacological potential of venoms. Individually or combined, proteomic and transcriptomic studies have demonstrated their feasibility to explore in depth the molecular diversity of venoms. In the absence of genome sequence, transcriptomes represent also valuable searchable databases for proteomic projects. Results The venom gland transcriptomes of 8 Costa Rican taxa from 5 genera (Crotalus, Bothrops, Atropoides, Cerrophidion, and Bothriechis of pitvipers were investigated using high-throughput 454 pyrosequencing. 100,394 out of 330,010 masked reads produced significant hits in the available databases. 5.165,220 nucleotides (8.27% were masked by RepeatMasker, the vast majority of which corresponding to class I (retroelements and class II (DNA transposons mobile elements. BLAST hits included 79,991 matches to entries of the taxonomic suborder Serpentes, of which 62,433 displayed similarity to documented venom proteins. Strong discrepancies between the transcriptome-computed and the proteome-gathered toxin compositions were obvious at first sight. Although the reasons underlaying this discrepancy are elusive, since no clear trend within or between species is apparent, the data indicate that individual mRNA species may be translationally controlled in a species-dependent manner. The minimum number of genes from each toxin family transcribed into the venom gland transcriptome of each species was calculated from multiple alignments of reads matched to a full-length reference sequence of each toxin family. Reads encoding ORF regions of Kazal-type inhibitor-like proteins were uniquely found in Bothriechis schlegelii and B. lateralis transcriptomes, suggesting a genus-specific recruitment event during the early-Middle Miocene. A transcriptome-based cladogram supports the large

  9. Acidente ofídico no estado de Goiás

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    Pinho Fábia Maria Oliveira

    2004-01-01

    Full Text Available OBJETIVO: Avaliar aspectos epidemiológicos dos acidentes por serpentes peçonhentas ocorridos no Estado de Goiás. MÉTODOS: Foram analisadas ''Fichas de investigação de acidentes por animais peçonhentos'', pertencentes ao sistema de notificação da Secretaria de Saúde do Estado de Goiás, no triênio 1998-2000. RESULTADOS: Foram notificados, neste período, 3.261 acidentes por serpentes peçonhentas, com coeficiente de incidência variando entre 20 e 23/100.000 habitantes. O maior número de casos ocorreu entre os meses de outubro e abril. Dentre os 2.350 casos em que houve referência ao gênero da serpente, 78,6% foram causados por Bothrops, 20,8% por Crotalus e 6% por Micrurus. Houve predominância do sexo masculino (78,5% e com faixa etária entre 20 e 39 anos de idade. As regiões anatômicas mais freqüentemente picadas foram: pé (43,6%, pernas (23,2% e mãos (20,1%. Em relação ao tempo de atendimento, mais de 80% dos envenenamentos foram atendidos com menos de 6 horas da picada. Os envenenamentos foram classificados, de acordo com a gravidade, em leves (31,6%, moderados (47,5% ou graves (9,6%. As complicações mais comuns foram necrose tecidual no local da picada (31,8% nos envenenamentos botrópicos e insuficiência renal aguda (1,2% nos envenenamentos crotálicos. A letalidade geral foi de 0,46%, sendo a maior taxa observada entre acidentes crotálicos (1%. CONCLUSÕES: Acidentes por serpentes peçonhentas no Estado de Goiás acometem principalmente a população jovem do sexo masculino e têm sido causa de óbito.

  10. 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

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    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

  11. Ecological Monitoring and Compliance Program 2014 Report

    Energy Technology Data Exchange (ETDEWEB)

    Hall, Derek B. [National Security Technologies, LLC, Las Vegas, Nevada (United States); Anderson, David C. [National Security Technologies, LLC, Las Vegas, Nevada (United States); Greger, Paul D. [National Security Technologies, LLC, Las Vegas, Nevada (United States); Ostler, W. Kent [National Security Technologies, LLC, Las Vegas, Nevada (United States)

    2015-05-12

    The Ecological Monitoring and Compliance Program (EMAC), funded through the U.S. Department of Energy, National Nuclear Security Administration Nevada Field Office (NNSA/NFO, formerly Nevada Site Office), monitors the ecosystem of the Nevada National Security Site (NNSS) and ensures compliance with laws and regulations pertaining to NNSS biota. This report summarizes the program’s activities conducted by National Security Technologies, LLC (NSTec), during calendar year 2014. Program activities included (a) biological surveys at proposed activity sites, (b) desert tortoise compliance, (c) ecosystem monitoring, (d) sensitive plant species monitoring, (e) sensitive and protected/regulated animal monitoring, and (f) habitat restoration monitoring. During 2014, all applicable laws, regulations, and permit requirements were met, enabling EMAC to achieve its intended goals and objectives. Sensitive and protected/regulated species of the NNSS include 42 plants, 1 mollusk, 2 reptiles, 236 birds, and 27 mammals. These species are protected, regulated, or considered sensitive according to state or federal regulations and natural resource agencies and organizations. The desert tortoise (Gopherus agassizii) and the western yellow-billed cuckoo (Coccyzus americanus) are the only species on the NNSS protected under the Endangered Species Act, both listed as threatened. However, only one record of the cuckoo has ever been documented on the NNSS, and there is no good habitat for this species on the NNSS. It is considered a rare migrant. Biological surveys for the presence of sensitive and protected/regulated species and important biological resources on which they depend were conducted for 18 projects. A total of 199.18 hectares (ha) was surveyed for these projects. Sensitive and protected/regulated species and important biological resources found during these surveys included a predator burrow, one sidewinder rattlesnake (Crotalus cerastes), two mating speckled rattlesnakes