WorldWideScience

Sample records for crotalus durissus collilineatus

  1. Random amplified polymorphisms between two South American subspecies of rattlesnakes (Crotalus durissus collilineatus and Crotalus durissus terrificus)

    OpenAIRE

    Echeverrigaray, Sergio; Grazziotin, Gilberto; Grazziotin, Felipe; Agostini, Gustavo

    2001-01-01

    The present work was made to determine the suitability of RAPD analysis for the identification of specimens of two subspecies of South American rattlesnakes (Crotalus durissus terrificus and Crotalus durissus collilineatus). The 11 arbitrary primer sequence tested amplified a total of 161 bands, of which 31% were polymorphics when compared with Crotalus specimens. The combining results from different primers allowed the identification of all the specimens under analysis. Several characteristi...

  2. Random amplified polymorphisms between two South American subspecies of rattlesnakes (Crotalus durissus collilineatus and Crotalus durissus terrificus

    Directory of Open Access Journals (Sweden)

    Sergio Echeverrigaray

    2001-09-01

    Full Text Available The present work was made to determine the suitability of RAPD analysis for the identification of specimens of two subspecies of South American rattlesnakes (Crotalus durissus terrificus and Crotalus durissus collilineatus. The 11 arbitrary primer sequence tested amplified a total of 161 bands, of which 31% were polymorphics when compared with Crotalus specimens. The combining results from different primers allowed the identification of all the specimens under analysis. Several characteristic bands of each subspecies were identified. Dendrogram, based on RAPD markers, show three groups that corresponded to the subspecies of Crotalus, and to Bothrops jararaca specimen.O presente trabalho foi desenvolvido visando determinar a aplicabilidade da análise de RAPD na identificação de indivíduos pertencentes a duas sub-espécies de cascavéis sul-americanas (Crotalus durissus terrificus e Crotalus durissus collilineatus. As 11 sequências arbitrárias inicializadoras testadas nestes experimentos, amplificaram um total de 161 bandas, das quais 31% foram polimórficas na comparação entre espécimes de Cratalus. Os resultados combinados obtidos com as distintas sequências inicializadoras, permitiram a identificação de todos os indivíduos avaliados. Várias bandas características de cada sub-espécie foram identificadas. O dendrogama, baseado em marcaodres de RAPD, mostra tres grupos que correspondem às duas sub-espécies de Crotalus e ao especimen de Bothrops jararaca.

  3. Anti-Leptospira spp. antibodies in Crotalus durissus collilineatus kept in captivity and its zoonotic relevance.

    Science.gov (United States)

    Rodrigues, T C S; Santos, A L Q; Lima, A M C; Gomes, D O; Brites, V L C

    2016-06-01

    Leptospirosis is a worldwide spread zoonosis that can affect all groups of vertebrates, including reptiles. Because it has been little studied in snakes, this study focused on determining the occurrence of anti-Leptospira spp. antibodies in 64 Crotalus durissus collilineatus kept in captivity and on identifying the most common serovars in these animals, using the microscopic agglutination test. Of these, almost 90% were positive and there were reactions to the 22 serovars used in the study. The most common serovar in these snakes was Javanica, Andamana and Patoc. Most frequent titers were 25 and 50, although high titers (such as 1600) were also recorded, despite the absence of clinical symptoms. The possibility should be considered of captive snakes serving as a serious source of leptospiral infection in humans, which is why it is essential to study, prevent and control the disease in breeding centers and serpentariums. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. Strategy for subduing prey in the predatory behavior of Crotalus durissus collilineatus Amaral, 1926, in captivity

    Directory of Open Access Journals (Sweden)

    Vera Lucia de Campos Brites

    2006-12-01

    Full Text Available Studies of the predatory behavior of six specimens of hatchlings from the same clutch of Crotalus durissus collilineatus, born and raised in captivity, were conducted. The animal focal technique (observing the snakes through a glass visor was used in studying the snakes in order to minimize stress. The snakes were fed mice sequentially and were observed until they no longer sought the prey. Different capture strategies were adopted by this subspecies after the first occasion on which prey was caught. Percentages of the total number of capture strategies observed indicate that 20% (n = 4 made use of seizure, and 80% (n = 16 made use of poisoning, as a mean of subduing prey. The strategy of seizing prey manifested itself on the third or fourth occasion that prey was offered and appeared to be linked to the small size of the prey. It was observed that in 95% (n = 20 of the cases, ingestion began with the cranium of the prey. Regardless of the body part that was ingested first by the serpent, all serpents displayed adjustment behavior of the cranium/head and the jaws (“yawning”.

  5. Identification of crotamine in the venom of Crotalus durissus collilineatus by three different methods.

    Science.gov (United States)

    de Oliveira, Sayonara Ay More; Magalhães, Marta Regina; Salazar, Vania Cristina Rodríguez; Valadares, Marize Campos; da Cunha, Luiz Carlos

    2015-03-01

    The use of mice for the identification of crotamine has been the motive of discussions of bioethical character and technical (efficiency), so that a reassessment of the use of animals in experiments is global trend the search for alternative tests. The objective of this study was to standardize a method for HPLC-PDA to identify the presence of crotamine in the venom of rattlesnakes, aiming to propose an alternative methodology to reduce or replace the use of animals. The Cdc was evaluated as to the presence of crotamine by 3 methods: traditional test lethality in mice (Mus musculus) swiss albino male, 18-22 g (i.p.), polyacrylamide gel electrophoresis (SDS-PAGE) and HPLC-PDA. The venoms of 50 specimens of Crotalus durissus collilineatus, held in the serpentarium CEPB/PUC Goiás, were obtained by manual massage of the gland, making the collection individually. To identify the band corresponding to crotamine, the venoms of specimens, analysis was performed on SDS-PAGE and references. Procedure in mice with 20% of the samples tested positive for crotamine, 24% negative and 56% uncertain outcome. With the SDS-PAGE was identified crotamine in 26% of samples, 26% negative and 48% continued with uncertain outcome. By HPLC method showed the presence myotoxin in 86% of samples, with 14% negative. The tests conducted in this study indicated that methodology which utilizes animals for identifying the presence of crotamine the venom of C. durissus can safely be replaced by the test SDS-PAGE and HPLC, since the methods are reproducible, and do not undergo any interference biological animal and mainly contribute to reducing the number of animals used for laboratory tests. Copyright © 2014 Elsevier Ltd. All rights reserved.

  6. Crotalus durissus collilineatus Venom Induces TNF-α and IL-10 Production in Human Peripheral Blood Mononuclear Cells

    Science.gov (United States)

    Ribeiro, Camila Bastos; dos Santos, Jéssica Cristina; Silva, Jacyelle Medeiros; de Godoi, Pedro Henrique Silva; Magalhães, Marta Regina; Spadafora-Ferreira, Mônica; Fonseca, Simone Gonçalves; Pfrimer, Irmtraut Araci Hoffman

    2014-01-01

    Snake venom has been the subject of numerous studies in an attempt to find properties and biological effects that may be beneficial to man. In this study we evaluated in vitro the effects of Crotalus durissus terrificus (Cdt) and Crotalus durissus collilineatus (Cdc) venom in human peripheral blood mononuclear cells (PBMCs). At 24 h, a significant decrease of viable cells was observed in cells stimulated with the Cdc venom at 0.0005 mg/mL and 0.005 mg/mL compared to the negative control. At 48 h, a significant decrease of viable cells was observed only in cells stimulated with Cdc venom at 0.005 mg/mL. A significant increase of TNF-α and IL-10 was detected 48 hours after culture of PBMC with Cdc, but not with Cdt venom. The expression of CD69 and PD1 (programmed death-1), activation and regulatory cell markers, on CD8+ and CD8− T cells did not change in the presence of Cdt and Cdc venom. Our results suggest the presence of proinflammatory and anti-inflammatory components in the Cdc venom. Further analysis should be done to identify those Cdc venom components as it has been done for the Cdt venom by other authors, indicating that modulatory components are found in the venom of different species of Crotalus snakes. PMID:24563803

  7. Identification of antinociceptive fraction of snake venom from Crotalus durissus collilineatus crotamine-negative and its acute toxicity evaluation.

    Science.gov (United States)

    de Oliveira, Sayonara Ay More; Magalhães, Marta Regina; de Oliveira, Lilibete P; da Cunha, Luiz Carlos

    2016-11-01

    The crude venom of the snake Crotalus durissus collilineatus (CDC) promotes neurological signs and symptoms in accidents involving humans and animals and the victims reports analgesia at the bite site, without tissue destruction. Studies shows that CDC has analgesic activity, among others. The crude venom is considered unsuitable for therapeutic purposes, with encouragement to the fractionation and purification of the same. Thus, the aim with CDC venom is: to perform fractionation by preparative HPLC; to test the antinociceptive activity of fractions and acute toxicity of active fractions. The CDC was fractionated on preparative HPLC-PDA (Oliveira et al., 2015) and the fractions were tested for their antinociceptive activity for writhing test by acetic acid (0.6%) in mice. For one of the fractions, which showed high analgesic effect both p.o. and i.p. routes, it evaluated the acute toxicity by the up and down method (OECD, 2001). In the fractionation by HPLC-PDA, CDC yielded 10 peaks (P1P10). SDS-PAGE showed that there was a good separation of components of the venom. All peaks were evaluated for their ability to reduce writhing, and the only one that apparently showed antinociceptive effect was Fr5 fraction (40 μg/kg). The Fr5 was able to reduce by 47% the number of contortions (i.p.) and 87% (p.o.), compared to control. The Fr5 fraction showed no morbidity and no mortality in the acute toxicity test (dose of 1000 μg/kg, p.o.); so it was not possible to estimate the LD50. According to the results, it can be stated that the venom and Fr5 of Crotalus durissus collilineatus snake of crotamine-negative type, may exhibit antinociceptive activity by suppressing nociception induced by acetic acid, suggesting it is related to effects on peripheral sites spinal and presents low acute toxicity values in experimental animals. Copyright © 2016 Elsevier Ltd. All rights reserved.

  8. Isolation and biochemical characterization of a γ-type phospholipase A2 inhibitor from Crotalus durissus collilineatus snake serum.

    Science.gov (United States)

    Gimenes, Sarah Natalie Cirilo; Ferreira, Francis Barbosa; Silveira, Ana Carolina Portella; Rodrigues, Renata Santos; Yoneyama, Kelly Aparecida Geraldo; Izabel Dos Santos, Juliana; Fontes, Marcos Roberto de Mattos; de Campos Brites, Vera Lúcia; Santos, André Luiz Quagliatto; Borges, Márcia Helena; Lopes, Daiana Silva; Rodrigues, Veridiana M

    2014-04-01

    In the present work, we describe the isolation and partial structural and biochemical characterization of the first phospholipase A2 inhibitor (γPLI) from Crotalus durissus collilineatus (Cdc) snake serum. Initially, the Cdc serum was subjected to a Q-Sepharose ion exchange column, producing six peaks at 280 nm absorbance (Q1-Q6). Subsequently, Q4 fraction was submitted to affinity chromatography with immobilized PLA2 BnSP-7, a step that resulted in two fractions (NHS-1 and NHS-2). The latter contained the inhibitor, denominated γCdcPLI. The molecular mass of γCdcPLI, determined by Matrix-Assisted Laser Desorption Ionization Time-of-Flight (MALDI-TOF), was 22,340 Da. Partial sequences obtained by Edman degradation and by mass spectrometry (MALDI-TOF/TOF), showed similarity, as expected, to other related inhibitors. Circular dichroism (CD) analysis showed the presence of approximately 22% alpha helices and 29% beta sheets in the protein secondary structure. Additionally, CD studies also indicated no significant changes in the secondary structure of γCdcPLI when it is complexed to BpPLA2-TXI. On the other hand, dynamic light scattering (DLS) assays showed a temperature-dependent oligomerization behavior for this inhibitor. Biochemical analyses showed γCdcPLI was able to inhibit the enzymatic, cytotoxic and myotoxic activities of PLA2s. Structural and functional studies performed on this inhibitor may elucidate the action mechanisms of PLA2 inhibitors. In addition, we hope this study may contribute to investigating the potential use of these inhibitors for the treatment of snakebite or inflammatory diseases in which PLA2s may be involved. Copyright © 2014 Elsevier Ltd. All rights reserved.

  9. Hepatozoon spp. (Apicomplexa: Hepatozoidae) infection and selected hematological values of the neotropical rattlesnake, Crotalus durissus collilineatus (Linnaeus, 1758) (Serpentes: Viperidae), from Brazil.

    Science.gov (United States)

    Motta, Rafael Otávio Cançado; Cunha, Lucas Maciel; Leite, Romário Cerqueira; da Silva, Israel José; Pinto, Ana Cristina Araújo; Braga, Erika Martins; da Cunha, Arildo Pinto; de Oliveira, Paulo Roberto

    2011-09-01

    This study aims to establish the hematological values of Crotalus durissus collilineatus snakes captured in Brazil as well as to verify the effects of hematozoan infection on these snakes. Eighty-three blood samples were drawn from C. d. collilineatus specimens for analysis. The sample set was composed of 30 males and 30 females, recently caught from the wild, and 11 males and 12 females bred in captivity. Blood samples were used to determine red blood cell counts, white blood cell counts, thrombocyte counts, hematocrit values, hemoglobin concentration, and total plasma protein. Blood smears were used to diagnose Hepatozoon spp. infection and to calculate the parasitic load in the sample as well as the percentage of immature red cells. Results obtained for the wild-caught animals, with and without parasites, were compared among themselves and with the values obtained for the captive-bred animals. Hematological values for C. durissus were established. Wild-caught snakes had an infection rate of 38.3%, while no Hepatozoon sp. infection was detected in the captive-bred animals. The snakes which were not infected by the Hepatozoon sp. exhibited average weight, length, and weight-length ratios higher than those of the infected animals. An increase in immature red cells was noted in the Hepatozoon-infected snakes.

  10. Expression of a new serine protease from Crotalus durissus collilineatus venom in Pichia pastoris and functional comparison with the native enzyme.

    Science.gov (United States)

    Boldrini-França, Johara; Santos Rodrigues, Renata; Santos-Silva, Ludier Kesser; de Souza, Dayane Lorena Naves; Gomes, Mário Sérgio Rocha; Cologna, Camila Takeno; de Pauw, Edwin; Quinton, Loïc; Henrique-Silva, Flávio; de Melo Rodrigues, Veridiana; Arantes, Eliane Candiani

    2015-12-01

    Snake venom serine proteases (SVSPs) act primarily on plasma proteins related to blood clotting and are considered promising for the treatment of several hemostatic disorders. We report the heterologous expression of a serine protease from Crotalus durissus collilineatus, named collinein-1, in Pichia pastoris, as well as the enzymatic comparative characterization of the toxin in native and recombinant forms. The complementary DNA (cDNA) encoding collinein-1 was amplified from cDNA library of C. d. collilineatus venom gland and cloned into the pPICZαA vector. The recombinant plasmid was used to transform cells of KM71H P. pastoris. Heterologous expression was induced by methanol and yielded 56 mg of recombinant collinein-1 (rCollinein-1) per liter of culture. The native collinein-1 was purified from C. d. collilineatus venom, and its identity was confirmed by amino acid sequencing. The native and recombinant enzymes showed similar effects upon bovine fibrinogen by releasing preferentially fibrinopeptide A. Although both enzymes have induced plasma coagulation, native Colinein-1 has shown higher coagulant activity. The serine proteases were able to hydrolyze the chromogenic substrates S-2222, S-2238, and S2302. Both enzymes showed high stability on different pH and temperature, and their esterase activities were inhibited in the presence of Zn2+ and Cu2+. The serine proteases showed similar k cat/K m values in enzyme kinetics assays, suggesting no significant differences in efficiency of these proteins to hydrolyze the substrate. These results demonstrated that rCollinein-1 was expressed with functional integrity on the evaluated parameters. The success in producing a functionally active recombinant SVSP may generate perspectives to their future therapeutic applications.

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

    Directory of Open Access Journals (Sweden)

    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.

  12. Snake venomic of Crotalus durissus terrificus--correlation with pharmacological activities.

    Science.gov (United States)

    Georgieva, Dessislava; Ohler, Michaela; Seifert, Jana; von Bergen, Martin; Arni, Raghuvir K; Genov, Nicolay; Betzel, Christian

    2010-05-07

    The snake venomic of Crotalus durissus terrificus was analyzed by 2-D and 1-D electrophoresis and subsequent MS/MS and enzymatic assays. The venomic of the South American rattlesnake comprises toxins from seven protein families: phospholipases A(2), serine proteinases, ecto-5'-nucleotidases, metalloproteinases, nerve growth factors, phosphodiesterases, and glutaminyl cyclase. The venom toxin composition correlates with the clinical manifestation of the crotalinae snake bites and explains pathological effects of the venom such as neurotoxicity, systemic myonecrosis, hemostatic disorders, myoglobinuria, and acute renal failure. The vast majority of toxins are potentially involved in neurotoxicity, myotoxicity, and coagulopathy. The predominant venom components are neurotoxic phospholipases A(2) and serine proteinases. The venom is a rich source of 5'-nucleotidases (7.8% of the identified toxins) inducing hemostatic disorders. Analysis of the venom protein composition provided a catalogue for secreted toxins. The venomic composition of Crotalus d. terrificus and venom gland transcriptome of the synonymous subspecies Crotalus d. collilineatus show differences in the occurrence of protein families and in the abundance of toxins. Some of the venom components identified by the proteomic analysis were not reported in the transcriptome of the Crotalus d. collilineatus venom gland. Enzymatic activities of the Crotalus d. terrificus venom were determined and correlated with the proteomic composition.

  13. Afibrinogenemia following snake bite (Crotalus durissus terrificus

    Directory of Open Access Journals (Sweden)

    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.

  14. Neuromuscular effects of venoms and crotoxin-like proteins from Crotalus durissus ruruima and Crotalus durissus cumanensis.

    Science.gov (United States)

    Cavalcante, Walter Luís Garrido; Ponce-Soto, Luis Alberto; Marangoni, Sérgio; Gallacci, Márcia

    2015-03-01

    A myographic study was performed to compare the neuromuscular effects of venoms and crotoxin-like proteins from Crotalus durissus ruruima and Crotalus durissus cumanensis in mice phrenic-diaphragm preparation. It was concluded that both venoms present neurotoxic activity as a consequence of their crotoxin content. Furthermore, crotoxin from C.d. cumanensis is more potent than that from C.d. ruruima venom. At the concentration range in which both venoms express neurotoxic activity, only C.d. cumanensis venom also manifest a direct myotoxic effect that probably involves the synergic participation of other components than crotoxin. Copyright © 2015 Elsevier Ltd. All rights reserved.

  15. Ophidascaris durissus sp. nov. (Nematoda Ascarididae parasitizing Crotalus durissus Linnaeus (Ophidia, Viperidae in Brazil

    Directory of Open Access Journals (Sweden)

    Marcia Helena Martins Panizzutti

    2003-03-01

    Full Text Available The species Ophidascaris durissus sp. nov. is proposed with basis on specimens recovered from the rattlesnake Crotalus durissus L., 1758 (type host captured in Foz do Iguaçu, Brazil (type locality. By the lack of interlabia, the new species can be compared only to O. natricis Yamaguti, 1935 from Japan and O. freitasi Hoa & Lien, 1970, from Vietnam. However, O. durissus sp. nov. differs from O. natricis mainly by the absence of internal lip papillae, location of the vulvar aperture and length of the spicules; from O. freitasi mostly by the greater number of pre-cloacal and distribution of post-cloacal papillae.

  16. Comparison of wildlife and captivity rattlesnakes (Crotalus durissus terrificus) microbiota

    OpenAIRE

    Ferreira Junior,Rui S.; Siqueira,Amanda K.; Campagner,Michelle V.; Salerno,Tatiana; Soares,Taíssa C.S.; Lucheis,Simone B.; Paes,Antonio C.; Barraviera,Benedito

    2009-01-01

    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%) microor...

  17. Microvesicles in the venom of Crotalus durissus terrificus (Serpentes, Viperidae).

    Science.gov (United States)

    Carneiro, Sylvia Mendes; Fernandes, Wilson; Sant'Anna, Sávio Stefanini; Yamanouye, Norma

    2007-01-01

    Microvesicles with electron-dense content are consistently observed by transmission electron microscopy on the luminal face of secretory cells of venom glands of viperid snakes. In this work, we evaluated their presence in Crotalus durissus terrificus venom glands and also in freshly collected venom. Microvesicles were found in the venom glands mainly in regions of exocytosis. They ranged from 40 to 80 nm in diameter. Freeze-fracture replicas of the glands revealed particles on the cytoplasmic leaflet (P-face) of these vesicles, suggesting that they carry transmembrane proteins. Vesicles separated by ultracentrifugation from cell-free venom were similar in size and structure to the microvesicles observed in the glands. A fine fuzzy coat surrounded each microvesicle. The function of these venom vesicles is still unknown, but they may contribute to inactivation of stored venom components, or their activation after the venom is released.

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

    Science.gov (United States)

    Duarte, Marcelo Ribeiro; Menezes, Frederico Alcântara

    2013-12-05

    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 Butantan Institute, in São Paulo, Brazil, from 1998 to 2012 show a declining tendency of snakes brought by donors. In addition, research shows no evidence of Crotalus durissus being an expanding species in the Brazilian territory.

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

    Science.gov (United States)

    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 Butantan Institute, in São Paulo, Brazil, from 1998 to 2012 show a declining tendency of snakes brought by donors. In addition, research shows no evidence of Crotalus durissus being an expanding species in the Brazilian territory. PMID:24314146

  20. Compartment syndrome after South American rattlesnake (Crotalus durissus terrificus) envenomation.

    Science.gov (United States)

    Bucaretchi, F; De Capitani, E M; Hyslop, S; Mello, S M; Fernandes, C B; Bergo, F; Nascimento, F B P

    2014-07-01

    In order to report the outcome of a patient who developed compartment syndrome after South American rattlesnake (Crotalus durissus terrificus) envenomation, confirmed by subfascial pressure measurement and magnetic resonance imaging (MRI). A 63-year-old male was admitted 1 h after being bitten on the right elbow by a "large" snake, which was not brought for identification. Physical and laboratory features upon admission revealed two fang marks, local tense swelling, paresthesia, intense local pain, hypertension, coagulopathy, and CK = 1530 U/L (RV < 170 U/L). The case was initially treated with bothropic antivenom (80 mL, intravenously), with no improvement. Evolution within 13-14 h post-bite revealed generalized myalgia, muscle weakness, palpebral ptosis, and severe rhabdomyolysis (CK = 126,160 U/L) compatible with envenoming by C. d. terrificus. The patient was then treated with crotalic antivenom (200 mL, intravenously), fluid replacement, and urine alkalinization. Twenty-four-hour post-bite MRI showed marked muscular edema in the anterior compartment of the right forearm, with a high subfascial pressure (40 mmHg) being detected 1 h later. ELISA of a blood sample obtained upon admission, before antivenom infusion, revealed a high serum concentration of C. d. terrificus venom. No fasciotomy was performed and the patient was discharged seven days later without sequelae. Snakebite by C. d. terrificus with subfascial venom injection may lead to increased intracompartmental pressure.

  1. 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...... 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...

  2. Megaselia scalaris (Diptera: Phoridae) causing myiasis in Crotalus durissus terrificus (Serpentes: Viperidae) in Brazil.

    Science.gov (United States)

    da Silva, R J; do Prado, A P; Rodrigues, R R; Lopes, C A; Godoy, W A

    1999-09-01

    We describe a case of myiasis in Crotalus durissus terrificus (Laurenti) caused by Megaselia scalaris (Loew). The snake was found in Anhembi, Sao Paulo, Brazil, with a lesion measuring 25 mm in diameter where the larvae of M. scalaris had penetrated the ribs. The opportunistic behavior of the larvae of M. scalaris is discussed.

  3. 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

  4. Histopathological evaluation in experimental envenomation of dogs with Crotalus durissus terrificus venom

    Directory of Open Access Journals (Sweden)

    F. Sangiorgio

    2008-01-01

    Full Text Available The present work evaluated histopathological aspects in experimental envenomation of dogs with Crotalus durissus terrificus venom. Twenty-eight mixed breed adult dogs were divided into three groups of seven animals each: Group I - only venom; Group II - venom + 50ml antiophidic serum + fluid therapy; Group III - venom + 50ml antiophidic serum + fluid therapy + urine alkalization. Lyophilized venom of Crotalus durissus terrificus was reconstituted in saline solution and inoculated subcutaneously at the dose of 1mg/kg body weight. Three animals of each group were subjected to euthanasia, and their muscular tissue, brain, spleen, kidneys, heart, lungs, stomach, small and large intestines, and popliteal lymph node fragments were collected for histopathological evaluation. There was myonecrosis in the inoculated limb, renal tubular degeneration, lymphoid hyperplasia of spleen, and unspecific reactive hepatitis. These results show the antigenicity and action of the venom on the immune system.

  5. Tetrodotoxin-insensitive electrical field stimulation-induced contractions on Crotalus durissus terrificus corpus cavernosum.

    Science.gov (United States)

    Campos, Rafael; Mónica, Fabíola Z; Rodrigues, Renata Lopes; Rojas-Moscoso, Julio Alejandro; Moreno, Ronilson Agnaldo; Cogo, José Carlos; de Oliveira, Marco Antonio; Antunes, Edson; De Nucci, Gilberto

    2017-01-01

    Reptiles are the first amniotes to develop an intromitent penis, however until now the mechanisms involved in the electrical field stimulation-induced contraction on corpora cavernosa isolated from Crotalus durissus terrificus were not investigated. Crotalus and rabbit corpora cavernosa were mounted in 10 mL organ baths for isometric tension recording. Electrical field stimulation (EFS)-induced contractions were performed in presence/absence of phentolamine (10 μM), guanethidine (30 μM), tetrodotoxin (1 μM and 1mM), A-803467 (10 μM), 3-iodo-L-Tyrosine (1 mM), salsolinol (3 μM) and a modified Krebs solution (equimolar substitution of NaCl by N-methyl-D-glucamine). Immuno-histochemistry for tyrosine hydroxylase was also performed. Electrical field stimulation (EFS; 8 Hz and 16 Hz) caused contractions in both Crotalus and rabbit corpora cavernosa. The contractions were abolished by previous incubation with either phentolamine or guanethidine. Tetrodotoxin (1 μM) also abolished the EFS-induced contractions of rabbit CC, but did not affect EFS-induced contractions of Crotalus CC. Addition of A-803467 (10 μM) did not change the EFS-induced contractions of Crotalus CC but abolished rabbit CC contractions. 3-iodo-L-Tyrosine and salsolinol had no effect on EFS-induced contractions of Crotalus CC and Rabbit CC. Replacement of NaCl by N- Methyl-D-glucamine (NMDG) abolished EFS-induced contractions of rabbit CC, but did not affect Crotalus CC. The presence of tyrosine hydroxylase was identified in endothelial cells only of Crotalus CC. Since the EFS-induced contractions of Crotalus CC is dependent on catecholamine release, insensitive to TTX, insensitive to A803467 and to NaCl replacement, it indicates that the source of cathecolamine is unlikely to be from adrenergic terminals. The finding that tyrosine hydroxylase is present in endothelial cells suggests that these cells can modulate Crotalus CC tone.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Science.gov (United States)

    Porto, Marcovan; de Oliveira, Marco Antonio; Pissinatti, Lorenzo; Rodrigues, Renata Lopes; Rojas-Moscoso, Julio Alejandro; Cogo, José Carlos; Metze, Konradin; Antunes, Edson; Nahoum, César; Mónica, Fabíola Z; De Nucci, Gilberto

    2013-01-01

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    National Research Council Canada - National Science Library

    Adriano B. Carregaro; Mariângela L. Cruz; André L. Cherubini; Stélio P.L. Luna

    2009-01-01

    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...

  10. The intrahippocampal infusion of crotamine from Crotalus durissus terrificus venom enhances memory persistence in rats.

    Science.gov (United States)

    Vargas, Liane S; Lara, Marcus V S; Gonçalves, Rithiele; Mandredini, Vanusa; Ponce-Soto, Luis Alberto; Marangoni, Sergio; Dal Belo, Cháriston A; Mello-Carpes, Pâmela B

    2014-07-01

    Previous research has shown that crotamine, a toxin isolated from the venom of Crotalus durissus terrificus, induces the release of acetylcholine and dopamine in the central nervous system of rats. Particularly, these neurotransmitters are important modulators of memory processes. Therefore, in this study we investigated the effects of crotamine infusion on persistence of memory in rats. We verified that the intrahippocampal infusion of crotamine (1 μg/μl; 1 μl/side) improved the persistence of object recognition and aversive memory. By other side, the intrahippocampal infusion of the toxin did not alter locomotor and exploratory activities, anxiety or pain threshold. These results demonstrate a future prospect of using crotamine as potential pharmacological tool to treat diseases involving memory impairment, although it is still necessary more researches to better elucidate the crotamine effects on hippocampus and memory. Copyright © 2014 Elsevier Ltd. All rights reserved.

  11. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  13. The inhibitory effect of Camellia sinensis extracts against the neuromuscular blockade of Crotalus durissus terrificus venom

    Science.gov (United States)

    de Jesus Reis Rosa, Luana; Silva, Gleidy Ana Araujo; Filho, Jorge Amaral; Silva, Magali Glauzer; Groppo, Francisco Carlos; Oshima-Franco, Yoko

    2010-01-01

    In geographically isolated populations where intensive medical care or serum therapy is not easily accessible snake envenomation is a major cause for concern. The aim of the present study was to test Camellia sinensis extracts, theaflavin and epigallocatechin (two of the main C. sinensis components) against the irreversible neuromuscular blockade induced by Crotalus durissus terrificus venom in mouse phrenic-nerve diaphragm preparations. A quantitative histological study was also performed. The venom (20µg/ml) completely decreased twitch tension after 70min and 5µg/ml venom abolished 50% of twitch amplitude after 60min. C. sinensis extract induced intense facilitatory effect in the preparation activity at 0.2mg/ml and slightly facilitatory effect at 0.05mg/ml. Both 0.05mg/ml C. sinensis extract and 0.05mg/ml commercial theaflavin maintained partial muscular activity in presence of 5µg/ml venom. The histological data confirms that Cs is able to protect the muscle from the myotoxic activity of the venom. Commercial epigallocatechin gallate did not show pre-synaptic nor post-synaptic activities. C. sinensis extract was able to protect the mouse phrenic-nerve diaphragm against the irreversible neuromuscular blockade induced by C. durissus terrificus venom. PMID:21544176

  14. Epidemiologia e quadro clínico do acidente por cascavel sul-americana (Crotalus durissus Epidemiology and clinical features of South American rattlesnakes (Crotalus durissus envenomation

    Directory of Open Access Journals (Sweden)

    Miguel Tanús Jorge

    1992-08-01

    Full Text Available Foram estudados 249 casos de acidentes por cascavel sul-americana (Crotalus durissus atendidos no HVB-IB, São Paulo, Brasil, de 1974 a 1990. Os acidentes foram mais comuns no período da tarde, nos meses de janeiro a abril e de outubro a dezembro. Dentre cem serpentes classificadas quanto à subespécie 99 eram C. d. terrificus. Pertenciam ao sexo masculino 80,7% dos pacientes. Os membros inferiores e superiores foram picados em, respectivamente, 66,4% e 29,2% dos casos. As manifestações clínicas mais freqüentes foram dor (61,0% e edema (55,0% no local da picada, ptose palpebral (75,9%, escurecimento da urina atribuível à mioglobinúria (38,6% e mialgia (36,1%. Nove pacientes foram submetidos a diálise devido a insuficiência renal aguda (3,6%, três apresentavam insuficiência respiratória que motivou intubação e/ou traqueostomia e um apresentou acidente vascular cerebral isquêmico. A alteração da coagulação sanguínea ocorreu em 48,1% dos pacientes. Oito amostras de sangue colhidas horas após a picada mostraram leucocitose sendo 6 com desvio a esquerda e, nos dias subseqüentes, tendência à normalização do número de leucócitos e aparecimento de eosinofilia. Atividade sérica da creatinoquinase apresentou-se aumentada em 20 dentre 21 pacientes, sendo maior no final das primeiras 24 horas após a picada, chegando a 2.377 vezes o valor de referência. A letalidade foi de 0,8%.Under study were two hundred and forty nine cases of accidents caused by South American rattlesnakes (Crotalus durissus admitted to the Vital Brasil Hospital, São Paulo, Brasil, from 1974 to 1990. The accidents were more common in the afternoon, from January to April and from October to December. Ninety nine out of 100 snakes classified as subspecies were C. d. terrificus. Most of the patients were males (80.7%. The inferior and superior limbs were bitten in 66.4% and 29.2% of the cases, respectively. The more frequent clinical manifestations were

  15. 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

  16. Intrahippocampal Infusion of Crotamine Isolated from Crotalus durissus terrificus Alters Plasma and Brain Biochemical Parameters †

    Science.gov (United States)

    Gonçalves, Rithiele; Vargas, Liane S.; Lara, Marcus V. S.; Güllich, Angélica; Mandredini, Vanusa; Ponce-Soto, Luis; Marangoni, Sergio; Dal Belo, Cháriston A.; Mello-Carpes, Pâmela B.

    2014-01-01

    Crotamine is one of the main constituents of the venom of the South American rattlesnake Crotalus durissus terrificus. Here we sought to investigate the inflammatory and toxicological effects induced by the intrahippocampal administration of crotamine isolated from Crotalus whole venom. Adult rats received an intrahippocampal infusion of crotamine or vehicle and were euthanized 24 h or 21 days after infusion. Plasma and brain tissue were collected for biochemical analysis. Complete blood count, creatinine, urea, glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), creatine-kinase (CK), creatine kinase-muscle B (CK-MB) and oxidative parameters (assessed by DNA damage and micronucleus frequency in leukocytes, lipid peroxidation and protein carbonyls in plasma and brain) were quantified. Unpaired and paired t-tests were used for comparisons between saline and crotamine groups, and within groups (24 h vs. 21 days), respectively. After 24 h crotamine infusion promoted an increase of urea, GOT, GPT, CK, and platelets values (p ≤ 0.01), while red blood cells, hematocrit and leukocytes values decreased (p ≤ 0.01). Additionally, 21 days after infusion crotamine group showed increased creatinine, leukocytes, TBARS (plasma and brain), carbonyl (plasma and brain) and micronucleus compared to the saline-group (p ≤ 0.01). Our findings show that crotamine infusion alter hematological parameters and cardiac markers, as well as oxidative parameters, not only in the brain, but also in the blood, indicating a systemic pro-inflammatory and toxicological activity. A further scientific attempt in terms of preserving the beneficial activity over toxicity is required. PMID:25380458

  17. Intrahippocampal Infusion of Crotamine Isolated from Crotalus durissus terrificus Alters Plasma and Brain Biochemical Parameters

    Directory of Open Access Journals (Sweden)

    Rithiele Gonçalves

    2014-11-01

    Full Text Available Crotamine is one of the main constituents of the venom of the South American rattlesnake Crotalus durissus terrificus. Here we sought to investigate the inflammatory and toxicological effects induced by the intrahippocampal administration of crotamine isolated from Crotalus whole venom. Adult rats received an intrahippocampal infusion of crotamine or vehicle and were euthanized 24 h or 21 days after infusion. Plasma and brain tissue were collected for biochemical analysis. Complete blood count, creatinine, urea, glutamic oxaloacetic transaminase (GOT, glutamic pyruvic transaminase (GPT, creatine-kinase (CK, creatine kinase-muscle B (CK-MB and oxidative parameters (assessed by DNA damage and micronucleus frequency in leukocytes, lipid peroxidation and protein carbonyls in plasma and brain were quantified. Unpaired and paired t-tests were used for comparisons between saline and crotamine groups, and within groups (24 h vs. 21 days, respectively. After 24 h crotamine infusion promoted an increase of urea, GOT, GPT, CK, and platelets values (p ≤ 0.01, while red blood cells, hematocrit and leukocytes values decreased (p ≤ 0.01. Additionally, 21 days after infusion crotamine group showed increased creatinine, leukocytes, TBARS (plasma and brain, carbonyl (plasma and brain and micronucleus compared to the saline-group (p ≤ 0.01. Our findings show that crotamine infusion alter hematological parameters and cardiac markers, as well as oxidative parameters, not only in the brain, but also in the blood, indicating a systemic pro-inflammatory and toxicological activity. A further scientific attempt in terms of preserving the beneficial activity over toxicity is required.

  18. Intrahippocampal infusion of crotamine isolated from Crotalus durissus terrificus alters plasma and brain biochemical parameters.

    Science.gov (United States)

    Gonçalves, Rithiele; Vargas, Liane S; Lara, Marcus V S; Güllich, Angélica; Mandredini, Vanusa; Ponce-Soto, Luis; Marangoni, Sergio; Dal Belo, Cháriston A; Mello-Carpes, Pâmela B

    2014-11-05

    Crotamine is one of the main constituents of the venom of the South American rattlesnake Crotalus durissus terrificus. Here we sought to investigate the inflammatory and toxicological effects induced by the intrahippocampal administration of crotamine isolated from Crotalus whole venom. Adult rats received an intrahippocampal infusion of crotamine or vehicle and were euthanized 24 h or 21 days after infusion. Plasma and brain tissue were collected for biochemical analysis. Complete blood count, creatinine, urea, glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), creatine-kinase (CK), creatine kinase-muscle B (CK-MB) and oxidative parameters (assessed by DNA damage and micronucleus frequency in leukocytes, lipid peroxidation and protein carbonyls in plasma and brain) were quantified. Unpaired and paired t-tests were used for comparisons between saline and crotamine groups, and within groups (24 h vs. 21 days), respectively. After 24 h crotamine infusion promoted an increase of urea, GOT, GPT, CK, and platelets values (p ≤ 0.01), while red blood cells, hematocrit and leukocytes values decreased (p ≤ 0.01). Additionally, 21 days after infusion crotamine group showed increased creatinine, leukocytes, TBARS (plasma and brain), carbonyl (plasma and brain) and micronucleus compared to the saline-group (p ≤ 0.01). Our findings show that crotamine infusion alter hematological parameters and cardiac markers, as well as oxidative parameters, not only in the brain, but also in the blood, indicating a systemic pro-inflammatory and toxicological activity. A further scientific attempt in terms of preserving the beneficial activity over toxicity is required.

  19. Snake venomics of the Central American rattlesnake Crotalus simus and the South American Crotalus durissus complex points to neurotoxicity as an adaptive paedomorphic trend along Crotalus dispersal in South America.

    Science.gov (United States)

    Calvete, Juan J; Sanz, Libia; Cid, Pedro; de la Torre, Pilar; Flores-Díaz, Marietta; Dos Santos, M Cristina; Borges, Adolfo; Bremo, Adolfo; Angulo, Yamileth; Lomonte, Bruno; Alape-Girón, Alberto; Gutiérrez, José María

    2010-01-01

    We report a comparative venomic and antivenomic characterization of the venoms of newborn and adult specimens of the Central American rattlesnake, Crotalus simus, and of the subspecies cumanensis, durissus, ruruima, and terrificus of South American Crotalus durissus. Neonate and adult C. simus share about 50% of their venom proteome. The venom proteome of 6-week-old C. simus is predominantly made of the neurotoxic heterodimeric phospholipase A(2) (PLA(2) crotoxin) (55.9%) and serine proteinases (36%), whereas snake venom Zn(2+)-metalloproteinases (SVMPs), exclusively of class PIII, represent only 2% of the total venom proteins. In marked contrast, venom from adult C. simus comprises toxins from 7 protein families. A large proportion (71.7%) of these toxins are SVMPs, two-thirds of which belong to the PIII class. These toxin profiles correlate well with the overall biochemical and pharmacological features of venoms from adult (hemorrhagic) and newborn (neurotoxic) C. simus specimens. The venoms of the South American Crotalus subspecies belong to one of two distinct phenotypes. C. d. cumanensis exhibits high levels of SVMPs and low lethal potency (LD(50)), whereas C. d. subspecies terrificus, ruruima, and durissus have low SVMP activity and high neurotoxicity to mice. Their overall toxin compositions explain the outcome of envenomation by these species. Further, in all C. simus and C. durissus venoms, the concentration of neurotoxins (crotoxin and crotamine) is directly related with lethal activity, whereas lethality and metalloproteinase activity show an inverse relationship. The similar venom toxin profiles of newborn C. simus and adult C. durissus terrificus, ruruima, and durissus subspecies strongly suggests that the South American taxa have retained juvenile venom characteristics in the adult form (paedomorphism) along their North-South stepping-stone dispersal. The driving force behind paedomorphism is often competition or predation pressure. The increased

  20. Biodegradable microparticles containing crotamine isolated from Crotalus durissus terrificus display antileishmanial activity in vitro.

    Science.gov (United States)

    Macedo, Sharon Rose A; de Barros, Neuza B; Ferreira, Amália S; Moreira-Dill, Leandro S; Calderon, Leonardo A; Soares, Andreimar M; Nicolete, Roberto

    2015-01-01

    To evaluate antileishmanial activity of crotamine, a toxin isolated from Crotalus durissus terrificus, in solution form and encapsulated in biodegradable microparticles in vitro. Particles were analyzed on-chip by surface plasmon resonance and characterized by testing their diameters, zeta potential and encapsulation rate. The viability of promastigotes as well as murine macrophages was assessed. Furthermore, the phagocytic index was determined for macrophages, and cell supernatants were collected for the determination of TNF-α levels. An infection assay using Leishmania amazonensis-infected macrophages was also conducted. The diameters and zeta potential of control particles (1.35 μm; -12.3 mV) and of those containing crotamine (3.09 μm; -20.9 mV) were adequate for the assays conducted. Crotamine-loaded particles were better captured by macrophages than control particles (increase of 12% in the phagocytic index), leading to increased TNF-α levels (196 pg/ml), and they also induced a significant decrease in the numbers of amastigotes compared to infected macrophages only. The approach presented here opens the possibility of working with safe concentrations of encapsulated toxins to reach antileishmanial effects. © 2015 S. Karger AG, Basel.

  1. Immunohistochemical and functional characterization of nitric oxide signaling pathway in isolated aorta from Crotalus durissus terrificus.

    Science.gov (United States)

    Mónica, Fabíola Z; Rojas-Moscoso, Julio; Porto, Marcovan; Schenka, André A; Antunes, Edson; Cogo, José Carlos; De Nucci, Gilberto

    2012-04-01

    We characterized the nitric oxide (NO)-cyclic GMP-phosphodiesterase-5 (PDE5) pathway in Crotalus durissus terrificus aorta. Concentration responses curves to acetylcholine (ACh), sodium nitroprusside (SNP), BAY41-2272 (soluble guanylyl cyclase [sGC] stimulator), BAY60-2770 (sGC activator) and tadalafil (PDE5 inhibitor) were constructed in phenylephrine (10 μM)-precontracted tissues with intact (E(+)) or denuded (E(-)) endothelium. ACh (0.0001-10 μM) and SNP (0.0001-10 μM) relaxed aorta, which were reduced by the NO synthase (L-NAME,100 μM) or the sGC inhibitors (ODQ, 10 μM). Tadalafil (0.0001-10 μM) relaxed E(+) rings with potency (pEC(50)) and maximal response (E(max)) values of 7.34±0.02 and 105±8%, respectively. E(-) or ODQ treatment significantly (PCrotalus sp. vessels, and has a remarkable degree of functional similarity to mammalian vessels. Copyright © 2011 Elsevier Inc. All rights reserved.

  2. Massive Muscular Infection by a Sarcocystis Species in a South American Rattlesnake (Crotalus durissus terrificus).

    Science.gov (United States)

    Roberts, J F; Wellehan, J F X; Weisman, J L; Rush, M; Childress, A L; Lindsay, David S

    2015-06-01

    Massive numbers of sarcocysts of a previously undescribed species of Sarcocystis were observed in the skeletal muscles throughout the body of an adult, female South American rattlesnake (Crotalus durissus terrificus). Examination of tissue sections by light microscopy demonstrated that sarcocysts were present in 20 to 40% of muscle fibers from 5 sampled locations. Sarcocysts were not present in cardiac muscle, smooth muscle, or other organs. Sarcocysts were 0.05-0.15 mm wide, had variable length depending on the viewed orientation and size of the muscle fiber, and had a sarcocyst wall less than 1-μm thick. Sarcocysts were subdivided by septa and had central degeneration in older sarcocysts. Host induced secondary encapsulation or an inflammatory response was not present. By transmission electron microscopy (TEM), the sarcocyst wall was Type I, with a parasitophorous membrane of approximately 100 nanometers in width arranged in an undulating pattern and intermittently folded inward in a branching pattern. The sarcocysts contained metrocytes in different stages of development and mature bradyzoites. The nucleic acid sequence from a section of the 18S small subunit rRNA gene was most closely related to S. mucosa that uses marsupials as intermediate hosts and has an unknown definitive host. This is apparently the third report of muscular Sarcocystis infection in snakes and is the first to describe the ultrastructure of the sarcocysts and use sequencing methods to aid in identification.

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

    Directory of Open Access Journals (Sweden)

    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.

  4. Inflammatory Mediators Release in Urine from Mice Injected with Crotalus durissus terrificus Venom

    Science.gov (United States)

    Hernández Cruz, A.; Barbosa Navarro, L.; Mendonça, R. Z.; Petricevich, V. L.

    2011-01-01

    In this study, we investigated in groups of female BALB/c mice injected with Crotalus durissus terrificus venom (Cdt) the renal function based on creatinine clearance, percentage of fractional excretion cytokines and histological examination of renal tissue. Cdt caused renal alterations that induced proteinuria during the initial hours post-venom and reduced creatinine clearance 15 min. up to 2 hours post-venom administration. In urine from mice injected with Cdt induced a decrease in IL-4 levels. More pronounced increments of IL-5, IL-6 and IFN-γ were observed after 15 and 30 min, respectively. The highest levels of TNF and IL-10 were observed at 1 and 4 hs, respectively. The ratios of pro- and anti-inflammatory cytokines in animals injected with Cdt, which may be manifested in the inflammatory status during the envenoming. In groups of animals treated with Cdt were observed a decreasing in creatinine clearance and its effect on glomerular filtration rate was accompanied by decreased fractional excretion of cytokines and morphologic disturbances. This loss of change selectively in envenomation could thus explain why the relatively excretion of cytokines is reduced while of total proteins increases. In conclusion the fractional excretion of cytokines is significantly reduced in mice injected with Cdt, despite proteinuria. PMID:22174490

  5. Multiple effects of toxins isolated from Crotalus durissus terrificus on the hepatitis C virus life cycle.

    Science.gov (United States)

    Shimizu, Jacqueline Farinha; Pereira, Carina Machado; Bittar, Cintia; Batista, Mariana Nogueira; Campos, Guilherme Rodrigues Fernandes; da Silva, Suely; Cintra, Adélia Cristina Oliveira; Zothner, Carsten; Harris, Mark; Sampaio, Suely Vilela; Aquino, Victor Hugo; Rahal, Paula; Jardim, Ana Carolina Gomes

    2017-01-01

    Hepatitis C virus (HCV) is one of the main causes of liver disease and transplantation worldwide. Current therapy is expensive, presents additional side effects and viral resistance has been described. Therefore, studies for developing more efficient antivirals against HCV are needed. Compounds isolated from animal venoms have shown antiviral activity against some viruses such as Dengue virus, Yellow fever virus and Measles virus. In this study, we evaluated the effect of the complex crotoxin (CX) and its subunits crotapotin (CP) and phospholipase A2 (PLA2-CB) isolated from the venom of Crotalus durissus terrificus on HCV life cycle. Huh 7.5 cells were infected with HCVcc JFH-1 strain in the presence or absence of these toxins and virus was titrated by focus formation units assay or by qPCR. Toxins were added to the cells at different time points depending on the stage of virus life cycle to be evaluated. The results showed that treatment with PLA2-CB inhibited HCV entry and replication but no effect on HCV release was observed. CX reduced virus entry and release but not replication. By treating cells with CP, an antiviral effect was observed on HCV release, the only stage inhibited by this compound. Our data demonstrated the multiple antiviral effects of toxins from animal venoms on HCV life cycle.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Science.gov (United States)

    Hernández Cruz, A.; Garcia-Jimenez, S.; Zucatelli Mendonça, R.; Petricevich, V. L.

    2008-01-01

    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. PMID:18604304

  8. American rattlesnake (Crotalus durissus terrificus bite accidents in dogs in Argentina

    Directory of Open Access Journals (Sweden)

    Koscinczuk P.

    2000-01-01

    Full Text Available The symptomatology and treatment of two dogs bitten by Crotalus durissus terrificus are described. Neurological signs were present few minutes after the accident with local anesthesia and ataxia of the affected limb and neurotoxic fascia. Alanine aminotransferase (ALT, aspartate aminotransferase (AST, creatinine kinase (CK, lactate dehydrogenase (LDH and calcium were evaluated in an attempt to investigate muscle damage. Renal failure was not observed but some alterations were detected in urine. Urine density was low and the urinary sediment contained granular clumps and small round cells. Muscle samples were obtained from both legs for histopathological study, showing edema and isolated necrotic fibers. Both dogs received treatment within four hours after the accident by intravenous route. The antivenom was administered diluted in 250ml of Ringer solution in a dose enough to inactivate more than 8mg of venom. Dexamethasone was applied previously to the antivenom. Clinical evolution was good and both patients were in good health condition on the second day after the accident.

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

    Science.gov (United States)

    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 (RN), 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 (CST) 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), RN 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.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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

  13. Cloning and characterization of an antibacterial L-amino acid oxidase from Crotalus durissus cumanensis venom.

    Science.gov (United States)

    Vargas, Leidy Johana; Quintana, Juan Carlos; Pereañez, Jaime Andrés; Núñez, Vitelbina; Sanz, Libia; Calvete, Juan

    2013-03-15

    An L-amino acid oxidase (LAAO) from Crotalus durissus cumanensis venom (CdcLAAO) was purified to homogeneity using a combination of size-exclusion and ion exchange chromatographies. CdcLAAO is a monomeric protein exhibiting an apparent molecular mass of 55 kDa and a calculated pI of 8. Its complete 498-amino-acid sequence was deduced through cDNA and protein sequencing. The enzyme oxidized L-Leu with K(m) and a V(Max) of 9.23 μM and 0.46 μM/min respectively, and exhibited Kcat and a Kcat/K(m) of 1.8 s(-1) and 195 mM(-1)s(-1). CdcLAAO inhibited in a dose-dependent manner the growth of Staphylococcus aureus and Acinetobacter baumannii. The inhibitory effect was more significant on S. aureus, with a Minimal Inhibitory Concentration (MIC) of 8 μg/mL and Minimal Bactericidal Concentration (MBC) of 16 μg/mL, than against A. baumannii, with a MIC of 16 μg/mL and MBC of 32 μg/mL. However, against Escherichia coli CdcLAAO did not show inhibitory capacity at the concentrations tested (2-128 μg/mL). CdcLAAO did not exhibit cytotoxic activity on the mouse myoblast cell line C(2)C(12) and on peripheral blood mononuclear cell (PBMC). Copyright © 2012 Elsevier Ltd. All rights reserved.

  14. Crotalus durissus terrificus Venom Interferes With Morphological, Functional, and Biochemical Changes in Murine Macrophage

    Science.gov (United States)

    Hernández Cruz, Anselmo; Z. Mendonça, Ronaldo; L. Petricevich, Vera

    2005-01-01

    Crotalus durissus terrificus venom (Cdt) is toxic for a variety of eukaryotic cells, especially at high concentrations. However its effects on host immune cells are not well known. The purpose of this study was to determine the effect of Cdt on functional status and the mediators production in peritoneal macrophages. The effects of Cdt were analyzed in vitro and were detected using functional status of macrophages as determined by the H2O2 release, spreading percentage, phagocytic index, vacuole formation, and mediators production. Several functional bioassays were employed: cytotoxicity was determined by taking the lyses percentage and the presence of hydrogen peroxide (H2O2) in macrophages, using the horseradish peroxidase-dependent oxidation of phenol red and nitric oxide (NO) in the supernatants of macrophages by the Griess reaction. The tumor necrosis factor (TNF) activity was detected by measuring its cytotoxic activity on L929 cells, and the production the level of other cytokines was assayed using enzyme-linked immunosorbent assay. In vitro studies revealed that Cdt produced (a) a discrete increase in the release of H2O2 and vacuole formation; (b) a decrease in spreading percentage and in the phagocytic index; and (c) an increment in the mediators production. More pronounced increments of IL-6 and TNF were observed after 24 and 48 hours, respectively. Maximum levels of IFN-γ and NO were observed after 96 hours. Interestingly, levels of all mediators presented a discreet decrease, as the amount of Cdt was increased. In contrast, the IL-10 levels observed for all doses studied here did not alter. The IL-6/IL-10 ratio may possibly reflect the balance of pro- and anti-inflammatory cytokines in macrophages, which may be manifested in the inflammatory status during the envenoming processes. Taken together, these data indicate that Cdt have a differential effect on macrophage activation and that this venom is a potent inhibitor of anti-inflammatory response. PMID

  15. Blood oxygen affinity increases during digestion in the South American rattlesnake, Crotalus durissus terrificus.

    Science.gov (United States)

    Bovo, Rafael P; Fuga, Adriana; Micheli-Campbell, Mariana A; Carvalho, José E; Andrade, Denis V

    2015-08-01

    Digesting snakes experience massive increases in metabolism that can last for many days and are accompanied by adjustments in the oxygen transport cascade. Accordingly, we examined the oxygen-binding properties of the blood in the South American rattlesnake (Crotalus durissus terrificus) during fasting and 24 and 48h after the snakes have ingested a rodent meal corresponding to 15% (±2%) of its own body mass. In general, oxygen-hemoglobin (Hb-O2) affinity was significantly increased 24h post-feeding, and then returned toward fasting values within 48h post-feeding. Content of organic phosphates ([NTP] and [NTP]/[Hb]), hemoglobin cooperativity (Hill's n), and Bohr Effect (ΔlogP50/ΔpH) were not affected by feeding. The postprandial increase in Hb-O2 affinity in the South American rattlesnake can be almost entirely ascribed by the moderate alkaline tide that follows meal ingestion. In general, digesting snakes were able to regulate blood metabolites at quite constant levels (e.g., plasma osmolality, lactate, glucose, and total protein levels). The level of circulating lipids, however, was considerably increased, which may be related to their mobilization, since lipids are known to be incorporated by the enterocytes after snakes have fed. In conclusion, our results indicate that the exceptional metabolic increment exhibited by C. d. terrificus during meal digestion is entirely supported by the aerobic pathways and that among the attending cardiorespiratory adjustments, pulmonary Hb-O2 loading is likely improved due to the increment in blood O2 affinity. Copyright © 2014 Elsevier Inc. All rights reserved.

  16. Effect of a biostimulatory homeopathic complex on venom production in captive rattlesnakes (Crotalus durissus).

    Science.gov (United States)

    Santa Rita, Paula Helena; Cleveland, Herbert Patric Kellermann; Pereira, Paula Laryssa Souza; Corrêa, William; Van Onselen, Valter Joost; Corrêa Filho, Ruy Alberto Caetano; Teixeira, Maria Araújo

    2016-11-01

    The purpose of this study was to evaluate the effect of two administration methods of a biostimulatory homeopathic complex (Convert H ® ) on the production of fresh and lyophilized venom of rattlesnakes (Crotalus durissus) under intensive captivity conditions. Sixty snakes were subjected to treatment following a randomized block design. The effects of sex and size were controlled for. Thirteen consecutive extractions were performed over 21 months. The first factor considered in the experiment was the origin of mice used as prey: a conventional colony (A1) or the Convert H colony (A2; mice receiving the homeopathic complex in water at 1%). The type of water given to snakes was the second factor: pure (B1) or amended with 5% of Convert H ® (B2). The experiment was structured in a factorial 2 × 2 design combining mouse and water types (A1B1, A1B2, A2B1, and A2B2). No consistent treatment effects on fresh venom production (mL) were observed when the experimental groups were compared with controls (A1B1). However, production of lyophilized venom (mg) was significantly higher (p < 0.05) in A2B2 animals than in controls in eight of 13 extractions performed, and also in aggregate. The results revealed that production of lyophilized venom, measured over multiple extractions, can be increased by administering the homeopathic complex simultaneously to rattlesnakes and prey. Copyright © 2016 The Faculty of Homeopathy. Published by Elsevier Ltd. All rights reserved.

  17. Kinetics of venom and antivenom serum levels, clinical evaluation and therapeutic effectiveness in dogs inoculated with Crotalus durissus terrificus venom

    Directory of Open Access Journals (Sweden)

    F. Sangiorgio

    2008-01-01

    Full Text Available This work evaluated the clinical and therapeutic aspects as well as serum levels of venom and antivenom IgG by enzyme-linked immunosorbent assay (ELISA in experimental envenomation of dogs with Crotalus durissus terrificus venom. Twenty-eight mixed breed adult dogs were divided into four groups of seven animals each, Group I: only venom; Group II, venom + 50 ml of anti-bothropic-crotalic serum (50mg + fluid therapy; Group III, venom + 50 ml of anti-bothropic-crotalic serum + fluid therapy + urine alkalination; Group IV, 50 ml of anti-bothropic-crotalic serum. The lyophilized venom of Crotalus durissus terrificus was reconstituted in saline solution and subcutaneously inoculated at the dose of 1mg/kg body weight. The dogs presented clinical signs of local pain, weakness, mandibular ptosis, mydriasis, emesis and salivation. The venom levels detected by ELISA ranged from 0 to 90ng/ml, according to the severity of the clinical signs. Serum antivenom ranged from 0 to 3ug/ml and was detected for up to 138h after treatment. ELISA results showed the effectiveness of the serum therapy for the venom neutralization.

  18. Liver dysfunction in patients bitten by Crotalus durissus terrificus (Laurenti, 1768 snakes in Botucatu (State of São Paulo, Brazil Alterações hepáticas em doentes picados por serpentes Crotalus durissus terrificus (Laurenti, 1768 na região de Botucatu, Estado de São Paulo, Brasil

    Directory of Open Access Journals (Sweden)

    Benedito Barra Viera

    1995-02-01

    Full Text Available Thirty-two patients bitten by venomous snakes sixteen by Bothrops spp. and sixteen by Crotalus durissus terrificus were studied. The group comprised thirty males and two females, aged eight to sixty-three years (mean 33±15. Bromsulphalein tests were increased in the majority of patients bitten by Crotalus durissus terrificus. The correlation coefficient of Spearman was positive between bromsulphalein tests and alanine aminotransferase levels, and between alanine aminotransferase and aspartate aminotransferase levels only in the Crotalus group. The only patient who died was bitten by Crotalus durissus terrificus and showed hydropic degeneration and mitochondrial injury in the liver. It was concluded that the hepatic damage might have been caused by at least two possible mechanisms: venom effect on liver mitochondria and cytokine effects on hepatocyte, specially interleukin-6.Os autores estudaram 32 doentes picados por serpentes venenosas, sendo 16 picados por Bothrops spp. e 16 por Crotalus durissus terrificus. Trinta doentes eram do sexo masculino e dois do feminino com idades variando entre 8 e 63 anos (méda 33±15. A prova da retenção da bromosulfaleína apresentou-se aumentada na maioria dos doentes picados por serpentes Crotalus durissus terrificus. Houve correlação positiva entre a retenção da bromosulfaleína e os níveis séricos de alanina aminotransferase e entre alanina e aspartato aminotransferase apenas nos doentes do grupo Crotalus. Um dos doentes evoluiu para o óbito e apresentou no exame anatomopatológico do fígado degeneração hidrópica e lesões mitocondriais. Os autores concluem que as alterações hepáticas são causadas por pelo menos dois mecanismos a saber: lesão mitocondrial por efeito do veneno crotálico; efeito das citoquinas, especialmente a interleucina-6.

  19. Biochemical and biological characterization of two serine proteinases from Colombian Crotalus durissus cumanensis snake venom.

    Science.gov (United States)

    Patiño, Arley Camilo; Pereañez, Jaime Andrés; Gutiérrez, José María; Rucavado, Alexandra

    2013-03-01

    Two clotting serine proteinases, named Cdc SI and Cdc SII, were isolated and characterized for the first time from Colombian Crotalus durissus cumanensis snake venom. The enzymes were purified using two chromatographic steps: molecular exclusion on Sephacryl S-200 and RP-HPLC on C8 Column. The molecular masses of the proteins, determined by MALDI-TOF mass spectrometry, were 28,561.4 and 28,799.2 Da for Cdc SI and Cdc SII, respectively. The aim of the present study was to evaluate enzymatic, coagulant and toxic properties of the two enzymes. The serine proteinases hydrolyzed specific chromogenic substrate (BaPNA) and exhibited a Michaelis-Menten behavior. Cdc SI had V(max) of 0.038 ± 0.003 nmol/min and K(M) of 0.034 ± 0.017 mM, while Cdc SII displayed values of V(max) of 0.267 ± 0.011 nmol/min and K(M) of 0.145 ± 0.023 mM. N-terminal sequences were VIGGDEXNIN and VIGGDICNINEHNFLVALYE for Cdc SI and Cdc SII, respectively. Molecular masses, N-terminal sequences, inhibition assays, and enzymatic profile suggest that Cdc SI and Cdc SII belong to the family of snake venom thrombin-like enzymes. These serine proteinases differed in their clotting activity on human plasma, showing a minimum coagulant dose of 25 μg and 0.571 μg for Cdc SI and Cdc SII, respectively. Enzymes also showed coagulant activity on bovine fibrinogen and degraded chain α of this protein. Toxins lack hemorrhagic and myotoxic activities, but are capable to induce defibrin(ogen)ation, moderate edema, and an increase in vascular permeability. These serine proteinases may contribute indirectly to the local hemorrhage induced by metalloproteinases, by causing blood clotting disturbances, and might also contribute to cardiovascular alterations characteristic of patients envenomed by C. d. cumanensis in Colombia. Copyright © 2012 Elsevier Ltd. All rights reserved.

  20. 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)

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

    Directory of Open Access Journals (Sweden)

    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.

  2. Vagal tone regulates cardiac shunts during activity and at low temperatures in the South American rattlesnake, Crotalus durissus.

    Science.gov (United States)

    Filogonio, Renato; Wang, Tobias; Taylor, Edwin W; Abe, Augusto S; Leite, Cléo A C

    2016-12-01

    The undivided ventricle of non-crocodilian reptiles allows for intracardiac admixture of oxygen-poor and oxygen-rich blood returning via the atria from the systemic circuit and the lungs. The distribution of blood flow between the systemic and pulmonary circuits may vary, based on differences between systemic and pulmonary vascular conductances. The South American rattlesnake, Crotalus durissus, has a single pulmonary artery, innervated by the left vagus. Activity in this nerve controls pulmonary conductance so that left vagotomy abolishes this control. Experimental left vagotomy to abolish cardiac shunting had no effect on long-term survival and failed to identify a functional role in determining metabolic rate, growth or resistance to food deprivation. Accordingly, the present investigation sought to evaluate the extent to which cardiac shunt patterns are actively controlled during changes in body temperature and activity levels. We compared hemodynamic parameters between intact and left-vagotomized rattlesnakes held at different temperatures and subjected to enforced physical activity. Increased temperature and enforced activity raised heart rate, cardiac output, pulmonary and systemic blood flow in both groups, but net cardiac shunt was reversed in the vagotomized group at lower temperatures. We conclude that vagal control of pulmonary conductance is an active mechanism regulating cardiac shunts in C. durissus.

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

    Directory of Open Access Journals (Sweden)

    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.

  4. Phospholipase A2 Inhibitor from Crotalus durissus terrificus rattlesnake: Effects on human peripheral blood mononuclear cells and human neutrophils cells.

    Science.gov (United States)

    Xavier, Caroline V; da S Setúbal, Sulamita; Lacouth-Silva, Fabianne; Pontes, Adriana S; Nery, Neriane M; de Castro, Onassis Boeri; Fernandes, Carla F C; Soares, Andreimar M; Fortes-Dias, Consuelo L; Zuliani, Juliana P

    2017-12-01

    Crotalus Neutralizing Factor (CNF) is an inhibitor of phospholipase A2 (PLA2), present in the blood plasma of Crotalus durissus terrificus snake. This inhibitor neutralizes the lethal and enzymatic activity of crotoxin, the main neurotoxin from this venom. In this study, we investigated the effects of CNF on the functionality of human peripheral blood mononuclear cells (PBMCs) and human neutrophils. The following parameters were evaluated: viability and proliferation, chemotaxis, cytokines and LTB4 production, cytosolic PLA2s activity, myeloperoxidase (MPO) and superoxide anion (O2-) production. CNF showed no toxicity on PBMCs or neutrophils, and acts by stimulating the release of TNF-α and LTB4, but neither stimulates IL-10 and IL-2 nor affects PBMCs proliferation and O2- release. In neutrophils, CNF induces chemotaxis but does not induce the release of both MPO and O2-. However, it induces LTB4 and IL-8 production. These data show the influence of CNF on PBMCs' function by inducing TNF-α and LTB4 production, and on neutrophils, by stimulating chemotaxis and LTB4 production, via cytosolic PLA2 activity, and IL-8 release. The inflammatory profile produced by CNF is shown for the first time. Our present results suggest that CNF has a role in activation of leukocytes and exert proinflammatory effects on these cell. Copyright © 2017 Elsevier B.V. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    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

  6. 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

    Directory of Open Access Journals (Sweden)

    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

  7. Allopurinol Reduces the Lethality Associated with Acute Renal Failure Induced by Crotalus durissus terrificus Snake Venom: Comparison with Probenecid

    Science.gov (United States)

    Frezzatti, Rodrigo; Silveira, Paulo Flavio

    2011-01-01

    Background Acute renal failure is one of the most serious complications of envenoming resulting from Crotalus durissus terrificus bites. This study evaluated the relevance of hyperuricemia and oxidative stress and the effects of allopurinol and probenecid in renal dysfunction caused by direct nephrotoxicity of C. d. terrificus venom. Methodology/Principal Findings Hematocrit, protein, renal function and redox status were assessed in mice. High ratio of oxidized/reduced glutathione and hyperuricemia induced by C. d. terrificus venom were ameliorated by both, allopurinol or probenecid, but only allopurinol significantly reduced the lethality caused by C. d. terrificus venom. The effectiveness of probenecid is compromised probably because it promoted hypercreatinemia and hypocreatinuria and worsed the urinary hypo-osmolality in envenomed mice. In turn, the highest effectiveness of allopurinol might be due to its ability to diminish the intracellular formation of uric acid. Conclusions/Significance Data provide consistent evidences linking uric acid with the acute renal failure induced by C. d. terrificus venom, as well as that this envenoming in mice constitutes an attractive animal model suitable for studying the hyperuricemia and that the allopurinol deserves to be clinically evaluated as an approach complementary to anti-snake venom serotherapy. PMID:21909449

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

    Directory of Open Access Journals (Sweden)

    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.

  9. Biochemical, functional, structural and phylogenetic studies on Intercro, a new isoform phospholipase A2 from Crotalus durissus terrificus snake venom.

    Science.gov (United States)

    Vieira, Lara F; Magro, Angelo J; Fernandes, Carlos A H; de Souza, Bibiana M; Cavalcante, Walter L G; Palma, Mário S; Rosa, José C; Fuly, André L; Fontes, Marcos R M; Gallacci, Márcia; Butzke, Diana S; Calderon, Leonardo A; Stábeli, Rodrigo G; Giglio, José R; Soares, Andreimar M

    2013-12-01

    Crotoxin is a neurotoxin from Crotalus durissus terrificus venom that shows immunomodulatory, anti-inflammatory, antimicrobial, antitumor and analgesic activities. Structurally, this toxin is a heterodimeric complex composed by a toxic basic PLA2 (Crotoxin B or CB) non-covalently linked to an atoxic non-enzymatic and acidic component (Crotapotin, Crotoxin A or CA). Several CA and CB isoforms have been isolated and characterized, showing that the crotoxin venom fraction is, in fact, a mixture of different molecules derived from the combination of distinct subunit isoforms. Intercro (IC) is a protein from the same snake venom which presents high similarity in primary structure to CB, indicating that it could be an another isoform of this toxin. In this work, we compare IC to the crotoxin complex (CA/CB) and/or CB in order to understand its functional aspects. The experiments with IC revealed that it is a new toxin with different biological activities from CB, keeping its catalytic activity but presenting low myotoxicity and absence of neurotoxic activity. The results also indicated that IC is structurally similar to CB isoforms, but probably it is not able to form a neurotoxic active complex with crotoxin A as observed for CB. Moreover, structural and phylogenetic data suggest that IC is a new toxin with possible toxic effects not related to the typical CB neurotoxin. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  10. Allopurinol reduces the lethality associated with acute renal failure induced by Crotalus durissus terrificus snake venom: comparison with probenecid.

    Science.gov (United States)

    Frezzatti, Rodrigo; Silveira, Paulo Flavio

    2011-09-01

    Acute renal failure is one of the most serious complications of envenoming resulting from Crotalus durissus terrificus bites. This study evaluated the relevance of hyperuricemia and oxidative stress and the effects of allopurinol and probenecid in renal dysfunction caused by direct nephrotoxicity of C. d. terrificus venom. Hematocrit, protein, renal function and redox status were assessed in mice. High ratio of oxidized/reduced glutathione and hyperuricemia induced by C. d. terrificus venom were ameliorated by both, allopurinol or probenecid, but only allopurinol significantly reduced the lethality caused by C. d. terrificus venom. The effectiveness of probenecid is compromised probably because it promoted hypercreatinemia and hypocreatinuria and worsed the urinary hypo-osmolality in envenomed mice. In turn, the highest effectiveness of allopurinol might be due to its ability to diminish the intracellular formation of uric acid. Data provide consistent evidences linking uric acid with the acute renal failure induced by C. d. terrificus venom, as well as that this envenoming in mice constitutes an attractive animal model suitable for studying the hyperuricemia and that the allopurinol deserves to be clinically evaluated as an approach complementary to anti-snake venom serotherapy.

  11. Allopurinol reduces the lethality associated with acute renal failure induced by Crotalus durissus terrificus snake venom: comparison with probenecid.

    Directory of Open Access Journals (Sweden)

    Rodrigo Frezzatti

    2011-09-01

    Full Text Available BACKGROUND: Acute renal failure is one of the most serious complications of envenoming resulting from Crotalus durissus terrificus bites. This study evaluated the relevance of hyperuricemia and oxidative stress and the effects of allopurinol and probenecid in renal dysfunction caused by direct nephrotoxicity of C. d. terrificus venom. METHODOLOGY/PRINCIPAL FINDINGS: Hematocrit, protein, renal function and redox status were assessed in mice. High ratio of oxidized/reduced glutathione and hyperuricemia induced by C. d. terrificus venom were ameliorated by both, allopurinol or probenecid, but only allopurinol significantly reduced the lethality caused by C. d. terrificus venom. The effectiveness of probenecid is compromised probably because it promoted hypercreatinemia and hypocreatinuria and worsed the urinary hypo-osmolality in envenomed mice. In turn, the highest effectiveness of allopurinol might be due to its ability to diminish the intracellular formation of uric acid. CONCLUSIONS/SIGNIFICANCE: Data provide consistent evidences linking uric acid with the acute renal failure induced by C. d. terrificus venom, as well as that this envenoming in mice constitutes an attractive animal model suitable for studying the hyperuricemia and that the allopurinol deserves to be clinically evaluated as an approach complementary to anti-snake venom serotherapy.

  12. Modulation of the Pharmacological Activities of Secretory Phospholipase A2 from Crotalus durissus cascavella Induced by Naringin

    Directory of Open Access Journals (Sweden)

    Marcos H. Toyama

    2011-01-01

    Full Text Available In this work we have characterized the action of the naringin, a flavonoid found in grapefruit and known for its various pharmacological effects, which include antioxidant blood lipid lowering and anticancer activity, on the structure and biochemical activities of a secretory phospholipase A (sPLA2 from Crotalus durissus cascavella, an important protein involved in the releasinge of arachidonic acid in phospholipid membranes. sPLA2 was incubated with naringin (mol:mol at 37 °C and a discrete reduction in the UV scanning signal and a modification of the circular dichroism spectra were observed after treatment with naringin, suggesting modifications of the secondary structure of the protein. This flavonoid was able to decrease enzymatic activity and some pharmacological effects, such as myonecrosis, platelet aggregation, and neurotoxic activity caused by sPLA2, however, the inflammatory effect was not affected by naringin. In addition, small angle X-ray scattering (SAXS data were collected for sPLA2 and naringin-treated sPLA2 to evaluate possible modifications of the protein structure. These structural investigations have shown that sPLA2 is an elongated dimer in solution and after treatment with naringin a conformational change in the dimeric configuration was observed. Our results suggest that structural modification may be correlated with the loss of enzymatic activity and alterations in pharmacological properties.

  13. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Science.gov (United States)

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

    2016-06-01

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

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

    Directory of Open Access Journals (Sweden)

    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.

  18. A taxonomic bibliography of the South American snakes of the Crotalus durissus complex (Serpentes, Viperidae

    Directory of Open Access Journals (Sweden)

    VANZOLINI PAULO E.

    2002-01-01

    Full Text Available A survey is made of the taxonomic literature on South American rattlesnakes (genus Crotalus, family Viperidae. Two main areas are emphasized: the attribution of the names proposed in the eighteenth century by Linnaeus and Laurenti, and the current scheme of division in subspecies. The attribution of names is examined based on the original descriptions and on relevant previous and contemporary literature. The presently adopted scheme, proposed by Klauber (1941, 1972 is found not entirely satisfactory, but reasonable enough - besides being hallowed by use. The scheme of geographical differentiation, intrinsically important and with broad practical implications (differentiation of the venom is found to be the culmination of a long series of deficient analyses, and in urgent need of proper investigation.

  19. 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

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

    Science.gov (United States)

    Almeida, Caroline de Souza; Andrade-Oliveira, Vinicius; Câmara, Niels Olsen Saraiva; Jacysyn, Jacqueline F; Faquim-Mauro, Eliana L

    2015-01-01

    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 induction of a

  1. 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

    Directory of Open Access Journals (Sweden)

    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

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

    Directory of Open Access Journals (Sweden)

    CARLOS H.M. OLIVEIRA

    Full Text Available ABSTRACT 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.

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

    Science.gov (United States)

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

    2016-01-01

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

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

    Directory of Open Access Journals (Sweden)

    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.

  5. Preliminary X-ray crystallographic studies of a tetrameric phospholipase A{sub 2} formed by two isoforms of crotoxin B from Crotalus durissus terrificus venom

    Energy Technology Data Exchange (ETDEWEB)

    Marchi-Salvador, D. P.; Corrêa, L. C.; Salvador, G. H. M.; Magro, A. J. [Departamento de Física e Biofísica, Instituto de Biociências, UNESP, CP 510, 18618-000 Botucatu-SP (Brazil); Oliveira, C. Z. [Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, FCFRP, USP, Ribeirão Preto-SP (Brazil); Iulek, J. [Departamento de Química, UEPG, Ponta Grossa-PR (Brazil); Soares, A. M. [Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, FCFRP, USP, Ribeirão Preto-SP (Brazil); Fontes, M. R. M., E-mail: fontes@ibb.unesp.br [Departamento de Física e Biofísica, Instituto de Biociências, UNESP, CP 510, 18618-000 Botucatu-SP (Brazil)

    2007-12-01

    Crotoxin B is a basic phospholipase A{sub 2} found in the venom of C. durissus terrificus and is one of the subunits that constitute crotoxin. Here, the crystallization, X-ray diffraction data collection and molecular-replacement solution of a novel tetrameric complex formed by two dimers of crotoxin B isoforms are presented. Crotoxin B is a basic phospholipase A{sub 2} found in the venom of Crotalus durissus terrificus and is one of the subunits that constitute crotoxin. This heterodimeric toxin, which is the main component of C. d. terrificus venom, is completed by an acidic, nontoxic and non-enzymatic component (crotoxin A) and is involved in important envenomation effects, such as neurological disorders, myotoxicity and renal failure. Although crotoxin was first crystallized in 1938, no crystal structure is currently available for crotoxin, crotoxin A or crotoxin B. In this work, the crystallization, X-ray diffraction data collection to 2.28 Å resolution and molecular-replacement solution of a novel tetrameric complex formed by two dimers of crotoxin B isoforms (CB1 and CB2) is presented.

  6. 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.

  7. Inhibitory effect of Crotalus durissus terrificus venom on chronic edema induced by injection of bacillus Calmette-Guérin into the footpad of mice.

    Science.gov (United States)

    da Silva, Nancy Gimenes; Sampaio, Sandra Coccuzzo; Gonçalves, Luís Roberto C

    2013-03-01

    In this study, we evaluated the effect of the Crotalus durissus terrificus (Cdt) venom on the chronic paw edema induced by the injection of bacillus Calmette-Guérin (BCG) into the footpad of mice. The BCG injection evoked chronic edema, which was significantly diminished in animals treated subcutaneously (s.c.) with Cdt venom 1 h before or after the BCG injection. This inhibition persisted throughout the evaluation period (15 days). In mice injected with Cdt venom 6 or 11 days after injection of BCG, we observed a significant reduction in edema only in the period after the venom injection. While studying possible mechanisms involved in this inhibition, we observed that pre-treatment with dexamethasone, zileuton or Boc2 (a selective antagonist of formyl peptide receptors), but not with indomethacin, canceled out the inhibitory effect of Cdt venom on the edema induced by BCG. These results strongly suggest that this rattlesnake venom can stimulate the generation of mediators from the lipoxygenase pathway, which can down-regulate this chronic inflammatory edema. Using fractionated venom, the results indicated that crotoxin was the only component of Cdt venom responsible for this inhibitory effect. These results indicated that crotoxin, the main toxin of the C. durissus terrificus venom, has a significant inhibitory effect on BCG-induced chronic edema, possibly by generating anti-inflammatory mediators from the lipoxygenase pathway. Copyright © 2012 Elsevier Ltd. All rights reserved.

  8. 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

  9. Thalidomide and pentoxifylline block the renal effects of supernatants of macrophages activated with Crotalus durissus cascavella venom

    Directory of Open Access Journals (Sweden)

    A.M.C. Martins

    2004-10-01

    Full Text Available Because thalidomide and pentoxifylline inhibit the synthesis and release of tumor necrosis factor-alpha (TNF-alpha, we determined the effect of these drugs on the renal damage induced by supernatants of macrophages activated with Crotalus durissus cascavella venom in order to identify the role of TNF-alpha in the process. Rat peritoneal macrophages were collected with RPMI medium and stimulated in vitro with C.d. cascavella venom (10 µg/ml in the absence and presence of thalidomide (15 µM or pentoxifylline (500 µM for 1 h and washed and kept in culture for 2 h. Supernatant (1 ml was tested on an isolated perfused rat kidney (N = 6 for each group. The first 30 min of each experiment were used as control. The supernatant was added to the perfusion system. All experiments lasted 120 min. The toxic effect of the preparation of venom-stimulated macrophages on renal parameters was determined. At 120 min, thalidomide (Thalid and pentoxifylline (Ptx inhibited (P < 0.05 the increase in perfusion pressure caused by the venom (control = 114.0 ± 1.3; venom = 137.1 ± 1.5; Thalid = 121.0 ± 2.5; Ptx = 121.4 ± 4.0 mmHg, renal vascular resistance (control = 4.5 ± 0.2; venom = 7.3 ± 0.6; Thalid = 4.5 ± 0.9; Ptx = 4.8 ± 0.6 mmHg/ml g-1 min-1, urinary flow (control = 0.23 ± 0.001; venom = 0.44 ± 0.01; Thalid = 0.22 ± 0.007; Ptx = 0.21 ± 0.009 ml g-1 min-1, glomerular filtration rate (control = 0.72 ± 0.06; venom = 1.91 ± 0.11; Thalid = 0.75 ± 0.04; Ptx = 0.77 ± 0.05 ml g-1 min-1 and the decrease in percent tubular sodium transport (control = 77.0 ± 0.9; venom = 73.9 ± 0.66; Thalid = 76.6 ± 1.1; Ptx = 81.8 ± 2.0%, percent tubular chloride transport (control = 77.1 ± 1.2; venom = 71.4 ± 1.1; Thalid = 77.6 ± 1.7; Ptx = 76.8 ± 1.2%, and percent tubular potassium transport (control = 72.7 ± 1.1; venom = 63.0 ± 1.1; Thalid = 72.6 ± 1.0; Ptx = 74.8 ± 1.0%, 30 min before and during the stimulation of macrophages with C.d. cascavella venom

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

    Directory of Open Access Journals (Sweden)

    MICHELE TAKAOKA

    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

  11. Pulmonary, microbiological and hematological changes in Crotalus durissus terrificus (Serpentes, Viperidae parasitized by nematodes of the genus Rhabdias (Nematoda, Rhabdiasidae Alterações Pulmonares, microbiológicas e hematológicas em Crotalus durissus terrificus (Serpentes, Viperidae parasitadas pelos nematódeos do gênero Rhabdias (Nematoda, Rhabdiasidae

    Directory of Open Access Journals (Sweden)

    K.R. Santos

    2008-06-01

    Full Text Available This study reported the pulmonary, microbiological, and hematological alterations in Crotalus durissus terrificus parasitized by nematodes of the genus Rhabdias. Histological, microbiological, and hematological analysis were performed on parasitized (n=6 and non-parasitized (n=6 snakes. Granulocytic and mononuclear cell infiltrates in the pulmonary parenchyma and epithelium were also observed during the histological analysis of parasitized snakes. Microbiological analysis of parasitized animals revealed the following Gram-negative bacteria: Citrobacter divergens, Burkholderia cepacia, Stenotrophomonas maltophilia, Proteus vulgaris, Enterobacter sakazakii, Enterobacter ammnigenus, Pseudomonas aeruginosa, Pantoea sp. and Providencia rettgeri. In non-parasitized snakes, the following species were identified: B. cepacia, Pseudomonas fluorescens, and Acinetobacter baumanii. Increased levels of plasmatic protein, decreased lymphocyte counts, and normal red blood cell values were observed in parasitized animals.Este trabalho relata as alterações pulmonares, microbiológicas e hematológicas em Crotalus durissus terrificus parasitadas pelo nematódeo do gênero Rhabdias. As análises histológicas, microbiológicas e hematológicas foram realizadas em serpentes parasitadas (n=6 e não parasitadas (n=6. Foram observados infiltrados de células granulocíticas e mononucleares no parênquima pulmonar durante a análise histopatológica das serpentes parasitadas. A análise microbiológica revelou as seguintes bactérias Gram-negativas; Citrobacter divergens, Burkholderia cepacia, Stenotrophomonas maltophilia, Proteus vulgaris, Enterobacter sakazakii, Enterobacter ammnigenus, Pseudomonas aeruginosa, Pantoea sp. e Providencia rettgeri. Nas serpentes não parasitadas foram identificadas: B. cepacia, Pseudomonas fluorescens e Acinetobacter baumanii. Nos animais parasitados observaram-se: aumento da concentração de proteína plasmática, diminuição da

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

    Directory of Open Access Journals (Sweden)

    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.

  13. An Evaluation of 3-Rhamnosylquercetin, a Glycosylated Form of Quercetin, against the Myotoxic and Edematogenic Effects of sPLA2 from Crotalus durissus terrificus

    Directory of Open Access Journals (Sweden)

    Daniela de Oliveira Toyama

    2014-01-01

    Full Text Available This paper shows the results of quercitrin effects on the structure and biological activity of secretory phospholipase (sPLA2 from Crotalus durissus terrificus, which is the main toxin involved in the pharmacological effects of this snake venom. According to our mass spectrometry and circular dichroism results, quercetin was able to promote a chemical modification of some amino acid residues and modify the secondary structure of C. d. terrificus sPLA2. Moreover, molecular docking studies showed that quercitrin can establish chemical interactions with some of the crucial amino acid residues involved in the enzymatic activity of the sPLA2, indicating that this flavonoid could also physically impair substrate molecule access to the catalytic site of the toxin. Additionally, in vitro and in vivo assays showed that the quercitrin strongly diminished the catalytic activity of the protein, altered its Vmax and Km values, and presented a more potent inhibition of essential pharmacological activities in the C. d. terrificus sPLA2, such as its myotoxicity and edematogenic effect, in comparison to quercetin. Thus, we concluded that the rhamnose group found in quercitrin is most likely essential to the antivenom activities of this flavonoid against C. d. terrificus sPLA2.

  14. An evaluation of 3-rhamnosylquercetin, a glycosylated form of quercetin, against the myotoxic and edematogenic effects of sPLA 2 from Crotalus durissus terrificus.

    Science.gov (United States)

    Toyama, Daniela de Oliveira; Gaeta, Henrique Hessel; de Pinho, Marcus Vinícius Terashima; Ferreira, Marcelo José Pena; Romoff, Paulete; Matioli, Fábio Filippi; Magro, Angelo José; Fontes, Marcos Roberto de Mattos; Toyama, Marcos Hikari

    2014-01-01

    This paper shows the results of quercitrin effects on the structure and biological activity of secretory phospholipase (sPLA2) from Crotalus durissus terrificus, which is the main toxin involved in the pharmacological effects of this snake venom. According to our mass spectrometry and circular dichroism results, quercetin was able to promote a chemical modification of some amino acid residues and modify the secondary structure of C. d. terrificus sPLA2. Moreover, molecular docking studies showed that quercitrin can establish chemical interactions with some of the crucial amino acid residues involved in the enzymatic activity of the sPLA2, indicating that this flavonoid could also physically impair substrate molecule access to the catalytic site of the toxin. Additionally, in vitro and in vivo assays showed that the quercitrin strongly diminished the catalytic activity of the protein, altered its Vmax and Km values, and presented a more potent inhibition of essential pharmacological activities in the C. d. terrificus sPLA2, such as its myotoxicity and edematogenic effect, in comparison to quercetin. Thus, we concluded that the rhamnose group found in quercitrin is most likely essential to the antivenom activities of this flavonoid against C. d. terrificus sPLA2.

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

    Science.gov (United States)

    Muller, Vanessa Danielle; Soares, Ricardo Oliveira; dos Santos, Nilton Nascimento; Trabuco, Amanda Cristina; Cintra, Adelia Cristina; Figueiredo, Luiz Tadeu; Caliri, Antonio; Sampaio, Suely Vilela; Aquino, Victor Hugo

    2014-01-01

    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. Crotoxin from Crotalus durissus terrificus snake venom induces the release of glutamate from cerebrocortical synaptosomes via N and P/Q calcium channels.

    Science.gov (United States)

    Lomeo, Rosangela da Silva; Gonçalves, Ana Paula de Faria; da Silva, Carolina Nunes; de Paula, André Tunes; Costa Santos, Danielle Oliveira; Fortes-Dias, Consuelo Latorre; Gomes, Dawidson Assis; de Lima, Maria Elena

    2014-07-01

    Crotoxin (Crtx), the main toxin in the venom of Crotalus durissus terrificus snake, is a heterodimer with a basic subunit, CB, and an acidic subunit, CA. CB is a phospholipase A2 that depends on CA to specifically bind to the cell membrane. This toxin acts in the central nervous system (CNS) causing chronic seizure effects and other cytotoxic effects. Here, we report its action on glutamate release in rat cerebral cortex synaptosomes. Aiming at a better understanding of the mechanism of action of Crtx, calcium channel blockers were used and internalization studies were performed in cerebellar granule neurons. Our results show that Crtx induces calcium-dependent glutamate release via N and P/Q calcium channels. In addition, the CB subunit of Crtx is shown to be internalized. This internalization does not depend on the presence of CA subunit neither on the PLA2 activity of CB. A correlation between CB internalization and glutamate release remains to be established. Copyright © 2014 Elsevier Ltd. All rights reserved.

  17. Ablation of the ability to control the right-to-left cardiac shunt does not affect oxygen uptake, specific dynamic action or growth in the rattlesnake Crotalus durissus.

    Science.gov (United States)

    Leite, Cleo A C; Taylor, Edwin W; Wang, Tobias; Abe, Augusto S; de Andrade, Denis O V

    2013-05-15

    The morphologically undivided ventricle of the heart in non-crocodilian reptiles permits the mixing of oxygen-rich blood returning from the lungs and oxygen-poor blood from the systemic circulation. A possible functional significance for this intra-cardiac shunt has been debated for almost a century. Unilateral left vagotomy rendered the single effective pulmonary artery of the South American rattlesnake, Crotalus durissus, unable to adjust the magnitude of blood flow to the lung. The higher constant perfusion of the lung circulation and the incapability of adjusting the right-left shunt in left-denervated snakes persisted over time, providing a unique model for investigation of the long-term consequences of cardiac shunting in a squamate. Oxygen uptake recorded at rest and during spontaneous and forced activity was not affected by removing control of the cardiac shunt. Furthermore, metabolic rate and energetic balance during the post-prandial metabolic increment, plus the food conversion efficiency and growth rate, were all similarly unaffected. These results show that control of cardiac shunting is not associated with a clear functional advantage in adjusting metabolic rate, effectiveness of digestion or growth rates.

  18. Effect of chlorogenic acid (5-Caffeoylquinic Acid) isolated from Baccharis oxyodonta on the structure and pharmacological activities of secretory phospholipase A2 from Crotalus durissus terrificus.

    Science.gov (United States)

    Toyama, Daniela O; Ferreira, Marcelo J P; Romoff, Paulete; Fávero, Oriana A; Gaeta, Henrique H; Toyama, Marcos H

    2014-01-01

    The aim of this paper was to investigate the effect of chlorogenic acid (5-caffeoylquinic acid, 5CQA), isolated from Baccharis oxyodonta, on the structure and pharmacological effect of secretory phospholipase A2 (sPLA2) from Crotalus durissus terrificus. All in vitro and in vivo experiments were conducted using a purified sPLA2 compared under the same experimental conditions with sPLA2 : 5CQA. 5CQA induced several discrete modifications in the secondary structure and the hydrophobic characteristics of native sPLA2 that induced slight changes in the α-helical content, increase in the random coil structure, and decrease of fluorescence of native sPLA2. Moreover, 5CQA significantly decreased the enzymatic activity and the oedema and myonecrosis induced by native sPLA2. As the catalytic activity of sPLA2 plays an important role in several of its biological and pharmacological properties, antibacterial activity was used to confirm the decrease in its enzymatic activity by 5CQA, which induced massive bacterial cell destruction. We found that 5CQA specifically abolished the enzymatic activity of sPLA2 and induced discrete protein unfolding that mainly involved the pharmacological site of sPLA2. These results showed the potential application of 5CQA in the snake poisoning treatment and modulation of the pathological effect of inflammation induced by secretory PLA2.

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

    Science.gov (United States)

    Floriano, Rafael Stuani; Nogueira, Rosa Maria Barilli; Sakate, Michiko; Laposy, Cecília Braga; da Motta, Yudney Pereira; Sangiorgio, Fabíola; David, Heloísa Costa; Nabas, João Marcelo

    2009-12-01

    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 clinical 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.

  20. Characterization of ribonucleic acids from the venom glands of Crotalus durissus terrificus (Ophidia, Reptilia) after manual extraction of the venom. Studies on template activity and base composition

    Science.gov (United States)

    De Lucca, F. L.; Imaizumi, M. T.; Haddad, A.

    1974-01-01

    RNA synthesis in the venom glands of Crotalus durissus terrificus was stimulated by the manual extraction of the venom (milking). RNA was extracted from venom glands activated by milking and fractionated by centrifugation through sucrose density gradients. Template activity for protein synthesis and base composition of the RNA fractions were studied. RNA fractions that sediment between 18S and 4S had the highest template activity. The base composition analysis indicated that the 28S and 18S rRNA have a C+G content of 65.4 and 58% respectively. The `melting' temperature (Tm) of DNA in 0.15m-NaCl–0.015m-trisodium citrate, pH7.0, was 85°C, corresponding to a C+G content of 38%. The base ratio of the RNA fractions that showed a high template activity was intermediate between that of rRNA and homologous DNA. The possible role of these fractions in the synthesis of the two main toxins (crotoxin and crotamine) of the South American rattlesnake's venom is discussed. ImagesPLATE 1 PMID:4463939

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

    Directory of Open Access Journals (Sweden)

    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

  2. Insights on the structure of native CNF, an endogenous phospholipase A2 inhibitor from Crotalus durissus terrificus, the South American rattlesnake.

    Science.gov (United States)

    Fortes-Dias, Consuelo Latorre; Ortolani, Paula Ladeira; Fernandes, Carlos Alexandre H; Lobo, Kelli Roberta; Amaral de Melo, Lutiana; Borges, Márcia Helena; Pazin, Wallance Moreira; Neto, Mário de Oliveira; Fernandez, Roberto Morato; Fontes, Marcos Roberto M

    2014-09-01

    Several snake species possess endogenous phospholipase A2 inhibitors (sbPLIs) in their blood plasma, the primary role of which is protection against an eventual presence of toxic phospholipase A2 (PLA2) from their venom glands in the circulation. These inhibitors have an oligomeric structure of, at least, three subunits and have been categorized into three classes (α, β and γ) based on their structural features. SbγPLIs have been further subdivided into two subclasses according to their hetero or homomeric nature, respectively. Despite the considerable number of sbγPLIs described, their structures and mechanisms of action are still not fully understood. In the present study, we focused on the native structure of CNF, a homomeric sbγPLI from Crotalus durissus terrificus, the South American rattlesnake. Based on the results of different biochemical and biophysical experiments, we concluded that, while the native inhibitor occurs as a mixture of oligomers, tetrameric arrangement appears to be the predominant quaternary structure. The inhibitory activity of CNF is most likely associated with this oligomeric conformation. In addition, we suggest that the CNF tetramer has a spherical shape and that tyrosinyl residues could play an important role in the oligomerization. The carbohydrate moiety, which is present in most sbγPLIs, is not essential for the inhibitory activity, oligomerization or complex formation of the CNF with the target PLA2. A minor component, comprising no more than 16% of the sample, was identified in the CNF preparations. The amino-terminal sequence of that component is similar to the B subunits of the heteromeric sbγPLIs; however, the role played by such molecule in the functionality of the CNF, if any, remains to be determined. Copyright © 2014 Elsevier B.V. All rights reserved.

  3. Individual venom profiling of Crotalus durissus terrificus specimens from a geographically limited region: crotamine assessment and captivity evaluation on the biological activities.

    Science.gov (United States)

    Lourenço, Airton; Zorzella Creste, Camila Fernanda; de Barros, Luciana Curtolo; Delazari dos Santos, Lucilene; Pimenta, Daniel C; Barraviera, Benedito; Ferreira, Rui Seabra

    2013-07-01

    Crotalus durissus terrificus (Cdt) venom major components comprise crotoxin, crotamine, gyroxin and convulxin. Crotamine exerts a myotoxic action, among others, but its expression varies even amid snakes from the same region. Biochemical, enzymatic and pharmacological variations of venoms may be associated with the geography, climate, gender, age, and diet, as well as captivity time and venom extraction intervals. The present study aimed to characterize the Cdt venom from the Botucatu region, (SP, Brazil), by assessing its biochemical, pharmacological and enzymatic properties. Venoms from newly captured snakes and already-captured animals were characterized comparatively to verify the sexual, environmental (length of captivity) and ontogenetic variations that could influence the venom composition. Protein concentration, SDS-PAGE and RP-HPLC were performed and the coagulant, toxic (LD50) and crotamine activities were assayed. Individual SDS-PAGE analyses (315 samples) were performed and the biological activities of the venom of 60 adults (captive and newly captured males and females) and 18 newborns were compared with the Brazilian Reference Venom. Crotamine was found in 39.7% (125/315) of the samples, as determined by SDS-PAGE and RP-HPLC. Protein concentration differed significantly between adults (75%) and newborns (60%). RP-HPLC and SDS-PAGE analyses showed highly variable protein concentration and copious crotoxin isoforms; however, the LD50 values decreased during the captivity time. Cdt venom biological activities were similar among adult groups, but diminished during the captivity period. The current findings demonstrate that venoms vary significantly in terms activity and protein concentration, despite originating from the same specie and region. Copyright © 2013 Elsevier Ltd. All rights reserved.

  4. Avaliação em camundongo da eficácia do antiveneno administrado no local da inoculação intramuscular do veneno de Crotalus durissus terrificus Evaluation in mice, of the antivenom efficacy injected at the same place of the intramuscular inoculation of the Crotalus durissus terrificus venom

    Directory of Open Access Journals (Sweden)

    Lindioneza Adriano Ribeiro

    1993-02-01

    Full Text Available A eficácia do antiveneno crotálico por via intramuscular (im no local da inoculação, também im, do veneno de Crotalus durissus terrificus foi avaliada em camundongos. Em três experimentos inocularam-se duas DLSO do veneno por via im e administrou-se o antiveneno de três formas: metade da DE50 por via intraperitoneal (ip e metade por via im, no mesmo local, imediatamente após (1º e 30' após (2º a inoculação do veneno; quatro quintos de DE50 por via ip e um quinto por via im, no mesmo local e 30' após a inoculação do veneno (3º. O antiveneno ofereceu, por via ip, maior proteção aos camundongos (menor taxa de óbito em 48 horas do que quando foi administrado, em parte, por via im, no local da inoculação do veneno (pThe efficacy of the Crotalus durissus terrificus antivenom administration by intramuscular (im injection at the same place of the im inoculation, of the C. d. terrificus venom was evaluated in mice. In three experiments two DL50 of the venom were inoculated and the antivenom was administered in three differents ways: half of the ED50 by intraperitoneal (ip administration and half by injection, at the same place, immediatelly after the venom inoculation and thirty minutes after the im venom inoculation; four fifth of ED50 by ip administration and one fifth by injection, at the same place and thirty minutes after the venom inoculation. The antivenom that was administred by intraperitoneal route provided a higher protection to mice (a lower death rate in a 48 hours period than when it was administred in parts, by intramuscular injection, at the same place of the venom inoculation (p<0,05. Therefore, it is concluded that this should not be used in human beings bitten by snakes.

  5. Comparación entre las técnicas de neutralización en ratón y E.L.I.S.A para determinar la potencia del anti-veneno crotalus durissus terrificus

    Directory of Open Access Journals (Sweden)

    María Cristina Forero CH.

    1984-12-01

    Full Text Available Las técnicas de neutralización en ratón y ELISA se compararon como métodos para determinar la potencia del antiveneno Crotalus durissus terrificus. lnformes en la literatura(3, (12, (13, sugerían la posibilidad de cambiar la técnica in vivo muy costosa por la técnica in vitro. Se utilizaron antivenenos procesados (Gamma-globulinas purificadas y no procesados. No se encontró correlación entre los títulos de neutralización y las lecturas de absorvancia obtenidas por ELISA para ninguna de las muestras. La falta de correlación se debe probablemente a que las técnicas miden dos poblaciones de anticuerpos diferentes. Mientras que con ELlSA se miden anticuerpos contra todos los antígenos del veneno, con la técnica de neutralización solo se miden anticuerpos contra la fracción letal. Se sugiere verificar nuevamente la correlación entre las dos técnicas usando como antígeno para ELISA la neurotoxina purificada. No se recomienda, en contra a lo afirmado por otros autores (3, el empleo de la técnica de ELlSA usando veneno total como antígeno, para determinar la potencia del antiveneno Crotalus durissus terrificus.

  6. Resposta imune e capacidade de neutralização de anticorpos produzidos em ovinos jovens imunizados com veneno de Crotalus durissus terrificus nativo e irradiado com Cobalto 60

    Directory of Open Access Journals (Sweden)

    Rui Seabra Ferreira Junior

    2009-06-01

    Full Text Available A técnica de Elisa foi utilizada para avaliar e comparar a resposta imune humoral de ovinos jovens para a produção de soro anticrotálico. Durante o processo de soroprodução, foi realizada a avaliação clínica dos animais. A capacidade de neutralização do soro produzido a partir de veneno de serpente Crotalus durissus terrificus, nativo (VN e irradiado (VIr com Cobalto-60 foi verificada por meio de desafios in vitro. Um grupo de seis animais recebeu veneno nativo, o segundo grupo recebeu veneno irradiado e o terceiro grupo foi o controle. Os animais receberam seis imunizações durante 84 dias com intervalo de 14 dias. Houve diferença significativa (p<5% no teste de ELISA do perfil de anticorpos produzidos pelos grupos experimentais (VN < VIr. O grupo imunizado com veneno irradiado apresentou perfil de anticorpos maior que o grupo imunizado com veneno nativo. A capacidade de neutralização do soro produzido a partir do VIr foi cinco vezes maior quando comparado ao soro produzido com VN. Estes resultados justificam o uso da radiação gama na destoxicação do veneno de Crotalus durissus terrificus como alternativa na produção de antiveneno.

  7. Enzyme specificity and effects of gyroxin, a serine protease from the venom of the South American rattlesnake Crotalus durissus terrificus, on protease-activated receptors.

    Science.gov (United States)

    Yonamine, Camila M; Kondo, Marcia Y; Nering, Marcela B; Gouvêa, Iuri E; Okamoto, Débora; Andrade, Douglas; da Silva, José Alberto A; Prieto da Silva, Alvaro R B; Yamane, Tetsuo; Juliano, Maria A; Juliano, Luiz; Lapa, Antônio J; Hayashi, Mirian A F; Lima-Landman, Maria Teresa R

    2014-03-01

    Gyroxin is a serine protease displaying a thrombin-like activity found in the venom of the South American rattlesnake Crotalus durissus terrificus. Typically, intravenous injection of purified gyroxin induces a barrel rotation syndrome in mice. The serine protease thrombin activates platelets aggregation by cleaving and releasing a tethered N-terminus peptide from the G-protein-coupled receptors, known as protease-activated receptors (PARs). Gyroxin also presents pro-coagulant activity suggested to be dependent of PARs activation. In the present work, the effects of these serine proteases, namely gyroxin and thrombin, on PARs were comparatively studied by characterizing the hydrolytic specificity and kinetics using PARs-mimetic FRET peptides. We show for the first time that the short (sh) and long (lg) peptides mimetizing the PAR-1, -2, -3, and -4 activation sites are all hydrolyzed by gyroxin exclusively after the Arg residues. Thrombin also hydrolyzes PAR-1 and -4 after the Arg residue, but hydrolyzes sh and lg PAR-3 after the Lys residue. The kcat/KM values determined for gyroxin using sh and lg PAR-4 mimetic peptides were at least 2150 and 400 times smaller than those determined for thrombin, respectively. For the sh and lg PAR-2 mimetic peptides the kcat/KM values determined for gyroxin were at least 6500 and 2919 times smaller than those determined for trypsin, respectively. The kcat/KM values for gyroxin using the PAR-1 and -3 mimetic peptides could not be determined due to the extreme low hydrolysis velocity. Moreover, the functional studies of the effects of gyroxin on PARs were conducted in living cells using cultured astrocytes, which express all PARs. Despite the ability to cleavage the PAR-1, -2, -3, and -4 peptides, gyroxin was unable to activate the PARs expressed in astrocytes as determined by evaluating the cytosolic calcium mobilization. On the other hand, we also showed that gyroxin is able to interfere with the activation of PAR-1 by thrombin or

  8. 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)

  9. 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

    Directory of Open Access Journals (Sweden)

    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

  10. 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.

  11. Loss of the ability to control right-to-left shunt does not influence the metabolic responses to temperature change or long-term fasting in the South American Rattlesnake Crotalus durissus.

    Science.gov (United States)

    Leite, Cleo A C; Wang, Tobias; Taylor, Edwin W; Abe, Augusto S; Leite, Gabrielle S P C; de Andrade, Denis O V

    2014-01-01

    Abstract In the undivided ventricle of noncrocodilian reptiles, the blood perfusing the systemic circulation is a controlled combination of oxygenated pulmonary blood, flowing from left to right across the heart, and relatively deoxygenated systemic blood, flowing from right to left. A clear inverse correlation has been experimentally established between metabolic demand and the magnitude of right-to-left cardiac shunt in several reptile groups. Unilateral left vagotomy renders the single effective pulmonary artery of the South American rattlesnake (Crotalus durissus) unable to adjust the magnitude of blood flow to the lung. This provides a unique model for investigation of the long-term consequences of abolition of the cardiac shunt in a squamate reptile. Rattlesnakes-vagotomized on the left or right side or sham operated-were exposed to long-term food deprivation or temperature change. Loss of control of the cardiac shunt following selective vagotomy did not change the progressive decrease in body mass or the onset of identifiable fasting stages. Resting metabolic rate and the increase in oxygen uptake measured during spontaneous or forced activity were also unchanged. The responses to reductions in temperature (from 30° to 20° or 15°C) in adult snakes or juvenile snakes were similarly unaffected by vagal transection. These data support rejection of the hypothesis that adjustment of the cardiac shunt is central to the control metabolic rate in squamate reptiles.

  12. 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.

  13. 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, six...... rats treated with a 2% solution of methylene blue (single dose: 2 mg/kg, IV) and forty min later, TV was injected as in group I. Before the onset of the experiments, each rat was anesthetized with azaper - one:ketamine. The femoral artery and vein were cannulated to record blood pressure (mean arterial...... of methylene blue. Forty min later, TV was injected causing a reduction in MAP (73.0 ± 6.6 to 27.0 ± 2.8mmHg) and HR (360.0 ± 9.9 to 180.0 ± 9.0 l/min). After 10 min, the MAP and HR returned to baseline levels, reaching 72.6 ± 7.9 mmHg and 360.0 ± 11.0 lat/min, respectively, with a 100% survival at 24h...

  14. Isolation, characterization and screening of the in vitro cytotoxic activity of a novel L-amino acid oxidase (LAAOcdt) from Crotalus durissus terrificus venom on human cancer cell lines.

    Science.gov (United States)

    Teixeira, Tuila Leveghim; Oliveira Silva, Viviane Aline; da Cunha, Daniel Batista; Polettini, Flávia Lino; Thomaz, Camila Daniele; Pianca, Ariana Aparecida; Zambom, Fabiana Letícia; da Silva Leitão Mazzi, Denise Pimenta; Reis, Rui Manuel; Mazzi, Maurício Ventura

    2016-09-01

    An L-amino acid oxidase (LAAOcdt) from Crotalus durissus terrificus venom was purified to homogeneity in a two-step procedure using molecular exclusion on Sephadex G-75, followed by Phenyl Sepharose FF chromatography. The molecular mass of the purified enzyme was 113 kDa, as determined by SDS-PAGE under reducing conditions. LAAOcdt showed amino acid homology to other L-amino acid oxidases isolated from different snake venoms. The comparative analysis of the internal peptide sequences of the NNPGILEYPVKPSEEGK fragments by LC-MS/MS spectrometry revealed 100% identity with C. durissus cumanensis LAAO. The purified protein catalyzed the oxidative deamination of L-amino acids, and the most specific substrates were L-Tyr and L-Phe. The enzyme presented optimum activity at pH 7.4 and at 44 °C. LAAOcdt also showed hemolytic activity (0.6-20 μg/μL) and induced both the formation plasma clots (5-100 μg/μL) and platelet aggregation (2.5 × 10(-3), 5.0 × 10(-3) and 10 × 10(-3) μg/mL), as well as bactericidal activity (2.5-10 μg/μL) against Staphylococcus aureus. Moreover, LAAOcdt exhibited cytotoxicity in distinct cancer cell lines, which presented a heterogeneous response profile. The mean IC50 value was 10.5 μg/mL. Glioma and pancreatic carcinoma cells were the most sensitive cell lines; they showed mean IC50 values of 7.2 μg/mL and 7.4 μg/mL, respectively. The exposure of the drug-sensitive cells to LAAOcdt for 24 h upregulated activated p-H2AX and efficiently decreased P42/P44 (ERK) activation in glioma cells (HCB151), which suggested an anti-proliferative effect. In addition, increased p21 expression was observed in SiHa cells, which showed a resistant phenotype. On the other hand, the flow cytometry and immunoblotting analyses showed that the enzyme did not induce cancer cell apoptosis. These results suggest that another cell death mechanism might contribute to the LAAOcdt-induced cytotoxicity. Taken together, this work may help to elucidate

  15. Neuromuscular paralysis by the basic phospholipase A2subunit 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.

  16. Fresh frozen plasma associated to the antiophidic serum in the treatment of dogs intoxicated experimentally by the poison of the serpent Crotalus durissus terrificus

    OpenAIRE

    Brunholi, Gisele Sartori Cavalare

    2010-01-01

    Acidentes ofídicos acarretam um problema médico relevante em nosso país devido às altas toxicidades e letalidades dos venenos. Cerca de 8% dos acidentes estão relacionados ao gênero Crotalus estando em segundo lugar no número de acidentes e em primeiro em relação ao índice de óbito. O estudo teve como objetivo avaliar e comparar a indução da intoxicação experimental e terapia por veneno crotálico em cães, com uso de soro antiofídico isolado ou associado a plasma congelado. Os cães foram divid...

  17. P9a(Cdt-PLA2) from Crotalus durissus terrificus as good immunogen to be employed in the production of crotalic anti-PLA2 IgG.

    Science.gov (United States)

    Fusco, Luciano S; Rodríguez, Juan Pablo; Torres-Huaco, Frank; Huancahuire-Vega, Salomón; Teibler, Pamela; Acosta, Ofelia; Marangoni, Sergio; Ponce-Soto, Luis Alberto; Leiva, Laura C

    2015-10-01

    Four proteins with phospholipase A2 (PLA2) activity, designated P9a(Cdt-PLA2), P9b(Cdt-PLA2), P10a(Cdt-PLA2) and P10b(Cdt-PLA2) were purified from the venom of Crotalus durissus terrificus by two chromatographic steps: a gel filtration and reversed phase HPLC. The profile obtained clearly shows that three of them have a similar abundance. The molecular mass, 14193.8340Da for P9a(Cdt-PLA2), 14134.9102Da for P9b(Cdt-PLA2), 14242.6289Da for P10a(Cdt-PLA2) and 14183.8730Da for P10b(Cdt-PLA2), were initially evaluated by SDS-PAGE and confirmed by ESI-Q-TOF spectrometry, and all of them displayed a monomeric conformation. Also, partial amino acid sequence of each protein was obtained and their alignments with other crotalic PLA2 revealed a high degree of identity among them. Additionally, we studied some pharmacological activities like neurotoxicity, myotoxicity and lethality, which prompted us to pick two of them, P9a(Cdt-PLA2) and P10a(Cdt-PLA2) that resulted to be less toxic that the others, and further characterize them to be used as immunogen. We next injected these last proteins in mice to produce antitoxins against them and ELISA and dot blots reveled that both toxins do not show immunogenic differences, unlike those other pharmacologic activities tested. Furthermore, the antibodies produced cross-reacted with all the isoforms purified demonstrating the feasibility of using only one of them and ensuring the cross-reaction of all. The results obtained show that P9a(Cdt-PLA2) isoform has the lowest toxicity and also a good purification performance; thus this protein may be a promising candidate to be employed in the production of crotalic antitoxins. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  18. 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

  19. 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.

  20. Envenomations by Bothrops and Crotalus Snakes Induce the Release of Mitochondrial Alarmins

    Science.gov (United States)

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

    2012-01-01

    Skeletal muscle necrosis is a common manifestation of viperid snakebite envenomations. Venoms from snakes of the genus Bothrops, such as that of B. asper, induce muscle tissue damage at the site of venom injection, provoking severe local pathology which often results in permanent sequelae. In contrast, the venom of the South American rattlesnake Crotalus durissus terrificus, induces a clinical picture of systemic myotoxicity, i.e., rhabdomyolysis, together with neurotoxicity. It is known that molecules released from damaged muscle might act as ‘danger’ signals. These are known as ‘alarmins’, and contribute to the inflammatory reaction by activating the innate immune system. Here we show that the venoms of B. asper and C. d. terrificus release the mitochondrial markers mtDNA (from the matrix) and cytochrome c (Cyt c) from the intermembrane space, from ex vivo mouse tibialis anterior muscles. Cyt c was released to a similar extent by the two venoms whereas B. asper venom induced the release of higher amounts of mtDNA, thus reflecting hitherto some differences in their pathological action on muscle mitochondria. At variance, injection of these venoms in mice resulted in a different time-course of mtDNA release, with B. asper venom inducing an early onset increment in plasma levels and C. d. terrificus venom provoking a delayed release. We suggest that the release of mitochondrial ‘alarmins’ might contribute to the local and systemic inflammatory events characteristic of snakebite envenomations. PMID:22363828

  1. 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.

  2. A Novel Phospholipase A(2) (D49) from the Venom of the Crotalus oreganus abyssus (North American Grand Canyon Rattlesnake)

    OpenAIRE

    Martins, W; Baldasso, PA; Honorio, KM; Maltarollo, VG; Ribeiro, RIMA; Carvalho, BMA; Soares, AM; Calderon, LA; Stabeli, RG; Caballol, MAO; Acosta, G; Oliveira, E; Marangoni, S; Albericio, F; Da Silva, SL

    2014-01-01

    Currently, Crotalus viridis was divided into two species: Crotalus viridis and Crotalus oreganus. The current classification divides 'the old' Crotalus viridis into two new and independent species: Crotalus viridis (subspecies: viridis and nuntius) and Crotalus oreganus (subspecies: abyssus, lutosus, concolor, oreganus, helleri, cerberus, and caliginis). The analysis of a product from cDNA (E6d), derived from the gland of a specie Crotalus viridis viridis, was found to produce an acid phospho...

  3. Quadro clínico do envenenamento crotálico experimental em bovinos (Crotalus durissus terrificus - crotamina positivo

    Directory of Open Access Journals (Sweden)

    Luiz Alberto do Lago

    2000-01-01

    Full Text Available Com o objetivo de descrever o quadro clínico de envenenamento crotálico em bovinos, cinco fêmeas, mestiças, com idade variando entre dois e três anos, foram inoculadas com veneno crotálico do tipo crotamina positivo. A dose foi de 0,03 mg/kg de peso vivo. Os animais apresentaram, cronologicamente, o seguinte quadro clínico: na 2ª hora de evolução, apatia, cabeça baixa, letargia profunda e edemaciação discreta no local da inoculação; na 6ª hora, o edema local desapareceu e mioclonias foram observadas nas grandes massas musculares; na 10ª hora, houve diminuição do tônus muscular, de reflexos superficiais, aparecimento de incoordenação motora e decúbito esternal; na 14ª hora, observaram-se movimentos de "pedalagem" e diminuição da sensibilidade profunda; na 16ª hora, paralisia flácida dos membros pélvicos; e, na 20ª hora, surgiram dispnéia, sialorréia e morte de todos os animais. A temperatura retal não se alterou até a 20ª hora, porém, a partir daí, ocorreu diminuição constante até a morte. A freqüência respiratória se manteve dentro da faixa de valores normais para bovinos, apesar da dispnéia observada ao atingir a 20ª hora. A duração do curso clínico foi de 28 ± 9 horas e, muito embora estas alterações clínicas sejam comuns a outras doenças, julga-se que a rapidez e a ordem com que aparecem seja um fator determinante no diagnóstico diferencial.

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

    OpenAIRE

    Kindlovits,Lívia M.; Miranda,Farlen J.B.; João Claudio Damasceno-Sá; Renato A. Da Matta; Nádia R.P. Almosny

    2017-01-01

    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 reconhe...

  5. Clinical safety evaluation of F(ab')₂ antivenom (Crotalus durissus-Bothrops asper) administration in dogs.

    Science.gov (United States)

    Woods, Craig; Young, David

    2011-10-01

    Antivenoms consisting of selective antigen binding antibody fragments, or F(ab')(2), are becoming more popular in human and veterinary medicine, owing to their preferred kinetics, tissue distribution, and removal of the Fc binding portion of IgG. Consequences of antivenom administration can include acute and delayed reactions, dependent, in part, on the antivenom's donor source, purity, and composition. This study evaluated an equine-derived polyvalent F(ab')(2) pit viper antivenom in healthy dogs of various size, age, and breed under controlled conditions. Dogs were allocated into 6 treatment groups (n = 10 per group) based on weight (3 weight groups) and dose (2 dose groups) and administered F(ab')(2) antivenom over 1 hour by IV infusion. Dogs were observed for adverse events at 3, 6, 12, and 24 hours after administration and blood was collected for CBC and serum biochemistry before and at 24 hours postadministration. No abnormalities were noted in the 3-vial group. In the 6-vial group, 3 dogs developed mild self-limiting edema around the head or neck, suggestive of a type 1 hypersensitivity, while another dog vomited and developed mild hypocalcemia at 24 hours, reflecting a 13% reaction rate at high doses. However, no dogs exhibited clinical signs of hypocalcemia or had any severe adverse events. CBC remained normal in all groups. This study demonstrated that the polyvalent F(ab')(2) pit viper antivenom is well tolerated in dogs given large doses in a short period of time. © Veterinary Emergency and Critical Care Society 2011.

  6. Inhibition of L-glutamate and GABA synaptosome uptake by crotoxin, the major neurotoxin from Crotalus durissus terrificus venom

    Directory of Open Access Journals (Sweden)

    A.L. Cecchini

    2004-01-01

    Full Text Available This paper describes a brief study on the crotoxin mechanism of action, regarding the transport of GABA and L-glutamate in rats cortico-cerebral synaptosomes and in heterologous systems, such as COS-7 cells expressing gabaergic transporters, and C6 glioma cells and Xenopus oocytes expressing glutamatergic transporters. Crotoxin concentrations over 1 µM caused an inhibitory effect of ³H-L-glutamate and ³H-GABA, and reversibly inhibited L-glutamate uptake by C6 glioma cells. When COS-7 cells were assayed, no inhibition of the ³H-GABA transport could be evidenced. Crotoxin kept its inhibitory effect on neurotransmitters uptake even when Ca2+ ions were removed from the medium, therefore, independently of its PLA2 activity. In addition, high concentrations (2 mM of BPB did not avoid the action of crotoxin on the neurotransmitters uptake. Crotoxin also inhibited ³H-L-glutamate, independently on Na+ channel blockade by TTX. In addition, an evaluation of the lactic dehydrogenase activity indicated that uptake inhibition does not involve a hydrolytic action of crotoxin upon the membrane. We may also suggest that crotoxin acts, at least partially, altering the electrogenic equilibrium, as evidenced by confocal microscopy, when a fluorescent probe was used to verify cell permeability on C6 glioma cells in presence of crotoxin.

  7. 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

  8. The genesis of an exceptionally lethal venom in the timber rattlesnake (Crotalus horridus) revealed through comparative venom-gland transcriptomics.

    Science.gov (United States)

    Rokyta, Darin R; Wray, Kenneth P; Margres, Mark J

    2013-06-12

    Snake venoms generally show sequence and quantitative variation within and between species, but some rattlesnakes have undergone exceptionally rapid, dramatic shifts in the composition, lethality, and pharmacological effects of their venoms. Such shifts have occurred within species, most notably in Mojave (Crotalus scutulatus), South American (C. durissus), and timber (C. horridus) rattlesnakes, resulting in some populations with extremely potent, neurotoxic venoms without the hemorrhagic effects typical of rattlesnake bites. To better understand the evolutionary changes that resulted in the potent venom of a population of C. horridus from northern Florida, we sequenced the venom-gland transcriptome of an animal from this population for comparison with the previously described transcriptome of the eastern diamondback rattlesnake (C. adamanteus), a congener with a more typical rattlesnake venom. Relative to the toxin transcription of C. adamanteus, which consisted primarily of snake-venom metalloproteinases, C-type lectins, snake-venom serine proteinases, and myotoxin-A, the toxin transcription of C. horridus was far simpler in composition and consisted almost entirely of snake-venom serine proteinases, phospholipases A2, and bradykinin-potentiating and C-type natriuretic peptides. Crotalus horridus lacked significant expression of the hemorrhagic snake-venom metalloproteinases and C-type lectins. Evolution of shared toxin families involved differential expansion and loss of toxin clades within each species and pronounced differences in the highly expressed toxin paralogs. Toxin genes showed significantly higher rates of nonsynonymous substitution than nontoxin genes. The expression patterns of nontoxin genes were conserved between species, despite the vast differences in toxin expression. Our results represent the first complete, sequence-based comparison between the venoms of closely related snake species and reveal in unprecedented detail the rapid evolution of

  9. Arizona black rattlesnake (Crotalus viridis cerberus)

    Science.gov (United States)

    Nowak, Erika M.

    2006-01-01

    The Arizona black rattlesnake makes its home at higher elevations in Arizona and far western New Mexico. The snake's use of high-altitude habitat and its black coloration as an adult distinguishes it from other subspecies of the western rattlesnake (Crotalus viridis), which prefer lower elevations and range from tan to reddish in color as adults. These physical and habitat differences are also reflected in genetic differences that suggest that the Arizona black rattlesnake may be a new species of rattlesnake. Despite the species's limited range, basic biological information needed to make management decisions is lacking for most Arizona black rattlesnake populations. To address this need, U.S. Geological Survey (USGS) scientists conducted research on the species in Arizona national park units from 2003 to 2005. The research examined relative population abundance, movement patterns, range requirements, dietary habits, and winter and summer habitat. Research in Arizona national parks was made possible through the support of the Western National Parks Association, Tonto National Monument, and the USGS Science Internships for Workforce Diversity Program. Importantly, the park-based research was used to augment a long-term mark-recapture study of the species that has been conducted by USGS biologists at sites near Flagstaff, Arizona, since 1999. USGS researchers were the first to conduct extensive studies of this species in the wild.

  10. A novel phospholipase A2 (D49) from the venom of the Crotalus oreganus abyssus (North American Grand canyon rattlesnake).

    Science.gov (United States)

    Martins, W; Baldasso, P A; Honório, K M; Maltarollo, V G; Ribeiro, R I M A; Carvalho, B M A; Soares, A M; Calderon, L A; Stábeli, R G; Caballol, M A O; Acosta, G; Oliveira, E; Marangoni, S; Albericio, F; Da Silva, S L

    2014-01-01

    Currently, Crotalus viridis was divided into two species: Crotalus viridis and Crotalus oreganus. The current classification divides "the old" Crotalus viridis into two new and independent species: Crotalus viridis (subspecies: viridis and nuntius) and Crotalus oreganus (subspecies: abyssus, lutosus, concolor, oreganus, helleri, cerberus, and caliginis). The analysis of a product from cDNA (E6d), derived from the gland of a specie Crotalus viridis viridis, was found to produce an acid phospholipase A2. In this study we isolated and characterized a PLA2 (D49) from Crotalus oreganus abyssus venom. Our studies show that the PLA2 produced from the cDNA of Crotalus viridis viridis (named E6d) is exactly the same PLA2 primary sequence of amino acids isolated from the venom of Crotalus oreganus abyssus. Thus, the PLA2 from E6d cDNA is actually the same PLA2 presented in the venom of Crotalus oreganus abyssus and does not correspond to the venom from Crotalus viridis viridis. These facts highlight the importance of performing more studies on subspecies of Crotalus oreganus and Crotalus viridis, since the old classification may have led to mixed results or mistaken data.

  11. A Novel Phospholipase A2 (D49 from the Venom of the Crotalus oreganus abyssus (North American Grand Canyon Rattlesnake

    Directory of Open Access Journals (Sweden)

    W. Martins

    2014-01-01

    Full Text Available Currently, Crotalus viridis was divided into two species: Crotalus viridis and Crotalus oreganus. The current classification divides “the old” Crotalus viridis into two new and independent species: Crotalus viridis (subspecies: viridis and nuntius and Crotalus oreganus (subspecies: abyssus, lutosus, concolor, oreganus, helleri, cerberus, and caliginis. The analysis of a product from cDNA (E6d, derived from the gland of a specie Crotalus viridis viridis, was found to produce an acid phospholipase A2. In this study we isolated and characterized a PLA2 (D49 from Crotalus oreganus abyssus venom. Our studies show that the PLA2 produced from the cDNA of Crotalus viridis viridis (named E6d is exactly the same PLA2 primary sequence of amino acids isolated from the venom of Crotalus oreganus abyssus. Thus, the PLA2 from E6d cDNA is actually the same PLA2 presented in the venom of Crotalus oreganus abyssus and does not correspond to the venom from Crotalus viridis viridis. These facts highlight the importance of performing more studies on subspecies of Crotalus oreganus and Crotalus viridis, since the old classification may have led to mixed results or mistaken data.

  12. The complete mitochondrial DNA sequence of Crotalus horridus (timber rattlesnake).

    Science.gov (United States)

    Hall, Jacob B; Cobb, Vincent A; Cahoon, A Bruce

    2013-04-01

    The complete mitogenome of the timber rattlesnake (Crotalus horridus) was completed using Sanger sequencing. It is 17,260 bp with 13 protein-coding genes, 21 tRNAs, two rRNAs and two control regions. Gene synteny is consistent with other snakes with the exception of a missing redundant tRNA (Ser) . This mitogenome should prove to be a useful addition of a well-known member of the Viperidae snake family.

  13. Comparison of venom composition and biological activities of the subspecies Crotalus lepidus lepidus, Crotalus lepidus klauberi and Crotalus lepidus morulus from Mexico.

    Science.gov (United States)

    Martínez-Romero, Gerardo; Rucavado, Alexandra; Lazcano, David; Gutiérrez, José María; Borja, Miguel; Lomonte, Bruno; Garza-García, Yolanda; Zugasti-Cruz, Alejandro

    2013-09-01

    The rock rattlesnakes Crotalus lepidus comprise a group (lepidus, klauberi, morulus and maculosus) of poorly known mountain cold-tolerant snakes in Mexico. In particular, Crotalus lepidus morulus is a snake endemic of the northeast of Mexico, whereas Crotalus lepidus klauberi and C. l. lepidus are distributed in some regions of the north and central Mexico and southern U. S. Until now very little data are available from C. lepidus subspecies from Mexico, as the terrain inhabited by these snakes is generally steep and rugged. In this work, we have determined some biochemical and biological properties of C. l. morulus, C. l. klauberi and C. l. lepidus crude venoms. Some minor differences in venoms were noted in SDS-PAGE, HPLC profile and MALDI-TOF mass spectrometry analysis. Partial sequences of metalloproteinases, phospholipases A₂ (PLA₂) and galactose-specific lectins were identified in the venoms. Venoms of C. l. klauberi and C. l. lepidus had significantly higher hemorrhagic and lethal activities than C. l. morulus venom. Proteolytic activity in azocasein was higher in C. l. morulus venom, whereas gelatin hydrolysis was higher in C. l. klauberi. Fibrinogenolytic and PLA₂ activities were very similar in all venoms tested. The histological observations in the gastrocnemius muscle damaged by venoms from all the subspecies confirmed myonecrotic and hemorrhagic activities (at 3 and 24 h), which resulted in a poor regenerative response after 14 days. However, C. l. lepidus and C. l. klauberi venom induced a higher increase in the plasma activity of creatine kinase (CK), evidencing higher myotoxicity, whereas paw edema-inducing activity was higher in C. l. lepidus venom. The results indicate that the venoms from the three subspecies have similar protein profiles in electrophoresis, HPLC and molecular weight determinations. However, differences were found in the biological activities in mice. Notably, the venoms of C. l. lepidus and C. l. klauberi present higher

  14. Isolation and characterisation of insulin-releasing compounds from Crotalus adamanteus, Crotalus vegrandis and Bitis nasicornis venom.

    Science.gov (United States)

    Moore, Sara W M; Bhat, Vikas K; Flatt, Peter R; Gault, Victor A; McClean, Stephen

    2015-07-01

    Crude venom from three venomous snakes, Crotalus adamanteus, Crotalus vegrandis and Bitis nasicornis was fractionated by gel filtration chromatography, and selected fractions screened for in-vitro insulinotropic activity using clonal pancreatic BRIN-BD11 cells. Nineteen fractions stimulated insulin secretion and the structural identity of bioactive compounds responsible was probed using MALDI-ToF MS and N-terminal Edman degradation sequencing. Partial N-terminal sequences were determined and their homology to existing sequences identified using BLAST searching. The main insulinotropic peptide families identified were made up of snake venom serine proteinases, phospholipases A2 (PLA2) and disintegrins. Snake venom constituents may have therapeutic potential for diabetes, with each of the three viper venoms tested providing insulinotropic compounds from a range of different toxin families. Copyright © 2015 Elsevier Ltd. All rights reserved.

  15. Absence of phospholipase A(2) in most Crotalus horridus venom due to translation blockage: comparison with Crotalus horridus atricaudatus venom.

    Science.gov (United States)

    Wang, Ying-Ming; Parmelee, Jeffrey; Guo, Yaw-Wen; Tsai, Inn-Ho

    2010-08-01

    To investigate the peculiar absence of phospholipases A(2) (PLA(2)s) in most Crotalus horridus (CH) venom, we cloned and sequenced the venom PLA(2)s of three CH specimens from different regions. The results revealed that all the venom glands contained mRNAs that encoded an acidic PLA(2) (designated as either CH-E6 or CH-E6'). The predicted CH-E6 from the Iowan CH and CH-E6' from the South Carolinian CH differed by only one amino acid residue, while the PLA(2) cDNA cloned from the Kentuckian CH contained an early stop codon instead of a Tyr(22) codon. Only the individual South Carolinian CH venom was found to contain the CH-E6' protein whose mass was confirmed by MALDI-TOF analysis. Our results suggest that low PLA(2) expression levels in most CH venom can be attributed to translation blockage. We also purified two acidic PLA(2)s and canebrake toxin from the pooled venom of Crotalus horridus atricaudatus (neurotoxic CH subspecies). One of the acidic PLA(2)s was identical to CH-E6 and showed high lipolytic activity and weak anti-platelet activities. The possibility that C. h. atricaudatus could be a hybrid between CH and Crotalus scutulatus is discussed. Copyright 2010 Elsevier Ltd. All rights reserved.

  16. 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...

  17. A comparative analysis of hemolithic activity between Cortalus molossus and Crotalus molossus nigrescens subespecies

    Directory of Open Access Journals (Sweden)

    Martínez-Martínez, A.

    2014-01-01

    Full Text Available Blacktailed rattlesnake (Crotalus molossus has four subspecies distributed in México; two of them are distributed in Chihuahua. Literature shows that few studies have been done in order to analyze the composition of their venom. Therefore, this study was carried out to evaluate protein content and hemolytic damage by agarose-human erythrocyte substrate between the Crotalus molossus nigrescens and Crotalus molossus molossus subspecies. Mann Whitney’s nonparanteric test was used in order to get the statistics analysis at an IC95. Protein concentration of Crotalus m. nigrescens was 466 ± 35.0 mg mL-1 and Crotalus m. molossus of 449 ± 90.0 mg mL-1 there was no statistical difference detected at IC95 (p>0.05. The protein profile of the venom of Crotalus m. nigrescens showed two protein bands with elevated masses approximately 230 kDa in comparison with Crotalus m. molossus that was of 16 kDa approximately. Crotalus m. nigrescens’s venom was 18 % more hemolytic (156.7±23.33 UHD than Crotalus m. molossus’s (128±23.32 UHD at the first hour and 20 % more hemolytic at the second hour, with UHD values of 116.7 ± 12.3 and 94 ± 9.8, respectively. The venom of the two subspecies did not show differences statistically at an IC95 at the first hour (p>0.05 and second hour (p>0.05. It could be concluded that two subspecies samples showed iqual proteomic characteristics. These differences can be attributed to the different eating habits due to the altitude, geographical distribution and the season. It can also be concluded that more investigation regarding venom of rattlesnakes is needed.

  18. Final Critical Habitat for the New Mexican ridge-nosed rattlesnake (Crotalus willardi obscurus)

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — To provide the user with a general idea of areas where final critical habitat for New Mexican ridge-nosed rattlesnake (Crotalus willardi obscurus) occur based on the...

  19. Anaphylatic reaction after Crotalus envenomation treatment in a dog: case report

    Directory of Open Access Journals (Sweden)

    L. G. Conceição

    2007-01-01

    Full Text Available Crotalus envenomation is the second most frequent condition caused by venomous snakes attacking dogs and cats in Brazil. Affected animals show severe neurological and muscular involvement, lower motor neuron signs and myalgia due to extensive rabdomiolysis. The present case report is about Crotalus envenomation in a dog which showed severe clinical and laboratory abnormalities. After treatment with antivenom (specific antidote and clinical support, the dog totally recovered from that condition.

  20. Cryptic sociality in rattlesnakes (Crotalus horridus) detected by kinship analysis.

    Science.gov (United States)

    Clark, Rulon W; Brown, William S; Stechert, Randy; Greene, Harry W

    2012-08-23

    Research on social behaviour has largely concentrated on birds and mammals in visually active, cooperatively breeding groups (although such systems are relatively rare) and focused much less on species that rarely interact other than for mating and parental care. We used microsatellite markers to characterize relatedness among aggregations of timber rattlesnakes (Crotalus horridus), a putatively solitary reptile that relies heavily on chemical cues, and found that juveniles and pregnant females preferentially aggregate with kin under certain conditions. The ability to recognize kin and enhance indirect fitness thus might be far more widespread than implied by studies of animals whose behaviour is primarily visually and/or acoustically mediated, and we predict that molecular markers will reveal many additional examples of 'cryptic' sociality.

  1. 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

  2. Ultrastructural description and cytochemical study of the spermatozoon of Crotallus durissus (Squamata, Serpentes).

    Science.gov (United States)

    Cunha, L D; Tavares-Bastos, L; Báo, S N

    2008-10-01

    The present study was undertaken to elucidate some aspects about the nature of the spermatozoon ultrastructure of Crotallus durissus using cytochemical methods. We also provide for the first time the ultrastructural description of this species spermatozoon. Cytochemical studies of spermatozoa have not been performed so far in the Serpentes, and species spermatozoon may prove helpful to better understand the reproductive biology of this group. Besides the synapomorphies of the Squamata and Serpentes, the C. durissus spermatozoon possess the following: circular acrosome tip; rounded perforatorium tip with a stopper-like basal modification; bilateral stratified laminar structures; central electron-dense structure within the proximal centriole; fibrous sheath extending until the level of the second mitochondrial ring; rounded mitochondria in cross-section, but with variable shape and organization in longitudinal and oblique sections, respectively; linear annulus; developed multilaminar membranes in the nuclear region and the midpiece. The formation of membrane filipin-sterol complexes occur sparsely along the head region, specially around the nucleus; the complexes were also present in the midpiece membrane and scarcely lining the flagellum. The complexes were present in the different layers of the multilaminar membranes. The ethanol-phosphotungstic acid (E-PTA) treatment relieved the presence of basic proteins in acrosome vesicle, pericentriolar material, peripheral fibers of the axoneme and fibrous sheath. The tannic acid technique revealed the microtubules of the centrioles and the axoneme; the extracellular tubules encircling the spermatozoa and those spread in the epididymal lumen were also observed. However, the immunocytochemistry assay using antibodies against alpha-tubulin and beta-tubulin, the primary microtubule monomers, does not support the existence of composition similarity between these tubular structures, since the extracellular tubules were not

  3. Characterization of microsatellite loci isolated in midget-faded rattlesnakes (Crotalus viridis concolor)

    Science.gov (United States)

    Oyler-McCance, Sara J.; St. John, J.; Parker, J.M.; Anderson, S.H.

    2005-01-01

    Primers for five polymorphic microsatellite loci were developed for the midget faded rattlesnake (Crotalus viridis concolor), a rare subspecies of western rattlesnake (Crotalus viridus) found only in parts of Wyoming, Colorado, and Utah. Five polymorphic microsatellites were isolated, four of which had relatively high levels of diversity (eight to nine alleles). We found only two departures from Hardy–Weinberg equilibrium and they occurred in different loci, so null alleles are likely not a problem. Moreover, we found that no two loci were linked. These loci will be applicable for population genetic analysis and perhaps analysis of paternity and mating systems.

  4. Diet of Crotalus lepidus klauberi (Banded Rock Rattlesnake)

    Science.gov (United States)

    Holycross, A.T.; Painter, C.W.; Prival, D.B.; Swann, D.E.; Schroff, M.J.; Edwards, T.; Schwalbe, C.R.

    2002-01-01

    We describe the diet of Crotalus lepidus klauberi (Banded Rock Rattlesnake) using samples collected in the field and from museum specimens, as well as several records from unpublished reports. Most records (approximately 91%) were from the northern Sierra Madrean Archipelago. Diet consisted of 55.4% lizards, 28.3% scolopendromorph centipedes, 13.8% mammals, 1.9% birds, and 0.6% snakes. Sceloporus spp. comprised 92.4% of lizards. Extrapolation suggests that Sceloporus jarrovii represents 82.3% of lizard records. Diet was independent of geographic distribution (mountain range), sex, source of sample (stomach vs. intestine/feces), and age class. However, predator snout-vent length differed significantly among prey types; snakes that ate birds were longest, followed in turn by those that ate mammals, lizards, and centipedes. Collection date also differed significantly among prey classes; the mean date for centipede records was later than the mean date for squamate, bird, or mammal records. We found no difference in the elevation of collection sites among prey classes.

  5. Response of western diamondback rattlesnakes Crotalus atrox to airborne sounds.

    Science.gov (United States)

    Young, Bruce A; Aguiar, Amie

    2002-10-01

    In order to test the hypothesis that snakes can not only perceive airborne sounds, but also respond to them, an acoustic isolation chamber was designed and constructed to perform best within the 150-450 Hz range in which snakes perceive sound. Suspended within this acoustic chamber was a steel mesh basket designed to minimize the potential for groundborne vibrations. A synthesized tone was created out of 20 different 150 ms sounds, each separated by a 50 ms period of silence; the acoustic energy of each of the 20 sounds was concentrated between 200-400 Hz, and each sound included frequency modulation. The trial stimuli were presented to western diamondback rattlesnakes Crotalus atrox at a level 5-10 dB above their perception threshold. Four significant behavioural responses were observed upon stimulus presentation: cessation of body movements, reduction or cessation of tongue flicking, rapid jerks of the head and rattling. At least one significant behavioural response was observed in 92% of the behavioural trials. This study provides the first experimental evidence that snakes can respond behaviourally to airborne sounds.

  6. Seasonal variation in hematology and blood plasma chemistry values of the timber rattlesnake (Crotalus horridus).

    Science.gov (United States)

    LaGrange, Seth M; Kimble, Steven J A; MacGowan, Brian J; Williams, Rod N

    2014-10-01

    Hematology, biochemical analyses, and body condition indices are useful tools for describing animal health, especially when making management decisions for species of conservation concern. We report hematologic, biochemical, and body condition index data for 13 free-ranging timber rattlesnakes (Crotalus horridus) sampled repeatedly over an active season in Indiana, USA.

  7. 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...

  8. Arboreal behavior in the timber rattlesnake, Crotalus horridus, in eastern Texas

    Science.gov (United States)

    D. Craig Rudolph; R. R. Schaefer; D. Saenz; R. N. Conner

    2004-01-01

    There have been several recent reports, and anecdotal observations extending back at least to J. J. Audubon, suggesting that the timber rattlesnake (Crotalus horridus) is one of the most arboreal members of the genus. Most previous records are of snakes located at heights of less than 5 m. Telemetry studies in eastern Texas have documented more...

  9. Comparison of the protective effect of a commercially available western diamondback rattlesnake toxoid vaccine for dogs against envenomation of mice with western diamondback rattlesnake (Crotalus atrox), northern Pacific rattlesnake (Crotalus oreganus oreganus), and southern Pacific rattlesnake (Crotalus oreganus helleri) venom.

    Science.gov (United States)

    Cates, Charles C; Valore, Erika V; Couto, Marcelo A; Lawson, Gregory W; McCabe, James G

    2015-03-01

    To evaluate effectiveness of a commercially available toxoid manufactured from western diamondback (WD) rattlesnake (Crotalus atrox) venom against envenomation of mice with WD, northern Pacific (NP) rattlesnake (Crotalus oreganus oreganus), and southern Pacific (SP) rattlesnake (Crotalus oreganus helleri) venom. 90 specific pathogen-free female mice. Mice were allocated into 3 cohorts (30 mice/cohort). Mice received SC injections of C atrox toxoid (CAT) vaccine (n = 15/group) or adjuvant (15/group) at day 0 and again at 4 weeks. At 8 weeks, mice were challenge-exposed with 1 of 3 venoms. Survival until 48 hours was evaluated by use of log-rank analysis of survival curves and the z test for proportions. 6 of 15 WD-challenged CAT-vaccinated mice, 3 of 15 NP-challenged CAT-vaccinated mice, and 0 of 15 SP-challenged CAT-vaccinated mice survived until 48 hours. All adjuvant-only vaccinates survived ≤ 21 hours. Mean survival time of CAT vaccinates was longer than that of adjuvant-only vaccinates for all venoms (1,311 vs 368 minutes for WD, 842 vs 284 minutes for NP, and 697 vs 585 minutes for SP). Results of the z test indicated a significantly increased survival rate for vaccinates exposed to WD rattlesnake venom but not for vaccinates exposed to NP or SP rattlesnake venom. Log-rank analysis revealed a significant difference between survival curves of vaccinated versus unvaccinated mice exposed to NP but not WD or SP venom. CAT vaccination improved survival rate and survival time after challenge exposure with WD rattlesnake venom and may offer limited protection against NP rattlesnake venom but did not provide significant cross-protection against SP rattlesnake venom.

  10. Body Size Evolution in Insular Speckled Rattlesnakes (Viperidae: Crotalus mitchellii)

    Science.gov (United States)

    Meik, Jesse M.; Lawing, A. Michelle; Pires-daSilva, André

    2010-01-01

    Background 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. Methodology/Principal Findings Insular speckled rattlesnakes show considerable variation in body size when compared to mainland source subspecies. In addition to previously known instances of gigantism on Ángel 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. Conclusions/Significance 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

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

    Directory of Open Access Journals (Sweden)

    Jesse M Meik

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

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

    Science.gov (United States)

    Meik, Jesse M; Lawing, A Michelle; Pires-daSilva, André

    2010-03-04

    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 what are

  13. Crotalus aquilus in the Mexican state of Mexico consumes a diverse summer diet

    Science.gov (United States)

    Mociño-deloya, E.; Setser, K.; Peurach, S.C.; Meik, J.M.

    2008-01-01

    We report observations of the summer diet of Crotalus aquilus (Queretaro dusky rattlesnake) from an agricultural region near San Pedro de los Metates, municipality of Acambay, state of Mexico, Mexico. We recovered the remains of 12 individual prey items from 11 different snakes. Eleven of 38 (29%) snakes observed contained prey remains, including 6 mammals, 3 lizards, and 3 snakes. These observations suggest that C. aquilus consumes a diverse diet and that they may be more ophiophagous than many other rattlesnakes.

  14. Clinical presentation and management of an Aruban rattlesnake bite in the Netherlands

    NARCIS (Netherlands)

    Dijkman, Marieke A; Damhuis, Dorien E M; Meulenbelt, Jan; de Vries, Irma|info:eu-repo/dai/nl/41398625X

    Bites by Aruban Rattlesnake (Crotalus durissus unicolor) are rare and not known to induce severe envenomations. Here, we present a case of a 57 year-old man bitten by his pet Aruban Rattlesnake (Crotalus durissus unicolor). He was admitted to hospital within 15 min. Three and a half hours later his

  15. SNAKE VENOMICS OF Crotalus tigris: THE MINIMALIST TOXIN ARSENAL OF THE DEADLIEST NEARTIC RATTLESNAKE VENOM

    Science.gov (United States)

    CALVETE, Juan J.; PÉREZ, Alicia; LOMONTE, Bruno; SÁNCHEZ, Elda E.; SANZ, Libia

    2012-01-01

    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 PLA2, 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 represents 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 towards neurotoxicity, which has been also reported for the South American rattlesnakes, may have resulted by paedomorphism. The ability of an experimental antivenom to effectively immunodeplete proteins from the type II venoms of C. tigris, C. horridus, C. oreganus helleri, C. scutulatus scutulatus, and S. 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 American and North American Crotalus. PMID:22181673

  16. 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

  17. 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.

  18. Ophidiomyces ophiodiicola dermatitis in eight free-ranging timber rattlesnakes (Crotalus horridus) from Massachusetts.

    Science.gov (United States)

    McBride, Michael P; Wojick, Kimberlee B; Georoff, Timothy A; Kimbro, Jason; Garner, Michael M; Wang, Xiaoling; Childress, April L; Wellehan, James F X

    2015-03-01

    Eight free-ranging timber rattlesnakes (Crotalus horridus) from two geographically isolated Massachusetts populations were observed with skin lesions located primarily on the head but occasionally also on the lateral and ventral surfaces of the body. The snakes underwent health assessments that included physical examination, clinical pathology, full body radiographs, and full thickness biopsies of skin lesions. Each snake had fungal elements present histologically in tissue sections from skin lesions. Ophidiomyces ophiodiicola was identified from skin lesions using polymerase chain reaction in all eight snakes.

  19. Response of Western Diamondback Rattlesnakes ("Crotalus atrox") to Chemical Cues of Mice ("Mus musculus") of Different Genders and Reproductive Status

    Science.gov (United States)

    Saviola, Anthony J.; Chiszar, David; Bealor, Matthew T.; Smith, Hobart M.

    2010-01-01

    Eight western diamondback rattlesnakes ("Crotalus atrox") were exposed to 6 stimuli: (1) clean, unused bedding; (2) an adult male mouse; (3) an adult lactating female mouse; (4) an adult lactating female mouse with a litter; (5) 2 adult nonlactating female mice, to control for the extra surface area in Condition 4; and (6) a litter of newborn…

  20. A Comparison of the Crystal Structures of Phospholipase A2 from Bovine Pancreas and Crotalus atrox Venom

    NARCIS (Netherlands)

    RENETSEDER, R; BRUNIE, S; DRENTH, J; SIGLER, PB

    1985-01-01

    The refined high resolution crystal structure of the bovine phospholipase A2 was compared with its counterpart from the venom of Crotalus atrox, the western diamondbacked rattlesnake. The strong similarity in their backbone conformations forms the basis of a common numbering system for the amino

  1. 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

  2. Iron and carbon monoxide attenuate degradation of plasmatic coagulation by Crotalus atrox venom.

    Science.gov (United States)

    Nielsen, Vance G; Boyer, Leslie V

    2016-07-01

    Hypofibrinogenemia is an important clinical consequence following envenomation by Crotalus species, usually attenuated or prevented by administration of antivenom. It has been determined that iron and carbon monoxide (CO) enhance fibrinogen as a thrombin substrate, likely secondary to conformational changes in molecular structure. We tested the hypothesis that pretreatment of plasma with iron and CO could attenuate the effects of exposure to Crotalus atrox venom. Human plasma was exposed to 0 to 10 μmol/l ferric chloride (iron source) and 0 to 100 μmol/l CO-releasing molecule-2 (CO source) followed by exposure to 0 to 0.5 μg/ml venom for 5 to 20 min. Changes in coagulation kinetics were determined with thrombelastography. Iron and CO significantly attenuated venom-mediated degradation of plasmatic coagulation in terms of onset time, velocity of clot growth and final clot strength. Further preclinical investigation of iron and CO administration as a 'bridge-to-antivenom' to preserve plasmatic coagulation is justified.

  3. Proteomic, toxicological and immunogenic characterization of Mexican west-coast rattlesnake (Crotalus basiliscus) venom and its immunological relatedness with the venom of Central American rattlesnake (Crotalus simus).

    Science.gov (United States)

    Segura, Álvaro; Herrera, María; Reta Mares, Francisco; Jaime, Claudia; Sánchez, Andrés; Vargas, Mariángela; Villalta, Mauren; Gómez, Aarón; Gutiérrez, José María; León, Guillermo

    2017-03-31

    The venom of the Mexican west-coast rattlesnake (Crotalus basiliscus) was characterized for its protein composition, toxicological profile and immunogenic properties. This venom is composed of 68% Zn 2+ -dependent metalloproteinases (SVMPs), 14% phospholipases A 2 (PLA 2 s), 11% serine proteinases, 4% SVMPs-inhibitor tripeptides (SVMP-ITs), 2% bradykinin-potentiating peptides (BPPs), 0.6% cysteine-rich secretory proteins (CRISPs), and 0.2% l-amino acid oxidases (LAAOs). SVMPs present in the venom are responsible for azocasein hydrolysis and hemorrhagic activity, but their contribution to the lethal activity of the venom in mice is masked by the neurotoxic activity of PLA 2 s, which in addition are also responsible for myotoxic activity. Despite its relatively high content of serine proteinases, the venom of C. basiliscus did not exert in vitro coagulant or in vivo defibrinogenating activities. The ability of antivenoms raised against the venoms of C. basiliscus and C. simus (from Costa Rica) to neutralize homologous and heterologous venoms revealed antigenic similarities between toxins of both venoms. Preclinical evaluation of an antivenom produced by using the venom of C. basiliscus as immunogen demonstrated that it is able to neutralize not only the most relevant toxic activities of C. basiliscus venom, but also those exerted by Costa Rican C. simus venom, including coagulant and defibrinogenating activities. The Central American rattlesnake (Crotalus simus) is widely distributed from Mexico to west central Costa Rica, and induces an important number of envenomations in this region. On the other hand, the immunogenic mixture used by Laboratorios de Biológicos y Reactivos de Mexico S.A. (Birmex) to produce the snake antivenom more frequently used in Mexico does not include the venom of C. simus. This immunogenic mixture is composed by the venoms of the Fer-de-lance (Bothrops asper) and the Mexican west-coast rattlesnake (Crotalus basiliscus). We studied the

  4. Preparation of crotalus venom radiolabeled with technetium-99m as a tool for biodistribution study

    Directory of Open Access Journals (Sweden)

    Priscilla Brunelli Pujatti

    2005-10-01

    Full Text Available Technetium-99m (99mTc has been the radionuclide of choice for nuclear medicine procedures and experimental research. Because of its optimal nuclear properties, 99mTc 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 99mTc 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 99mTc-CV can be a useful tool for biodistribution studies.Tecnécio-99m tem sido o radioisótopo de escolha para procedimentos médicos e pesquisas experimentais. Em decorrência de suas propriedades nucleares, 99mTc é adequado para detecção de alta eficiência com a vantagem do baixo risco radiológico. O veneno de Crotalus (CV apresentou propriedades antitumorais em estudos clínicos e estudos de biodistribuição são fundamentais em pesquisas clínicas. Esse trabalho teve como objetivo obter um análogo de veneno de Crotalus marcado com 99mTc que preservasse sua atividade biológica. Após a marcação, a atividade biológica foi avaliada através do ensaio de atividade hemolítica. Veneno nativo e marcado apresentaram atividade hemolítica indireta quando incubados em um meio contendo uma fonte exógena de lecitina. Obteve-se um alto rendimento de marcação e a atividade biológica das moléculas foi preservada. Nossos resultados sugerem que 99mTc-CV pode representar uma ferramenta muito útil para estudos de biodistribuição.

  5. A case of neurotoxicity following envenomation by the Sidewinder rattlesnake, Crotalus cerastes.

    Science.gov (United States)

    Bosak, Adam R; Ruha, Anne-Michelle; Graeme, Kimberlie A

    2014-06-01

    North American rattlesnake envenomations typically result in local tissue injury and hematologic derangements. Neurotoxicity is uncommon but when present often manifests as fasciculations and paresthesias. Neurotoxicity following Sidewinder (Crotalus cerastes) envenomation has not been previously reported. A 56-year-old man bitten on the right foot developed painful paresthesias, weakness and fasciculations of the right lower extremity, and involuntary muscle contractions of the anterior thigh. Local tissue effects and hemotoxicity never developed. The patient was discharged 5 days after the bite with resolution of fasciculations but continued to have right-sided weakness. The snake was identified as a Sidewinder, C. cerastes, by the patient and two independent herpetologists. This is the first reported case of a Sidewinder rattlesnake envenomation resulting in neurotoxicity.

  6. Ecology and behavior of the Midget Faded Rattlesnake (Crotalus oreganus concolor) in Wyoming

    Science.gov (United States)

    Parker, J.M.; Anderson, S.H.

    2007-01-01

    We conducted a three-year study to describe the ecology and behavior of the Midget Faded Rattlesnake, Crotalus organus concolor. We encountered 426 and telemetered 50 C. o. concolor between 2000 and 2002. We found that their primary diet was lizards (associated with rock outcrops), though they will consume small mammals and birds. They den in aggregations, although in low numbers when compared to other subspecies. Movements and activity ranges were among the largest reported for rattlesnakes. Minimum convex polygon area was 117.8 ha for males, 63.9 ha for nongravid females, and 4.8 ha for gravid females. Mean distances traveled per year were 2122.0 m for males, 1956.0 m for nongravid females, and 296.7 m for gravid and postpartum females. Following emergence from hibernation, they spent several weeks shedding, often in aggregations before migration, and migrations occurred in early summer. Most snakes made straight-line movements to and from discrete summer activity ranges where short, multidirectional movements ensued, although others made multidirectional movements throughout the active season. We observed mating behavior between 21 July and 12 August. Gravid females gave birth during the third week of August. Mean clutch size was 4.17 (range 2-7). We found that the sex ratio was skewed favoring females 1:1.24, and they were sexually dimorphic in size (males SVL = 44.1 cm; females SVL = 40.8 cm). Our data further illustrate the diversity within the large group of Western Rattlesnakes (Crotalus viridis). Copyright 2007 Society for the Study of Amphibians and Reptiles.

  7. Phylogenetic relationships of the enigmatic longtailed rattlesnakes (Crotalus ericsmithi, C. lannomi, and C. stejnegeri).

    Science.gov (United States)

    Reyes-Velasco, Jacobo; Meik, Jesse M; Smith, Eric N; Castoe, Todd A

    2013-12-01

    The longtailed rattlesnakes of western Mexico represent an enigmatic group of poorly known venomous snake species: Crotalus ericsmithi, C. lannomi, and C. stejnegeri. In the 120 years since their discovery, fewer than twenty individuals have been deposited in natural history collections worldwide. These three species share similar morphological traits, including a particularly long tail that has been interpreted as either an ancestral condition among rattlesnakes or as derived within the longtailed group. An understanding of the phylogenetic distinctiveness and relationships among the longtailed rattlesnakes, and their relationships to other rattlesnake groups, has previously been hampered by a dearth of comparative material and tissues for collection of DNA sequence data. Facilitated by the recent availability of tissue samples from multiple individuals of each species, we estimate the phylogenetic relationships among the longtailed rattlesnakes and their placement among other rattlesnake groups, using DNA sequence data from three mitochondrial and three nuclear gene fragments. We explore phylogenetic signal in our data using Bayesian and maximum likelihood methods, species tree analyses and hypothesis testing. Our results strongly support the monophyly of longtailed rattlesnakes and suggest the three species diverged from each other during the mid to late Pliocene or early Pleistocene (~1.5-5.6 mya). Contrary to prevailing hypotheses, we find no evidence for an early or basal divergence of the longtailed clade within the rattlesnake tree, and instead estimate that it diverged relatively recently (~6.8 mya) from its sister lineage, composed of the diamondback rattlesnakes (C. atrox group) and the prairie rattlesnakes (C. viridis group). With our added sampling of lineages and identification of previously used problematic sequences, we provide a revised hypothesis for relationships among Crotalus species, yet underscore the need for future studies and new data to

  8. 76 FR 77006 - Endangered Species; Receipt of Applications for Permit; Correction

    Science.gov (United States)

    2011-12-09

    ... 50 CFR 17.21(g) to include the Aruba island rattlesnake (Crotalus durissus unicolor), to enhance...) in the wild in Madagascar for the purpose of scientific research. Multiple Applicants The following...

  9. Radio Transmitter Implantation and Movement in the Wild Timber Rattlesnake ( Crotalus horridus ).

    Science.gov (United States)

    Hale, Vanessa L; MacGowan, Brian; Corriveau, Lorraine; Huse, David C; Currylow, Andrea F T; Thompson, Steve

    2017-07-01

    Radiotelemetry transmitters have become critical to studies of wildlife ecology. However, little is known about how transmitter implantation surgery affects the mobility of some species, including the timber rattlesnake, Crotalus horridus . Tracking snake movement can provide insights into the effects of transmitter implantation. During 2007-11, 71 radio transmitters were surgically implanted intracoelomically in 47 timber rattlesnakes. Over 20 of these snakes underwent surgery at least twice in 5 yr to replace old transmitters. Surgeries were performed under general anesthesia with a local nerve block at the site of implantation, 20 cm cranial to the cloaca. Snakes were also administered postsurgical meloxicam and enrofloxacin every 24 h for three doses. Two to five days after surgery, snakes were released at their original locations and radiotracked regularly during the active seasons (April-October 2007-11). Average daily movement data (distance traveled) were compiled for each snake. Snakes undergoing transmitter surgery in a given year did not differ significantly in distance traveled compared to snakes that had transmitters but did not have surgery in that year. Distance traveled for each snake did not differ before or after surgery or between weeks 1 and 2 postsurgery, indicating that the transmitter implantation did not alter snake movement.

  10. VENOM VARIATION IN HEMOSTASIS OF THE SOUTHERN PACIFIC RATTLESNAKE (Crotalus oreganus helleri): ISOLATION OF HELLERASE

    Science.gov (United States)

    Salazar, Ana Maria; Guerrero, Belsy; Cantu, Bruno; Cantu, Esteban; Rodríguez-Acosta, Alexis; Pérez, John C.; Galán, Jacob A.; Tao, Andy; Sánchez, Elda E.

    2009-01-01

    Envenomations by the Southern Pacific Rattlesnake (Crotalus oreganus helleri) are the most common snakebite accidents in southern California. Intraspecies venom variation may lead to unresponsiveness of antivenom therapy. Even in a known species, venom toxins are recognized as diverse in conformity with interpopulational, seasonal, ontogenetic and individual factors. Five venoms of individual C. o. helleri located in Riverside and San Bernardino counties of southern California were studied for their variation in their hemostasis activity. The results demonstrated that Riverside 2 and San Bernardino 1 venoms presented the highest lethal activity without hemorrhagic activity. In contrast, San Bernardino 2 and 3 venoms had the highest hemorrhagic and fibrinolytic activities with low lethal and coagulant activities. Riverside 1, Riverside 2 and San Bernardino 1 venoms presented a significant thrombin-like activity. San Bernardino 2 and 3 venoms presented an insignificant thrombin-like activity. In relation to the fibrinolytic activity, San Bernardino 3 venom was the most active on fibrin plates, which was in turn neutralized by metal chelating inhibitors. These results demonstrate the differences amongst C. o helleri venoms from close localities. A metalloproteinase, hellerase, was purified by anionic and cationic exchange chromatography from San Bernardino 3 venom. Hellerase exhibited the ability to break fibrin clots in vitro, which can be of biomedically importance in the treatment of heart attacks and strokes. PMID:18804187

  11. Incipient speciation with biased gene flow between two lineages of the Western Diamondback Rattlesnake (Crotalus atrox).

    Science.gov (United States)

    Schield, Drew R; Card, Daren C; Adams, Richard H; Jezkova, Tereza; Reyes-Velasco, Jacobo; Proctor, F Nicole; Spencer, Carol L; Herrmann, Hans-Werner; Mackessy, Stephen P; Castoe, Todd A

    2015-02-01

    We used mitochondrial DNA sequence data from 151 individuals to estimate population genetic structure across the range of the Western Diamondback Rattlesnake (Crotalus atrox), a widely distributed North American pit viper. We also tested hypotheses of population structure using double-digest restriction site associated DNA (ddRADseq) data, incorporating thousands of nuclear genome-wide SNPs from 42 individuals. We found strong mitochondrial support for a deep divergence between eastern and western C. atrox populations, and subsequent intermixing of these populations in the Inter-Pecos region of the United States and Mexico. Our nuclear RADseq data also identify these two distinct lineages of C. atrox, and provide evidence for nuclear admixture of eastern and western alleles across a broad geographic region. We identified contrasting patterns of mitochondrial and nuclear genetic variation across this genetic fusion zone that indicate partially restricted patterns of gene flow, which may be due to either pre- or post-zygotic isolating mechanisms. The failure of these two lineages to maintain complete genetic isolation, and evidence for partially-restricted gene flow, imply that these lineages were in the early stages of speciation prior to secondary contact. Copyright © 2014 Elsevier Inc. All rights reserved.

  12. Zebrin II / aldolase C expression in the cerebellum of the western diamondback rattlesnake (Crotalus atrox).

    Science.gov (United States)

    Aspden, Joel W; Armstrong, Carol L; Gutierrez-Ibanez, Cristian I; Hawkes, Richard; Iwaniuk, Andrew N; Kohl, Tobias; Graham, David J; Wylie, Douglas R

    2015-01-01

    Aldolase C, also known as Zebrin II (ZII), is a glycolytic enzyme that is expressed in cerebellar Purkinje cells of the vertebrate cerebellum. In both mammals and birds, ZII is expressed heterogeneously, such that there are sagittal stripes of Purkinje cells with high ZII expression (ZII+), alternating with stripes of Purkinje cells with little or no expression (ZII-). The patterns of ZII+ and ZII- stripes in the cerebellum of birds and mammals are strikingly similar, suggesting that it may have first evolved in the stem reptiles. In this study, we examined the expression of ZII in the cerebellum of the western diamondback rattlesnake (Crotalus atrox). In contrast to birds and mammals, the cerebellum of the rattlesnake is much smaller and simpler, consisting of a small, unfoliated dome of cells. A pattern of alternating ZII+ and ZII- sagittal stripes cells was not observed: rather all Purkinje cells were ZII+. This suggests that ZII stripes have either been lost in snakes or that they evolved convergently in birds and mammals.

  13. Responses of infrared-sensitive tectal units of the pit viper Crotalus atrox to moving objects.

    Science.gov (United States)

    Kaldenbach, Felix; Bleckmann, Horst; Kohl, Tobias

    2016-06-01

    Rattlesnakes perceive IR radiation with their pit organs. This enables them to detect and strike towards warm-blooded prey even in the dark. In addition, the IR sense allows rattlesnakes to find places for thermoregulation. Animate objects (e.g., prey) tend to move and thus cause moving IR images across the pit membrane. Even when an object is stationary, scanning head movements of rattlesnakes will result in moving IR images across the pit membrane. We recorded the neuronal activity of IR-sensitive tectal neurons of the rattlesnake Crotalus atrox while stimulating the snakes with an IR source that moved horizontally at various velocities. As long as object velocity was low (angular velocity of ~5°/s) IR-sensitive tectal neurons hardly showed any responses. With increasing object velocity though, neuronal activity reached a maximum at ~50°/s. A further increase in object velocity up to ~120°/s resulted in a slight decrease of neuronal activity. Our results demonstrate the importance of moving stimuli for the snake's IR detection abilities: in contrast to fast moving objects, stationary or slowly moving objects will not be detected when the snake is motionless, but might be detected by scanning head movements.

  14. Short Note. Chemosensory response in stunted prairie rattle snakes Crotalus viridis viridis

    Directory of Open Access Journals (Sweden)

    Anthony J. SAVIOLA, David CHISZAR, Hobart M. SMITH,Stephen P. MACKESSY

    2013-04-01

    Full Text Available Rattlesnakes use chemical stimuli in ambush site selection and for relocation of envenomated prey through strike-induced chemosensory searching. Shifts in responsiveness to prey chemicals have been documented in many snakes, and often correlate with prey commonly taken as snakes increase in age and size as well as geographical locations of the species. For instance, neonate rattlesnakes that prey primarily on ectotherms responded most strongly to chemical cues of commonly taken lizard prey, whereas adult rattlesnakes that prey primarily on small mammals responded significantly to chemical cues of commonly taken rodents. In the current study, 11 Prairie Rattlesnakes Crotalus viridis viridis which were classified as large neonates based on measures of snout-vent length (SVL and body mass, yet chronologically were at or near adulthood, were tested for their responsiveness to chemical extracts of natural and non-natural prey items. Although the snakes had eaten only neonate lab mice (Mus musculus, they responded significantly more to chemical cues of natural prey items and particularly to chemical cues of prey normally taken by subadults (Peromyscus mice and Sceloporus lizard. These results suggest that ontogenetic shifts in responsiveness to natural prey chemical cues are innately programmed and are not based on body size or feeding experience in C. v. viridis. This does not imply, however, that growth and experience are without effects, especially with novel prey or rare prey that have experienced recent population expansion [Current Zoology 59 (2: 175-179, 2013].

  15. Natural variation in steroid hormone profiles of male Timber Rattlesnakes, Crotalus horridus, in northwest Arkansas.

    Science.gov (United States)

    Lind, Craig M; Beaupre, Steven J

    2014-09-15

    We describe the seasonal profile of circulating steroid hormones (testosterone and corticosterone) in relation to the breeding season in free ranging male Timber Rattlesnakes, Crotalus horridus, over the course of three active seasons. In addition, we examine variation in steroid concentrations across years and in relation to body condition. We found that seasonal profiles of plasma testosterone were different compared to other crotalines with similar mating patterns. Concentrations of testosterone were elevated above baseline in the three months leading up to the single late summer breeding season. Testosterone peaked in July at the onset of the breeding season and dropped to baseline during the peak months of breeding (August and September). Testosterone concentrations also varied annually. Although the exact cause of annual variation could not be established, our results indicate that weather patterns may have driven observed differences. Testosterone concentrations were positively related to body condition, indicating that testosterone production is modulated according to energetic status (particularly in the two months prior to the breeding season). Corticosterone did not vary seasonally or with any measured variable, a result similar to other studied crotalines. Our results highlight the importance of long-term descriptive studies of the regulatory mechanisms that underlie behavior and physiology in diverse taxa, as these mechanisms can vary greatly within and among populations and are valuable in elucidating the intrinsic and extrinsic sources of such variation. Copyright © 2014 Elsevier Inc. All rights reserved.

  16. Metagenomic analysis of the gut microbiota of the Timber Rattlesnake, Crotalus horridus.

    Science.gov (United States)

    McLaughlin, Richard William; Cochran, Philip A; Dowd, Scot E

    2015-07-01

    Snakes are capable of surviving long periods without food. In this study we characterized the microbiota of a Timber Rattlesnake (Crotalus horridus), devoid of digesta, living in the wild. Pyrosequencing-based metagenomics were used to analyze phylogenetic and metabolic profiles with the aid of the MG-RAST server. Pyrosequencing of samples taken from the stomach, small intestine and colon yielded 691696, 957756 and 700419 high quality sequence reads. Taxonomic analysis of metagenomic reads indicated Eukarya was the most predominant domain, followed by bacteria and then viruses, for all three tissues. The most predominant phylum in the domain Bacteria was Proteobacteria for the tissues examined. Functional classifications by the subsystem database showed cluster-based subsystems were most predominant (10-15 %). Almost equally predominant (10-13 %) was carbohydrate metabolism. To identify bacteria in the colon at a finer taxonomic resolution, a 16S rRNA gene clone library was created. Proteobacteria was again found to be the most predominant phylum. The present study provides a baseline for understanding the microbial ecology of snakes living in the wild.

  17. Metabolic cost of venom replenishment by Prairie Rattlesnakes (Crotalus viridis viridis).

    Science.gov (United States)

    Smith, Matthew T; Ortega, Jason; Beaupre, Steven J

    2014-08-01

    Snakes demonstrate a great deal of variation in the amount of venom injected in both predatory and defensive strikes. There is some evidence that snakes can adaptively meter venom dosage. An underlying assumption in the evolution of venom metering is that the production of venom is energetically costly. To date there has been very little research that has quantified the metabolic costs associated with venom production. We used open-flow respirometry to test for significant differences between Prairie Rattlesnakes (Crotalus v. viridis) that had venom extracted and control snakes that did not. Although previous studies demonstrated high metabolic costs for venom production, we found that snakes that had venom extracted did not have significantly higher metabolic rates than control snakes. The pattern of metabolic deviation from baseline measurements was similar for both treatments, and on average snakes that had venom extracted only exhibited a 1.1% increase over baseline compared to a 2.5% increase in control snakes. Our data suggest that venom is not energetically costly to produce and that perhaps other costs associated with venom can better explain the variability in venom expenditure. Copyright © 2014 Elsevier Ltd. All rights reserved.

  18. Directional sensitivity in the thermal response of the facial pit in western diamondback rattlesnakes (Crotalus atrox).

    Science.gov (United States)

    Kohl, Tobias; Colayori, Samantha E; Westhoff, Guido; Bakken, George S; Young, Bruce A

    2012-08-01

    Recent work published in the accompanying paper used a combination of 3D morphological reconstruction to define optical spread functions and heat transfer physics to study how external heat energy would reach the sensory membrane within the facial pit of pitvipers. The results from all of the species examined indicated asymmetric directional sensitivity, e.g. the pit would preferentially respond to stimuli located below and behind the snake. The present study was intended as a test of these findings through a quantitative neurophysiological analysis of directional sensitivity in the facial pit of the western diamondback rattlesnake, Crotalus atrox. An infrared emitter was positioned through a coordinate system (with varying angular orientations and distances) and the response it evoked measured through neurophysiological recordings of a trigeminal nerve branch composed of the afferents from the sensory membrane of the facial pit. Significant differences were found in the strength of the membrane's neural response to a constant stimulus presented at different orientations (relative to the facial pit opening) and over different distances. The peak sensitivity (at 12 deg above and 20 deg in front of the facial pit opening) was in good agreement with the predicted directional sensitivities based on optical spread functions and 3D topography. These findings support the hypothesis that the topography, and functional performance, of the facial pit has undergone an adaptive radiation within the pit vipers, and that differences in the behavioral ecology of the pit vipers (i.e. terrestrial versus arboreal) are reflected within the facial pits.

  19. Shallow phylogeographic structure in the declining Mexican Lance-headed Rattlesnake, Crotalus polystictus (Serpentes: Viperidae

    Directory of Open Access Journals (Sweden)

    Jesse M. Meik

    2012-06-01

    Full Text Available We investigated matrilineal relationships among populations of the Mexican lance-headed rattlesnake (Crotalus polystictus, a pitviper inhabiting high-elevation valleys of the densely populated southernMexican Plateau. A fragment of the mitochondrial ATPase 8 and 6 genes (589 base pairs revealed comparatively low levels of genetic diversity, with few nucleotide polymorphisms across the portion of the geographic distribution sampled. The shallow intraspecificsequence divergence (1.0% in C. polystictus ATPase 8 and 6 genes contrasts with deep divergences (~1.0–14.1% observed within other montane rattlesnake lineages from the Mexican highlands, and is more typical of intraspecific variation observed in lowland rattlesnake species with similar distributional extents (e.g., C. tigris. We posit that the low genetic diversity in C. polystictus relative to that of other highland rattlesnakes may reflect ecological differences resulting in a different evolutionary response to Pleistocene climatic events. Our finding of apparently low genetic diversity in C. polystictus highlights the importance of conservation initiatives to protect high elevation grasslands in central Mexico.

  20. Distribución y representatividad de las especies del género Crotalus en las áreas naturales protegidas de México Distribution and representation of Crotalus species within the natural protected areas of Mexico

    OpenAIRE

    Dinorah Malinaly Paredes-García; Aurelio Ramírez-Bautista; Miguel Angel Martínez-Morales

    2011-01-01

    Se realizó un análisis sistemático de la distribución de las especies del género Crotalus en México para conocer los sitios de mayor concentración y evaluar su nivel de representatividad dentro del esquema federal de áreas naturales protegidas (ANP). A partir de registros georreferenciados se construyeron los modelos de distribución potencial de las especies con un número suficiente de registros, empleando los algoritmos GARP y MaxEnt. Con estos mapas de distribución se dedujeron las áreas de...

  1. Linking the transcriptome and proteome to characterize the venom of the eastern diamondback rattlesnake (Crotalus adamanteus).

    Science.gov (United States)

    Margres, Mark J; McGivern, James J; Wray, Kenneth P; Seavy, Margaret; Calvin, Kate; Rokyta, Darin R

    2014-01-16

    Understanding the molecular basis of the phenotype is key to understanding adaptation, and the relationship between genes and specific traits is represented by the genotype-phenotype map. The specialization of the venom-gland towards toxin production enables the use of transcriptomics to identify a large number of loci that contribute to a complex phenotype (i.e., venom), while proteomic techniques allow verification of the secretion of the proteins produced by these loci, creating a genotype-phenotype map. We used the extensive database of mRNA transcripts generated by the venom-gland transcriptome of Crotalus adamanteus along with proteomic techniques to complete the genotype-phenotype map for the C. adamanteus venom system. Nanospray LC/MS(E) analysis of a whole venom sample identified evidence for 52 of the 78 unique putative toxin transcript clusters, including 44 of the 50 most highly expressed transcripts. Tandem mass spectrometry and SDS-PAGE of reversed-phase high-performance liquid chromatography fractions identified 40 toxins which clustered into 20 groups and represented 10 toxin families, creating a genotype-phenotype map. By using the transcriptome to understand the proteome we were able to achieve locus-specific resolution and provide a detailed characterization of the C. adamanteus venom system. Identifying the mechanisms by which genetic variation presents itself to the sieve of selection at the phenotypic level is key to understanding the molecular basis of adaptation, and the first step in understanding this relationship is to identify the genetic basis of the phenotype through the construction of a genotype-phenotype map. We used the high-throughput venom-gland transcriptomic characterization of the eastern diamondback rattlesnake (C. adamanteus) and proteomic techniques to complete and confirm the genotype-phenotype map, providing a detailed characterization of the C. adamanteus venom system. Copyright © 2013 Elsevier B.V. All rights reserved.

  2. Behavioral thermal tolerances of free-ranging rattlesnakes (Crotalus oreganus) during the summer foraging season.

    Science.gov (United States)

    Putman, Breanna J; Clark, Rulon W

    2017-04-01

    Increasing temperature due to climate change is one of the greatest challenges for wildlife worldwide. Behavioral data on free-ranging individuals is necessary to determine at what temperatures animals modify activity as this would determine their capacity to continue to move, forage, and mate under altered thermal regimes. In particular, high temperatures could limit available surface activity time and time spent on fitness-related activities. Conversely, performance, such as feeding rate, can increase with temperature potentially having positive fitness effects. Here, we examine how the hunting behaviors of free-ranging Northern Pacific Rattlesnakes (Crotalus oreganus) associate with air temperature and body temperature. We continuously recorded snakes in the field using videography, capturing behaviors rarely considered in past studies such as movements in and out of refuge and strikes on prey. We found that as mean daily air temperature increased, hunting activity and the likelihood of hunting at night decreased, while the number of movements and distance moved per day increased. Snakes typically retreated to refuge before body temperatures reached 31°C. Body temperatures of snakes hunting on the surface were lower compared to temperatures of non-hunting snakes in refuge in the morning, while this relationship was inverted in the afternoon. Snake body size influenced the disparity of these temperatures. Finally, strike initiation and success occurred across a wide range of body temperatures, indicating hunting performance may not be strongly constrained by temperature. These results on the temperatures at which free-ranging rattlesnakes exhibit fitness-related behaviors could be valuable for understanding their vulnerabilities to future climates. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. Crotalus oreganus lutosus (Great Basin Rattlesnake). Defensive behavior/head hiding and tongue display

    Science.gov (United States)

    Johnson, Alexander K.; Medica, Phil A.

    2017-01-01

    A variety of snake species employ head hiding as defensive behavior (Greene 1973. J. Herpetol. 7:143–161), but such behavior seems to be rarely observed in rattlesnakes. A recent report (Medica 2009. Herpetol. Rev. 40:95–97) presented observations on Crotalus scutulatus scutulatus and cited cases of similar behavior involving C. atrox, C. ruber, and C. viridis viridis. Here we report an encounter on 28 May 2016 with a C. oreganus lutosus (total length ca. 900 mm) observed 16 km S of Fallon, Churchill County, Nevada, USA. At 0925 h this snake was spotted by AKJ near the edge of the pavement on U.S. Hwy. 95. When approached, the snake presented typical active defensive behavior, with the rattle elevated (although not rattling), striking occasionally, and frequently flicking its conspicuous lavender and black tongue. As AKJ continued to take pictures the snake coiled more tightly (Fig. 1A), maintaining its rattle concealed and finally hiding its head beneath its coils (Fig. 1B). After photographing this pose AKJ made his first physical contact with the snake by scooping it up with a small shovel and placing it off the road. At this point the snake recovered its feisty attitude and would not retreat despite attempts to move it further away from the road. The snake did not rattle during the entire encounter (which lasted less than 5 min). Head hiding as a “last resort” in response to a persistent unavoidable threat may be more widespread within the family Crotalidae than previously documented.

  4. Organotopic organization of the primary Infrared Sensitive Nucleus (LTTD) in the western diamondback rattlesnake (Crotalus atrox).

    Science.gov (United States)

    Kohl, Tobias; Bothe, Maximilian S; Luksch, Harald; Straka, Hans; Westhoff, Guido

    2014-12-15

    Pit vipers (Crotalinae) have a specific sensory system that detects infrared radiation with bilateral pit organs in the upper jaw. Each pit organ consists of a thin membrane, innervated by three trigeminal nerve branches that project to a specific nucleus in the dorsal hindbrain. The known topographic organization of infrared signals in the optic tectum prompted us to test the implementation of spatiotopically aligned sensory maps through hierarchical neuronal levels from the peripheral epithelium to the first central site in the hindbrain, the nucleus of the lateral descending trigeminal tract (LTTD). The spatial organization of the anatomical connections was revealed in a novel in vitro whole-brain preparation of the western diamondback rattlesnake (Crotalus atrox) that allowed specific application of multiple neuronal tracers to identified pit-organ-supplying trigeminal nerve branches. After adequate survival times, the entire peripheral and central projections of fibers within the pit membrane and the LTTD became visible. This approach revealed a morphological partition of the pit membrane into three well-defined sensory areas with largely separated innervations by the three main branches. The peripheral segregation of infrared afferents in the sensory epithelium was matched by a differential termination of the afferents within different areas of the LTTD, with little overlap. This result demonstrates a topographic organizational principle of the snake infrared system that is implemented by maintaining spatially aligned representations of environmental infrared cues on the sensory epithelium through specific neuronal projections at the level of the first central processing stage, comparable to the visual system. © 2014 Wiley Periodicals, Inc.

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

    Science.gov (United States)

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

    2017-04-01

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

  7. Phenotypic integration in the feeding system of the eastern diamondback rattlesnake (Crotalus adamanteus).

    Science.gov (United States)

    Margres, Mark J; Wray, Kenneth P; Seavy, Margaret; McGivern, James J; Sanader, Dragana; Rokyta, Darin R

    2015-07-01

    Selection can vary geographically across environments and temporally over the lifetime of an individual. Unlike geographic contexts, where different selective regimes can act on different alleles, age-specific selection is constrained to act on the same genome by altering age-specific expression. Snake venoms are exceptional traits for studying ontogeny because toxin expression variation directly changes the phenotype; relative amounts of venom components determine, in part, venom efficacy. Phenotypic integration is the dependent relationship between different traits that collectively produce a complex phenotype and, in venomous snakes, may include traits as diverse as venom, head shape and fang length. We examined the feeding system of the eastern diamondback rattlesnake (Crotalus adamanteus) across environments and over the lifetime of individuals and used a genotype-phenotype map approach, protein expression data and morphological data to demonstrate that: (i) ontogenetic effects explained more of the variation in toxin expression variation than geographic effects, (ii) both juveniles and adults varied geographically, (iii) toxin expression variation was a result of directional selection and (iv) different venom phenotypes covaried with morphological traits also associated with feeding in temporal (ontogenetic) and geographic (functional) contexts. These data are the first to demonstrate, to our knowledge, phenotypic integration between multiple morphological characters and a biochemical phenotype across populations and age classes. We identified copy number variation as the mechanism driving the difference in the venom phenotype associated with these morphological differences, and the parallel mitochondrial, venom and morphological divergence between northern and southern clades suggests that each clade may warrant classification as a separate evolutionarily significant unit. © 2015 John Wiley & Sons Ltd.

  8. Diversity-dependent cladogenesis throughout western Mexico: Evolutionary biogeography of rattlesnakes (Viperidae: Crotalinae: Crotalus and Sistrurus).

    Science.gov (United States)

    Blair, Christopher; Sánchez-Ramírez, Santiago

    2016-04-01

    Rattlesnakes (Crotalus and Sistrurus) represent a radiation of approximately 42 species distributed throughout the New World from southern Canada to Argentina. Interest in this enigmatic group of snakes continues to accrue due, in part, to their ecomorphological diversity, contributions to global envenomations, and potential medicinal importance. Although the group has garnered substantial attention from systematists and evolutionary biologists for decades, little is still known regarding patterns of lineage diversification. In addition, few studies have statistically quantified broad-scale biogeographic patterns in rattlesnakes to ascertain how dispersal occurred throughout the New World, particularly among the different major biomes of the Americas. To examine diversification and biogeographic patterns in this group of snakes we assemble a multilocus data set consisting of over 6700bp encompassing three nuclear loci (NT-3, RAG-1, C-mos) and seven mitochondrial genes (12S, 16S, ATPase6, ATPase8, ND4, ND5, cytb). Fossil-calibrated phylogenetic and subsequent diversification rate analyses are implemented using maximum likelihood and Bayesian inference, to examine their evolutionary history and temporal dynamics of diversity. Based on ancestral area reconstructions we explore dispersal patterns throughout the New World. Cladogenesis occurred predominantly during the Miocene and Pliocene with only two divergences during the Pleistocene. Two different diversification rate models, advocating diversity-dependence, are strongly supported. These models indicate an early rapid radiation followed by a recent speciation rate decline. Biogeographic analyses suggest that the high elevation pine-oak forests of western Mexico served as a major speciation pump for the majority of lineages, with the desert biome of western North America colonized independently at least twice. All together, these results provide evidence for rapid diversification of rattlesnakes throughout the

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

    Science.gov (United States)

    Castañeda-Gaytán, Gamaliel; Castañeda-Gaytán, Juan; Ponce-López, Roberto; Olvera-Rodríguez, Alejandro; Alagón, Alejandro; Pérez-Morales, Rebeca

    2018-01-01

    Rattlesnake venoms may be classified according to the presence/absence and relative abundance of the neurotoxic phospholipases A2s (PLA2s), 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 LD50 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. PMID:29316683

  10. 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 Junior, Geraldo B; Lima, Juliana B; Veras, Maria do Socorro B; Daher, Elizabeth F

    2013-01-01

    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.

  11. Proteomic analysis of the rare Uracoan rattlesnake Crotalus vegrandis venom: Evidence of a broad arsenal of toxins.

    Science.gov (United States)

    Viala, Vincent Louis; Hildebrand, Diana; Fucase, Tamara Mieco; Sciani, Juliana Mozer; Prezotto-Neto, José Pedro; Riedner, Maria; Sanches, Leonardo; Nishimura, Paula Juliana; Oguiura, Nancy; Pimenta, Daniel Carvalho; Schlüter, Hartmut; Betzel, Christian; Arni, Raghuvir Krishnaswami; Spencer, Patrick Jack

    2015-12-01

    The investigation of venoms has many clinical, pharmacological, ecological and evolutionary outcomes. The Crotalus spp. venom can cause hemorrhage, neurotoxicity, myotoxicity, coagulopathy and hypotension. Although neurotoxicity and hemorrhage usually does not occur for the same species, the rare Venezuelan species Crotalus vegrandis presents both characteristic. Different from the other species it has a restricted ecological niche and geographical distribution. Nevertheless, it has a raising medical importance as this rattlesnake population is increasing. Few works describe its neurotoxic and hemorrhagic features, but other toxins might play an important role in envenomation. We combined proteomic methods to identify for the first time the main components of it venom: 2D SDS-PAGE and gel-filtration chromatography for protein mixture decomplexation; LC-MS(2) of low molecular mass fractions and tryptic peptides; bioinformatic identification of toxin families and specific protein species based on unique peptide analysis and sequence database enriched with species-specific venom gland transcripts; and finally polyclonal anti-crotamine Western-blotting. Our results point to a broad arsenal of toxins in C. vegrandis venom: PIII and PII metalloproteases, crotoxin subunits, other phospholipases, isoforms of serine proteases and lectins, l-amino-acid oxidase, nerve growth factor, as well as other less abundant toxins. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. Effect of iron and carbon monoxide on fibrinogenase-like degradation of plasmatic coagulation by venoms of four Crotalus species.

    Science.gov (United States)

    Nielsen, Vance G; Redford, Daniel T; Boyle, Patrick K

    2017-01-01

    Annually, thousands suffer poisonous snake bite, often from defibrinogenating species. Iron and carbon monoxide (CO) improve coagulation kinetics by modulation of fibrinogen as demonstrated in various Agkistrodon species and Crotalus atrox. Thus, we sought to determine whether pretreatment of plasma with iron and CO could attenuate venom-mediated catalysis of fibrinogen obtained from four common Crotalus species with known fibrinogenase activity. Human plasma was pretreated with ferric chloride (0-10 μmol/l) and CO-releasing molecule-2 (0-100 μmol/l) prior to exposure to venom from a Northern Pacific rattlesnake, Arizona black rattlesnake, prairie rattlesnake, or red diamond rattlesnake. The concentration of venom used decreased coagulation function of one or more kinetic parameters by at least 50% of normal values. Coagulation kinetics were determined with thrombelastography.Three snake venoms significantly degraded plasmatic coagulation kinetics, prolonging the onset to clot formation, diminishing velocity of clot growth and decreasing clot strength. However, red diamond rattlesnake venom exposure resulted in mixed coagulation kinetics, significantly decreasing the time to onset of coagulation without decreasing the velocity of clot growth. Iron and CO attenuated these coagulation kinetic changes in a species-specific manner. Further in vitro investigation of other fibrinogenolytic venoms is indicated to determine if iron and CO can attenuate venom compromised coagulation.

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

    Directory of Open Access Journals (Sweden)

    Polianna L.M.M. Albuquerque

    2013-09-01

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

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

    Directory of Open Access Journals (Sweden)

    Ezequiel Capeletti

    2016-09-01

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

  15. 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.

  16. The venom-gland transcriptome of the eastern diamondback rattlesnake (Crotalus adamanteus)

    Science.gov (United States)

    2012-01-01

    Background Snake venoms have significant impacts on human populations through the morbidity and mortality associated with snakebites and as sources of drugs, drug leads, and physiological research tools. Genes expressed by venom-gland tissue, including those encoding toxic proteins, have therefore been sequenced but only with relatively sparse coverage resulting from the low-throughput sequencing approaches available. High-throughput approaches based on 454 pyrosequencing have recently been applied to the study of snake venoms to give the most complete characterizations to date of the genes expressed in active venom glands, but such approaches are costly and still provide a far-from-complete characterization of the genes expressed during venom production. Results We describe the de novo assembly and analysis of the venom-gland transcriptome of an eastern diamondback rattlesnake (Crotalus adamanteus) based on 95,643,958 pairs of quality-filtered, 100-base-pair Illumina reads. We identified 123 unique, full-length toxin-coding sequences, which cluster into 78 groups with less than 1% nucleotide divergence, and 2,879 unique, full-length nontoxin coding sequences. The toxin sequences accounted for 35.4% of the total reads, and the nontoxin sequences for an additional 27.5%. The most highly expressed toxin was a small myotoxin related to crotamine, which accounted for 5.9% of the total reads. Snake-venom metalloproteinases accounted for the highest percentage of reads mapping to a toxin class (24.4%), followed by C-type lectins (22.2%) and serine proteinases (20.0%). The most diverse toxin classes were the C-type lectins (21 clusters), the snake-venom metalloproteinases (16 clusters), and the serine proteinases (14 clusters). The high-abundance nontoxin transcripts were predominantly those involved in protein folding and translation, consistent with the protein-secretory function of the tissue. Conclusions We have provided the most complete characterization of the genes

  17. The venom-gland transcriptome of the eastern diamondback rattlesnake (Crotalus adamanteus).

    Science.gov (United States)

    Rokyta, Darin R; Lemmon, Alan R; Margres, Mark J; Aronow, Karalyn

    2012-07-16

    Snake venoms have significant impacts on human populations through the morbidity and mortality associated with snakebites and as sources of drugs, drug leads, and physiological research tools. Genes expressed by venom-gland tissue, including those encoding toxic proteins, have therefore been sequenced but only with relatively sparse coverage resulting from the low-throughput sequencing approaches available. High-throughput approaches based on 454 pyrosequencing have recently been applied to the study of snake venoms to give the most complete characterizations to date of the genes expressed in active venom glands, but such approaches are costly and still provide a far-from-complete characterization of the genes expressed during venom production. We describe the de novo assembly and analysis of the venom-gland transcriptome of an eastern diamondback rattlesnake (Crotalus adamanteus) based on 95,643,958 pairs of quality-filtered, 100-base-pair Illumina reads. We identified 123 unique, full-length toxin-coding sequences, which cluster into 78 groups with less than 1% nucleotide divergence, and 2,879 unique, full-length nontoxin coding sequences. The toxin sequences accounted for 35.4% of the total reads, and the nontoxin sequences for an additional 27.5%. The most highly expressed toxin was a small myotoxin related to crotamine, which accounted for 5.9% of the total reads. Snake-venom metalloproteinases accounted for the highest percentage of reads mapping to a toxin class (24.4%), followed by C-type lectins (22.2%) and serine proteinases (20.0%). The most diverse toxin classes were the C-type lectins (21 clusters), the snake-venom metalloproteinases (16 clusters), and the serine proteinases (14 clusters). The high-abundance nontoxin transcripts were predominantly those involved in protein folding and translation, consistent with the protein-secretory function of the tissue. We have provided the most complete characterization of the genes expressed in an active snake

  18. The venom-gland transcriptome of the eastern diamondback rattlesnake (Crotalus adamanteus

    Directory of Open Access Journals (Sweden)

    Rokyta Darin R

    2012-07-01

    Full Text Available Abstract Background Snake venoms have significant impacts on human populations through the morbidity and mortality associated with snakebites and as sources of drugs, drug leads, and physiological research tools. Genes expressed by venom-gland tissue, including those encoding toxic proteins, have therefore been sequenced but only with relatively sparse coverage resulting from the low-throughput sequencing approaches available. High-throughput approaches based on 454 pyrosequencing have recently been applied to the study of snake venoms to give the most complete characterizations to date of the genes expressed in active venom glands, but such approaches are costly and still provide a far-from-complete characterization of the genes expressed during venom production. Results We describe the de novo assembly and analysis of the venom-gland transcriptome of an eastern diamondback rattlesnake (Crotalus adamanteus based on 95,643,958 pairs of quality-filtered, 100-base-pair Illumina reads. We identified 123 unique, full-length toxin-coding sequences, which cluster into 78 groups with less than 1% nucleotide divergence, and 2,879 unique, full-length nontoxin coding sequences. The toxin sequences accounted for 35.4% of the total reads, and the nontoxin sequences for an additional 27.5%. The most highly expressed toxin was a small myotoxin related to crotamine, which accounted for 5.9% of the total reads. Snake-venom metalloproteinases accounted for the highest percentage of reads mapping to a toxin class (24.4%, followed by C-type lectins (22.2% and serine proteinases (20.0%. The most diverse toxin classes were the C-type lectins (21 clusters, the snake-venom metalloproteinases (16 clusters, and the serine proteinases (14 clusters. The high-abundance nontoxin transcripts were predominantly those involved in protein folding and translation, consistent with the protein-secretory function of the tissue. Conclusions We have provided the most complete

  19. Preparation of crotalus venom radiolabeled with technetium{sup 99m} as a tool for biodistribution study

    Energy Technology Data Exchange (ETDEWEB)

    Pujatti, Priscilla Brunelli; Santos, Raquel Gouvea dos [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN), Belo Horizonte, MG (Brazil)]. E-mail: santosr@cdtn.br; Simal, Carlos Jorge Rodrigues [Universidade Federal de Minas Gerais, Belo Horizonte, MG (Brazil). Faculdade de Medicina

    2005-10-15

    Technetium-{sup 99m} ({sup 99m} Tc) has been the radionuclide of choice for nuclear medicine procedures and experimental research. Because of its optimal nuclear properties, {sup 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 {sup 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 {sup 99m} Tc-CV can be a useful tool for biodistribution studies. (author)

  20. Inhibition of Lung Tumor Colonization and Cell Migration with the Disintegrin Crotatroxin 2 Isolated from the Venom of Crotalus atrox

    Science.gov (United States)

    Galán, Jacob A.; Sánchez, Elda E.; Rodríguez-Acosta, Alexis; Soto, Julio G.; Bashir, Sajid; McLane, Mary Ann; Paquette-Straub, Carrie; Pérez, John C.

    2009-01-01

    Disintegrins are low molecular weight proteins (4-15 kDa) with an RGD binding region at their binding loop. Disintegrin and disintegrin-like proteins are found in the venom of four families of snakes: Atractaspididae, Elapidae, Viperidae and Colubridae. This report describes the biological activity of a disintegrin, crotatroxin 2, isolated by a three-step chromatography procedure from the venom of the Western diamondback rattlesnake (Crotalus atrox). The intact molecular mass for crotatroxin 2 was 7.384 kDa and 71 amino acids. Crotatroxin 2 inhibited human whole blood platelet aggregation with an IC50 of 17.5 nM, inhibited cell (66.3p) migration by 63%, and inhibited experimental lung tumor colonization in BALB/c mice at 1000 μg/kg. Our data suggest that while crotatroxin 2 inhibits platelet aggregation, cancer cell migration, and lung tumor colonization it is done via different integrins. PMID:18387648

  1. Type specimens of Crotalus scutulatus (Chordata: Reptilia: Squamata: Viperidae) re-examined, with new evidence after more than a century of confusion

    Science.gov (United States)

    Cardwell, Michael D.; Gotte, Steve W.; McDiarmid, Roy W.; Gilmore, Ned; Poindexter, James A.

    2013-01-01

    The original description of Crotalus scutulatus (Chordata: Reptilia: Squamata: Viperidae) was published in 1861 by Robert Kennicott, who did not identify a type specimen or a type locality. We review the history of specimens purported to be the type(s) and various designations of type locality. We provide evidence that ANSP 7069 (formerly one of two specimens of USNM 5027) is the holotype and that the appropriate type locality is Fort Buchanan, near present-day Sonoita, in Santa Cruz County, Arizona.

  2. Additional observations and notes on the natural history of the prairie rattlesnake (Crotalus viridis) in Colorado.

    Science.gov (United States)

    Fitzgerald, Kevin T; Shipley, Bryon K; Newquist, Kristin L; Vera, Rebecca; Flood, Aryn A

    2013-11-01

    On account of their unique anatomy, physiology, natural history, ecology, and behavior, rattlesnakes make ideal subjects for a variety of different scientific disciplines. The prairie rattlesnake (Crotalus viridis) in Colorado was selected for investigation of its relationship to colonies of black-tailed prairie dogs (Cynomys ludovicianus) with regard to spatial ecology. A total of 31 snakes were anesthetized and had radiotransmitters surgically implanted. In addition, at the time of their capture, all snakes underwent the following: (1) they had bacterial culture taken from their mouths for potential isolation of pathogenic bacteria; (2) similarly, they had cloacal bacterial cultures taken to assess potentially harmful bacteria passed in the feces; and (3) they had blood samples drawn to investigate the presence of any zoonotic agents in the serum of the snakes. The results of the study and their implications are discussed here. Traditionally, a low incidence of bacterial wound infection has been reported following snakebite. Nevertheless, the oral cavity of snakes has long been known to house a wide variety of bacterial flora. In our study, 10 different bacterial species were isolated from the mouths of the rattlesnakes, 6 of which are capable of being zoonotic pathogens and inducing human disease. More studies are necessary to see why more rattlesnake bites do not become infected despite the presence of such pathogenic bacteria. The results of fecal bacteria isolated revealed 13 bacterial species, 12 of which can cause disease in humans. Of the snakes whose samples were cultured, 26% were positive for the presence of the pathogen Salmonella arizonae, one of the causative agents of reptile-related salmonellosis in humans. It has long been reported that captive reptiles have a much higher incidence than wild, free-ranging species. This study shows the incidence of Salmonella in a wild, free-ranging population of rattlesnakes. In addition, Stenotrophomonas

  3. Distribución y representatividad de las especies del género Crotalus en las áreas naturales protegidas de México Distribution and representation of Crotalus species within the natural protected areas of Mexico

    Directory of Open Access Journals (Sweden)

    Dinorah Malinaly Paredes-García

    2011-06-01

    Full Text Available Se realizó un análisis sistemático de la distribución de las especies del género Crotalus en México para conocer los sitios de mayor concentración y evaluar su nivel de representatividad dentro del esquema federal de áreas naturales protegidas (ANP. A partir de registros georreferenciados se construyeron los modelos de distribución potencial de las especies con un número suficiente de registros, empleando los algoritmos GARP y MaxEnt. Con estos mapas de distribución se dedujeron las áreas de mayor concentración de especies y se estimó su presencia en las ANP. La mayor riqueza de especies se concentra en las regiones de la sierra Madre Occidental, el noroeste del Altiplano Mexicano, la porción norte de la sierra Madre Oriental y el extremo oeste de la Faja Volcánica Transmexicana. Se estima que el 84.4% de las ANP incluyen al menos 1 especie de Crotalus y que el 88.2% de las especies que se distribuyen en México están presentes al menos dentro de 1 ANP. En este contexto de alta representatividad, el plantear la incorporación de ANP adicionales para la protección y conservación de crotalinos se dificulta, por lo que debe optarse por estrategias alternativas, tales como educación ambiental, conservación de hábitats mediante el uso sustentable de recursos, y el manejo y reproducción en cautiverio de especies en riesgo; además, se debe fomentar la investigación y el monitoreo de poblaciones.We performed a systematic analysis of the distribution of the Crotalus species in Mexico to depict the places of high species concentration and to evaluate its representation within the federal natural protected areas (NPAs scheme. Based on geo-referenced records, we built distribution models for species with enough records using the GARP and MaxEnt algorithms. We then deduced areas of high species concentration and estimated its presence in the NPAs. The regions with the highest species richness are the Sierra Madre Occidental, the

  4. Lung surface-active fraction as a model system for macromolecular ultrastructural studies with Crotalus atrox venom.

    Science.gov (United States)

    Frosolono, M F; Pawlowski, R; Charms, B L; Corbusier, C; Abrams, M; Jones, J

    1973-01-01

    The dog lung surface-active fraction and phosphatidylcholine constituents were subjected to hydrolysis by Crotalus atrox phospholipase A(2). Relative rates of hydrolysis were: dipalmitoyl glycerophosphorylcholine > phosphatidylcholine isolated from the surface-active fraction > phosphatidylcholine as an integral component of the intact surface-active macromolecular structure. Cholesterol markedly inhibited, whereas tripalmitin increased, the rate of hydrolysis with both pure phosphatidylcholine substrates. The effect of temperature on the velocity indicated the enzyme was most active when the substrates were in the gel state. These kinetic results, in conjunction with surface chemistry studies, can be interpreted to indicate that the phosphatidylcholine in the intact surface-active macromolecular particle is liquid crystalline due to molecular interactions with other constituents. Gas-liquid chromatographic analysis of the 2-lysophosphatidylcholines and fatty acids produced from the enzymatic hydrolysis of the intact surface-active fraction indicated that palmitoyl residues were more accessible to the enzyme, perhaps because they occupied positions near the surface of the particle.

  5. Isolation and biochemical characterization of rubelase, a non-hemorrhagic elastase from Crotalus ruber ruber (Red Rattlesnake) venom.

    Science.gov (United States)

    Komori, Yumiko; Sakai, Kaname; Masuda, Katsuyoshi; Nikai, And Toshiaki

    2011-07-01

    A novel non-hemorrhagic basic metalloprotease, rubelase, was isolated from the venom of Crotalus ruber ruber. Rubelase hydrolyzes succinyl-L-alanyl-L-alanyl-L-alanyl p-nitroanilide (STANA), a specific substrate for elastase, and the hydrolytic activity was inhibited by chelating agents. It also hydrolyzes collagen and fibrinogen. However, hemorrhagic activity was not observed. By ESI/Q-TOF and MALDI/TOF mass spectrometry combined with Edman sequencing procedure, the molecular mass of rubelase was determined to be 23,266 Da. Although its primary structure was similar to rubelysin (HT-2), a hemorrhagic metalloprotease isolated from the same snake venom, the circumstances surrounding putative zinc binding domain HEXXHXXGXXH were found to be different when the three-dimensional computer models of both metalloproteases were compared. The cytotoxic effects of rubelase and rubelysin on cultured endothelial and smooth muscle cells were also different, indicating that the substitution of several amino acid residues causes the changes of active-site conformation and cell preference.

  6. The transcriptomic and proteomic basis for the evolution of a novel venom phenotype within the Timber Rattlesnake (Crotalus horridus).

    Science.gov (United States)

    Rokyta, Darin R; Wray, Kenneth P; McGivern, James J; Margres, Mark J

    2015-05-01

    The genetics underlying adaptive trait evolution describes the intersection between the probability that particular types of mutation are beneficial and the rates they arise. Snake venoms can vary in a directly meaningful manner through coding mutations and regulatory mutations. The amounts of different components determine venom efficacy, but point mutations in coding sequences can also change efficacy and function. The Timber Rattlesnake (Crotalus horridus) has populations that have evolved neurotoxic venom from the typical hemorrhagic rattlesnake venom present throughout most of its range. We identified only a handful of nonsynonymous differences in just five loci between animals with each venom type, and these differences affected lower-abundance toxins. Expression of at least 18 loci encoding hemorrhagic toxins was severely reduced in the production of neurotoxic venom. The entire phospholipase A2 toxin family was completely replaced in the neurotoxic venom, possibly through intergeneric hybridization. Venom paedomorphosis could, at best, explain only some of the loss of expression of hemorrhagic toxins. The number of potential mechanisms for altering venom composition and the patterns observed for C. horridus suggest that rapid venom evolution should occur primarily through changes in venom composition, rather than point mutations affecting coding sequences. Copyright © 2015 Elsevier Ltd. All rights reserved.

  7. Multilocus species delimitation in the Crotalus triseriatus species group (Serpentes: Viperidae: Crotalinae), with the description of two new species.

    Science.gov (United States)

    Bryson, Robert W; Linkem, Charles W; Dorcas, Michael E; Lathrop, Amy; Jones, Jason M; Alvarado-Díaz, Javier; Grünwald, Christoph I; Murphy, Robert W

    2014-07-01

    Members of the Crotalus triseriatus species group of montane rattlesnakes are widely distributed across the highlands of Mexico and southwestern USA. Although five species are currently recognized within the group, species limits remain to be tested. Genetic studies suggest that species may be paraphyletic and that at least one cryptic species may be present. We generate 3,346 base pairs of DNA sequence data from seven nuclear loci to test competing models of species delimitation in the C. triseriatus group using Bayes factor delimitation. We also examine museum specimens from the Trans-Mexican Volcanic Belt for evidence of cryptic species. We find strong support for a nine-species model and genetic and morphological evidence for recognizing two new species within the group, which we formally describe here. Our results suggest that the current taxonomy of the C. triseriatus species group does not reflect evolutionary history. We suggest several conservative taxonomic changes to the group, but future studies are needed to better clarify relationships among species and examine genetic patterns and structure within wide-ranging lineages.

  8. Constipation associated with brumation? Intestinal obstruction caused by a fecalith in a wild red diamond rattlesnake (Crotalus ruber).

    Science.gov (United States)

    Corbit, A G; Person, C; Hayes, W K

    2014-02-01

    This report describes the fecalith-induced intestinal obstruction of a free-ranging red diamond rattlesnake (Crotalus ruber) and the snake's subsequent history following surgical removal of the fecalith. The captured snake exhibited an abnormally distended abdomen and an extremely hard mass, detected via palpation, near its vent. Coeliotomy yielded a 2.5-cm, 5-g fecalith from the large intestine. Microscopic dissection of the fecalith revealed no evidence of gastrointestinal parasitic worms. Subsequently, we implanted a radio-transmitter that allowed us to track the snake's movements for 7 months (until the radio signal vanished), indicating normal behaviour, complete recovery and good health apart from the obstruction. This observation suggests that fecalith development and intestinal obstruction represent potential risks of long-term faecal retention, an unusual physiological trait well documented among rattlesnakes and other stout, heavy-bodied terrestrial viperid snakes. Dehydration and decreased gut motility associated with brumation (≈hibernation) may predispose temperate snakes to fecalith formation. Regional drought and a small mammal diet with indigestible hairs might have also promoted fecalith formation in this specimen. © 2013 Blackwell Verlag GmbH.

  9. Genetic Basis for Variation of Metalloproteinase-Associated Biochemical Activity in Venom of the Mojave Rattlesnake (Crotalus scutulatus scutulatus).

    Science.gov (United States)

    Dagda, Ruben K; Gasanov, Sardar; De La Oiii, Ysidro; Rael, Eppie D; Lieb, Carl S

    2013-01-01

    The metalloproteinase composition and biochemical profiles of rattlesnake venom can be highly variable among rattlesnakes of the same species. We have previously shown that the neurotoxic properties of the Mojave rattlesnake (Crotalus scutulatus scutulatus) are associated with the presence of the Mojave toxin A subunit suggesting the existence of a genetic basis for rattlesnake venom composition. In this report, we hypothesized the existence of a genetic basis for intraspecies variation in metalloproteinase-associated biochemical properties of rattlesnake venom of the Mojave rattlesnake. To address this question, we PCR-amplified and compared the genomic DNA nucleotide sequences that code for the mature metalloproteinase domain of fourteen Mojave rattlesnakes captured from different geographical locations across the southwest region of the United States. In addition, the venoms from the same rattlesnakes were tested for their ability to hydrolyze fibrinogen, fibrin, casein, and hide powder azure and for induction of hemorrhage in mice. Overall, based on genomic sequencing and biochemical data, we classified Mojave rattlesnake venom into four distinct groups of metalloproteinases. These findings indicate that differences in nucleotide sequences encoding the mature proteinase domain and noncoding regions contribute to differences in venom metalloproteinase activities among rattlesnakes of the same species.

  10. Genetic Basis for Variation of Metalloproteinase-Associated Biochemical Activity in Venom of the Mojave Rattlesnake (Crotalus scutulatus scutulatus

    Directory of Open Access Journals (Sweden)

    Ruben K. Dagda

    2013-01-01

    Full Text Available The metalloproteinase composition and biochemical profiles of rattlesnake venom can be highly variable among rattlesnakes of the same species. We have previously shown that the neurotoxic properties of the Mojave rattlesnake (Crotalus scutulatus scutulatus are associated with the presence of the Mojave toxin A subunit suggesting the existence of a genetic basis for rattlesnake venom composition. In this report, we hypothesized the existence of a genetic basis for intraspecies variation in metalloproteinase-associated biochemical properties of rattlesnake venom of the Mojave rattlesnake. To address this question, we PCR-amplified and compared the genomic DNA nucleotide sequences that code for the mature metalloproteinase domain of fourteen Mojave rattlesnakes captured from different geographical locations across the southwest region of the United States. In addition, the venoms from the same rattlesnakes were tested for their ability to hydrolyze fibrinogen, fibrin, casein, and hide powder azure and for induction of hemorrhage in mice. Overall, based on genomic sequencing and biochemical data, we classified Mojave rattlesnake venom into four distinct groups of metalloproteinases. These findings indicate that differences in nucleotide sequences encoding the mature proteinase domain and noncoding regions contribute to differences in venom metalloproteinase activities among rattlesnakes of the same species.

  11. Isolamento e caracterização da crotoxina símile do veneno de Crotalus vegrandis com atividade antitumoral

    OpenAIRE

    Paula Juliana Nishimura

    2016-01-01

    Diversos estudos de serpentes têm mostrado que algumas substâncias componentes de seus venenos possuem eficiência na atividade antitumoral nos ensaios realizados em laboratório em células in vitro e in vivo. O veneno da cascavel Crotalus vegrandis possuem atividades neurotoxica, miotoxica e hemorrágica. A crotoxina simile é um importante componente desse veneno sendo formada por uma proteína que possui uma unidade ácida (Crotapotina) ligada a uma subunidade básica (PLA2). Devido ao fato de ex...

  12. Gape size, its morphological basis, and the validity of gape indices in western diamond-backed rattlesnakes (Crotalus atrox).

    Science.gov (United States)

    Hampton, Paul M; Moon, Brad R

    2013-02-01

    Maximum gape is important to the ecology and evolution of many vertebrates, particularly gape-limited predators, because it can restrict the sizes and shapes of prey that can be eaten. Although many cranial elements probably contribute to gape, it is typically estimated from jaw length or jaw width, or occasionally from a combination of these two measures. We measured maximum gape directly for 18 individuals of the western diamond-backed rattlesnake, Crotalus atrox. We measured each individual's body length, several external cranial dimensions, several cranial osteological dimensions from cleaned skeletons, and we calculated gape index values from two published gape indices (GI). Cranial bone lengths and gape circumference showed negative allometry with snout-vent length (SVL), indicating that small individuals have relatively larger heads and gapes than their larger conspecifics. We then used Akaike's Information Criterion to determine which external and osteological measurements were the best predictors of gape. Body size (SVL) was the best predictor of maximum gape overall; however, when SVL was excluded from the analysis, quadrate (QL) and mandible lengths (MdLs) were the best predictors of maximum gape using both external and osteological measurements. Quadrate length probably contributes directly to gape; however, the importance of MdL to gape is less clear and may be due largely to its allometric relationships with head length and SVL. The two published GI did not prove to be better indicators of actual gape than the jaw and QLs in this study, and the gape values they produced differed significantly from our empirically determined gapes. For these reasons, we urge caution with the use and interpretation of computed GI in future studies. The extensive variation in quadrate and mandible morphology among lineages suggest that these bones are more important to variation in gape among species and lineages than within a single species. Copyright © 2012 Wiley

  13. Iron and carbon monoxide attenuate Crotalus atrox venom-enhanced tissue-type plasminogen activator-initiated fibrinolysis.

    Science.gov (United States)

    Nielsen, Vance G; Boyer, Leslie V; Matika, Ryan W; Amos, Quinlan; Redford, Daniel T

    2016-07-01

    In addition to degrading fibrinogen as a source of consumptive coagulopathy, rattlesnake venom has also been demonstrated to enhance fibrinolysis and degrade alpha-2-antiplasmin. The goals of this investigation was to characterize the kinetic fibrinolytic profile of Crotalus atrox venom in the absence and presence of tissue-type plasminogen activator (tPA), and to also ascertain if iron and carbon monoxide (CO, a positive modulator of alpha-2-antiplasmin) could attenuate venom-enhanced fibrinolysis. Utilizing thrombelastographic methods, the coagulation and fibrinolytic kinetic profiles of human plasma exposed to C. atrox venom (0-2 μg/ml) were determined in the absence or presence of tPA (0-100 IU/ml). Then, either separately or in combination, plasma was exposed to iron (ferric chloride, 10 μmol/l) or CO (carbon monoxide-releasing molecule-2, 100 μmol/l) prior to incubation with venom; the plasma sample was subsequently subjected to thrombelastographic analysis with addition of tPA. Venom exposure in the absence of tPA did not result in detectable fibrinolysis. In the presence of tPA, venom markedly enhanced fibrinolysis. Iron and CO, markedly attenuated venom enhancement of fibrinolysis. C. atrox venom enhances tPA-mediated fibrinolysis, and interventions that enhance/protect alpha-2-antiplasmin activity significantly attenuate venom-enhanced fibrinolysis. Future preclinical investigation is required to determine if iron and CO can attenuate venom-mediated degradation of alpha-2-antiplasmin-dependent fibrinolytic resistance.

  14. Deconstructing a Species-Complex: Geometric Morphometric and Molecular Analyses Define Species in the Western Rattlesnake (Crotalus viridis).

    Science.gov (United States)

    Davis, Mark A; Douglas, Marlis R; Collyer, Michael L; Douglas, Michael E

    2016-01-01

    Morphological data are a conduit for the recognition and description of species, and their acquisition has recently been broadened by geometric morphometric (GM) approaches that co-join the collection of digital data with exploratory 'big data' analytics. We employed this approach to dissect the Western Rattlesnake (Crotalus viridis) species-complex in North America, currently partitioned by mitochondrial (mt)DNA analyses into eastern and western lineages (two and seven subspecies, respectively). The GM data (i.e., 33 dorsal and 50 lateral head landmarks) were gleaned from 2,824 individuals located in 10 museum collections. We also downloaded and concatenated sequences for six mtDNA genes from the NCBI GenBank database. GM analyses revealed significant head shape differences attributable to size and subspecies-designation (but not their interactions). Pairwise shape distances among subspecies were significantly greater than those derived from ancestral character states via squared-change parsimony, with the greatest differences separating those most closely related. This, in turn, suggests the potential for historic character displacement as a diversifying force in the complex. All subspecies, save one, were significantly differentiated in a Bayesian discriminant function analysis (DFA), regardless of whether our priors were uniform or informative (i.e., mtDNA data). Finally, shape differences among sister-clades were significantly greater than expected by chance alone under a Brownian model of evolution, promoting the hypothesis that selection rather than drift was the driving force in the evolution of the complex. Lastly, we combine head shape and mtDNA data so as to derived an integrative taxonomy that produced robust boundaries for six OTUs (operational taxonomic units) of the C. viridis complex. We suggest these boundaries are concomitant with species-status and subsequently provide a relevant nomenclature for its recognition and representation.

  15. Experimentally altered navigational demands induce changes in the cortical forebrain of free-ranging northern pacific rattlesnakes (Crotalus o. oreganus).

    Science.gov (United States)

    Holding, Matthew L; Frazier, Julius A; Taylor, Emily N; Strand, Christine R

    2012-01-01

    The hippocampus of birds and mammals plays a crucial role in spatial memory and navigation. The hippocampus exhibits plasticity in adulthood in response to diverse environmental factors associated with spatial demands placed on an animal. The medial and dorsal cortices of the telencephalon of squamate reptiles have been implicated as functional homologues to the hippocampus. This study sought to experimentally manipulate the navigational demands placed on free-ranging northern Pacific rattlesnakes (Crotalus o. oreganus) to provide direct evidence of the relationship between spatial demands and neuroplasticity in the cortical telencephalon of the squamate brain. Adult male rattlesnakes were radio-tracked for 2 months, during which time 1 of 3 treatments was imposed weekly, namely 225-meter translocation in a random direction, 225-meter walk and release at that day's capture site (handling control) or undisturbed (control). Snakes were then sacrificed and the brains were removed and processed for histological analysis of cortical features. The activity range was larger in the translocated (Tr) group compared to the handled (Hd) and undisturbed control (Cn) groups when measured via 95% minimum convex polygon (MCP). At the 100% MCP level, Tr snakes had larger activity ranges than the Cn snakes only. The volume of the medial cortex (MC) was larger in the Tr group compared to the Cn group. The MC of Hd snakes was not significantly different from that of either of the other groups. No differences in dorsal cortex (DC) or lateral cortex volumes were detected among the groups. Numbers of 5-bromo-2'-deoxyuridine (BrdU)-labeled cells in the MC and DC 3 weeks after BrdU injection were not affected by treatment. This study establishes a causal relationship between navigational demands and greater MC volume in a free-ranging reptile. Copyright © 2012 S. Karger AG, Basel.

  16. Molecular models of the Mojave rattlesnake (Crotalus scutulatus scutulatus) venom metalloproteinases reveal a structural basis for differences in hemorrhagic activities.

    Science.gov (United States)

    Dagda, Ruben K; Gasanov, Sardar E; Zhang, Boris; Welch, William; Rael, Eppie D

    2014-03-01

    Rattlesnake venom can differ in composition and in metalloproteinase-associated activities. The molecular basis for this intra-species variation in Crotalus scutulatus scutulatus (Mojave rattlesnake) remains an enigma. To understand the molecular basis for intra-species variation of metalloproteinase-associated activities, we modeled the three-dimensional structures of four metalloproteinases based on the amino acid sequence of four variations of the proteinase domain of the C. s. scutulatus metalloproteinase gene (GP1, GP2, GP3, and GP4). For comparative purposes, we modeled the atrolysin metalloproteinases of C. atrox as well. All molecular models shared the same topology. While the atrolysin metalloproteinase molecular models contained highly conserved substrate binding sites, the Mojave rattlesnake metalloproteinases showed higher structural divergence when superimposed onto each other. The highest structural divergence among the four C. s. scutulatus molecular models was located at the northern cleft wall and the S'1-pocket of the substrate binding site, molecular regions that modulate substrate selectivity. Molecular dynamics and field potential maps for each C. s. scutulatus metalloproteinase model demonstrated that the non-hemorrhagic metalloproteinases (GP2 and GP3) contain highly basic molecular and field potential surfaces while the hemorrhagic metalloproteinases GP1 and atrolysin C showed extensive acidic field potential maps and shallow but less dynamic active site pockets. Hence, differences in the spatial arrangement of the northern cleft wall, the S'1-pocket, and the physico-chemical environment surrounding the catalytic site contribute to differences in metalloproteinase activities in the Mojave rattlesnake. Our results provide a structural basis for variation of metalloproteinase-associated activities in the rattlesnake venom of the Mojave rattlesnake.

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

    Science.gov (United States)

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

    2012-08-01

    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 family. The NP family comprises three members, ANP (atrial natriuretic peptide), BNP (b-type natriuretic peptide) and CNP (c-type natriuretic peptide), and has an important role in blood pressure regulation and electrolyte homeostasis. The aim of the present study was to characterize a novel natriuretic-like peptide (Coa_NP2), isolated from C. o. abyssus venom. The Coa_NP2 presents an average molecular mass of 3419.88Da (theoretical average molecular mass 3418.94Da, monoisotopic molecular mass 3416.66Da and theoretical PI 7.78) and its amino acid sequence presents the loop region that is characteristic of natriuretic peptides. The peptide has 32 amino acids and its complete sequence is SYGISSGCFGLKLDRIGTMSGLGCWRLLQDSP. Coa_NP2 is a natriuretic peptide of the ANP/BNP-like family, since the carboxyterminal region of CNP has its own NP domain. We demonstrate, herein, that Coa_NP2 produces a dose-dependent decrease in mean arterial pressure in rats, followed by significant increases in concentrations of markers of nitric oxide formation measured in the plasma and vasorelaxation in a thoracic aortic ring bath. The structural and biological aspects confirm Coa_NP2 as a new natriuretic peptide, isolated from snake venom. Copyright © 2012 Elsevier Inc. All rights reserved.

  18. Deconstructing a Species-Complex: Geometric Morphometric and Molecular Analyses Define Species in the Western Rattlesnake (Crotalus viridis.

    Directory of Open Access Journals (Sweden)

    Mark A Davis

    Full Text Available Morphological data are a conduit for the recognition and description of species, and their acquisition has recently been broadened by geometric morphometric (GM approaches that co-join the collection of digital data with exploratory 'big data' analytics. We employed this approach to dissect the Western Rattlesnake (Crotalus viridis species-complex in North America, currently partitioned by mitochondrial (mtDNA analyses into eastern and western lineages (two and seven subspecies, respectively. The GM data (i.e., 33 dorsal and 50 lateral head landmarks were gleaned from 2,824 individuals located in 10 museum collections. We also downloaded and concatenated sequences for six mtDNA genes from the NCBI GenBank database. GM analyses revealed significant head shape differences attributable to size and subspecies-designation (but not their interactions. Pairwise shape distances among subspecies were significantly greater than those derived from ancestral character states via squared-change parsimony, with the greatest differences separating those most closely related. This, in turn, suggests the potential for historic character displacement as a diversifying force in the complex. All subspecies, save one, were significantly differentiated in a Bayesian discriminant function analysis (DFA, regardless of whether our priors were uniform or informative (i.e., mtDNA data. Finally, shape differences among sister-clades were significantly greater than expected by chance alone under a Brownian model of evolution, promoting the hypothesis that selection rather than drift was the driving force in the evolution of the complex. Lastly, we combine head shape and mtDNA data so as to derived an integrative taxonomy that produced robust boundaries for six OTUs (operational taxonomic units of the C. viridis complex. We suggest these boundaries are concomitant with species-status and subsequently provide a relevant nomenclature for its recognition and representation.

  19. Is Hybridization a Source of Adaptive Venom Variation in Rattlesnakes? A Test, Using a Crotalus scutulatus × viridis Hybrid Zone in Southwestern New Mexico.

    Science.gov (United States)

    Zancolli, Giulia; Baker, Timothy G; Barlow, Axel; Bradley, Rebecca K; Calvete, Juan J; Carter, Kimberley C; de Jager, Kaylah; Owens, John Benjamin; Price, Jenny Forrester; Sanz, Libia; Scholes-Higham, Amy; Shier, Liam; Wood, Liam; Wüster, Catharine E; Wüster, Wolfgang

    2016-06-16

    Venomous snakes often display extensive variation in venom composition both between and within species. However, the mechanisms underlying the distribution of different toxins and venom types among populations and taxa remain insufficiently known. Rattlesnakes (Crotalus, Sistrurus) display extreme inter- and intraspecific variation in venom composition, centered particularly on the presence or absence of presynaptically neurotoxic phospholipases A₂ such as Mojave toxin (MTX). Interspecific hybridization has been invoked as a mechanism to explain the distribution of these toxins across rattlesnakes, with the implicit assumption that they are adaptively advantageous. Here, we test the potential of adaptive hybridization as a mechanism for venom evolution by assessing the distribution of genes encoding the acidic and basic subunits of Mojave toxin across a hybrid zone between MTX-positive Crotalus scutulatus and MTX-negative C. viridis in southwestern New Mexico, USA. Analyses of morphology, mitochondrial and single copy-nuclear genes document extensive admixture within a narrow hybrid zone. The genes encoding the two MTX subunits are strictly linked, and found in most hybrids and backcrossed individuals, but not in C. viridis away from the hybrid zone. Presence of the genes is invariably associated with presence of the corresponding toxin in the venom. We conclude that introgression of highly lethal neurotoxins through hybridization is not necessarily favored by natural selection in rattlesnakes, and that even extensive hybridization may not lead to introgression of these genes into another species.

  20. Genetic diversity and genetic structure of an endemic Mexican Dusky Rattlesnake (Crotalus triseriatus) in a highly modified agricultural landscape: implications for conservation.

    Science.gov (United States)

    Sunny, Armando; Monroy-Vilchis, Octavio; Zarco-González, Martha M; Mendoza-Martínez, Germán David; Martínez-Gómez, Daniel

    2015-12-01

    It is necessary to determine genetic diversity of fragmented populations in highly modified landscapes to understand how populations respond to land-use change. This information will help guide future conservation and management strategies. We conducted a population genetic study on an endemic Mexican Dusky Rattlesnake (Crotalus triseriatus) in a highly modified landscape near the Toluca metropolitan area, in order to provide crucial information for the conservation of this species. There was medium levels of genetic diversity, with a few alleles and genotypes. We identified three genetically differentiated clusters, likely as a result of different habitat cover type. We also found evidence of an ancestral genetic bottleneck and medium values of effective population size. Inbreeding coefficients were low and there was a moderate gene flow. Our results can be used as a basis for future research and C. triseriatus conservation efforts, particularly considering that the Trans-Mexican Volcanic Belt is heavily impacted by destructive land-use practices.

  1. The rattling sound of rattlesnakes (Crotalus viridis) as a communicative resource for ground squirrels (Spermophilus beecheyi) and burrowing owls (Athene cunicularia).

    Science.gov (United States)

    Owings, Donald H; Rowe, Matthew P; Rundus, Aaron S

    2002-06-01

    Animal communication involves very dynamic processes that can generate new uses and functions for established communicative activities. In this article, the authors describe how an aposematic signal, the rattling sound of rattlesnakes (Crotalus viridis), has been exploited by 2 ecological associates of rattlesnakes: (a) California ground squirrels (Spermophilus beecheyi) use incidental acoustic cues in rattling sounds to assess the danger posed by the rattling snake, and (b) burrowing owls (Athene cunicularia) defend themselves against mammalian predators by mimicking the sound of rattling. The remarkable similarity between the burrowing owl's defensive hiss and the rattlesnake's rattling reflects both exaptation and adaptation. Such exploitation of the rattling sound has favored alternations in both the structure and the deployment of rattling by rattlesnakes.

  2. 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

  3. 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

  4. 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

    2013-03-15

    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, Crotalus 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 IC(50) of 1 nM, collagen- and ADP-induced platelet aggregation with the estimated IC(50)s 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. Published by Elsevier Ltd.

  5. Physiological importance of the coronary arterial blood supply to the rattlesnake heart

    DEFF Research Database (Denmark)

    Hagensen, Mette; Abe, Augusto S.; Falk, Erling

    2008-01-01

    remains unknown. In the present study we investigate the effects of permanent coronary artery occlusion in the South American rattlesnake (Crotalus durissus) on the ability to maintain heart rate and blood pressure at rest and during short term activity. We used colored silicone rubber (Microfil...

  6. A new serovar and a new serological variant belonging to Salmonella enterica subspecies diarizonae

    Directory of Open Access Journals (Sweden)

    CA Solari

    2003-06-01

    Full Text Available Description of a new serovar (S. IIIb 16:k:e,n,x,z15 and a new serological variant (S. IIIb 42:z10:e,n,x,z15:z60 belonging to the genus Salmonella isolated from stool specimens of Brazilian snakes (Crotalus durissus.

  7. 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

    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...

  8. 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.

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

    Directory of Open Access Journals (Sweden)

    Rulon W Clark

    Full Text Available 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.

  10. Disintegrins of Crotalus simus tzabcan venom: Isolation, characterization and evaluation of the cytotoxic and anti-adhesion activities of tzabcanin, a new RGD disintegrin.

    Science.gov (United States)

    Saviola, Anthony J; Modahl, Cassandra M; Mackessy, Stephen P

    2015-09-01

    Disintegrins are small non-enzymatic proteins common in the venoms of many viperid snakes. These proteins have received significant attention due to their ability to inhibit platelet aggregation and cell adhesion, making them model compounds in drug development and design investigations. The present study used a combination of molecular and proteomic techniques to screen the venom of the Middle American Rattlesnake (Crotalus simus tzabcan) for novel disintegrins. Six disintegrin isoforms were identified, and the most abundant, named tzabcanin, was further isolated and characterized. Tzabcanin consists of 71 amino acids, has a mass of 7105 Da (by MALDI-TOF mass spectrometry) and contains the canonical RGD binding domain. Tzabcanin was not cytotoxic to MCF-7 cells but showed weak cytotoxicity to Colo-205 cells following a 24 h incubation period. Tzabcanin inhibited cell adhesion of both cell lines to immobilized fibronectin and vitronectin, and cell adhesion to immobilized tzabcanin was inhibited when cells were incubated with a cation chelator (EDTA), indicating that integrin-tzabcanin binding is specific. This study provides a detailed analysis of the purification and characterization of tzabcanin and provides sequence and mass data for the multiple disintegrins present in the venom of C. s. tzabcan. Copyright © 2015 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

  11. 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

    2016-09-01

    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. Copyright © 2016 Elsevier Ltd. All rights reserved.

  12. 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

  13. Biochemical and functional studies of ColTx-I, a new myotoxic phospholipase A2 isolated from Crotalus oreganus lutosus (Great Basin rattlesnake) snake venom.

    Science.gov (United States)

    Almeida, J R; Resende, L M; Silva, A G; Ribeiro, R I M A; Stábeli, R G; Soares, A M; Calderon, L A; Marangoni, S; Da Silva, S L

    2016-07-01

    Commonly, phospholipases A2 (PLA2s) play key roles in the pathogenesis of the local tissue damage characteristic of crotaline and viperine snake envenomations. Crotalus oreganus lutosus snake venom has not been extensively studied; therefore, the characterization of its components represents a valuable biotechnological tool for studying pathophysiological processes of envenoming and for gaining a deeper understanding of its biological effects. In this study, for the first time, a basic PLA2 myotoxin, ColTx-I, was purified from C. o. lutosus through two chromatographic steps. ColTx-I is monomeric with calculated molecular mass weight (Mw) of 14,145 Da and a primary structure closely related to basic PLA2s from viperid venoms. The pure enzyme has a specific activity of 15.87 ± 0.65 nmol/min/mg at optimal conditions (pH 8.0 and 37 °C). ColTx-I activity was found to be dependent on Ca(2+), as its substitution by other ionic species as well as the addition of chelating agents significantly reduced its phospholipase activity. In vivo, ColTx-I triggered dose-dependent inflammatory responses, measured using the paw edema model, with an increase in IL-6 levels, systemic and local myotoxicity, characterized by elevated plasma creatine kinase activity. ColTx-I induced a complex series of degenerative events associated with edema, inflammatory infiltrate and skeletal muscle necrosis. These biochemical and functional results suggest that ColTx-I, a myotoxic and inflammatory mediator, plays a relevant role in C. o. lutosus envenomation. Thus, detailed studies on its mechanism of action, such as evaluating the synergism between ColTx-I and other venom components may reveal targets for the development of more specific and effective therapies. Copyright © 2016 Elsevier Ltd. All rights reserved.

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

    Science.gov (United States)

    Samy, Ramar Perumal; Kandasamy, Matheswaran; Gopalakrishnakone, Ponnampalam; Stiles, Bradley G; Rowan, Edward G; Becker, David; Shanmugam, Muthu K; Sethi, Gautam; Chow, Vincent T K

    2014-01-01

    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.

  15. Retrospective evaluation of the effect of antivenom administration on hospitalization duration and treatment cost for dogs envenomated by Crotalus viridis: 113 dogs (2004-2012).

    Science.gov (United States)

    Katzenbach, Julia E; Foy, Daniel S

    2015-01-01

    To compare the effect of antivenom administration on mortality, hospitalization duration, and cost of hospitalization for dogs envenomated by Crotalus viridis. Retrospective study (January 2004-December 2012). Private veterinary emergency and referral center. One hundred thirteen dogs with confirmed C. viridis envenomation. None. Dogs were divided into groups treated with either supportive care only (group 1) or supportive care plus antivenin crotalidae polyvalent (group 2). A modified snakebite severity score was used to compare patients from group 1 and group 2. Patients in group 2 received one 10 mL vial of antivenin crotalidae polyvalent over 4-6 hours. The mortality rate was 1.8% (2/113).  Group 1 had a median duration of hospitalization of 20 hours (range, 8-50 hours), while group 2 had a median duration of hospitalization of 24 hours (range, 1.5-74 hours). Group 1 had a median cost of hospitalization of 1050.00 USD (range, 423.52-2266.09 USD) while group 2 had a median cost of hospitalization of 2002.19 USD (range, 1139.91-6908.01 USD). Both the duration of hospitalization (P < 0.01) and the cost of hospitalization (P < 0.01) were significantly greater in the group of dogs receiving antivenom. Both the cost and the duration of hospitalization were significantly greater in the group of dogs that received antivenom. The difference in cost between the groups was approximately the cost of the antivenom vial and administration. Amongst the parameters evaluated, no significant benefit was associated with antivenom administered to dogs envenomated by C. viridis. © Veterinary Emergency and Critical Care Society 2015.

  16. 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.

  17. Integrated "omics" profiling indicates that miRNAs are modulators of the ontogenetic venom composition shift in the Central American rattlesnake, Crotalus simus simus.

    Science.gov (United States)

    Durban, Jordi; Pérez, Alicia; Sanz, Libia; Gómez, Aarón; Bonilla, Fabián; Rodríguez, Santos; Chacón, Danilo; Sasa, Mahmood; Angulo, Yamileth; Gutiérrez, José M; Calvete, Juan J

    2013-04-10

    Understanding the processes that drive the evolution of snake venom is a topic of great research interest in molecular and evolutionary toxinology. Recent studies suggest that ontogenetic changes in venom composition are genetically controlled rather than environmentally induced. However, the molecular mechanisms underlying these changes remain elusive. Here we have explored the basis and level of regulation of the ontogenetic shift in the venom composition of the Central American rattlesnake, Crotalus s. simus using a combined proteomics and transcriptomics approach. Proteomic analysis showed that the ontogenetic shift in the venom composition of C. s. simus is essentially characterized by a gradual reduction in the expression of serine proteinases and PLA2 molecules, particularly crotoxin, a β-neurotoxic heterodimeric PLA2, concominantly with an increment of PI and PIII metalloproteinases at age 9-18 months. Comparison of the transcriptional activity of the venom glands of neonate and adult C. s. simus specimens indicated that their transcriptomes exhibit indistinguisable toxin family profiles, suggesting that the elusive mechanism by which shared transcriptomes generate divergent venom phenotypes may operate post-transcriptionally. Specifically, miRNAs with frequency count of 1000 or greater exhibited an uneven distribution between the newborn and adult datasets. Of note, 590 copies of a miRNA targeting crotoxin B-subunit was exclusively found in the transcriptome of the adult snake, whereas 1185 copies of a miRNA complementary to a PIII-SVMP mRNA was uniquely present in the newborn dataset. These results support the view that age-dependent changes in the concentration of miRNA modulating the transition from a crotoxin-rich to a SVMP-rich venom from birth through adulthood can potentially explain what is observed in the proteomic analysis of the ontogenetic changes in the venom composition of C. s. simus. Existing snake venom toxins are the result of early

  18. Integrated “omics” profiling indicates that miRNAs are modulators of the ontogenetic venom composition shift in the Central American rattlesnake, Crotalus simus simus

    Science.gov (United States)

    2013-01-01

    Background Understanding the processes that drive the evolution of snake venom is a topic of great research interest in molecular and evolutionary toxinology. Recent studies suggest that ontogenetic changes in venom composition are genetically controlled rather than environmentally induced. However, the molecular mechanisms underlying these changes remain elusive. Here we have explored the basis and level of regulation of the ontogenetic shift in the venom composition of the Central American rattlesnake, Crotalus s. simus using a combined proteomics and transcriptomics approach. Results Proteomic analysis showed that the ontogenetic shift in the venom composition of C. s. simus is essentially characterized by a gradual reduction in the expression of serine proteinases and PLA2 molecules, particularly crotoxin, a β-neurotoxic heterodimeric PLA2, concominantly with an increment of PI and PIII metalloproteinases at age 9–18 months. Comparison of the transcriptional activity of the venom glands of neonate and adult C. s. simus specimens indicated that their transcriptomes exhibit indistinguisable toxin family profiles, suggesting that the elusive mechanism by which shared transcriptomes generate divergent venom phenotypes may operate post-transcriptionally. Specifically, miRNAs with frequency count of 1000 or greater exhibited an uneven distribution between the newborn and adult datasets. Of note, 590 copies of a miRNA targeting crotoxin B-subunit was exclusively found in the transcriptome of the adult snake, whereas 1185 copies of a miRNA complementary to a PIII-SVMP mRNA was uniquely present in the newborn dataset. These results support the view that age-dependent changes in the concentration of miRNA modulating the transition from a crotoxin-rich to a SVMP-rich venom from birth through adulhood can potentially explain what is observed in the proteomic analysis of the ontogenetic changes in the venom composition of C. s. simus. Conclusions Existing snake venom

  19. Survival of timber rattlesnakes (Crotalus horridus) estimated by capture-recapture models in relation to age, sex, color morph, time, and birthplace

    Science.gov (United States)

    Brown, W.S.; Kery, M.; Hines, J.E.

    2007-01-01

    Juvenile survival is one of the least known elements of the life history of many species, in particular snakes. We conducted a mark–recapture study of Crotalus horridus from 1978–2002 in northeastern New York near the northern limits of the species' range. We marked 588 neonates and estimated annual age-, sex-, and morph-specific recapture and survival rates using the Cormack-Jolly-Seber (CJS) model. Wild-caught neonates (field-born, n  =  407) and neonates produced by captive-held gravid females (lab-born, n  =  181) allowed comparison of the birthplace, or lab treatment effect, in estimated survival. Recapture rates declined from about 10–20% over time while increasing from young to older age classes. Estimated survival rates (S ± 1 SE) in the first year were significantly higher among field-born (black morph: S  =  0.773 ± 0.203; yellow morph: S  =  0.531 ± 0.104) than among lab-born snakes (black morph: S  =  0.411 ± 0.131; yellow morph: S  =  0.301 ± 0.081). Lower birth weights combined with a lack of field exposure until release apparently contributed to the lower survival rate of lab-born snakes. Subsequent survival estimates for 2–4-yr-old snakes were S  =  0.845 ± 0.084 for the black morph and S  =  0.999 (SE not available) for the yellow morph, and for ≥5-yr-old snakes S  =  0.958 ± 0.039 (black morph) and S  =  0.822 ± 0.034 (yellow morph). The most parsimonious model overall contained an independent time trend for survival of each age, morph, and lab-treatment group. For snakes of the first two age groups (ages 1 yr and 2–4 yr), survival tended to decline over the years for both morphs, while for adult snakes (5 yr and older), survival was constant or even slightly increased. Our data on survival and recapture are among the first rigorous estimates of these parameters in a rattlesnake and among the few yet available for any viperid snake. These data are useful for analyses of the life

  20. Recombinant disintegrin (r-Cam-dis) from Crotalus adamanteus inhibits adhesion of human pancreatic cancer cell lines to laminin-1 and vitronectin.

    Science.gov (United States)

    Suntravat, Montamas; Barret, Henriquez S; Jurica, Cameron A; Lucena, Sara E; Perez, John C; Sánchez, Elda E

    2015-01-01

    Pancreatic cancer is a malignant cancer common worldwide having poor prognosis, even when diagnosed at its early stage. Cell adhesion plays a critical role in cancer invasion and metastasis. Integrins are major mediators of cell adhesion and play an important role in invasion and metastatic growth of human pancreatic cancer cells. Snake disintegrins are the most potent ligands of several integrins and have potential therapeutic applications for cancers. We have previously cloned and expressed a new recombinant RGD-disintegrin from Crotalus adamanteus (r-Cam-dis). This recently published r-Cam-dis has an extra nine amino acids derived from the vector (SPGARGSEF) at the N-terminus end and has strong anti-platelet activity. However, this r-Cam-dis contains the contamination of the cleavage of the N-terminal end of the pET-43.1a cloning vector. In this study, we have cloned r-Cam-dis in a different cloning vector (pGEX-4T-1) showing five different amino acids (GSPEF) at the N-terminal part. This new r-Cam-dis was expressed and tested for inhibition of platelet aggregation, specific binding activity with seven different integrins, and inhibition of adhesion of three different pancreatic cancer cell lines on laminin-1 and vitronectin. The r-Cam-dis showed potent binding to αvβ3 integrin, but was moderate to weak with αvβ5, αvβ6, α2β1, and α6β1. Interestingly, the inhibition of r-Cam-dis on pancreatic cancer cell lines adhesion to laminin-1 was more effective than that to vitronectin. Based on our binding results to integrin receptors and previous adhesion studies using function-blocking monoclonal antibodies, it is suggested that r-Cam-dis could be inhibiting adhesion of pancreatic cancer cell lines through integrins α2β1, α6β1, αvβ5, and αvβ6.

  1. 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

  2. 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

  3. Self-association and active enzyme forms of Naja naja naja and Crotalus atrox phospholipase A2 studied by analytical ultracentrifugation.

    Science.gov (United States)

    Bukowski, T; Teller, D C

    1986-12-02

    The dimerization of phospholipase A2 (PLPA2) from Naja naja naja (Pakistani cobra) and Crotalus atrox (Western Diamondback rattlesnake) has been studied from pH 2.5 to 11 at 20 degrees C in 1 mM CaCl2, 0.21 M ionic strength. For the C. atrox enzyme, it was found necessary to use a combination of sedimentation equilibrium and fluorescence yield data to analyze the association. Sedimentation equilibrium in the analytical ultracentrifuge sufficed for the study of the N. naja PLPA2. In the region of enzymatic activity at pH 8, the dimerization association constants found were k2 = 2.8 X 10(6) L/mol and k2 = 6.9 X 10(4) L/mol for the C. atrox and N. naja enzymes, respectively. Analytical linked functions are presented which describe the data. Because the associations are linked to Ca2+ as well as the hydrogen ion, no attempt was made to interpret the ionization of residues in terms of the molecular structure. Active-enzyme sedimentation velocity experiments have been used to study the relation between enzymatic activity and association for both the C. atrox and N. naja enzymes. The substrate 1,2-dibutyryl-sn-glycero-3-phosphocholine (diC4PC) did not dissociate the C. atrox PLPA2. The substrate 1,2-dihexanoyl-sn-glycero-3-phosphocholine (diC6PC) at 7.5 mM dissociated the C. atrox PLPA2 when monitored either as the active enzyme or as the Sr2+-inhibited enzyme. At low enzyme concentrations, 40 mM diC4PC had no effect on N. naja PLPA2 dimerization. However, the sedimentation coefficients observed at enzyme concentrations above 0.2 mg/mL in active-enzyme sedimentation velocity experiments were larger than the values predicted from the thermodynamic studies. Sedimentation coefficients observed for the N. naja PLPA2 acting on diC6PC were larger than those of the monomeric protein, which was the form layered on this substrate. The dissociation of the C. atrox PLPA2 effected by diC6PC was analyzed by the thermodynamics of association and the kinetic Michaelis constant. The

  4. Hemogregarinas en serpientes de Costa Rica

    OpenAIRE

    Moreno, E.; Bolaños, R.

    2016-01-01

    Thirty-three percent of 216 venomous and non-venomous captive snake of Costa Rica were parasitized with hemogregarines (Protozoa: Sporozoa). The species most affected were Lachesis muta, Crotalus durissus and Boa constrictor with scores above 50%. None of the arboreal species harbored hemogregarines.Five morphological blood types were demonstrated and frequently more than one type occurred in a single host.Lung and liver sections of the parasitized snakes showed cysts similar to those of schy...

  5. Contribuição à biologia de serpentes da Bahia, Brasil. I. vivíparas

    OpenAIRE

    Silva, Rejane Maria Lira da; Silva, Luciana Lyra Casais e; Queiroz, Ilka Biondi de; Nunes, Tânia Brazil

    1994-01-01

    P. 187 - 193. 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) ...

  6. Contribution to reproductive biology of snakes in Bahia, Brazil: I. viviparous

    OpenAIRE

    Lira-da-Silva, Rejane Maria; Casais-e-Silva, Luciana Lyra; Queiroz, Ilka Biondi de; Nunes, Tânia Brazil

    1994-01-01

    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...

  7. Anti-inflammatory potential of zootherapeutics derived from animals used in Brazilian traditional medicine.

    Science.gov (United States)

    Ferreira, Felipe S; Brito, Samuel V; Sales, Débora L; Menezes, Irwin R A; Coutinho, Henrique D M; Souza, Emmanuel P; Almeida, Waltécio O; Alves, Rômulo R N

    2014-11-01

    Animals are used for the treatment of diseases caused by inflammatory processes, although few studies evaluate their potential for these purposes. To evaluate the anti-inflammatory potential of zootherapeutic products derived from vertebrates used in Brazilian traditional medicine. The species analyzed were Tupinambis merianae, Iguana iguana, Crotalus durissus, Boa constrictor, and Euphractus sexcinctus. The methods used in anti-inflammatory assays were ear edema (topical) and paw (systemic). With regard to topical anti-inflammatory activity, the fat from T. merianae, C. durissus, I. iguana, B. constrictor, and E. sexcinctus reduced inflammation, while for systemic anti-inflammatory activity, only the fat and the skin of C. durissus, the skin of I. iguana and the fat from B. constrictor reduced inflammation. Studies should be conducted to evaluate the mechanisms of action for each product that demonstrated anti-inflammatory activity as well as against other inflammatory processes.

  8. Non-Clinical Studies for Evaluation of 8-C-Rhamnosyl Apigenin Purified from Peperomia obtusifolia against Acute Edema

    Directory of Open Access Journals (Sweden)

    Cinthia I. Tamayose

    2017-09-01

    Full Text Available Compound 8-C-rhamnosyl apigenin (8CR induced a moderate reduction in the enzymatic activity of secretory phospholipase A2 (sPLA2 from Crotalus durissus terrificus and cytosolic phospholipase A2 (cPLA2, but the compound also significantly inhibited the enzymatic activity of the enzyme cyclooxygenase. In vitro assays showed that the compound induced a slight change in the secondary structure of sPLA2 from Crotalus durissus terrificus snake venom. In vivo assays were divided into two steps. In the first step, the 8CR compound was administered by intraperitoneal injections 30 min prior to administration of sPLA2. In this condition, 8CR inhibited edema and myonecrosis induced by the sPLA2 activity of Crotalus durissus terrificus in a dose-dependent manner by decreasing interleukin-1β (IL-1β, tumor necrosis factor α (TNF-α, prostaglandin E2 (PGE2, and lipid peroxidation. This has been demonstrated by monitoring the levels of malondialdehyde (MDA in rat paws after the course of edema induced by sPLA2. These results, for the first time, show that sPLA2 of Crotalus durissus terrificus venom induces massive muscle damage, as well as significant edema by mobilization of cyclooxygenase enzymes. Additionally, its pharmacological activity involves increased lipid peroxidation as well as TNF-α and IL-1β production. Previous administration by the peritoneal route has shown that dose-dependent 8CR significantly decreases the enzymatic activity of cyclooxygenase enzymes. This resulted in a decrease of the amount of bioactive lipids involved in inflammation; it also promoted a significant cellular protection against lipid peroxidation. In vivo experiments performed with 8CR at a concentration adjusted to 200 μg (8 mg/kg of intraperitoneal injection 15 min after sPLA2 injection significantly reduced sPLA2 edema and the myotoxic effect induced by sPLA2 through the decrease in the enzymatic activity of cPLA2, cyclooxygenase, and a massive reduction of lipid

  9. 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 (female

  10. 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

  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. Ocorrência de alguns endo e ectoparasitos no serpentário da UNIFENAS - Universidade de Alfenas - MG

    Directory of Open Access Journals (Sweden)

    Terezinha Araújo

    1999-01-01

    Full Text Available Specimens of Crotalus durissus terrificus and Bothrops alternatus held in captivity by University of Alfenas- MG were examined with the aim of studying the parasitological fauna of the snakes. These animals showed anorexia, dehydration, diarrhea and anemia. Three rattlesnakes and two Bothrops alternatus died about two months after the beginning of the clinical signs. It were identified helmints of the genus Kalicephalus, Ophidascaris, Rhabdias and Oxyuris; protozoans of the genus Haemogregarina and mites of the genus Ophionysus. Ivermectin was administrated to the survivors.

  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. Atividade antiinflamatória do extrato aquoso de Arrabidaea chica (Humb. & Bonpl. B. Verl. sobre o edema induzido por venenos de serpentes amazônicas

    Directory of Open Access Journals (Sweden)

    Denys Paixão Costa de Oliveira

    Full Text Available O estudo investigou o efeito antiinflamatório do extrato aquoso da Arrabidaea chica (Humb. & Bonpl. B. Verl., Bignoniaceae, popularmente conhecida como "crajiru", sobre o edema induzido por venenos de serpentes amazônicas dos gêneros Brothrops e Crotalus, em camundongos albinos, por via oral, intraperitoneal e subcutânea. O efeito anti-edematogênico foi avaliado pela medição do diâmetro dos coxins das patas posteriores, sendo medidos as 1, 3, 6, 12 e 24 horas, para B. atrox e 1, 3 e 6 horas para Crotalus durissus ruruima, e também avaliado por histopatologia. O estudo mostrou que o efeito inibitório do extrato aquoso para o gênero Bothrops, pelas vias subcutânea e intraperitoneal (12 horas foi de 55,87% e 65,70%, respectivamente. Para o gênero Crotalus o efeito inibitório do extrato pela via subcutânea após 3 horas foi de 33,55% e após 6 horas de 79,81%. Pela via intraperitoneal após 3 horas foi de 48,02% e após 6 horas de 92,52%. Na análise histopatológica, o infiltrado de granulócitos e a miocitólise foram os efeitos inflamatórios inibidos mais significativamente. Os resultados sugerem a presença no extrato aquoso de A. chica de substâncias com atividade inibitória sobre os efeitos inflamatórios dos venenos das serpentes Bothrops atrox e Crotalus durissus ruruima.

  15. Crotalidae polyvalent immune Fab (ovine) antivenom is effective in the neutralization of South American viperidae venoms in a murine model.

    Science.gov (United States)

    Richardson, William H; Tanen, David A; Tong, Tri C; Betten, David P; Carstairs, Shaun D; Williams, Saralyn R; Cantrell, Frank L; Clark, Richard F

    2005-06-01

    Crotalidae polyvalent immune Fab (ovine) (CroFab; FabAV) is used in the treatment of symptomatic crotaline envenomations in North America. Unlike Antivenin (Crotalidae) Polyvalent, which is approved for treatment of crotaline envenomation in North and South America, FabAV is manufactured using only venoms from crotaline snakes native to the United States. This study was designed to evaluate the efficacy of FabAV in the neutralization of venom from 2 South American crotaline snakes: Crotalus durissus terrificus (tropical rattlesnake) and Bothrops atrox (fer-de-lance). A randomized, blinded, placebo-controlled murine model of intraperitoneal venom injection was used. Venom potency was determined in preliminary median lethal dose (LD 50) dosing studies. Study animals were then divided into 7 groups: (1) C durissus terrificus venom (Sigma-Aldrich Co.)+FabAV, (2) C durissus terrificus venom (Sigma-Aldrich Co.)+0.9% normal saline solution, (3) C durissus terrificus venom (Biotoxins Inc.)+FabAV, (4) C durissus terrificus venom (Biotoxins Inc.)+normal saline solution, (5) B atrox venom+FabAV, (6) B atrox venom+normal saline solution, and (7) FabAV+normal saline solution. Twice the estimated LD 50 was the chosen venom dose, and the amount of FabAV injected was 10 times the amount needed for venom neutralization. Statistical analysis included Fisher's exact test and log-rank testing to compare survival rates and times. The venom LD 50 was found in preliminary studies to be 0.9 mg/kg and 1.35 mg/kg for the C durissus terrificus venom obtained from Sigma-Aldrich Co. and Biotoxins Inc., respectively. The LD 50 for B atrox venom was 5.0 mg/kg. All animals receiving venom only and saline solution died. Animals receiving FabAV together with either venom survived to the end of the 24-hour observation period ( P <.001). Comparison of survival times between groups demonstrated a significant difference in time to death between venom-only control groups and the FabAV+venom groups (P

  16. Crystallization and preliminary X-ray crystallographic analysis of the heterodimeric crotoxin complex and the isolated subunits crotapotin and phospholipase A{sub 2}

    Energy Technology Data Exchange (ETDEWEB)

    Santos, K. F.; Murakami, M. T. [Department of Physics, IBILCE/UNESP, Cristóvão Colombo 2265, CEP 15054-000, São José do Rio Preto, SP (Brazil); Cintra, A. C. O. [Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas, Universidade de São Paulo, Ribeirão Preto, SP (Brazil); Toyama, M. H.; Marangoni, S. [Departamento de Bioquímica, Universidade de Campinas, Campinas, SP (Brazil); Forrer, V. P.; Brandão Neto, J. R. [Laboratório Nacional de Luz Síncrotron, Campinas, SP (Brazil); Polikarpov, I. [Instituto de Física de São Carlos, Universidade de São Paulo, São Carlos, SP (Brazil); Arni, R. K., E-mail: arni@ibilce.unesp.br [Department of Physics, IBILCE/UNESP, Cristóvão Colombo 2265, CEP 15054-000, São José do Rio Preto, SP (Brazil); Center for Applied Toxinology, CEPID (Brazil)

    2007-04-01

    Crotoxin, a potent neurotoxin from the venom of the South American rattlesnake Crotalus durissus terrificus, exists as a heterodimer formed between a phospholipase A{sub 2} and a catalytically inactive acidic phospholipase A{sub 2} analogue (crotapotin). Large single crystals of the crotoxin complex and of the isolated subunits have been obtained. Crotoxin, a potent neurotoxin from the venom of the South American rattlesnake Crotalus durissus terrificus, exists as a heterodimer formed between a phospholipase A{sub 2} and a catalytically inactive acidic phospholipase A{sub 2} analogue (crotapotin). Large single crystals of the crotoxin complex and of the isolated subunits have been obtained. The crotoxin complex crystal belongs to the orthorhombic space group P2{sub 1}2{sub 1}2, with unit-cell parameters a = 38.2, b = 68.7, c = 84.2 Å, and diffracted to 1.75 Å resolution. The crystal of the phospholipase A{sub 2} domain belongs to the hexagonal space group P6{sub 1}22 (or its enantiomorph P6{sub 5}22), with unit-cell parameters a = b = 38.7, c = 286.7 Å, and diffracted to 2.6 Å resolution. The crotapotin crystal diffracted to 2.3 Å resolution; however, the highly diffuse diffraction pattern did not permit unambiguous assignment of the unit-cell parameters.

  17. The venom of South American rattlesnakes inhibits macrophage functions and is endowed with anti-inflammatory properties

    Directory of Open Access Journals (Sweden)

    Maria C. C. de Sousa e Silva

    1996-01-01

    Full Text Available The injection of Crotalus durissus terrificus venom into the foot pad of mice did not induce a significant inflammatory response as evaluated by oedema formation, increased vascular permeability and cell migration. The subcutaneous injection of the venom, or its addition to cell cultures, had an inhibitory effect on the spreading and phagocytosis of resident macrophages, without affecting the viability of the cells. This effect was not observed when the venom was added to cultures of thioglycollate elicited macrophages, but it was able to inhibit these macrophage functions when the cells were obtained from animals injected simultaneously with the venom and thioglycollate. These observations suggest that the venom interferes with the mechanisms of macrophage activation. Leukocyte migration induced by intraperitoneal injection of thioglycollate was also inhibited by previous venom injection. This down-regulatory activity of the venom on macrophage functions could account for the mild inflammatory response observed in the site of the snake bite in Crotalus durissus terrificus envenomation in man.

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

    Energy Technology Data Exchange (ETDEWEB)

    Soares, Marcella Araugio; Pujatti, Priscilla Brunelli; Santos, Raquel Gouvea dos [Centro de Desenvolvimento da Tecnologia Nuclear CDTN/CNEN-MG, Belo Horizonte, MG (Brazil)]. E-mails: maso@cdtn.br; santosr@cdtn.br; Dias, Consuelo Latorre Fortes [Fundacao Ezequiel Dias FUNED, Belo Horizonte, MG (Brazil); Chavez Olortegui, Carlos Delfin [Universidade Federal de Minas Gerais UFMG, Belo Horizonte, MG (Brazil). Inst. de Ciencias Biologicas; Santos, Wagner Gouvea dos [Medical College of Virginia, Richmond, VA (United States). Neurosurgery Dept.

    2007-07-01

    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 {sup 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 {sup 125}I-Crtx and {sup 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)

  19. {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)

  20. 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)

  1. 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.

  2. A proposed 3D structure for crotamine based on homology building, molecular simulations and circular dichroism.

    Science.gov (United States)

    Siqueira, A M; Martins, N F; De Lima, M E; Diniz, C R; Cartier, A; Brown, D; Maigret, B

    2002-03-01

    Crotamine, isolated from the venom of the South American rattlesnake Crotalus durissus terrificus is a strongly basic 42-amino acid polypeptide belonging to the small basic myotoxin family. As no tridimensional structure is available for this myotoxin subfamily, despite its important pharmacological interest, we propose in this paper a theoretical 3D model for crotamine. Starting from a homology modelling procedure, followed by intensive molecular dynamics (MD) simulations in water and complementary CD experiments, the designed 3D model is the first example of a tridimensional structure in this family of small basic myotoxins. Crotamine, therefore, belongs to a newly identified structural family presenting a common fold also found in beta-defensin and antopleurine-B. The proposed 3D model will be used for future calculations about crotamine aggregation and interaction with membranes.

  3. [Snake envenomation in French Guiana].

    Science.gov (United States)

    Chippaux, J P

    2002-01-01

    French Guiana is a French Overseas Department in South America. Ninety-five percent of the territory is a tropical rainforest. Its rich fauna includes seven families of snakes but only 3 are potentially venomous. Less than 12% of species and, depending on biotope, 10 to 30% of specimens collected are dangerous for humans. The annual incidence of snakebite is less than 50 bites per 100,000 inhabitants overall but increases to 600 per 100,000 for persons active in the rainforest where the risk is highest. The most common envenomation by Viperidae such as Bothrops, which is abundant throughout French Guiana, induces inflammation, necrosis and hemorrhage. Crotalus durissus, a rattlesnake living in coastal savannah, or Micrurus sp cause neuromuscular poisoning. Coral snakes are encountered throughout French Guiana, but envenomation is very rare. Antivenom therapy must be administered by the intravenous route in association with symptomatic treatment and, if necessary, resuscitation in a specialized care unit.

  4. Oxygenation properties and isoform diversity of snake hemoglobins

    DEFF Research Database (Denmark)

    Storz, Jay F.; Natarajan, Chandrasekhar; Moriyama, Hideaki

    2015-01-01

    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 - and -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...

  5. 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)

  6. 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

  7. 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

  8. South american rattlesnake venom: its hemolytic power

    Directory of Open Access Journals (Sweden)

    Édimo Garcia de Lima

    1989-12-01

    Full Text Available The hemolytic power of rattlesnake venom (Crotalus durissus terrificus was Studied. A high percentage of sample with negative hemolytic power was detected when sheep red blood cells were used. A large number of venoms with hemolytic power, though with a low hemolysis percentage, were detected when liquid, recently extracted venom was used. When crystallized venom was used under the same experimental conditions, a higher percentage ofpositivityfor hemolysis was obtained. When the results obtained on agar plates were compared to those obtained in test tubes, a large number of animals with a higher percentage of hemolysis were detected, though this value was not proportional to the number of animals showing positive plate hemolysis. When the hemolytic power of these venoms was tested on human red blood cells, a large percentage of animals with venoms having a low hemolytic power was also detected. Hemolytic power was much greater when human red blood cells were tested with crystallized venom. The preparation of red blood cells also had an important effect and the use of red blood cells from defibrinated blood is recommended. We conclude that rattlesnake venom has hemolytic power that increases when the venom is crystallized. Red blood cells should be properly preparedfor the lysis reactions. We suggest that the lytic power of the venom is related to venom concentration and to the purity of its fractions.Foi estudado o poder hemolitico do veneno da cascavel (Crotalus durissus terrificus. Encontrou-se grande número de suas frações sem capacidade de hemolisar eritrócitos de carneiro. O veneno "in natura", recentemente extraído, e em estado líquido tem pouca atividade litica. A cristalização do veneno aumenta sua concentração e poder lítico. Os resultados de hemólise do sangue de carneiro obtidos em placas e tubos foram comparados evidenciando um grande número de animais com venenos com alto poder hemolítico. Os valores não foram

  9. In vitro evaluation of copaiba oil as a kojic acid skin enhancer

    Directory of Open Access Journals (Sweden)

    Robson Vicente Machado de Oliveira

    2010-06-01

    Full Text Available The capacity of copaíba oil to act as a skin penetration enhancer for the depigmenting agent kojic acid was evaluated using an in vitro diffusion system with static flux and shed rattlesnake skin membrane, Crotalus durissus terrificus, in saline solution at 34±2 ºC as the fluid receptor. The quantities of kojic acid liberated into the fluid receptor were determined by spectrophotometry at 268 nm with intervals of one and a half hours. The membranes, pretreated with copaíba oil at 25% and 50% v/v, gave flux values of 8.0 and 12.7 µg/cm²/h, permeability values of 2.0 and 3.3 cm×10-4/h, and promotion factors of 4.1 and 3.7, respectively. These results indicate that copaíba oil, at the two concentrations studied, has the capacity to promote penetration of kojic acid.A propriedade do óleo de copaíba como agente promotor de penetração cutânea do despigmentante ácido kójico foi avaliada utilizando-se sistema de difusão in vitro com fluxo estático, membrana de pele da serpente cascavel - Crotalus durissus terrificus e solução salina a 34±2 ºC como fluido receptor. As quantidades liberadas do ácido kójico no fluido receptor foram determinadas por espectrofotometria em 268 nm em intervalos de 1:30 h. As membranas pré-tratadas com óleo de copaíba a 25 e 50% v/v apresentaram valores de fluxo de 8,0 e 12,7 µg/cm²/h, permeabilidade de 2,0 e 3,3 cm×10-4/h, e fatores de promoção de 4,1 e 3,7, respectivamente. Os resultados obtidos indicaram que o óleo de copaíba, nas duas concentrações estudadas, apresentou capacidade de promoção da penetração do ácido kójico.

  10. 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

  11. Gopherus agassizii (desert tortoise) and Crotalus ruber (red diamond rattlesnake). Burrow co-occupancy

    Science.gov (United States)

    Lovich, Jeffrey E.

    2011-01-01

    I observed an adult Desert Tortoise and an adult Red Diamond Rattlesnake (sexes unknown) in a shallow tortoise burrow on 6 January 1997 at a wind energy generation facility near Palm Springs, Riverside Co., California, USA (33.9599°N, 116.6613°W).

  12. 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...

  13. Tertiary Structural studies of Myotoxin a from Crotalus viridis viridis Venom by Nuclear Magnetic Resonance

    Science.gov (United States)

    1993-05-01

    electrophorus electri... 47 47 47 Sw:Ibb2SWheat P09864 wheat (triticum aestivum). proteina ... 46 46 51 Sw:Pa29$Pseau P20253 mulga snake (pseudechis...Sw:Pa2bSPsepo P20259 red-bellied black snake (pseudechis... 34 46 37 Sw:IbblSWheat P09863 wheat (triticum aestivum). proteina ... 46 46 52 Sw:Pa2a$Psepo P20258 red

  14. Notizen zur Fortpflanzungsbiologie und Jugendentwicklung zweier Grubenottern (Serpentes: Crotalidae: Crotalus atrox und Agkistrodon p. piscivorus)

    NARCIS (Netherlands)

    Petzold, Hans Günter

    1963-01-01

    Über die postembryonale Entwicklung — besonders die Gewichtszunahme — junger Giftschlangen (Viperiden, Crotaliden, Elapiden) gibt es aus naheliegenden Gründen nur wenige eingehendere Untersuchungen. KLAUBER (1956) bringt an Hand eines großen Materials eine Zusammenfassung einschlägiger

  15. 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.

  16. Heterologous fibrin sealant derived from snake venom: from bench to bedside - an overview.

    Science.gov (United States)

    Ferreira, Rui Seabra; de Barros, Luciana Curtolo; Abbade, Luciana Patrícia Fernandes; Barraviera, Silvia Regina Catharino Sartori; Silvares, Maria Regina Cavariani; de Pontes, Leticia Gomes; Dos Santos, Lucilene Delazari; Barraviera, Benedito

    2017-01-01

    Hemostatic and adhesive agents date back to World War II, when homologous fibrin sealant came onto scene. Considering that infectious diseases can be transmitted via human blood, a new heterologous fibrin sealant was standardized in the 1990s. Its components were a serine protease (a thrombin-like enzyme) extracted from the venom of Crotalus durissus terrificus snakes and a fibrinogen-rich cryoprecipitate extracted from the blood of Bubalus bubalis buffaloes. This new bioproduct has been used as a coagulant, sealant, adhesive and recently as a candidate scaffold for mesenchymal stem cells and bone and cartilage repair. This review discusses the composition of a new heterologous fibrin sealant, and cites published articles related to its preclinical applications aiming at repairing nervous system traumas and regenerating bone marrow. Finally, we present an innovative safety trial I/II that found the product to be a safe and clinically promising candidate for treating chronic venous ulcers. A multicenter clinical trial, phase II/III, with a larger number of participants will be performed to prove the efficacy of an innovative biopharmaceutical product derived from animal venom.

  17. Influence of radiolysis products of water in cytotoxicity assay

    Energy Technology Data Exchange (ETDEWEB)

    Camillo, Maria A.P.; Luly, Juliana H.; Silva, Jose A. Alves da; Magalhaes, Vanessa D.; Higa, Olga Z. [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil). Centro de Biotecnologia]. E-mail: mcamillo@ipen.br

    2007-07-01

    The ionizing radiation has shown to be an useful tool in the improvement of immunogens such as toxins; venoms; protozoa among others. A recent study has demonstrated that hydroxyl radical is the main radiolysis product of water involved in the protein irradiation. It promotes important structural modifications in the toxins meanwhile the immunogenic properties are preserved. Tests of biological activity are essential to determine the detoxification of the samples and the safety of the inoculum. The in vitro cytotoxicity assay, determining the cellular viability, was chosen for this analysis. This test has good reproducibility; it can be carried out with little amount of sample and has been useful for biological evaluation of biomaterials. Herein are presented the results gotten with gyroxin (a serine protease, glycoprotein with 26 kDa) isolated from the rattlesnake Crotalus durissus terrificus venom. The toxin was irradiated with 2 kGy of gamma ray of cobalt-60 in saline solution. The evaluation of the biological activity was carried out with different cell lines, using MTS dye or {sup 3}H-thymidine incorporation as indicators of cells viability. The best correlation between the cellular viability assay and lethality assay in vivo was obtained with endothelial cells (HUVECs) and {sup 3}H-thymidine incorporation. The MTS evaluation presented a false-positive result of cytotoxicity, probably due to the reaction with free radicals formed by radiolysis of water during the irradiation process. (author)

  18. 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)

  19. 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)

  20. 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)

  1. 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.

  2. Evaluación comparativa de dos métodos para determinar la actividad de fosfolipasa A en venenos de serpientes

    Directory of Open Access Journals (Sweden)

    Fanny Lazo

    2014-06-01

    Full Text Available Se ha evaluado la actividad de fosfolipasa A en seis venenos de serpientes comparando las ventajas de un método espectrofotométrico, previamente adaptado a nuestras condiciones de laboratorio, frente al método macroscópico de retardo en la coagulación de una emulsión lipoproteica. En ambos casos, la actividad de fosfolipasa A estuvo presente en los venenos de Micrurus spixii, Crotalus durissus, Bothrops brazili, Lachesis muta y Bothrops atrox y la actividad decreció en orden. En cambio, la actividad de la enzima en el veneno de Micrurus surinamensis sólo fue detectada por el método espectrofotométrico. Así mismo, los resultados basados en la cantidad de veneno utilizado para medir la actividad, mostraron una mayor sensibilidad con el método espectrofotométrico en comparación con el método macroscópico ya que se requirieron cantidades menores en el orden de 2 a 10 veces.

  3. 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.

  4. 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)

  5. 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.

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

    Directory of Open Access Journals (Sweden)

    A Della Torre

    2011-01-01

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

  7. [Pharmacologic and enzymatic effects of snake venoms from Antioquia and Choco (Colombia)].

    Science.gov (United States)

    Otero, R; Guillermo Osorio, R; Valderrama, R; Augusto Giraldo, C

    1992-01-01

    We compared several pharmacological and enzymatic effects induced by 11 snake venoms from seven species, six of them from different geographic areas of Antioquia and Choco, north-west of Colombia, South America (Bothrops atrox, B. nasutus, B. schlegelii, B. punctatus, Lachesis muta, Micrurus mipartitus), and Crotalus durissus terrificus venom, from specimens captured in other provinces of the country (Tolima, Huila, Meta and Atlantico). Differences were observed in edema-forming, hemorrhage, defibrination, indirect hemolysis, myonecrosis, proteolysis and lethal activity between venoms from different genera or species, as well as according to the geographic area of origin in B. atrox and B. nasutus snake venoms. Bothrops venoms, in particular B. atrox and L. muta, produced major local effects. All of the venoms, including M. mipartitus, had myotoxic effects. The most defibrinating venoms were B. atrox, L. muta, B. punctatus and C. d. terrificus. All of the venoms had indirect hemolytic activity; the venom of M. mipartitus being greatest. The most lethal venoms were those of C. d. terrificus and M. mipartitus. Within Bothrops species, the venom of B. schlegelii was the least active in terms of local and systemic pathologic effects.

  8. 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.

  9. 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.

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

    Science.gov (United States)

    Moço, Tatiana Cristina; O'Dwyer, Lucia Helena; Vilela, Fabiana Custódio; Barrella, Thomaz Henrique; da Silva, Reinaldo José

    2002-12-01

    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 S o 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.

  11. 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.

  12. 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.

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

    Energy Technology Data Exchange (ETDEWEB)

    Igarashi, Tomoko; Oishi, Yuko [Department of Cardiac Physiology, National Cardiovascular Center Research Institute, 5-7-1 Fujishiro-dai, Suita, Osaka 565-8565 (Japan); Araki, Satohiko [Sugashima Marine Biological Laboratory, Graduate School of Science, Nagoya University, Toba, Mie 517-0004 (Japan); Mori, Hidezo [Department of Cardiac Physiology, National Cardiovascular Center Research Institute, 5-7-1 Fujishiro-dai, Suita, Osaka 565-8565 (Japan); Takeda, Soichi, E-mail: stakeda@ri.ncvc.go.jp [Department of Cardiac Physiology, National Cardiovascular Center Research Institute, 5-7-1 Fujishiro-dai, Suita, Osaka 565-8565 (Japan); Laboratory for Structural Biochemistry, Riken Harima Institute at SPring-8, 1-1-1 Kouto, Mikazuki, Sayo, Hyogo 679-5148 (Japan)

    2006-07-01

    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.

  14. 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

  15. 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

  16. Development of snake-directed antipredator behavior by wild white-faced capuchin monkeys: I. Snake-species discrimination.

    Science.gov (United States)

    Meno, Whitney; Coss, Richard G; Perry, Susan

    2013-03-01

    Young animals are known to direct alarm calls at a wider range of species than adults. Our field study examined age-related differences in the snake-directed antipredator behavior of infant, juvenile, and adult white-faced capuchin monkeys (Cebus capucinus) in terms of alarm calling, looking behavior, and aggressive behavior. In the first experiment, we exposed infant and juvenile white-faced capuchins to realistic-looking inflatable models of their two snake predators, the boa constrictior (Boa constrictor) and neotropical rattlesnake (Crotalus durissus) and a white airplane as a novel control. In the second experiment, infants, juveniles, and adults were presented photographic models of a coiled boa constrictor, rattlesnake, indigo snake (Drymarchon corais), a noncapuchin predator, and a white snake-like model. We found that antipredator behavior changed during the immature stage. Infants as young as 4 months old were able to recognize snakes and display antipredator behavior, but engaged in less snake-model discrimination than juveniles. All age classes exhibited a lower response to the white snake-like model, indicating that the absence of color and snake-scale patterns affected snake recognition. Infants also showed a higher level of vigilance after snake-model detection as exhibited by a higher proportion of time spent looking and head cocking at the models. Aggressive antipredator behavior was found in all age classes, but was more prevalent in juveniles and adults than infants. This study adds to the knowledge of development of antipredator behavior in primates by showing that, although alarm calling behavior and predator recognition appear at a very young age in capuchins, snake-species discrimination does not become apparent until the juvenile stage. © 2012 Wiley Periodicals, Inc.

  17. Novo método de dosagem de soros antipeçonhentos em camundongos lactentes: I - dosagem do soro anticrotálico

    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.

  18. 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.

  19. 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)

  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. 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.

  2. Anti-snake venom activities of extracts and fractions from callus cultures of Sapindus saponaria.

    Science.gov (United States)

    da Silva, Marcos L; Marcussi, Silvana; Fernandes, Renata S; Pereira, Paulo S; Januário, Ana Helena; França, Suzelei C; Da Silva, Saulo L; Soares, Andreimar M; Lourenço, Miriam V

    2012-03-01

    Sapindus saponaria L. (Sapindaceae) bark, root, and fruits are used as sedatives and to treat gastric ulcer and also demonstrate diuretic and expectorant effects. The anti-snake venom properties of callus of S. saponaria are investigated here for the first time. In vitro cultivated callus of Sapindus saponaria were lyophilized, and the extracts were prepared with different solvents, before submitting to phytochemical studies and evaluation of the anti-ophidian activity. Crude extracts were fractionated by liquid-liquid partition and the fractions were monitored by thin layer chromatography (TLC). Subsequently, anti-ophidian activities were analyzed toward Bothrops jararacussu Lacerda (Viperidae), B. moojeni Hoge (Viperidae), B. alternates Duméril (Viperidea) and Crotalus durissus terrificus Lineu (Viperidae) venoms and isolated myotoxins and phospholipase A(2) (PLA(2)). Fractions A1, A2 and the extract in MeOH:H(2)O (9:1) significantly inhibited the toxic and pharmacological activities induced by snake venoms and toxins, when compared to other extracts and fractions. The lethal, clotting, phospholipase, edema-inducing, hemorrhagic and myotoxic activities were partially inhibited by the different extracts and fractions. TLC profiles of the crude extracts (B and C) and fractions (A1 and A2) showed β-sitosterol and stigmasterol as their main compounds. Stigmasterol exhibited inhibitory effects on enzymatic and myotoxic activities of PLA(2). Sapindus saponaria extracts and fractions presented anti-ophidian activity and could be used as an adjuvant to serum therapy or for its supplementation, and in addition, as a rich source of potential inhibitors of enzymes involved in several pathophysiological human and animal diseases.

  3. 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)

  4. 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.

  5. Opioid neurotransmission modulates defensive behavior and fear-induced antinociception in dangerous environments.

    Science.gov (United States)

    Coimbra, Norberto Cysne; Calvo, Fabrício; Almada, Rafael Carvalho; Freitas, Renato Leonardo; Paschoalin-Maurin, Tatiana; Dos Anjos-Garcia, Tayllon; Elias-Filho, Daoud Hibrahim; Ubiali, Walter Adriano; Lobão-Soares, Bruno; Tracey, Irene

    2017-06-23

    The effects of endogenous opioid peptide antagonists on panic-related responses are controversial. Using elevated mazes and a prey-versus-predator paradigm, we investigated the involvement of the endogenous opioid peptide-mediated system in the modulation of anxiety- and panic attack-induced responses and innate fear-induced antinociception in the present work. Wistar rats were intraperitoneally pretreated with either physiological saline or naloxone at different doses and were subjected to either the elevated plus- or T-maze test or confronted by Crotalus durissus terrificus. The defensive behaviors of the rats were recorded in the presence of the predator and at 24h after the confrontation, when the animals were placed in the experimental enclosure without the rattlesnake. The peripheral non-specific blockade of opioid receptors had a clear anxiolytic-like effect on the rats subjected to the elevated plus-maze but not on those subjected to the elevated T-maze; however, a clear panicolytic-like effect was observed, i.e., the defensive behaviors decreased, and the prey-versus-predator interaction responses evoked by the presence of the rattlesnakes increased. A similar effect was noted when the rats were exposed to the experimental context in the absence of the venomous snake. After completing all tests, the naloxone-treated groups exhibited less anxiety/fear-induced antinociception than the control group, as measured by the tail-flick test. These findings demonstrate the anxiolytic and panicolytic-like effects of opioid receptor blockade. In addition, the fearlessness behavior displayed by preys treated with naloxone at higher doses enhanced the defensive behavioral responses of venomous snakes. Copyright © 2017 IBRO. Published by Elsevier Ltd. All rights reserved.

  6. Prospection, structural analysis and phylogenetic relationships of endogenous gamma-phospholipase A(2) inhibitors in Brazilian Bothrops snakes (Viperidae, Crotalinae).

    Science.gov (United States)

    Estevão-Costa, Maria Inácia; Rocha, Bruno Coelho; de Alvarenga Mudado, Maurício; Redondo, Rodrigo; Franco, Glória Regina; Fortes-Dias, Consuelo Latorre

    2008-07-01

    During the last 20 years, there have been an increasing number of reports on endogenous phospholipase A(2) inhibitors (PLIs) in the sera of snakes. These studies have demonstrated the existence of three different structural classes of PLIs (alpha, beta and gamma). The gamma class members are potent inhibitors of phospholipases A(2) (PLA(2)) from the venom of Viperidae snakes. These enzymes, together with the mammalian pro-inflammatory PLA(2), belong to the IIA class of the PLA(2)-superfamily. Although coming from distinct sources, these phospholipases A(2) share main structural features. For this reason, gammaPLIs have been considered as potential models for the development of selective inhibitors of pro-inflammatory PLA(2) in humans. In spite of the rich diversity of the ophidian fauna in Brazil, only two gammaPLI representatives, from Crotalus durissus terrificus and Lachesis muta, have been described in Brazilian snakes so far. Here we investigated the presence of transcripts of novel gammaPLIs in six Bothrops species (Viperidae, Crotalinae) commonly found in our country: Bothrops alternatus, Bothrops erythromelas, Bothrops jararaca, Bothrops jararacussu, Bothrops moojeni and Bothrops neuwiedi. gammaPLI transcripts were present in every species analysed. The deduced mature proteins possessed 181 amino acid residues following a 19-residue signal peptide, similar to the gammaPLIs from C. d. terrificus taken as our model, with the exception of the deduced proteins from B. erythromelas and B. neuwiedi snakes. In these particular cases, an insertion of 4-amino acid residues was consistently present. A Bayesian tree was obtained for the Brazilian Bothrops gammaPLIs, showing four clusters: (1) L. muta and B. jararacussu, (2) B. alternatus, (3) B. erythromelas and B. neuwiedi, (4) B. jararaca and B. moojeni. Detailed structural analysis and further comparisons of these novel Bothrops inhibitors with gammaPLIs from New and Old World snakes are provided.

  7. Crystallographic characterization of functional sites of crotoxin and ammodytoxin, potent β-neurotoxins from Viperidae venom.

    Science.gov (United States)

    Faure, Grazyna; Saul, Frederick

    2012-09-15

    This review will focus on a description of the three-dimensional structures of two β-neurotoxins, the monomeric PLA(2) ammodytoxin from Vipera ammodytes ammodytes, and heterodimeric crotoxin from Crotalus durissus terrificus, and a detailed structural analysis of their multiple functional sites. We have recently determined at high resolution the crystal structures of two natural isoforms of ammodytoxin (AtxA and AtxC) (Saul et al., 2010) which exhibit different toxicity profiles and different anticoagulant properties. Comparative structural analysis of these two PLA(2) isoforms, which differ only by two amino acid residues, allowed us to detect local conformational changes and delineate the role of critical residues in the anticoagulant and neurotoxic functions of these PLA(2) (Saul et al., 2010). We have also determined, at 1.35Å resolution, the crystal structure of heterodimeric crotoxin (Faure et al., 2011). The three-dimensional structure of crotoxin revealed details of the binding interface between its acidic (CA) and basic (CB) subunits and allowed us to identify key residues involved in the stability and toxicity of this potent heterodimeric β-neurotoxin (Faure et al., 2011). The precise spatial orientation of the three covalently linked polypeptide chains in the mature CA subunit complexed with CB helps us to understand the role played by critical residues of the CA subunit in the increased toxicity of the crotoxin complex. Since the CA subunit is a natural inhibitor of the catalytic and anticoagulant activities of CB, identification of the CA-CB binding interface describes residues involved in this inhibition. We propose future research directions based on knowledge of the recently reported 3D structures of crotoxin and ammodytoxin. Copyright © 2012 Elsevier Ltd. All rights reserved.

  8. A study on the membrane depolarization of skeletal muscles caused by a scorpion toxin, sea anemone toxin II and crotamine and the interaction between toxins.

    Science.gov (United States)

    Chang, C. C.; Hong, S. J.; Su, M. J.

    1983-01-01

    Quinquestriatus toxin (QTX) isolated from the venom of a scorpion (Leiurus quinquestriatus) and sea anemone (Anemonia sulcata) toxin II enhanced the twitch response of the rat and mouse diaphragms and like crotamine (isolated from the venom of Crotalus durissus terrificus) caused spontaneous fasciculation of the muscle. Trains of action potentials in muscles at 70-250 Hz, which could not be antagonized by (+)-tubocurarine, were triggered by single stimulation or occurred spontaneously after treatment with these toxins. QTX and toxin II prolonged the rat muscle action potential 3 to 4 fold whereas crotamine prolonged the action potential by only 30%. The membrane potential was depolarized from about -82 mV to -55 mV by crotamine 2 micrograms ml-1, -41 mV by toxin II 5 micrograms ml-1 and to -50 mV by QTX 1 microgram ml-1. The concentrations to induce 50% maximal depolarization (K0.5) were 0.07, 0.15 and greater than 0.4 microgram ml-1, respectively, for QTX, crotamine and toxin II, whereas the rates of depolarization were in the order toxin II greater than or equal to crotamine greater than QTX. The depolarizing effects of crotamine and QTX, but not of toxin II, were saturable. The depolarizing effects of all three toxins were irreversible whereas the membrane potential could be restored by tetrodotoxin non-competitively. Simultaneous treatment with crotamine and QTX or crotamine and toxin II at concentrations below K0.5 caused only additive effects on depolarization. When the muscle was depolarized by pretreating with a saturating concentration of crotamine, the onset of depolarization by QTX was greatly retarded whereas that by toxin II was unaffected.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:6197125

  9. 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

  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. 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.

  12. 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.

  13. 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

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

    Directory of Open Access Journals (Sweden)

    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

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

    Energy Technology Data Exchange (ETDEWEB)

    Santos, Raquel Gouvea dos; Soares, Marcella Araugio; Andrade, Henrique Martins de, E-mail: santosr@cdtn.br [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil); Santos, Marcos Antonio da Cunha [Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, MG (Brazil). Departamento de Estatistica

    2008-07-01

    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 {sup 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 {sup 99m}Tc-Cdt and {sup 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 {sup 99m}Tc-Cdt or {sup 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 {sup 131}I-Crtx was calculated based on non penetrating radiation in the mouse model. Biokinetics data from {sup 99m}Tc-Cdt after i.v. injection in mice tumor model showed rapid blood clearance (T{sub 1/2}= 36.1 ± 2.4 min.), slow tumor clearance (T{sub 1/2}: 108.3 ± 19.5 min.) and indicated the kidneys as the main excretion pathway. Interaction studies in vitro demonstrated that {sup 125}I-Crtx recognize specific sites on Erlich tumor cell membrane. Upon intravenous and intratumor administration of {sup 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 {sup 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). {sup 131}I control was run in a parallel experiment and showed no significant tumor

  16. 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

  17. Assessing the influence of mechanical ventilation on blood gases and blood pressure in rattlesnakes.

    Science.gov (United States)

    Bertelsen, Mads F; Buchanan, Rasmus; Jensen, Heidi M; Leite, Cleo A C; Abe, Augusto S; Nielsen, Søren S; Wang, Tobias

    2015-07-01

    To characterize the impact of mechanical positive pressure ventilation on heart rate (HR), arterial blood pressure, blood gases, lactate, glucose, sodium, potassium and calcium concentrations in rattlesnakes during anesthesia and the subsequent recovery period. Prospective, randomized trial. Twenty one fasted adult South American rattlesnakes (Crotalus durissus terrificus). Snakes were anesthetized with propofol (15 mg kg(-1)) intravenously, endotracheally intubated and assigned to one of four ventilation regimens: Spontaneous ventilation, or mechanical ventilation at a tidal volume of 30 mL kg(-1) at 1 breath every 90 seconds, 5 breaths minute(-1), or 15 breaths minute(-1). Arterial blood was collected from indwelling catheters at 30, 40, and 60 minutes and 2, 6, and 24 hours following induction of anesthesia and analyzed for pH, PaO2, PaCO2, and selected variables. Mean arterial blood pressure (MAP) and HR were recorded at 30, 40, 60 minutes and 24 hours. Spontaneous ventilation and 1 breath every 90 seconds resulted in a mild hypercapnia (PaCO2 22.4 ± 4.3 mmHg [3.0 ± 0.6 kPa] and 24.5 ± 1.6 mmHg [3.3 ± 0.2 kPa], respectively), 5 breaths minute(-1) resulted in normocapnia (14.2 ± 2.7 mmHg [1.9 ± 0.4 kPa]), while 15 breaths minute(-1) caused marked hypocapnia (8.2 ± 2.5 mmHg [1.1 ± 0.3 kPa]). Following recovery, blood gases of the four groups were similar from 2 hours. Anesthesia, independent of ventilation was associated with significantly elevated glucose, lactate and potassium concentrations compared to values at 24 hours (p < 0.0001). MAP increased significantly with increasing ventilation frequency (p < 0.001). HR did not vary among regimens. Mechanical ventilation had a profound impact on blood gases and blood pressure. The results support the use of mechanical ventilation with a frequency of 1-2 breaths minute(-1) at a tidal volume of 30 mL kg(-1) during anesthesia in fasted snakes. © 2014 Association of Veterinary Anaesthetists and the American

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

    Science.gov (United States)

    Barboza, Raynner R D; Souto, Wedson de M S; Mourão, José da S

    2007-09-07

    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. 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. 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 mentioned. Fat from various animals constituted

  19. 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

  20. 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

  1. 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

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

    Directory of Open Access Journals (Sweden)

    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

  3. 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

  4. 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

    Directory of Open Access Journals (Sweden)

    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

  5. Environmental Impact Statement. Disposal and Reuse of Williams Air Force Base, Arizona.

    Science.gov (United States)

    1994-06-01

    sidewinder (Crotalus cerestes). Washes and rocky areas provide suitable habitat for the side- blotched lizard (Uta stansburiana), lesser earless lizard...Thoteforcum shwteicessI utility limed associated With each altterntI*ive1 the This ha? rs hn the estinated Unader of yea? 201). Far manle. with the proposed... blotched lizard Uta stansburiana Arid and semi-arid areas Sidewinder Crotalus cerastes Creosote bush, desert S!,ckled rattlesnake Crotalus mitchell/i

  6. Structure of an Engineered Porcine Phospholipase A2 with Enhanced Activity at 2.1 Å Resolution. Comparison with the Wild-type Porcine and Crotalus atrox Phospholipase A2

    NARCIS (Netherlands)

    Thunnissen, Marjolein M.G.M.; Kalk, Kor H.; Drenth, Jan; Dijkstra, Bauke W.

    1990-01-01

    The crystal structure of an engineered phospholipase A2 with enhanced activity has been refined to an R-factor of 18.6% at 2.1 Å resolution using a combination of molecular dynamics refinement by the GROMOS package and least-squares refinement by TNT. This mutant phospholipase was obtained

  7. 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

  8. Aspectos clínico-patológicos e laboratoriais do envenenamento crotálico experimental em equinos

    Directory of Open Access Journals (Sweden)

    Cinthia Távora A. Lopes

    2012-09-01

    Full Text Available Descrevem-se os quadros clínico-patológicos e laboratoriais de equinos inoculados experimentalmente com a peçonha de Caudisona durissa terrificus (Crotalus durissus terrificus na antiga nomenclatura, com a finalidade de fornecer subsídios que favoreçam a compreensão desse tipo de acidente ofídico em equinos. O veneno liofilizado foi diluído em 1ml de solução salina a 0,9% e inoculado por via subcutânea em cinco equinos, nas doses de 0,12mg/kg (um animal, 0,066mg/kg (dois animais e 0,03mg/kg (dois animais. O veneno causou a morte do equino que recebeu a dose de 0,12mg/kg e de um dos dois que receberam a dose de 0,066mg/kg, com evolução de 27h27min e 52h29min, respectivamente. O segundo animal que recebeu a dose de 0,066mg/kg também adoeceu, mas recuperou-se após 12 dias da inoculação. A dose de 0,03mg/kg determinou quadros não fatais do envenenamento, com período de evolução que variou entre 6 e 10 dias. O quadro clínico caracterizou-se por considerável aumento de volume no local de inoculação (escápula que se estendeu por todo o membro, apatia e cabeça baixa, alterações locomotoras evidenciadas pelo arrastar das pinças no solo, decúbito e dificuldade para levantar, redução dos reflexos auricular, palatal, do lábio superior e de ameaça, e aumento das frequências cardíaca e respiratória. Os exames laboratoriais revelaram leucocitose por neutrofilia e linfocitose em apenas dois animais. Houve aumento das enzimas creatina quinase (CK, dehidrogenase láctica (DHL e da ureia, e também redução nos níveis séricos de cálcio, fósforo e magnésio. O tempo de tromboplastina parcial ativada (TTPA aumentou nos equinos que morreram. Os achados de necropsia foram edema do tecido subcutâneo em todo o membro em que foi aplicado o veneno, sufusões no epicárdio dos ventrículos cardíacos esquerdo e direito, e bexiga com áreas hemorrágicas em grande parte da mucosa. Ao exame histopatológico observaram-se f

  9. The substrate specificity of phospholipase A

    NARCIS (Netherlands)

    Deenen, L.L.M. van; Haas, Gerard H. de

    1963-01-01

    Investigations on variously modified analogues of phospholipids elucidated the following substrate characteristics for phospholipase A (Crotalus adamanteus). 1. 1. Within the class of α-phosphoglycerides -isomers are readily hydrolysed, while -α-phospholipids appeared not to be attacked. 2.

  10. The site of action of phosphatide acyl-hydrolase (phospholipase A) on mixed-acid phosphatides containing a poly-unsaturated fatty acid

    NARCIS (Netherlands)

    Haas, Gerard H. de; Daemen, F.J.M.; Deenen, L.L.M. van

    1962-01-01

    In order to establish the positional specificity of phospholipase A, two structural isomeric -α-phosphatidyl ethanolamines were prepared by a synthesis de novo. The phospholipase present in Crotalus adamantheus venom liberated nearly one equivalent palmitic acid from

  11. The Response of Specialized Biological Tissue to Infrared Radiation

    National Research Council Canada - National Science Library

    Welch, Ashley J

    1998-01-01

    .... The two pit organs were extracted from an in vitro Western Diamondback rattlesnake (Crotalus atrox) heads. This consided of discussing along the maxillary bone just under the back surface of the pit organ between the eye and nostril...

  12. In vitro evaluation of copaiba oil as a kojic acid skin enhancer

    National Research Council Canada - National Science Library

    Robson Vicente Machado de Oliveira; Mitsuko Taba Ohara; Marta Maria Duarte Carvalho Vila; Marcos Moisés Gonçalves

    2010-01-01

    The capacity of copaíba oil to act as a skin penetration enhancer for the depigmenting agent kojic acid was evaluated using an in vitro diffusion system with static flux and shed rattlesnake skin membrane, Crotalus...

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

    Directory of Open Access Journals (Sweden)

    R.S. Ferreira Júnior

    2004-01-01

    Full Text Available Snake envenoming is a major problem both to veterinary and human medicine in tropical countries due to high incidence, severity, and sequelae. In Brazil, most envenomings involving animals are caused by Bothrops and Crotalus snakes; these are the highest risk to animals. This study reports on Bothrops and Crotalus envenomings in dogs, the main species responsible for epidemiology, pathogenesis, venom action, clinical signs, sequelae and complications, clinical pathology, necropsy findings, diagnosis, and treatment. Veterinarians must be capable of identifying the snake not only by observing its characteristics but also symptom evolution.

  14. PRODUCTION OF AN EFFECTIVE ANTI-Bothrops-TETANUS MIXED HYPERIMMUNE SERUM OF EQUINE ORIGIN

    OpenAIRE

    GUIDOLIN, R.; STEPHANO, M. A.; MORAIS, J. F.; SAKAGUTI, E. H.; PRADO, S. M. A.; FRATELLI, F.; VANCETTO, M. D. C.; HIGASHI, H. G.

    1998-01-01

    The present investigation reveals the possibility of simultaneous immunization of horses with Bothrops or Crotalus snake venoms and Tetanus antigens for the production of anti-Bothrops-Tetanus or anti-Crotalus-Tetanus mixed serum, with high titers of the respective specific antibodies. Bothrops antivenoms with an average neutralizing titer of 4.16 mg venom/ml were obtained from plasma of horses with titers lower than 0.5 mg venom/ml when Tetanus antigens were not used. This suggests the exist...

  15. Organization of phospholipids in human red cell membranes as detected by the action of various purified phospholipases

    NARCIS (Netherlands)

    Zwaal, R.F.A.; Roelofsen, B.; Comfurius, P.; Deenen, L.L.M. van

    1975-01-01

    1. 1. The action of eight purified phospholipases on intact human erythrocytes has been investigated. Four enzymes, e.g. phospholipases A2 from pancreas and Crotalus adamanteus, phospholipase C from Bacillus cereus, and phospholipase D from cabbage produce neither haemolysis nor hydrolysis of

  16. Synthetic lecithins containing one short-chain fatty acid and their breakdown by phospholipase A

    NARCIS (Netherlands)

    Bird, P.H.R.; Haas, Gerard H. de; Heemskerk, C.H.Th.; Deenen, L.L.M. van

    1965-01-01

    The synthesis is described of two structurally isomericL -α-lecithins containing as fatty acid residues butyric acid and oleic acid. Phospholipase A (EC 3.1.1.4) from Crotalus adamanteus hydrolysed the β-fatty acid linkage irrespective of the nature of the fatty acid constituent, though

  17. Three-dimensional structures of human phospholipase A2 from pancreas and synovial fluid by model building

    DEFF Research Database (Denmark)

    Christensen, I T; Jørgensen, Flemming Steen; Svensson, L A

    1993-01-01

    on the X-ray structure of the highly homologous bovine pancreatic PLA2 (Type I) by amino acid substitution and modification of the C-terminal part followed by geometry relaxation. The structure of the PLA2 from human synovial fluid was constructed from the X-ray structure of PLA2 from Crotalus atrox (Type...

  18. Nellis and Creech Air Force Bases Capital Improvements Program Environmental Assessment

    Science.gov (United States)

    2008-09-01

    this document should be directed to: 99 ABW/PA 4430 Grissom Ave, Suite 107 Nellis AFB NV 89191 ATTN: Mr. Mike Estrada In addition, the document... boa (Lichanura trivirgata), gopher snake (Pituophis melanoleucus), and Mojave rattlesnake (Crotalus scutulatus). Bird species that include a

  19. 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...

  20. Environmental Assessment for Kirtland Air Force Base Arsenic Compliance System

    Science.gov (United States)

    2003-09-01

    Texas horned lizard Phrynosoma FSC cornu tum BIRDS Neotrophic Phalacrocorax ST cormorant brasilianus white-faced ibis Plegadis chihi FSC bald eagle...Bufo woodhousii), New Mexico spadefoot (Spea multiplicata), coachwhip snake (Masticophis flagellum), whiptail lizards (Cnemidophorus spp.), lesser...earless lizard (Holbrookia maculata), and the western rattlesnake (Crotalus viridis). Several of these species have extensive periods of dormancy during

  1. Don't tread on me

    Science.gov (United States)

    N. Schiff

    2009-01-01

    Always be careful when you turn over a piece of wood that is lying on the ground. You might find something horrid there. Last month, under an insect· rearing cage in the Delta Experimental Forest, a research associate discovered a Canebrake rattlesnake, Crotalus horrid us, coiled up and ready to strike. The associate carefully collected the snake; we took some pictures...

  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. Grassland Birds Wintering at U.S. Navy Facilities in Southern Texas

    Science.gov (United States)

    2010-07-01

    brevis Texas horned lizard* Phrynosoma cornutum Texas spiny lizard Sceloporus olivaceus Texas indigo snake * Drymarchon corais Southwestern rat snake ...Elaphe guttata Ribbon snake Thamnophis proximus Western diamondback rattlesnake Crotalus atrox Structural and Floristic Characteristics of Native and...months. Dry periods increase nutritional stress and, in turn, reduce the production of offspring, which commonly cause small mammal popula- tions to

  4. Final Environmental Assessment, Family Camp Facility Buckey Air Force Base, Colorado

    Science.gov (United States)

    2008-02-07

    northern prairie lizard (Sceloporus undulates garmani), bullsnake (Pituophis catenifir), western hognose snake (Heterodon nasicus), plains garter... snake (Thamnophis radix), and prairie rattlesnake (Crotalus viridis viridis) (BAFB 2004b). 3.4.2.2 Impacts This section analyzes potential impacts to...Region 8 1595 Wynkoop Street Denver, CO 80202-1129 Mr. Brent Bibles Wildlife Researcher Colorado Division of Wildlife Wildlife Research Center

  5. Final Environmental Assessment: Construction of Maintenance and Storage Facility, Perimeter Fence Upgrade and Demolition of Three Buildings and Two Structures Gila River Air Force Space Surveillance Station Arizona

    Science.gov (United States)

    2012-11-01

    are listed in the Table 6. Typical species that have been observed include lizards, jackrabbits, ground squirrels, desert mice, snakes , scorpions...Reptiles Arizona coral snake1 Micruroides euryxanthus Tuscon baded gecko Coleonyx variegatus bogerti Zebra-tailed lizard Callisaurus draconoides...Sidewinder Crotalus cerastes Western shovel-nose snake Chionactis occipatilis Mammals Coyote Canis latrans Kit fox1,2 Vulpes macrotis Badger1,2

  6. Rattlesnake Envenomation in Three Dairy Goats

    OpenAIRE

    Joseph Smith; David Kovalik; Anita Varga

    2015-01-01

    Cases of rattlesnake envenomation in dairy goats are lacking. These cases present three dairy goats presented to a veterinary referral hospital for envenomation of Northern Pacific Rattlesnake (Crotalus oreganus). Treatments and clinical characteristics reported are similar to those for llamas, alpacas, and horses. These cases suggest that quick treatment in the event of a bite may have a more favorable clinical response. Existing rattlesnake bite scoring systems applicable to other species m...

  7. Rattlesnake Envenomation in Three Dairy Goats

    Directory of Open Access Journals (Sweden)

    Joseph Smith

    2015-01-01

    Full Text Available Cases of rattlesnake envenomation in dairy goats are lacking. These cases present three dairy goats presented to a veterinary referral hospital for envenomation of Northern Pacific Rattlesnake (Crotalus oreganus. Treatments and clinical characteristics reported are similar to those for llamas, alpacas, and horses. These cases suggest that quick treatment in the event of a bite may have a more favorable clinical response. Existing rattlesnake bite scoring systems applicable to other species may be applicable to goats, and existing respiratory pathology may predispose goats to a less favorable outcome.

  8. Comparative study of anticoagulant and procoagulant properties of 28 snake venoms from families Elapidae, Viperidae, and purified Russell's viper venom-factor X activator (RVV-X).

    Science.gov (United States)

    Suntravat, Montamas; Nuchprayoon, Issarang; Pérez, John C

    2010-09-15

    Snake venoms consist of numerous molecules with diverse biological functions used for capturing prey. Each component of venom has a specific target, and alters the biological function of its target. Once these molecules are identified, characterized, and cloned; they could have medical applications. The activated clotting time (ACT) and clot rate were used for screening procoagulant and anticoagulant properties of 28 snake venoms. Crude venoms from Daboia russellii siamensis, Bothrops asper, Bothrops moojeni, and one Crotalus oreganus helleri from Wrightwood, CA, had procoagulant activity. These venoms induced a significant shortening of the ACT and showed a significant increase in the clot rate when compared to the negative control. Factor X activator activity was also measured in 28 venoms, and D. r. siamensis venom was 5-6 times higher than those of B. asper, B. moojeni, and C. o. helleri from Wrightwood County. Russell's viper venom-factor X activator (RVV-X) was purified from D. r. siamensis venom, and then procoagulant activity was evaluated by the ACT and clot rate. Other venoms, Crotalus atrox and two Naja pallida, had anticoagulant activity. A significant increase in the ACT and a significant decrease in the clot rate were observed after the addition of these venoms; therefore, the venoms were considered to have anticoagulant activity. Venoms from the same species did not always have the same ACT and clot rate profiles, but the profiles were an excellent way to identify procoagulant and anticoagulant activities in snake venoms.

  9. No safety in the trees: Local and species-level adaptation of an arboreal squirrel to the venom of sympatric rattlesnakes.

    Science.gov (United States)

    Pomento, Abby M; Perry, Blair W; Denton, Robert D; Gibbs, H Lisle; Holding, Matthew L

    2016-08-01

    Within some species, squirrels respond to variable selection from venomous snake predators by showing population-level variation in resistance, while between species, some rattlesnakes possess venom that is more effective at overcoming venom resistance in different species of squirrels. A functional evaluation of resistance variation to venom within and between species of squirrels and snakes can link resistance variation to its evolutionary causes across these different evolutionary scales. To do this, we compared the effectiveness of squirrel sera in inhibiting rattlesnake (Crotalus spp.) venom metalloproteinase activity between populations and between species to test for a response to local variation in selection from a single rattlesnake predator and for specialization of two resistant squirrel species to each of their distinct sympatric snake predators. We found that Timber Rattlesnake (Crotalus horridus) venom inhibition by Eastern gray squirrels (Sciurus carolinensis) is higher at a site where the rattlesnakes are present, which suggests selection may maintain venom resistance in populations separated by short distances. Next, we performed a reciprocal cross of venoms and sera from two rattlesnake and two squirrel species. This showed that squirrel resistance is lower when tested against venom from allopatric compared to sympatric rattlesnake species, demonstrating that squirrel inhibitors are specialized to sympatric venom and suggesting a tradeoff in terms of specialization to the venom of a specific species of rattlesnake predator. This pattern can be explained if inhibitors must recognize venom proteins and resistance evolution tracks venom evolution. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. The binding effectiveness of anti-r-disintegrin polyclonal antibodies against disintegrins and PII and PIII metalloproteases: An immunological survey of type A, B and A+B venoms from Mohave rattlesnakes.

    Science.gov (United States)

    Cantú, Esteban; Mallela, Sahiti; Nyguen, Matthew; Báez, Raúl; Parra, Victoria; Johnson, Rachel; Wilson, Kyle; Suntravat, Montamas; Lucena, Sara; Rodríguez-Acosta, Alexis; Sánchez, Elda E

    2017-01-01

    Snake venoms are known to have different venom compositions and toxicity, but differences can also be found within populations of the same species contributing to the complexity of treatment of envenomated victims. One of the first well-documented intraspecies venom variations comes from the Mohave rattlesnake (Crotalus scutulatus scutulatus). Initially, three types of venoms were described; type A venom is the most toxic as a result of ~45% Mojave toxin in the venom composition, type B lacks the Mojave toxin but contains over 50% of snake venom metalloproteases (SVMPs). Also, type A+B venom contains a combination of Mojave toxin and SVMP. The use of an anti-disintegrin antibody in a simple Enzyme-Linked Immunosorbent Assay (ELISA) can be used to identify the difference between the venoms of the type A, B, and A+B Mohave rattlesnakes. This study implements the use of an anti-recombinant disintegrin polyclonal antibody (ARDPA) for the detection of disintegrins and ADAMs (a disintegrin and metalloproteases) in individual crude snake venoms of Mohave rattlesnakes (Crotalus scutulatus scutulatus) of varying geographical locations. After correlation with Western blots, coagulation activity and LD50 data, it was determined that the antibody allows for a quick and cost-efficient identification of venom types. Copyright © 2016 Elsevier Inc. All rights reserved.

  11. Current management of copperhead snakebite.

    Science.gov (United States)

    Walker, J Patrick; Morrison, Ray L

    2011-04-01

    Several thousand snakebites occur annually in the US, but fewer than 10 deaths occur. Most deaths are from envenomations by rattlesnakes (Crotalus species), but deaths from copperhead and water moccasin (Agkistrodon species) are rare. All snakebites presented to East Texas Medical Center, Crockett, a level III trauma center, from 1995 to 2010 were reviewed. A total of 142 snakebites were treated. Ninety-four were of the Agkistrodon species-contortrix contortrix (copperhead) or piscivorus leukostoma (water moccasin). Three were rattlesnakes, and 3 were from the Texas coral snake (Micrurus fulvius tener). Forty-two were unidentified pit vipers. The following results are of the 88 copperhead bites. The most common presenting symptoms were pain and swelling. Eighty-five percent were of grade 1 envenomations. Ten patients had laboratory abnormalities secondary to the snakebite. Forty-four were admitted for observation. The average length of stay for patients admitted was 2 days. No patients received antivenom, and no patients required surgical intervention. There were no deaths. One patient had edema and ecchymosis that persisted for more than 1 month. Accurate identification of the pit viper species involved in snakebites is essential. Although envenomation by a rattlesnake (Crotalus species) may require antivenom and uncommonly surgery, a bite by a copperhead (Agkistrodon contortrix) rarely requires any intervention other than observation. The unnecessary use of antivenom should be discouraged. Copyright © 2011 American College of Surgeons. Published by Elsevier Inc. All rights reserved.

  12. 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.

  13. XENOTRANSFUSION IN AN ISLAND FOX (UROCYON LITTORALIS CLEMENTAE) USING BLOOD FROM A DOMESTIC DOG (CANIS LUPUS FAMILIARIS).

    Science.gov (United States)

    Martony, Molly E; Krause, Kristian J; Weldy, Scott H; Simpson, Stephen A

    2016-09-01

    Successful xenotransfusion in an island fox ( Urocyon littoralis clementae) has not been previously reported but may be necessary in an emergency. An 11-yr-old male, intact, captive island fox was exhibiting clinical signs of rattlesnake envenomation including hypoperfusion, tachypnea, facial edema, and multifocal facial and cervical ecchymosis. Blood work revealed severe thrombocytopenia (18 K/μl) and anemia (Hct 15.8%). A presumptive diagnosis of rattlesnake ( Crotalus sp.) envenomation was made. Initial treatment included oxygen therapy, fluid therapy, antibiotics, antacids, pain medications, and polyvalent crotalid anti-venom. Emergency xenotransfusion using whole blood (45 ml) from a domestic dog was used due to worsening clinical signs from anemia. No acute or delayed transfusion reactions were observed in the fox and the patient made a full recovery 5 days later. Successful xenotransfusion in an island fox using whole blood from a domestic dog ( Canis lupus familiaris) is possible and may be lifesaving.

  14. Clinically Significant Envenomation From Postmortem Copperhead (Agkistrodon contortrix).

    Science.gov (United States)

    Emswiler, Michael P; Griffith, F Phillip; Cumpston, Kirk L

    2017-03-01

    Over 14,000 copperhead (Agkistrodon contortrix) bites were reported to United States poison centers between 1983 and 2008, and 1809 cases were reported to poison centers in 2014. The copperhead is primarily found in the southeastern United States and belongs to the pit viper subfamily Crotalinae, which also includes the water moccasin (Agkistrodon piscivorus) and rattlesnakes (Crotalus and Sistrurus genera). Postmortem rattlesnakes have been reported to cause clinically significant envenomation; we report a case of a postmortem copperhead causing clinically significant envenomation after inadvertent puncture with the deceased copperhead fang. The copperhead was transected twice, leaving the snake in 3 separate pieces. While handling the snake head, an inadvertent puncture occurred on the right index finger followed by pain and swelling in the affected extremity necessitating antivenom administration. Care should be taken when handling deceased pit vipers due to the continued risk of envenomation. Copyright © 2017 Wilderness Medical Society. Published by Elsevier Inc. All rights reserved.

  15. Isolation and characterization of three Ca2+-dependent beta-galactoside-specific lectins from snake venoms.

    Science.gov (United States)

    Gartner, T K; Ogilvie, M L

    1984-01-01

    Three lactose-inhibited lectins from the venoms of the snakes Agkistrodon contortrix contortrix (southern copperhead), Ancistrodon piscivorous leukostoma (western cottonmouth moccasin) and Crotalus atrox (western diamondback rattlesnake) have been isolated and newly characterized. The three lectins are similar to thrombolectin, a lectin isolated from the venom of Bothrops atrox (fer-de-lance) (Gartner, Stocker & Williams, 1980), with regard to sugar specificity, Mr, Ca2+ requirements and sensitivity to reducing agents. Each lectin is a dimer (Mr 28 000) consisting of monomers (Mr 14 000) indistinguishable on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. Haemagglutination activity is dependent on the presence of Ca2+ and is inhibited by reducing agents. The lectins are not identical and can be distinguished on the basis of relative affinities for inhibiting sugars, isoelectric points and immunoprecipitation assays using anti-(cottonmouth lectin) serum. Images Fig. 1. Fig. 2. Fig. 4. Fig. 5. PMID:6391472

  16. 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.

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

    Science.gov (United States)

    Araújo, M S; Martins, M

    2007-05-01

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

  18. Facial diplegia, pharyngeal paralysis, and ophthalmoplegia after a timber rattlesnake envenomation.

    Science.gov (United States)

    Madey, Jason J; Price, Amanda B; Dobson, Joseph V; Stickler, David E; McSwain, S David

    2013-11-01

    The timber rattlesnake, also known as Crotalus horridus, is well known to cause significant injury from toxins stored within its venom. During envenomation, toxic systemic effects immediately begin to cause damage to many organ systems including cardiovascular, hematologic, musculoskeletal, respiratory, and neurologic. One defining characteristic of the timber rattlesnake is a specific neurotoxin called crotoxin, or the "canebrake toxin," which is a potent β-neurotoxin affecting presynaptic nerves that can cause paralysis by inhibiting appropriate neuromuscular transmission. We present an unusual case of an 8-year-old boy bitten twice on his calf by a timber rattlesnake, who presented with a life-threatening envenomation and suffered multisystem organ failure as well as a prominent presynaptic neurotoxicity resulting in facial diplegia, pharyngeal paralysis, and ophthalmoplegia.

  19. Crystal structures of catrocollastatin/VAP2B reveal a dynamic, modular architecture of ADAM/adamalysin/reprolysin family proteins.

    Science.gov (United States)

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

    2007-05-29

    Catrocollastatin/vascular apoptosis-inducing protein (VAP)2B is a metalloproteinase from Crotalus atrox venom, possessing metalloproteinase/disintegrin/cysteine-rich (MDC) domains that bear the typical domain architecture of a disintegrin and metalloproteinase (ADAM)/adamalysin/reprolysin family proteins. Here we describe crystal structures of catrocollastatin/VAP2B in three different crystal forms, representing the first reported crystal structures of a member of the monomeric class of this family of proteins. The overall structures show good agreement with both monomers of atypical homodimeric VAP1. Comparison of the six catrocollastatin/VAP2B monomer structures and the structures of VAP1 reveals a dynamic, modular architecture that may be important for the functions of ADAM/adamalysin/reprolysin family proteins.

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

    Directory of Open Access Journals (Sweden)

    MS. Araújo

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

  1. Sidewinding with minimal slip: Snake and robot ascent of sandy slopes

    CERN Document Server

    Marvi, Hamidreza; Gravish, Nick; Astley, Henry; Travers, Matthew; Hatton, Ross L; Mendelson, Joseph R; Choset, Howie; Hu, David L; Goldman, Daniel I

    2014-01-01

    Limbless organisms such as snakes can navigate nearly all terrain. In particular, desert-dwelling sidewinder rattlesnakes (Crotalus cerastes) operate effectively on inclined granular media (such as sand dunes) that induce failure in field-tested limbless robots through slipping and pitching. Our laboratory experiments reveal that as granular incline angle increases, sidewinder rattlesnakes increase the length of their body in contact with the sand. Implementing this strategy in a physical robot model of the snake enables the device to ascend sandy slopes close to the angle of maximum slope stability. Plate drag experiments demonstrate that granular yield stresses decrease with increasing incline angle. Together, these three approaches demonstrate how sidewinding with contact-length control mitigates failure on granular media.

  2. Opossum peptide that can neutralize rattlesnake venom is expressed in Escherichia coli.

    Science.gov (United States)

    Komives, Claire F; Sanchez, Elda E; Rathore, Anurag S; White, Brandon; Balderrama, Michael; Suntravat, Montamas; Cifelli, Angela; Joshi, Varsha

    2017-01-01

    An eleven amino acid ribosomal peptide was shown to completely neutralize Western Diamondback Rattlesnake (Crotalus atrox) venom in mice when a lethal dose of the venom was pre-incubated with the peptide prior to intravenous injection. We have expressed the peptide as a concatenated chain of peptides and cleaved them apart from an immobilized metal affinity column using a protease. After ultrafiltration steps, the mixture was shown to partially neutralize rattlesnake venom in mice. Preliminary experiments are described here that suggest a potential life-saving therapy could be developed. To date, no recombinant therapies targeting cytotoxic envenomation have been reported. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:81-86, 2017. © 2016 American Institute of Chemical Engineers.

  3. Snake co-occurrence patterns are best explained by habitat and hypothesized effects of interspecific interactions.

    Science.gov (United States)

    Steen, David A; McClure, Christopher J W; Brock, Jean C; Craig Rudolph, D; Pierce, Josh B; Lee, James R; Jeffrey Humphries, W; Gregory, Beau B; Sutton, William B; Smith, Lora L; Baxley, Danna L; Stevenson, Dirk J; Guyer, Craig

    2014-01-01

    Snakes often occur in species-rich assemblages, and sympatry is thought to be facilitated primarily by low diet overlap, not interspecific interactions. We selected, a priori, three species pairs consisting of species that are morphologically and taxonomically similar and may therefore be likely to engage in interspecific, consumptive competition. We then examined a large-scale database of snake detection/nondetection data and used occupancy modelling to determine whether these species occur together more or less frequently than expected by chance while accounting for variation in detection probability among species and incorporating important habitat categories in the models. For some snakes, we obtained evidence that the probabilities that habitat patches are used are influenced by the presence of potentially competing congeneric species. Specifically, timber rattlesnakes (Crotalus horridus) were less likely than expected by chance to use areas that also contained eastern diamond-backed rattlesnakes (Crotalus adamanteus) when the proportion of evergreen forest was relatively high. Otherwise, they occurred together more often than expected by chance. Complex relationships were revealed between habitat use, detection probabilities and occupancy probabilities of North American racers (Coluber constrictor) and coachwhips (Coluber flagellum) that indicated the probability of competitive exclusion increased with increasing area of grassland habitat, although there was some model uncertainty. Cornsnakes (Pantherophis guttatus or Pantherophis slowinskii) and ratsnakes (Pantherophis alleghaniensis, Pantherophis spiloides, or Pantherophis obsoletus) exhibited differences in habitat selection, but we obtained no evidence that patterns of use for this species pair were influenced by current interspecific interactions. Overall, our results are consistent with the hypothesis that competitive interactions influence snake assemblage composition; the strength of these effects was

  4. 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

  5. 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

  6. 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

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

    Science.gov (United States)

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

    2009-11-02

    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.

  8. 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.

  9. Poisonous snakebite in central Texas. Possible indicators for antivenin treatment.

    Science.gov (United States)

    White, R R; Weber, R A

    1991-01-01

    Sixty-seven patients hospitalized for poisonous snakebite between 1975 and 1990 were managed by elevation, tetanus prophylaxis, intravenous fluids and antibiotics, and often by a limited excision of the bite site in the Emergency Department, with sequential laboratory studies as needed. Antivenin was used for systemic envenomation, and 23 of the 67 patients (34%) received 133 vials. Thirteen of the twenty-three patients (56%) had adverse reactions to the antivenin. Two significant observations arose. First age was an indicator. Eleven of eighteen patients 12 years or younger (61%) received antivenin, whereas 12 of 49 patients older than 12 years (24%) received antivenin (p = 0.0085, Fisher's exact test). Second species of snake was an indicator. Sixty-two snakes were identified (93%). Of 39 rattlesnake (Crotalus and Sistrurus) bites, 20 patients received antivenin (53%), but of 23 copperhead and water moccasin (Agkistrodon) bites, only three patients (12.5%) received antivenin (p = 0.0025). Antivenin may be indicated for use in systemic rattlesnake envenomation, especially in younger patients. PMID:2025067

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

    Directory of Open Access Journals (Sweden)

    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.

  11. Predation and the maintenance of color polymorphism in a habitat specialist squamate.

    Directory of Open Access Journals (Sweden)

    Vincent R Farallo

    Full Text Available Multiple studies have addressed the mechanisms maintaining polymorphism within a population. However, several examples exist where species inhabiting diverse habitats exhibit local population-specific polymorphism. Numerous explanations have been proposed for the maintenance of geographic variation in color patterns. For example, spatial variation in patterns of selection or limited gene flow can cause entire populations to become fixed for a single morph, resulting in separate populations of the same species exhibiting separate and distinct color morphs. The mottled rock rattlesnake (Crotalus lepidus lepidus is a montane species that exhibits among-population color polymorphism that correlates with substrate color. Habitat substrate in the eastern part of its range is composed primarily of light colored limestone and snakes have light dorsal coloration, whereas in the western region the substrate is primarily dark and snakes exhibit dark dorsal coloration. We hypothesized that predation on high contrast color and blotched patterns maintain these distinct color morphs. To test this we performed a predation experiment in the wild by deploying model snakes at 12 sites evenly distributed within each of the two regions where the different morphs are found. We employed a 2×2 factorial design that included two color and two blotched treatments. Our results showed that models contrasting with substrate coloration suffered significantly more avian attacks relative to models mimicking substrates. Predation attempts on blotched models were similar in each substrate type. These results support the hypothesis that color pattern is maintained by selective predation.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    CA Gallardo-Casas

    2012-01-01

    Full Text Available In Mexico, medicinal plants are widely used. The use of Randia aculeata by healers against snakebites has never been scientifically tested in relation to possible effects on blood parameters and muscle tissue damage. Interviews were carried out in Jamapa, Veracuz, Mexico, with local residents to collect information about the traditional use of Randia aculeata. In this locality, seven pieces of fruit from the plant are mixed in a liter of alcohol, and then administered orally against snakebites. By using histological techniques and a murine model, we explored its cytoprotective properties against the effects of Crotalus simus and Bothrops asper venoms. Possible protections provided by the plant against tissue damage to skeletal and cardiac muscles and against the typical loss of red blood cells were analyzed. Randia aculeata caused an increase in microhematocrit and total hemoglobin, parameters that are often decremented in association with the loss of red blood cells, which is a characteristic effect of animal venom. Randia aculeata was also shown to protect against the lowering of platelet levels caused by Bothrops asper venom. Finally, Randia aculeata produced a partial inhibition of necrosis following administration of snake venom in skeletal and myocardial muscles. The present results provide solid evidence for the traditional use of Randia aculeata against snakebites, as demonstrated by protection against muscular tissue damage and the diminution of red blood cells.

  14. Epidemiological and clinical aspects of snAkebite in Belo Horizonte, Southeast Brazil

    Directory of Open Access Journals (Sweden)

    CAIAFFA Waleska Teixeira

    1997-01-01

    Full Text Available Epidemiologic and clinical aspects of 310 hospitalized snakebite patients and 310 matched controls were described, over a seven years period, from an emergency hospital in Belo Horizonte, Southeast Brazil. The diagnosis was based upon clinical picture or actual snake identification. Fifty six percent of victims were bitten by the snakes of genus Bothrops, 32.0% by Crotalus, 1.0% by Lachesis and 10.0% undetermined. During the study period, stable number of cases and marked seasonal variation were noted. In comparing cases of snakebite and controls, those from a rural area or who were involved in agricultural labor activity were identified as a high risk group, with an odds ratio (OR of 14.7 and 6.7, respectively, in favor of being bitten. Upon treatment, snakebite patients were 13.5 times more likely to have had early anaphylactic reactions than their controls, with a higher association in the age group ³ 20 years (OR = 30.3. Increased risks were also detected for pyrexia (OR = 11.7, with a marked association in the group under 19 years old (OR = 16.6. Severe cases of snakebite are an important treatable cause of morbidity in Brazil but therapy may be potentially life threatening. The higher case-fatality ratio encountered, compared to national statistics may be due the representativeness of the more severe cases who sought hospitalization. Preventing snakebite and early referral of those who are bitten is proposed

  15. Snakebite cases in the municipalities of the State of Paraiba, Brazil

    Directory of Open Access Journals (Sweden)

    Hellyson Fidel Araujo De Oliveira

    2013-10-01

    Full Text Available Introduction This study investigated the epidemiological and clinical profile of snakebite cases reported from 2007 to 2012 in the municipalities of the Curimataú region, State of Paraíba, in northeastern Brazil. Methods Data were collected from the Health Department of the State of Paraíba using the Injury Notification Information System data banks of the Health Ministry. Results A total of 304 snakebite cases were studied. The cases occurred most frequently from April to June. The genera Bothrops, Crotalus, and Micrurus were responsible for 74.6%, 6.2%, and 1.3% of cases, respectively. Snakebite cases predominated in males living in rural areas and between 10 and 19 years old. The highest incidence of bites occurred on the feet. The majority of the victims received medical assistance within 1 to 3h after being bitten. With regard to severity, 48% of the cases were classified as mild, 26% as moderate, and 2.6% as severe. Successful cures predominated, and no deaths were reported. The average antivenom ampoule dose used for the treatment in some snakebite cases was lower than that recommended by the Health Ministry. Conclusions Although our results show that Paraíba has a good level of medical care, there are serious deficiencies in recording snakebite information. These data indicate the need to improve the recording process for snakebite cases. Further training for health professionals seems to be necessary to optimize their skills in treating snakebite victims.

  16. 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.

  17. 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.

  18. Envenomation: a real risk of keeping exotic house pets.

    Science.gov (United States)

    de Haro, Luc; Pommier, Philip

    2003-08-01

    The fashion of exotic animals maintained as pets is increasing in France. Cases of envenomation after exotic animals bites or stings were studied. All 54 non-native animal envenomations reported by hospitals at the Poison Centre of Marseilles (3 south-eastern regions of France) between 1997 and 2002 were surveyed. They involved 22 snakes, 18 fishes, 11 spiders, 1 scorpion and 2 marine invertebrates. The snakes belonged to crotalids (genus Crotalus, Sistrurus, Agkistrodon and Trimeresurus), viperids (genus Bitis, Echis and Macrovipera, responsible for extensive swelling and coagulation disturbances), elapids (Naja and Dendroaspis which induce severe neurological signs), and colubrids (Lampropeltis with pain, edema and lymphangitis). Nine of the 22 patients bitten by snakes needed Intensive Care Unit management, and 5 of them received antivenom. Fish stings produced severe pain and local swelling. An Amazonian Stingray Potamotrygon histrix case had extensive swelling and malaise, headache and tremor. Pain and lymphangitis developed from tarantulas bites, and a black widow bite produced a severe diffuse muscle pain and contractions, and blood pressure disturbances. Exotic pets can be dangerous for their owners and family. Since antivenom from foreign countries is not often available in France, these cases raise the question of society's responsibility for treatment costs.

  19. 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.

  20. Problems with mitigation translocation of herpetofauna.

    Science.gov (United States)

    Sullivan, Brian K; Nowak, Erika M; Kwiatkowski, Matthew A

    2015-02-01

    Mitigation translocation of nuisance animals is a commonly used management practice aimed at resolution of human-animal conflict by removal and release of an individual animal. Long considered a reasonable undertaking, especially by the general public, it is now known that translocated subjects are negatively affected by the practice. Mitigation translocation is typically undertaken with individual adult organisms and has a much lower success rate than the more widely practiced conservation translocation of threatened and endangered species. Nonetheless, the public and many conservation practitioners believe that because population-level conservation translocations have been successful that mitigation translocation can be satisfactorily applied to a wide variety of human-wildlife conflict situations. We reviewed mitigation translocations of reptiles, including our own work with 3 long-lived species (Gila monsters [Heloderma suspectum], Sonoran desert tortoises [Gopherus morafkai], and western diamond-backed rattlesnakes [Crotalus atrox]). Overall, mitigation translocation had a low success rate when judged either by effects on individuals (in all studies reviewed they exhibited increased movement or increased mortality) or by the success of the resolution of the human-animal conflict (translocated individuals often returned to the capture site). Careful planning and identification of knowledge gaps are critical to increasing success rates in mitigation translocations in the face of increasing pressure to find solutions for species threatened by diverse anthropogenic factors, including climate change and exurban and energy development. © 2014 Society for Conservation Biology.

  1. Heat in evolution's kitchen: evolutionary perspectives on the functions and origin of the facial pit of pitvipers (Viperidae: Crotalinae).

    Science.gov (United States)

    Krochmal, Aaron R; Bakken, George S; LaDuc, Travis J

    2004-11-01

    Pitvipers (Viperidae: Crotalinae) possess thermal radiation receptors, the facial pits, which allow them to detect modest temperature fluctuations within their environments. It was previously thought that these organs were used solely to aid in prey acquisition, but recent findings demonstrated that western diamondback rattlesnakes (Crotalus atrox) use them to direct behavioral thermoregulation, suggesting that facial pits might be general purpose organs used to drive a suite of behaviors. To investigate this further, we conducted a phylogenetic survey of viperine thermoregulatory behavior cued by thermal radiation. We assessed this behavior in 12 pitviper species, representing key nodes in the evolution of pitvipers and a broad range of thermal environments, and a single species of true viper (Viperidae: Viperinae), a closely related subfamily of snakes that lack facial pits but possess a putative thermal radiation receptor. All pitviper species were able to rely on their facial pits to direct thermoregulatory movements, while the true viper was unable to do so. Our results suggest that thermoregulatory behavior cued by thermal radiation is a universal role of facial pits and probably represents an ancestral trait among pitvipers. Further, they establish behavioral thermoregulation as a plausible hypothesis explaining the evolutionary origin of the facial pit.

  2. Coccidian parasites (Apicomplexa) from snakes in the southcentral and southwestern United States: new host and geographic records.

    Science.gov (United States)

    McAllister, C T; Upton, S J; Trauth, S E; Dixon, J R

    1995-02-01

    Four hundred thirty-five leptotyphlopid, colubrid, elapid, and viperid snakes were collected from various localities in Arkansas, New Mexico, Oklahoma, and Texas, and their feces were examined for coccidian parasites. Of these, 131 (30%) were passing oocysts or sporocysts of at least 1 coccidian; 88 (67%) of the infected snakes had only 1 species of coccidian when they were examined. Aquatic and semiaquatic snakes accounted for 48% of the infections, whereas strictly terrestrial snakes comprised the other 52%. There was more than a 2-fold difference in prevalence among these 2 groups as 63 of 129 (49%) of the aquatic and semiaquatic snakes versus 68 of 306 (22%) of the terrestrial snakes harbored coccidia. Most terrestrial snakes were infected by species of Caryospora and Sarcocystis that are either facultatively or obligatorily heteroxenous. The aquatic and semiaquatic species most often harbored eimerians. Attempts to transmit some of the Sarcocystis spp. experimentally from Crotalus atrox to Mus musculus, Peromyscus leucopus, Peromyscus maniculatus, or Microtus ochrogaster were unsuccessful. This report documents 27 new host and several distributional records for coccidians from snakes in the southcentral and southwestern United States.

  3. 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

  4. 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).

  5. 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

  6. [Biochemical studies on functional proteins in the brain nervous system. Neurobiochemical studies on nerve growth factor].

    Science.gov (United States)

    Hayashi, K

    1996-04-01

    Nerve growth factor (NGF) is a polypeptide that is necessary for the survival and growth of developing sympathetic and sensory neurons as well as basal forebrain cholinergic neurons in the brain. The effects of NGF are mediated by the binding of the factor to its specific receptor present on the surface of NGF-responsive cells. Since NGF-responsive basal forebrain cholinergic neurons are lost in Alzheimer's disease, treatment with NGF may be therapeutically beneficial for the patients with this disease. Our studies were focused on the purification, amino acid sequence, enzyme immunoassay (EIA), NGF-biosynthesizing cells, regulation of the biosynthesis, compounds stimulating NGF-biosynthesis/secretion, etc. The major results obtained are summarized as follows: 1) We purified NGFs from snake venoms and determined their amino acid sequences. Among them, Crotalus adamanteus-and Vipera russelli-NGFs were glycoproteins. 2) We developed highly sensitive (0.03-0.05 pg/tube or bead), simple, and reliable EIA systems for NGFs and NT-3. 3) Catecholamine, its derivatives, p-quinone derivatives, nicotine, nicotine derivatives, coenzymes, etc. enhanced the NGF-biosynthesis/secretion of fibroblasts and astrocytes by increasing their cellular content of NGF mRNA. 4) Basal forebrain-lesioned rats (an animal model of amnesia) administered 6-(4-hydroxybutyl)-2,3,5-trimethyl-1,4-benzoquinone (TMQ) orally showed improvement not only their NGF level and choline acetyltransferase (CAT) activity in the brain but also their memory and learning. 5) Interleukins (IL)-4 and -5 significantly increased the amounts of NGF secreted by mouse astrocytes; whereas IL-2, -3, and -6 had no significant effect. 6) Interferons-beta and -lambda inhibited the DNA- and NGF-synthesis in growing astrocytes cultured from neonatal mouse brain but they had no effect on the NGF-synthesis/secretion in quiescent astrocytes. 7) Alkylcatechols regulated the NGF gene expression in astrocytes via both protein kinase C

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    J. Vellard

    1929-01-01

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

  9. Prevalence of common thrombophilia markers and risk factors in Indian patients with primary venous thrombosis

    Directory of Open Access Journals (Sweden)

    Mahendra Narain Mishra

    Full Text Available CONTEXT AND OBJECTIVE: Venous thrombosis occurs as a result of interaction of genetic and acquired factors including activated protein C resistance (APC-R, fibrinogen levels, antithrombin, protein C, protein S, lupus anticoagulants and anticardiolipin antibodies. This study was aimed at determining the prevalence of these common thrombophilia markers in Asian Indians with primary venous thrombosis. DESIGN AND SETTING: This was a cross-sectional study carried out in Mumbai. METHODS: Samples from 78 patients with a confirmed diagnosis of venous thrombosis and 50 controls were tested. Semi-quantitative estimation (functional assays of protein C, protein S and antithrombin was performed. Quantitative estimation of fibrinogen was done using the Clauss method. Lupus anticoagulants were screened using lupus-sensitive activated partial thromboplastin time and β2-glycoprotein-I dependent anticardiolipin antibodies were estimated by ELISA. APC-R was measured using a clotting-based method with factor V deficient plasma and Crotalus viridis venom. Statistical analysis was performed using Epi-info (version 6. RESULTS: The popliteal vein was the most commonly involved site. Forty-four samples (56% gave abnormal results. The commonest were elevated fibrinogen and APC-R (17.9% each, followed by low protein S (16.6%. CONCLUSIONS: This study confirms the literature findings that fibrinogen level estimation and screening for APC-R are important for the work-up on venous thrombosis patients since these, singly or in combination, may lead to a primary thrombotic episode. The frequency of the other thrombophilia markers was higher among the patients than among the controls, but without statistically significant difference.

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

    Directory of Open Access Journals (Sweden)

    Ribeiro Lindioneza Adriano

    1995-01-01

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

  11. 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.

  12. 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.

  13. [Genetic origin of venom variability: impact on the preparation of antivenin serums].

    Science.gov (United States)

    Nkinin, S W; Chippaux, J P; Piétin, D; Doljansky, Y; Trémeau, O; Ménez, A

    1997-01-01

    One of the main possible origin of the biochemical variations of venoms could be genetic. We studied the venom of members of litters born in a snake farm (12 Crotalus atrox and 21 Naja haje). We first used the electrophoresis in cellulose acetate (AE). Then, variations were confirmed by immunoelectrophoresis (AIE) using an antivenom (IPSER Africa, Pasteur Mérieux Sérums & Vaccines) and immunsera prepared on rabbit from i) venom presenting the maximum of bands in electrophoresis (complete venom) and ii) pure toxins (neurotoxin-alpha and cardiotoxin-gamma). At last, the toxicity of some samples was measured and the ability of SAV to neutralise the corresponding sample was measured. The AE of C. atrox venoms showed a good homogeneity, probably due to a good genetic stability of the investigated group. On the other hand, N. haje venoms have revealed a great heterogeneity. The 13 samples were allocated to five groups according to the absence of some fractions compared to the complete venom. The AIE showed that the neurotoxin-alpha is present in every sample, but variable in quantity, even when it did not appear on AE. We suggest that these pattern variations are due either to relative variations of protein fractions in samples or to modifications of the chemical composition of the neurotoxin-alpha. However, the variation of toxicity between the different samples questioned the neutralisation ability of antivenoms. We propose that venom sample choice for SAV production should be based on biochemical criteria and toxicity of samples rather than random pooling.

  14. When less means more: dehydration improves innate immunity in rattlesnakes.

    Science.gov (United States)

    Brusch, George A; DeNardo, Dale F

    2017-06-15

    Immune function can vary based on availability of resources, and most studies of such influences have focused on the co-investment of energy into immune and other physiological functions. When energy resources are limited, trade-offs exist, which can compromise immunity for other functions. As with energy, water limitation can also alter various physiological processes, yet water has received little consideration for its possible role in modulating immune functions. We examined the relationship between immunocompetence and hydration state using the western diamond-backed rattlesnake ( Crotalus atrox ). This species is known to undergo substantial seasonal fluctuations in water availability with extreme limitations during the hot-dry season. We collected blood samples from free-ranging C. atrox to compare osmolality and innate immune function (lysis, agglutination and bacterial growth inhibition) during the milder and relatively moister early spring season, the hot-dry season and the hot-wet season. To isolate effects of dehydration from other possible seasonal influences, we complemented this field study with a laboratory study in which we withheld food and water from individually housed adult C. atrox for up to 16 weeks. We collected blood samples from each snake as it dehydrated and collected a final sample after the snake was given water ad libitum at the end of the experiment. Our results demonstrate that C. atrox experience significant dehydration during the hot-dry season, and that, in general, innate immune function is highly correlated with osmolality, whether natural or artificially manipulated. © 2017. Published by The Company of Biologists Ltd.

  15. Effects of PI and PIII Snake Venom Haemorrhagic Metalloproteinases on the Microvasculature: A Confocal Microscopy Study on the Mouse Cremaster Muscle.

    Science.gov (United States)

    Herrera, Cristina; Voisin, Mathieu-Benoit; Escalante, Teresa; Rucavado, Alexandra; Nourshargh, Sussan; Gutiérrez, José María

    2016-01-01

    The precise mechanisms by which Snake Venom Metalloproteinases (SVMPs) disrupt the microvasculature and cause haemorrhage have not been completely elucidated, and novel in vivo models are needed. In the present study, we compared the effects induced by BaP1, a PI SVMP isolated from Bothrops asper venom, and CsH1, a PIII SVMP from Crotalus simus venom, on cremaster muscle microvasculature by topical application of the toxins on isolated tissue (i.e., ex vivo model), and by intra-scrotal administration of the toxins (i.e., in vivo model). The whole tissue was fixed and immunostained to visualize the three components of blood vessels by confocal microscopy. In the ex vivo model, BaP1 was able to degrade type IV collagen and laminin from the BM of microvessels. Moreover, both SVMPs degraded type IV collagen from the BM in capillaries to a higher extent than in PCV and arterioles. CsH1 had a stronger effect on type IV collagen than BaP1. In the in vivo model, the effect of BaP1 on type IV collagen was widespread to the BM of arterioles and PCV. On the other hand, BaP1 was able to disrupt the endothelial barrier in PCV and to increase vascular permeability. Moreover, this toxin increased the size of gaps between pericytes in PCV and created new gaps between smooth muscle cells in arterioles in ex vivo conditions. These effects were not observed in the case of CsH1. In conclusion, our findings demonstrate that both SVMPs degrade type IV collagen from the BM in capillaries in vivo. Moreover, while the action of CsH1 is more directed to the BM of microvessels, the effects of BaP1 are widespread to other microvascular components. This study provides new insights in the mechanism of haemorrhage and other pathological effects induced by these toxins.

  16. Modulation of orthogonal body waves enables high maneuverability in sidewinding locomotion.

    Science.gov (United States)

    Astley, Henry C; Gong, Chaohui; Dai, Jin; Travers, Matthew; Serrano, Miguel M; Vela, Patricio A; Choset, Howie; Mendelson, Joseph R; Hu, David L; Goldman, Daniel I

    2015-05-12

    Many organisms move using traveling waves of body undulation, and most work has focused on single-plane undulations in fluids. Less attention has been paid to multiplane undulations, which are particularly important in terrestrial environments where vertical undulations can regulate substrate contact. A seemingly complex mode of snake locomotion, sidewinding, can be described by the superposition of two waves: horizontal and vertical body waves with a phase difference of ± 90°. We demonstrate that the high maneuverability displayed by sidewinder rattlesnakes (Crotalus cerastes) emerges from the animal's ability to independently modulate these waves. Sidewinder rattlesnakes used two distinct turning methods, which we term differential turning (26° change in orientation per wave cycle) and reversal turning (89°). Observations of the snakes suggested that during differential turning the animals imposed an amplitude modulation in the horizontal wave whereas in reversal turning they shifted the phase of the vertical wave by 180°. We tested these mechanisms using a multimodule snake robot as a physical model, successfully generating differential and reversal turning with performance comparable to that of the organisms. Further manipulations of the two-wave system revealed a third turning mode, frequency turning, not observed in biological snakes, which produced large (127°) in-place turns. The two-wave system thus functions as a template (a targeted motor pattern) that enables complex behaviors in a high-degree-of-freedom system to emerge from relatively simple modulations to a basic pattern. Our study reveals the utility of templates in understanding the control of biological movement as well as in developing control schemes for limbless robots.

  17. Structural correlates of speed and endurance in skeletal muscle: the rattlesnake tailshaker muscle

    Science.gov (United States)

    Schaeffer; Conley; Lindstedt

    1996-01-01

    The western diamondback rattlesnake Crotalus atrox can rattle its tail continuously for hours at frequencies approaching 90 Hz. We examined the basis of these fast sustainable contractions using electromyography, data on oxygen uptake and the quantitative ultrastructure of the tailshaker muscle complex. The tailshaker muscle has no apparent unique structures; rather, the relative proportions of the structures common to all skeletal muscles appear to be present (1) to minimize activation, contraction and relaxation times via an extremely high volume density of sarcoplasmic reticulum (26 %) as well as, (2) to maximize ATP resysnthesis via a high volume density of mitochondria (26 %). The high rate of ATP supply is reflected in the in vivo muscle mass-specific oxygen uptake of this group of muscles which, at 585 ml O2 kg-1 min-1 during rattling at 30 °C body temperature, exceeds that reported for other ectotherm and many endotherm muscles. Since the change in oxygen uptake paralleled that of the rattling frequency over the range of measured body temperatures, there was a nearly constant O2 cost per muscle contraction (0.139±0.016 µl O2 g-1). Electromyo-graphic analysis suggests that each of the six muscles that make up the shaker complex may be a single motor unit. Finally, the maximum rate of mitochondrial oxygen uptake is similar to that of various mammals, a hummingbird, a lizard, an anuran amphibian and of isolated mitochondria (at 10 000-40 000 molecules O2 s-1 µm2 of cristae surface area, when normalized to 30 °C), suggesting a shared principle of design of the inner mitochondrial membrane among the vertebrates.

  18. Thrombin-dependent Incorporation of von Willebrand Factor into a Fibrin Network*

    Science.gov (United States)

    Miszta, Adam; Pelkmans, Leonie; Lindhout, Theo; Krishnamoorthy, Ganeshram; de Groot, Philip G.; Hemker, Coenraad H.; Heemskerk, Johan W. M.; Kelchtermans, Hilde; de Laat, Bas

    2014-01-01

    Attachment of platelets from the circulation onto a growing thrombus is a process involving multiple platelet receptors, endothelial matrix components, and coagulation factors. It has been indicated previously that during a transglutaminase reaction activated factor XIII (FXIIIa) covalently cross-links von Willebrand factor (VWF) to polymerizing fibrin. Bound VWF further recruits and activates platelets via interactions with the platelet receptor complex glycoprotein Ib (GPIb). In the present study we found proof for binding of VWF to a fibrin monomer layer during the process of fibrinogen-to-fibrin conversion in the presence of thrombin, arvin, or a snake venom from Crotalus atrox. Using a domain deletion mutant we demonstrated the involvement of the C domains of VWF in this binding. Substantial binding of VWF to fibrin monomers persisted in the presence of the FXIIIa inhibitor K9-DON, illustrating that cross-linking via factor XIII is not essential for this phenomenon and suggesting the identification of a second mechanism through which VWF multimers incorporate into a fibrin network. Under high shear conditions, platelets were shown to adhere to fibrin only if VWF had been incorporated. In conclusion, our experiments show that the C domains of VWF and the E domain of fibrin monomers are involved in the incorporation of VWF during the polymerization of fibrin and that this incorporation fosters binding and activation of platelets. Fibrin thus is not an inert end product but partakes in further thrombus growth. Our findings help to elucidate the mechanism of thrombus growth and platelet adhesion under conditions of arterial shear rate. PMID:25381443

  19. Conflict and assessment in a predator-prey system: ground squirrels versus rattlesnakes.

    Science.gov (United States)

    Swaisgood; Owings; Rowe

    1999-05-01

    Adult California ground squirrels, Spermophilus beecheyi beecheyi, actively confront and harass northern Pacific rattlesnakes, Crotalus viridis oreganus, which are the principal predator of ground squirrel pups. In this report we examine the roles of risk (snake size) and context (location of encounter and squirrel reproductive category) in rattlesnake assessment by ground squirrels. In interpreting the results, we borrow heavily from the well-developed conceptual framework applied in the analogous case of intraspecific conflict. Large and small snakes were tethered near the home burrows of male ground squirrels, and maternal and nonmaternal female ground squirrels. Ground squirrels appeared to employ assessment strategies which served to mediate the level of risk associated with confronting larger snakes. The results suggest that ground squirrels exercise greater caution when dealing with large snakes and invest more in monitoring the snake from a safe distance. Maternal squirrels, which have more at stake reproductively, spent more time and effort in snake-directed activities than did squirrels from other reproductive categories. Mothers also differentiated more strongly between large and small snakes, perhaps reflecting the greater vulnerability of their pups to larger snakes. Finally, ground squirrels discriminated between snakes found close to their home burrow and those encountered further abroad. At the home burrow, squirrels monitored the snake from a closer distance, displayed a greater willingness to confront the snake, and escalated to more dangerous levels of harassment. This assessment strategy may reflect a higher payoff to squirrels that persist in driving snakes out of the home area, thereby reducing the risk of future ambush. Copyright 1999 The Association for the Study of Animal Behaviour.

  20. Screening for target toxins of the antiophidic protein DM64 through a gel-based interactomics approach.

    Science.gov (United States)

    Rocha, Surza L G; Neves-Ferreira, Ana G C; Trugilho, Monique R O; Angulo, Yamileth; Lomonte, Bruno; Valente, Richard H; Domont, Gilberto B; Perales, Jonas

    2017-01-16

    DM64 is a glycosylated protein with antivenom activity isolated from the serum of the opossum Didelphis aurita. It binds non-covalently to myotoxins I (Asp49) and II (Lys49) from Bothrops asper venom and inhibits their myotoxic effect. In this study, an affinity column with immobilized DM64 as bait was used to fish potential target toxins. All ten isolated myotoxins tested were able to effectively bind to the DM64 column. To better access the specificity of the inhibitor, crude venoms from Bothrops (8 species), Crotalus (2 species) and Naja naja atra were submitted to the affinity purification. Venom fractions bound and nonbound to the DM64 column were analyzed by two-dimensional gel electrophoresis and MALDI-TOF/TOF MS. Although venom fractions bound to the column were mainly composed of basic PLA2, a few spots corresponding to acidic PLA2 were also observed. Some unexpected protein spots were also identified: C-type lectins and CRISP may represent putative new targets for DM64, whereas the presence of serine peptidases in the venom bound fraction is likely a consequence of nonspecific binding to the column matrix. The present results contribute to better delineate the inhibitory potential of DM64, providing a framework for the development of more specific antivenom therapies. Local tissue damage induced by myotoxic PLA2 remains a serious consequence of snake envenomation, since it is only partially neutralized by traditional antivenom serotherapy. Myotoxin inhibition by highly specific molecules offers great promise in the treatment of snakebites, a health problem largely neglected by governments and pharmaceutical industries. Bioactive compounds such as DM64 can represent a valuable source of scaffolds for drug development in this area. The present study has systematically profiled the binding specificity of DM64 toward a variety of snake venom toxin classes and therefore can lead to a better understanding of the structure-function relationship of this important

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

    Directory of Open Access Journals (Sweden)

    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.

  2. Pentastomídeos de répteis do Brasil: revisão dos cephalobaenidae

    Directory of Open Access Journals (Sweden)

    A. Arandas Rêgo

    1983-12-01

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

  3. Structural and evolutionary insights into endogenous alpha-phospholipase A2 inhibitors of Latin American pit vipers.

    Science.gov (United States)

    Estevão-Costa, Maria Inácia; Fernandes, Carlos Alexandre H; Mudadu, Maurício de Alvarenga; Franco, Glória Regina; Fontes, Marcos Roberto M; Fortes-Dias, Consuelo Latorre

    2016-03-15

    Phospholipases A2 are major components of snake venoms (svPLA2s) and are able to induce multiple local and systemic deleterious effects upon envenomation. Several snake species are provided with svPLA2 inhibitors (sbPLIs) in their circulating blood, which confer a natural resistance against the toxic components of homologous and heterologous venoms. The sbPLIs belong to any of three structural classes named α, β and γ. In the present study, we identified, characterized and performed structural and evolutionary analyses of sbαPLIs transcripts and the encoded proteins, in the most common Latin American pit vipers belonging to Crotalus, Bothrops and Lachesis genera. Mutation data indicated that sbαPLIs from Latin American snakes might have evolved in an accelerated manner, similarly to that reported for sbαPLIs from Asian snakes, and possibly co-evoluted with svPLA2s in response to the diversity of target enzymes. The importance of sbαPLI trimerization for the effective binding and inhibition of acidic svPLA2s is discussed and conserved cationic residues located at the central pore of the inhibitor trimer are suggested to be a significant part of the binding site of sbαPLIs to acidic svPLA2s. Our data contribute to the current body of knowledge on the structural and evolutionary characteristics of sbPLIs, in general, and may assist in the future development of selective inhibitors for secretory PLA2 from several sources. Copyright © 2016 Elsevier Ltd. All rights reserved.

  4. Inhibition of hemorrhagic and edematogenic activities of snake venoms by a broad-spectrum protease inhibitor, murinoglobulin; the effect on venoms from five different genera in Viperidae family.

    Science.gov (United States)

    Ribeiro Filho, Wilker; Sugiki, Masahiko; Yoshida, Etsuo; Maruyama, Masugi

    2003-08-01

    In order to obtain basic data on the effect of broad-spectrum protease inhibitor against local symptoms of Viperidae snake envenomation, inhibitory capacity of rat murinoglobulin on local hemorrhagic and edematogenic activities of venoms from Crotalus atrox, Bothrops jararaca, Lachesis muta muta, Trimeresurus flavoviridis and Echis carinatus sochureki were examined. Murinoglobulin, pre-incubated with the crude venoms at 37 degrees C for 15 min, inhibited hemorrhagic activity of all five venoms to various extents. The activity of C. atrox was almost completely inhibited at the murinoglobulin/venom ratio (w/w) of 20. The activity of B. jararaca, Lachesis muta muta and T. flavoviridis venoms was considerably inhibited at the ratio of 20 (77.2, 80.0 and 86.2% inhibition, respectively), however some of the activity still remained even at the ratio of 40 (84.2, 79.8 and 86.2% inhibition, respectively). Among the five venoms, E. c. sochureki venom is quite resistant to murinoglobulin treatment and statistically significant inhibition was only found at the ratio of 40 (64.1% inhibition). Fibrinolytic and gelatinase activities were more susceptible to murinoglobulin inhibition. The treatment at the ratios of 10 and 20 almost completely inhibited respectively the fibrinolytic and the gelatinase activities of all the venoms. Murinoglobulin treatment also significantly inhibited the edematogenic activity of L. muta muta, T. flavoviridis and Echis carinatus sochureki. The treatment of murinoglobulin at the ratio of 40 considerably suppressed the swelling up to 60 min after subcutaneous injection of L. muta muta and E. c. sochureki venoms, and up to 30 min after T. flavoviridis venom injection. Murinoglobulin is a potent inhibitor against local effects of multiple snake venoms in Viperidae family.

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

    Science.gov (United States)

    Adade, Camila M; Carvalho, Ana Lúcia O; Tomaz, Marcelo A; Costa, Tatiana F R; Godinho, Joseane L; Melo, Paulo A; Lima, Ana Paula C A; Rodrigues, Juliany C F; Zingali, Russolina B; Souto-Padrón, Thaïs

    2014-10-01

    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.

  6. In vitro antimicrobial activity of natural toxins and animal venoms tested against Burkholderia pseudomallei

    Directory of Open Access Journals (Sweden)

    Chow Vincent TK

    2006-06-01

    Full Text Available Abstract Background Burkholderia pseudomallei are the causative agent of melioidosis. Increasing resistance of the disease to antibiotics is a severe problem in treatment regime and has led to intensification of the search for new drugs. Antimicrobial peptides are the most ubiquitous in nature as part of the innate immune system and host defense mechanism. Methods Here, we investigated a group of venoms (snakes, scorpions and honey bee venoms for antimicrobial properties against two strains of Gram-negative bacteria Burkholderia pseudomallei by using disc-diffusion assay for in vitro susceptibility testing. The antibacterial activities of the venoms were compared with that of the isolated L-amino acid oxidase (LAAO and phospholipase A2 (PLA2s enzymes. MICs were determined using broth dilution method. Bacterial growth was assessed by measurement of optical density at the lowest dilutions (MIC 0.25 mg/ml. The cell viability was measured using tetrazolium salts (XTT based cytotoxic assay. Results The studied venoms showed high antimicrobial activity. The venoms of C. adamanteus, Daboia russelli russelli, A. halys, P. australis, B. candidus and P. guttata were equally as effective as Chloramphenicol and Ceftazidime (30 μg/disc. Among those tested, phospholipase A2 enzymes (crotoxin B and daboiatoxin showed the most potent antibacterial activity against Gram-negative (TES bacteria. Naturally occurring venom peptides and phospholipase A2 proved to possess highly potent antimicrobial activity against Burkholderia pseudomallei. The XTT-assay results showed that the cell survival decreased with increasing concentrations (0.05–10 mg/mL of Crotalus adamanteus venom, with no effect on the cell viability evident at 0.5 mg/mL. Conclusion This antibacterial profile of snake venoms reported herein will be useful in the search for potential antibacterial agents against drug resistant microorganisms like B. pseudomallei.

  7. In vitro antimicrobial activity of natural toxins and animal venoms tested against Burkholderia pseudomallei

    Science.gov (United States)

    Perumal Samy, R; Pachiappan, A; Gopalakrishnakone, P; Thwin, Maung M; Hian, Yap E; Chow, Vincent TK; Bow, Ho; Weng, Joseph T

    2006-01-01

    Background Burkholderia pseudomallei are the causative agent of melioidosis. Increasing resistance of the disease to antibiotics is a severe problem in treatment regime and has led to intensification of the search for new drugs. Antimicrobial peptides are the most ubiquitous in nature as part of the innate immune system and host defense mechanism. Methods Here, we investigated a group of venoms (snakes, scorpions and honey bee venoms) for antimicrobial properties against two strains of Gram-negative bacteria Burkholderia pseudomallei by using disc-diffusion assay for in vitro susceptibility testing. The antibacterial activities of the venoms were compared with that of the isolated L-amino acid oxidase (LAAO) and phospholipase A2 (PLA2s) enzymes. MICs were determined using broth dilution method. Bacterial growth was assessed by measurement of optical density at the lowest dilutions (MIC 0.25 mg/ml). The cell viability was measured using tetrazolium salts (XTT) based cytotoxic assay. Results The studied venoms showed high antimicrobial activity. The venoms of C. adamanteus, Daboia russelli russelli, A. halys, P. australis, B. candidus and P. guttata were equally as effective as Chloramphenicol and Ceftazidime (30 μg/disc). Among those tested, phospholipase A2 enzymes (crotoxin B and daboiatoxin) showed the most potent antibacterial activity against Gram-negative (TES) bacteria. Naturally occurring venom peptides and phospholipase A2 proved to possess highly potent antimicrobial activity against Burkholderia pseudomallei. The XTT-assay results showed that the cell survival decreased with increasing concentrations (0.05–10 mg/mL) of Crotalus adamanteus venom, with no effect on the cell viability evident at 0.5 mg/mL. Conclusion This antibacterial profile of snake venoms reported herein will be useful in the search for potential antibacterial agents against drug resistant microorganisms like B. pseudomallei. PMID:16784542

  8. 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.

  9. Epidemiology of accidents with poisonous animals registered in Nova Xavantina – MT

    Directory of Open Access Journals (Sweden)

    Vivian Tallita Pinheiro Santana

    2015-07-01

    Full Text Available Background and Objective: Accidents with poisonous animals are a public health problem in Brazil, especially in countryside cities. Updated regional information is important for the development of epidemiological surveillance activities. Through this research, we determined the epidemiological aspects of the cases registered in Nova Xavantina, State of Mato Grosso, Brazil, from January 2007 to May 2013. Methods: This is a cross-sectional and descriptive epidemiological study. Results: In this period, 68 accidents occurred, of which 44 were caused by snakes, 19 by scorpions, and four by spiders. Most accidents with snakes were caused by the genera Bothrops (79.5%, Crotalus (11.5%, and Micrurus (2.3%. As for the victims, there was a prevalence of males (76.5%, aged between 30 and 49 years (41.2% of mixed race (72.1%, with basic education (55.9% involved in field activities, such as agriculture and livestock (41.2%.There was a predominance of accidents of moderate severity (48.5% and all serious cases were caused by snakes. In most injuries (75%, the relationship between the accidents and occupational activities was not recorded. The most affected body regions were the feet (39.7%, legs (20.6%, and hands (11.8%. The predominant local manifestations were pain, swelling, and bruising. The predominant systemic manifestations were hemorrhage, nausea and vomiting. Conclusion: The study period was characterized by the predominance of accidents involving snakes and male individuals in field activities and of moderate severity. Thus, educational campaigns and the adoption of preventive measures are essential to reduce the occurrence of these diseases in this city. KEYWORDS: Poisoning; snake bites; Scorpion Stings.

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

    Directory of Open Access Journals (Sweden)

    Lindioneza Adriano Ribeiro

    1995-10-01

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

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

    Directory of Open Access Journals (Sweden)

    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

  12. 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

    Directory of Open Access Journals (Sweden)

    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.

  13. Construction of concept maps as tool for Biochemistry learning

    Directory of Open Access Journals (Sweden)

    Silvia Lopes de Menezes

    2006-07-01

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

  14. Use of medicinal fauna in Mexican traditional medicine.

    Science.gov (United States)

    Alonso-Castro, Angel Josabad

    2014-02-27

    Mexico has great biodiversity of fauna. The use of fauna with medicinal properties is a common practice since pre-Hispanic times. In the last decade, there has been an interest in ethnozoological studies in Mexico. Therefore, more studies are needed in order to gather information regarding the use of fauna with medicinal properties in México. Ethnozoological studies are necessary in order to discover new medications for human health. This review presents current information in terms of ethnozoological, conservation status, trade, toxicological and pharmacological effects of fauna used for medicinal purposes in Mexican traditional medicine (MTM), based on scientific literature. Future prospects for research with medicinal fauna are discussed. Bibliographic investigation was carried out by analyzing recognized books and peer-reviewed papers, consulting worldwide accepted scientific databases from the last five decades. Reports included in this review complied with the three criteria cited as follows: (i) used in Mexican traditional medicine for medicinal and/or magical-religious purposes, (ii) with experimental studies regarding the toxicological or medicinal effects and/or with studies exploring mechanisms of medicinal effects, and (iii) with information obtained from a clear source. A total of 163 animal species, belonging to 79 families and 4 taxonomic categories, used for medicinal purposes are reported in this review. Medicinal fauna used in MTM come from birds (48), fishes (3), insects (22), mammals (49) and reptiles (41). The most versatile species which had the greatest number of medicinal properties were Mephitis macroura (21 uses), Crotalus atrox (17 uses), Dasypus novemcinctus (13 uses) and Didelphis virginiana (13 uses). However, 14 of the 161 species listed in this review are classified as endangered. Animal species are mainly used for the treatment of inflammatory, respiratory and gastrointestinal diseases. Furthermore, insects and reptiles are the

  15. Molecular basis for prey relocation in viperid snakes.

    Science.gov (United States)

    Saviola, Anthony J; Chiszar, David; Busch, Chardelle; Mackessy, Stephen P

    2013-03-01

    Vertebrate predators use a broad arsenal of behaviors and weaponry for overcoming fractious and potentially dangerous prey. A unique array of predatory strategies occur among snakes, ranging from mechanical modes of constriction and jaw-holding in non-venomous snakes, to a chemical means, venom, for quickly dispatching prey. However, even among venomous snakes, different prey handling strategies are utilized, varying from the strike-and-hold behaviors exhibited by highly toxic elapid snakes to the rapid strike-and-release envenomation seen in viperid snakes. For vipers, this mode of envenomation represents a minimal risk predatory strategy by permitting little contact with or retaliation from prey, but it adds the additional task of relocating envenomated prey which has wandered from the attack site. This task is further confounded by trails of other unstruck conspecific or heterospecific prey. Despite decades of behavioral study, researchers still do not know the molecular mechanism which allows for prey relocation. During behavioral discrimination trials (vomeronasal responsiveness) to euthanized mice injected with size-fractionated venom, Crotalus atrox responded significantly to only one protein peak. Assays for enzymes common in rattlesnake venoms, such as exonuclease, L-amino acid oxidase, metalloproteinase, thrombin-like and kallikrein-like serine proteases and phospholipase A(2), showed that vomeronasal responsiveness was not dependent on enzymatic activity. Using mass spectrometry and N-terminal sequencing, we identified the proteins responsible for envenomated prey discrimination as the non-enzymatic disintegrins crotatroxin 1 and 2. Our results demonstrate a novel and critical biological role for venom disintegrins far beyond their well-established role in disruption of cell-cell and cell-extracellular matrix interactions. These findings reveal the evolutionary significance of free disintegrins in venoms as the molecular mechanism in vipers allowing for

  16. Acidente ofídico no estado de Goiás

    Directory of Open Access Journals (Sweden)

    Pinho Fábia Maria Oliveira

    2004-01-01

    Full Text Available OBJETIVO: Avaliar aspectos epidemiológicos dos acidentes por 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.

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

    Science.gov (United States)

    2011-01-01

    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 divergence between A. mexicanus

  18. Epidemiologia dos acidentes ofídicos notificados pelo Centro de Assistência e Informação Toxicológica de Campina Grande (Ceatox-CG, Paraíba Epidemiologic profile of snakebites reported by the Poison Information Center of Campina Grande, Paraíba

    Directory of Open Access Journals (Sweden)

    Josiverton de Carvalho Lemos

    2009-03-01

    Full Text Available Os acidentes causados por animais peçonhentos ainda constituem problema de Saúde Pública no Brasil. Embora a produção e a distribuição dos soros no país encontrem-se estabilizadas, há problemas relacionados à notificação de acidentes deste tipo em várias regiões. Considerando esta realidade, foi realizado um estudo transversal entre janeiro e dezembro de 2005, utilizando documentação indireta sobre os acidentes ofídicos ocorridos em Campina Grande e 80 municípios adjacentes à região, com o objetivo de conhecer o perfil epidemiológico e clínico deste tipo de caso. Todos os pacientes atendidos tiveram diagnóstico médico de acidente por serpentes realizado pelo Centro de Atendimento Toxicológico de Campina Grande (Ceatox-CG. Os dados foram coletados através da ficha de notificação do Sinan (Sistema Nacional de Notificação de Agravo do Ministério da Saúde. Para análise dos dados, foi utilizada estatística descritiva e os dados foram tabulados no programa Microsoft Excel 2003. Dos 1.443 atendimentos no Centro, 737 foram causados por animais peçonhentos e, destes, 277 foram provocados por serpentes peçonhentas e não peçonhentas. Os acidentes predominaram no sexo masculino, na faixa etária de 10 a 29 anos, principalmente em agricultores na zona rural, entre maio e novembro de 2005. O gênero Bothrops foi responsável pelo maior número de casos (71,5%, e as extremidades superior e inferior do corpo foram os locais de maior predominância de ataques. Na extremidade inferior, foi atingido principalmente o pé. Em relação à gravidade, foram mais frequentes os acidentes classificados como leves causados pelo gênero Bothrops e apenas um paciente evoluiu para óbito. A média do tempo de atendimento, em horas, foi superior a 12 horas nos acidentes considerados graves, causados pelos gêneros Bothrops e Crotalus. Nossos resultados concordam com o perfil epidemiológico nacional dos acidentes ofídicos, atingindo com

  19. 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