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1.
J. venom. anim. toxins incl. trop. dis ; 27: e20200068, 2021. tab, graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1154772

Resumo

Maintenance of snakes at Butantan Institute started in the last century, intending to produce a different antivenom serum to reduce death caused by snakebites. Through a successful campaign coordinated by Vital Brazil, farmers sent venomous snakes to Butantan Institute by the railway lines with no cost. From 1908 to 1962, the snakes were kept in an outdoor serpentarium, where venom extraction was performed every 15 days. During this period, the snake average survival was 15 days. In 1963, the snakes were transferred to an adapted building, currently called Laboratory of Herpetology (LH), to be maintained in an intensive system. Although the periodicity of venom extraction remained the same, animal average survival increased to two months. With the severe serum crisis in 1983, the Ministry of Health financed remodeling for the three public antivenom producers, and with this support, the LH could be improved. Air conditioning and exhausting systems were installed in the rooms, besides the settlement of critical hygienic-sanitary managements to increase the welfare of snakes. In the early 1990s, snake survival was ten months. Over the years to the present day, several improvements have been made in the intensive serpentarium, as the establishment of two quarantines, feeding with thawed rodents, an interval of two months between venom extraction routines, and monitoring of snake health through laboratory tests. With these new protocols, average snake survival increased significantly, being eight years for the genus Bothrops, ten years for genus Crotalus and Lachesis, and four years for the genus Micrurus. Aiming the production of venoms of good quality, respect for good management practices is essential for the maintenance of snakes in captivity. New techniques and efficient management must always be sought to improve animal welfare, the quality of the venom produced, and the safety of those working directly with the venomous snakes.(AU)


Assuntos
Animais , Mordeduras de Serpentes , Viperidae , Venenos Elapídicos/biossíntese , Bem-Estar do Animal , Custos e Análise de Custo
2.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 27: e20200068, 2021. tab, graf
Artigo em Inglês | VETINDEX | ID: vti-31985

Resumo

Maintenance of snakes at Butantan Institute started in the last century, intending to produce a different antivenom serum to reduce death caused by snakebites. Through a successful campaign coordinated by Vital Brazil, farmers sent venomous snakes to Butantan Institute by the railway lines with no cost. From 1908 to 1962, the snakes were kept in an outdoor serpentarium, where venom extraction was performed every 15 days. During this period, the snake average survival was 15 days. In 1963, the snakes were transferred to an adapted building, currently called Laboratory of Herpetology (LH), to be maintained in an intensive system. Although the periodicity of venom extraction remained the same, animal average survival increased to two months. With the severe serum crisis in 1983, the Ministry of Health financed remodeling for the three public antivenom producers, and with this support, the LH could be improved. Air conditioning and exhausting systems were installed in the rooms, besides the settlement of critical hygienic-sanitary managements to increase the welfare of snakes. In the early 1990s, snake survival was ten months. Over the years to the present day, several improvements have been made in the intensive serpentarium, as the establishment of two quarantines, feeding with thawed rodents, an interval of two months between venom extraction routines, and monitoring of snake health through laboratory tests. With these new protocols, average snake survival increased significantly, being eight years for the genus Bothrops, ten years for genus Crotalus and Lachesis, and four years for the genus Micrurus. Aiming the production of venoms of good quality, respect for good management practices is essential for the maintenance of snakes in captivity. New techniques and efficient management must always be sought to improve animal welfare, the quality of the venom produced, and the safety of those working directly with the venomous snakes.(AU)


Assuntos
Animais , Mordeduras de Serpentes , Viperidae , Venenos Elapídicos/biossíntese , Bem-Estar do Animal , Custos e Análise de Custo
3.
J. venom. anim. toxins incl. trop. dis ; 26: e20190044, 2020. tab, graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1091017

Resumo

Abstract Background: Endogenous phospholipase A2 inhibitors from snake blood (sbPLIs) have been isolated from several species around the world, with the primary function of self-protection against the action of toxic phospholipases A2. In American snakes, sbPLIs were solely described in pit vipers, in which the natural protection role is justified. In this study, we described a sbPLI in Boa constrictor (popularly known as jiboia), a non-venomous snake species from America. Methods: PLA2 inhibitory activity was tested in the blood plasma of B. constrictor using C. d. terrificus venom as the enzyme source. Antibodies developed against CNF, a sbγPLI from Crotalus durissus terrificus, were used to investigate the presence of homologues in the blood plasma of B. constrictor. A CNF-like molecule with a PLA2 inhibitory activity was purified by column chromatography. The encoding gene for the inhibitor was cloned from B. constrictor liver tissue. The DNA fragment was cloned, purified and sequenced. The deduced primary sequence of interest was aligned with known sbγPLIs from the literature. Results: The blood plasma of B. constrictor displayed PLA2 inhibitory activity. A CNF-like molecule (named BcNF) was identified and purified from the blood plasma of B. constrictor. Basic properties such as molecular mass, composing amino acids, and pI were comparable, but BcNF displayed reduced specific activity in PLA2 inhibition. BcNF showed highest identity scores (ISs) with sbγPLIs from pit vipers from Latin America (90-100%), followed by gamma inhibitors from Asian viperid (80-90%). ISs below 70% were obtained for BcNF and non-venomous species from Asia. Conclusion: A functional sbγPLI (BcNF) was described in the blood plasma of B. constrictor. BcNF displayed higher primary identity with sbγPLIs from Viperidae than to sbγPLIs from non-venomous species from Asia. The physiological role played by sbγPLIs in non-venomous snake species remains to be understood. Further investigation is needed.(AU)


Assuntos
Animais , Serpentes , Viperidae , Venenos Elapídicos , Fosfolipases A2 , Inibidores de Fosfolipase A2
4.
J. venom. anim. toxins incl. trop. dis ; 26: e20200057, 2020. tab, graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1143218

Resumo

Certain environmental toxins permanently damage the thymic epithelium, accelerate immune senescence and trigger secondary immune pathologies. However, the exact underlying cellular mechanisms and pathways of permanent immune intoxication remain unknown. The aim of the present study was to demonstrate gene expressional changes of apoptosis-related cellular pathways in human thymic epithelial cells following exposure to snake venom from Bitis gabonica and Dendroaspis angusticeps. Methods: Snake venoms were characterized by analytical methods including reversed phase high-performance liquid chromatography and sodium dodecyl sulphate-polyacrylamide gel electrophoresis, then applied on human thymic epithelial cells (1889c) for 24 h at 10 μg/mL (as used in previous TaqMan Array study). Gene expressional changes restricted to apoptosis were assayed by TaqMan Array (Human Apoptosis Plate). Results: The most prominent gene expressional changes were shown by CASP5 (≈ 2.5 million-fold, confirmed by dedicated quantitative polymerase chain reaction) and CARD9 (0.016-fold) for B. gabonica, and BIRC7 (6.46-fold) and CASP1 (0.30-fold) for D. angusticeps. Conclusion: The observed apoptotic environment suggests that pyroptosis may be the dominant pathway through which B. gabonica and D. angusticeps snake venoms trigger thymic epithelial apoptosis following envenomation.(AU)


Assuntos
Animais , Venenos de Serpentes/efeitos adversos , Reação em Cadeia da Polimerase , Apoptose , Viperidae/genética , Células Epiteliais/química , Piroptose , Métodos de Análise Laboratorial e de Campo , Eletroforese em Gel de Poliacrilamida
5.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 26: e20200057, 2020. tab, graf
Artigo em Inglês | VETINDEX | ID: vti-32055

Resumo

Certain environmental toxins permanently damage the thymic epithelium, accelerate immune senescence and trigger secondary immune pathologies. However, the exact underlying cellular mechanisms and pathways of permanent immune intoxication remain unknown. The aim of the present study was to demonstrate gene expressional changes of apoptosis-related cellular pathways in human thymic epithelial cells following exposure to snake venom from Bitis gabonica and Dendroaspis angusticeps. Methods: Snake venoms were characterized by analytical methods including reversed phase high-performance liquid chromatography and sodium dodecyl sulphate-polyacrylamide gel electrophoresis, then applied on human thymic epithelial cells (1889c) for 24 h at 10 μg/mL (as used in previous TaqMan Array study). Gene expressional changes restricted to apoptosis were assayed by TaqMan Array (Human Apoptosis Plate). Results: The most prominent gene expressional changes were shown by CASP5 (≈ 2.5 million-fold, confirmed by dedicated quantitative polymerase chain reaction) and CARD9 (0.016-fold) for B. gabonica, and BIRC7 (6.46-fold) and CASP1 (0.30-fold) for D. angusticeps. Conclusion: The observed apoptotic environment suggests that pyroptosis may be the dominant pathway through which B. gabonica and D. angusticeps snake venoms trigger thymic epithelial apoptosis following envenomation.(AU)


Assuntos
Animais , Venenos de Serpentes/análise , Venenos de Serpentes/genética , Apoptose/genética , Células Epiteliais , Piroptose , Viperidae , Elapidae
6.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 25: e147018, Apr. 15, 2019. tab, graf
Artigo em Inglês | VETINDEX | ID: vti-19286

Resumo

Background:Lachesis muta rhombeata is one of the venomous snakes of medical importance in Brazil whose envenoming is characterized by local and systemic effects which may produce even shock and death. Its venom is mainly comprised of serine and metalloproteinases, phospholipases A2 and bradykinin-potentiating peptides. Based on a previously reported fractionation of L. m. rhombeata venom (LmrV), we decided to perform a subproteome analysis of its major fraction and investigated a novel component present in this venom.Methods:LmrV was fractionated through molecular exclusion chromatography and the main fraction (S5) was submitted to fibrinogenolytic activity assay and fractionated by reversed-phase chromatography. The N-terminal sequences of the subfractions eluted from reversed-phase chromatography were determined by automated Edman degradation. Enzyme activity of LmrSP-4 was evaluated upon chromogenic substrates for thrombin (S-2238), plasma kallikrein (S-2302), plasmin and streptokinase-activated plasminogen (S-2251) and Factor Xa (S-2222) and upon fibrinogen. All assays were carried out in the presence or absence of possible inhibitors. The fluorescence resonance energy transfer substrate Abz-KLRSSKQ-EDDnp was used to determine the optimal conditions for LmrSP-4 activity. Molecular mass of LmrSP-4 was determined by MALDI-TOF and digested peptides after trypsin and Glu-C treatments were analyzed by high resolution MS/MS using different fragmentation modes.Results:Fraction S5 showed strong proteolytic activity upon fibrinogen. Its fractionation by reversed-phase chromatography gave rise to 6 main fractions (S5C1-S5C6). S5C1-S5C5 fractions correspond to serine proteinases whereas S5C6 represents a C-type lectin. S5C4 (named LmrSP-4) had its N-terminal determined by Edman degradation up to the 53rd amino acid residue and was chosen for characterization studies. LmrSP-4 is a fibrinogenolytic...(AU)

7.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 25: e.20190009, Sep. 16, 2019. ilus, graf
Artigo em Inglês | VETINDEX | ID: vti-24668

Resumo

Background:Several studies have pointed out that certain snake venoms contain compounds presenting cytotoxic activities that selectively interfere with cancer cell metabolism. In this study, Pseudocerastes persicus venom and its fractions were investigated for their anticancer potential on lung cancer cells.Methods:Lung cancer cells (A549) and normal fibroblast cells (Hu02) were treated with the P. persicus venom and its HPLC fractions and the cell cytotoxic effects were analyzed using MTT and lactate dehydrogenase release assays. Apoptosis was determined in venom-treated cell cultures using caspase-3 and caspase-9 assay kits.Results:The treatment of cells with HPLC fraction 21 (25-35 kDa) of P. persicus venom resulted in high LDH release in normal fibroblast cells and high caspase-3 and caspase-9 activities in lung cancer cells. These results indicate that fraction 21 induces apoptosis in cancer cells, whereas necrosis is predominantly caused by cell death in the normal cells. Fraction 21 at the final concentration of 10 μg/mL killed approximately 60% of lung cancer cells, while in normal fibroblast cells very low cell cytotoxic effect was observed.Conclusion:HPLC fraction 21 at low concentrations displayed promising anticancer properties with apoptosis induction in the lung cancer cells. This fraction may, therefore, be considered a promising candidate for further studies.(AU)


Assuntos
Animais , Viperidae , Venenos de Víboras/análise , Venenos de Víboras/toxicidade , Citotoxinas , Neoplasias/terapia , Cromatografia Líquida de Alta Pressão , Apoptose
8.
J. venom. anim. toxins incl. trop. dis ; 25: e147018, 2019. tab, graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1002495

Resumo

Lachesis muta rhombeata is one of the venomous snakes of medical importance in Brazil whose envenoming is characterized by local and systemic effects which may produce even shock and death. Its venom is mainly comprised of serine and metalloproteinases, phospholipases A2 and bradykinin-potentiating peptides. Based on a previously reported fractionation of L. m. rhombeata venom (LmrV), we decided to perform a subproteome analysis of its major fraction and investigated a novel component present in this venom. Methods: LmrV was fractionated through molecular exclusion chromatography and the main fraction (S5) was submitted to fibrinogenolytic activity assay and fractionated by reversed-phase chromatography. The N-terminal sequences of the subfractions eluted from reversed-phase chromatography were determined by automated Edman degradation. Enzyme activity of LmrSP-4 was evaluated upon chromogenic substrates for thrombin (S-2238), plasma kallikrein (S-2302), plasmin and streptokinase-activated plasminogen (S-2251) and Factor Xa (S-2222) and upon fibrinogen. All assays were carried out in the presence or absence of possible inhibitors. The fluorescence resonance energy transfer substrate Abz-KLRSSKQ-EDDnp was used to determine the optimal conditions for LmrSP-4 activity. Molecular mass of LmrSP-4 was determined by MALDI-TOF and digested peptides after trypsin and Glu-C treatments were analyzed by high resolution MS/MS using different fragmentation modes. Results: Fraction S5 showed strong proteolytic activity upon fibrinogen. Its fractionation by reversed-phase chromatography gave rise to 6 main fractions (S5C1-S5C6). S5C1-S5C5 fractions correspond to serine proteinases whereas S5C6 represents a C-type lectin. S5C4 (named LmrSP-4) had its N-terminal determined by Edman degradation up to the 53rd amino acid residue and was chosen for characterization studies. LmrSP-4 is a fibrinogenolytic serine proteinase with high activity against S-2302, being inhibited by PMSF and benzamidine, but not by 1,10-phenantroline. In addition, this enzyme exhibited maximum activity within the pH range from neutral to basic and between 40 and 50 °C. About 68% of the LmrSP-4 primary structure was covered, and its molecular mass is 28,190 Da. Conclusions: Novel serine proteinase isoforms and a lectin were identified in LmrV. Additionally, a kallikrein-like serine proteinase that might be useful as molecular tool for investigating bradykinin-involving process was isolated and partially characterized.(AU)


Assuntos
Plasminogênio , Venenos de Serpentes , Lachesis muta , Serina Proteases , Calicreínas , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Fosfolipases A2
9.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 24: 32, Dec. 17, 2018. tab, graf
Artigo em Inglês | VETINDEX | ID: vti-19373

Resumo

Background: Lachesis muta rhombeata (Lmr) is the largest venomous snake in Latin America and its venom contains mainly enzymatic components, such as serine and metalloproteases, L-amino acid oxidase and phospholipases A2. Metalloproteases comprise a large group of zinc-dependent proteases that cleave basement membrane components such as fibronectin, laminin and collagen type IV. These enzymes are responsible for local and systemic changes, including haemorrhage, myonecrosis and inflammation. This study aimed the isolation and enzymatic characterization of the first metalloprotease (Lmr-MP) from Lmr venom (LmrV). Methods and results: Lmr-MP was purified through two chromatographic steps and submitted to enzymatic characterization. It showed proteolytic activity on azocasein with maximum activity at pH 7.0-9.0. It was inhibited by EDTA (a metal chelator that removes zinc, which is essential for enzymatic activity) and no effect was observed with PMSF, iodoacetic acid or pepstatin (inhibitors of serine, cysteine and aspartyl proteases, respectively). Ca2+, Mg2+ and Ba2+ ions increased its activity, while Al3+, Cu2+, Ni2+ and Zn2+ inhibited it. Additionally, ZnCl2 showed a dose dependent inhibition of the enzyme. Lmr-MP activity was also evaluated upon chromogenic substrates for plasma kallikrein (S-2302), plasmin and streptokinase-activated plasminogen (S-2251) and Factor Xa (S-2222) showing the highest activity on S-2302. The activity in different solutions (5 mM or 50 mM ammonium bicarbonate, pH 7.8; 0.1% trifluoroacetic acid + 50% acetonitrile; phosphate buffer saline, pH 7.4; 50 mM sodium acetate, pH 4.0 or ammonium acetate pH 4.5) was also evaluated and the results showed that its activity was abolished at acidic pHs. Its molecular mass (22,858 Da) was determined by MALDI-TOF and about 90% of its primary structure was verified by high-resolution mass spectrometry... (AU)


Assuntos
Animais , Viperidae , Venenos de Víboras/análise , Venenos de Víboras/química , Enzimas , Metaloproteases/química
10.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 24: 42, Jan. 24, 2018. ilus, mapas, tab, graf
Artigo em Inglês | VETINDEX | ID: vti-18969

Resumo

Background:The blunt-nosed viperMacrovipera lebetina(Linnaeus, 1758) is a medically important snake species inthe Middle East. Its nominate subspeciesMacrovipera l. lebetinais confined to Cyprus, where it is the only dangerouslyvenomous snake species and heavily pursued. Despite the vipers large size, data on its body mass and sex-specificmorphological differences are scarce. It is commonly believed thatM. l. lebetinaprefers freshwater proximity duringsummer. Hence, we aimed at investigatingM. l. lebetinasex-specific morphological differences and its possibleattraction to freshwater bodies in late summer.Methods:Morphometric characteristics, proximity to water and conservation status ofM. l. lebetinawere investigatedin Paphos district (Cyprus) in 2014, 2015 and 2017. Vipers were caught in different habitats, examined morphologicallyfor metric and meristic characters, and released back into their habitat. Additionally, local people were interviewedabout the conservation situation of the species.Results:Of 38 recorded blunt-nosed vipers, morphological characteristics were collected from 34 (10 adult males, 16adult females, eight unsexed juveniles). Rounded total length (ToL) ranged from 23.5 cm to 133.0 cm and weightbetween 10 g and 1456 g. Adult males significantly exceeded adult females in tail length (TaL), ToL and head length(HL). No significant sex-specific differences were found in snout-vent length (SVL), head width (HW), weight or bodycondition index (BCI), nor for the ratios TaL / SVL, TaL / ToL, HL / SVL or HL / HW. Adult females from late summer(2015) had a significantly lower mean BCI than those from spring (2014).Distances of blunt-nosed vipers to the nearest water bodies (natural and artificial...(AU)


Assuntos
Animais , Viperidae/anatomia & histologia , Comportamento Animal , Água Doce , Peso Corporal , Mordeduras de Serpentes , Pesos e Medidas Corporais/veterinária , Chipre , Conservação dos Recursos Naturais
11.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 24: 38, Jan. 24, 2018. ilus, graf
Artigo em Inglês | VETINDEX | ID: vti-19378

Resumo

Background: Bitis arietans is a venomous snake found in sub-Saharan Africa and in parts of Morocco and Saudi Arabia. The envenomation is characterized by local and systemic reactions including pain, blistering, edema and tissue damage, besides hemostatic and cardiovascular disturbances, which can cause death or permanent disabilities in its victims. However, the action mechanisms that provoke these effects remain poorly understood, especially the activities of purified venom components. Therefore, in order to elucidate the molecular mechanisms that make the Bitis arietans venom so potent and harmful to human beings, this study reports the isolation and biochemical characterization of a snake venom serine protease (SVSP). Methods: Solubilized venom was fractionated by molecular exclusion chromatography and the proteolytic activity was determined using fluorescent substrates. The peaks that showed serine protease activity were determined by blocking the proteolytic activity with site-directed inhibitors. In sequence, the fraction of interest was submitted to another cycle of molecular exclusion chromatography. The purified serine protease was identified by mass spectrometry and characterized biochemically and immunochemically. Results: A serine protease of 33 kDa with fibrinogen-degrading and kinin-releasing activities was isolated, described, and designated herein as Kn-Ba. The experimental Butantan Institute antivenom produced against Bitis arietans venom inhibited the Kn-Ba activity. Conclusions: The in vitro activities of Kn-Ba can be correlated with the capacity of the venom to provoke bleeding and clotting disorders as well as hypotension, which are common symptoms presented by envenomed victims. Obtaining satisfactory Kn-Ba inhibition through the experimental antivenom is important, given the WHO's recommendation of immunotherapy in cases of human accidents with venomous snakes.(AU)


Assuntos
Animais , Viperidae , Venenos de Víboras/análise , Venenos de Víboras/química , Serina Proteases/análise , Cininas , Fibrinogênio , Antivenenos
12.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 24: 43, Jan. 24, 2018. tab, graf
Artigo em Inglês | VETINDEX | ID: vti-18865

Resumo

Background:There are 6 species of venomous snakes in Taiwan. Two of them, Deinagkistrodon acutus (D. acutus) and Daboia siamensis (D. siamensis), can cause significant coagulopathy. However, a significant proportion of patients with snakebites cannot identify the correct snake species after envenomation, which hampers the application of antivenom. Hence, the differential diagnosis between the two snakebites by clinical presentations is important. This study aims to compare their clinical and laboratory features for the purpose of differential diagnosis between the two snakebites.Methods:We retrospectively reviewed the medical records of patients who arrived at the emergency department due to D. acutus or D. siamensis envenomation, between 2003 and 2016, in one medical center in eastern Taiwan. Since these snakebites are rare, we also included 3 cases reported from another hospital in central Taiwan.Results:In total, 15 patients bitten by D. acutus and 12 patients by D. siamensis were analyzed. Hemorrhagic bulla formation and the need for surgical intervention only presented for D. acutus envenomation cases (Both 53.3% vs. 0.0%, P= 0.003). As to laboratory features, lower platelet counts (20.0 × 103/μL [interquartile range, 14-66 × 103/μL] vs. 149.0 × 103/μL [102.3-274.3 × 103/μL], P = 0.001), lower D-dimer level (1423.4 μg/L [713.4-4212.3 μg/L] vs. 12,500.0 μg/L [2351.4-200,000 μg/L], P = 0.008), higher proportion of patients with moderate-to-severe thrombocytopenia (platelet count < 100 × 103/μL) (80% vs. 16.7%, odds ratio (OR) = 20.0, 95% CI, 2.77-144.31; P = 0.002), and lower proportion of patients with extremely high D-dimer (> 5000 ng/mL) (16.7% vs. 66.7%, adjusted OR = 0.1 (95% CI, 0.01-0.69; P = 0.036) were found a...(AU)


Assuntos
Humanos , Animais , Viperidae , Venenos de Víboras/análise , Venenos de Víboras/química , Mordeduras de Serpentes/cirurgia , Trombocitopenia , Transtornos da Coagulação Sanguínea , Taiwan
13.
J. venom. anim. toxins incl. trop. dis ; 23: 33, 2017. graf, ilus
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-954809

Resumo

Background: Venoms represent a still underexplored reservoir of bioactive components that might mitigate or cure diseases in conditions in which conventional therapy is ineffective. The bradykinin-potentiating peptides (BPPs) comprise a class of angiotensin-I converting enzyme (ACE) inhibitors. The BPPs usually consist of oligopeptides with 5 to 13 residues with a high number of proline residues and the tripeptide Ile-Pro-Pro (IPP-tripeptide) in the C-terminus region and have a conserved N-terminal pyroglutamate residue. As a whole, the action of the BPPs on prey and snakebite victims results in the decrease of the blood pressure. The aim of this work was to isolate and characterize novel BPPs from the venom of Bitis gabonica rhinoceros. Methods: The crude venom of B. g. rhinoceros was fractionated by size exclusion chromatography and the peptide fraction (<7 kDa) was separated by reverse phase chromatography (RP-HPLC) and analyzed by ESI-IT-TOF-MS/MS. One new BPP was identified, synthetized and assayed for ACE inhibition and, in vivo, for edema potentiation. Results: Typical BPP signatures were identified in three RP-HPLC fractions. CID fragmentation presented the usual y-ion of the terminal P-P fragment as a predominant signal at m/z 213.1. De novo peptide sequencing identified one Bothrops-like BPP and one new BPP sequence. The new BPP was synthesized and showed poor inhibition over ACE, but displayed significant bradykinin-induced edema potentiation. Conclusions: So far, few BPPs are described in Viperinae, and based on the sequenced peptides, two non-canonical sequences were detected. The possible clinical role of this new peptides remains unclear.(AU)


Assuntos
Animais , Oligopeptídeos , Peptídeos/isolamento & purificação , Bioquímica/classificação , Bradicinina , Viperidae , Bothrops
14.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 23: e33, 2017. graf, ilus
Artigo em Inglês | VETINDEX | ID: vti-33079

Resumo

Background: Venoms represent a still underexplored reservoir of bioactive components that might mitigate or cure diseases in conditions in which conventional therapy is ineffective. The bradykinin-potentiating peptides (BPPs) comprise a class of angiotensin-I converting enzyme (ACE) inhibitors. The BPPs usually consist of oligopeptides with 5 to 13 residues with a high number of proline residues and the tripeptide Ile-Pro-Pro (IPP-tripeptide) in the C-terminus region and have a conserved N-terminal pyroglutamate residue. As a whole, the action of the BPPs on prey and snakebite victims results in the decrease of the blood pressure. The aim of this work was to isolate and characterize novel BPPs from the venom of Bitis gabonica rhinoceros. Methods: The crude venom of B. g. rhinoceros was fractionated by size exclusion chromatography and the peptide fraction (<7 kDa) was separated by reverse phase chromatography (RP-HPLC) and analyzed by ESI-IT-TOF-MS/MS. One new BPP was identified, synthetized and assayed for ACE inhibition and, in vivo, for edema potentiation. Results: Typical BPP signatures were identified in three RP-HPLC fractions. CID fragmentation presented the usual y-ion of the terminal P-P fragment as a predominant signal at m/z 213.1. De novo peptide sequencing identified one Bothrops-like BPP and one new BPP sequence. The new BPP was synthesized and showed poor inhibition over ACE, but displayed significant bradykinin-induced edema potentiation. Conclusions: So far, few BPPs are described in Viperinae, and based on the sequenced peptides, two non-canonical sequences were detected. The possible clinical role of this new peptides remains unclear.(AU)


Assuntos
Animais , Oligopeptídeos , Peptídeos/isolamento & purificação , Bioquímica/classificação , Bradicinina , Viperidae , Bothrops
15.
Acta Sci. Biol. Sci. ; 39(3): 309-319, July.-Sept.2017. tab, graf, ilus
Artigo em Inglês | VETINDEX | ID: vti-716860

Resumo

Toxins and venoms produced by living organisms have exhibited a variety of biological activities against microorganisms. In this study, we tested seven snake venoms from the family Viperidae for antibacterial activity and the activities of reversal of antibiotic resistance and inhibition of biofilm formation against 22 clinical isolates of Staphylococcus aureus. Bothrops moojeni venom exhibited anti staphylococcal activity with the lowest mean value of minimum inhibitory concentration (MIC). Moreover, reversal of antibiotic resistance was observed for combinations of B. moojeni venom (½ x MIC) and norfloxacin or ampicillin (both ½ x MIC) for 86.4% and 50% of the isolates, respectively. B. moojeni venom alone at ½ MIC inhibited 90% of biofilm formation, whereas in combination with ciprofloxacin, both at ½ MIC, a reduction on the NorA efflux pump activity was observed. The detection of in vitro mutants colonies of S. aureus resistant to B. moojeni venom was low and they did not survive. A phospholipase A2 was purified from the venom of B. moojeni and displayed anti-staphylococcal activity when tested alone or in combination with ciprofloxacin. The results presented here will contribute to the search for new antimicrobial agents against resistant S. aureus.(AU)


Toxinas e venenos exibem uma variedade de atividades biológicas contra micro-organismos. Neste estudo, investigou-se a atividade de sete venenos de serpentes, da família Viperidae, sobre o crescimento de Staphylococcus aureus, na reversão fenotípica da resistência a antibióticos e inibição de formação de biofilme contra 22 isolados clínicos de S. aureus. O veneno de Bothrops moojeni apresentou a menor média de concentração inibitória mínima (CIM). Além disso, observou-se reversão da resistência a antibióticos para combinações do veneno de B. moojeni (½ x CIM) e norfloxacina ou ampicilina (ambos ½ x CIM) para 86,4% e 50% dos isolados, respectivamente. O veneno de B. moojeni na concentração de ½ CIM inibiu 90% de formação de biofilme, enquanto ele em combinação com ciprofloxacina, ambos na concentração de ½ CIM, diminuiu a atividade da bomba de efluxo NorA. A detecção in vitro de colônias mutantes de S. aureus resistente ao veneno de B. moojeni foi baixa e eles não sobreviveram. Uma fosfolipase A2 purificada a partir do veneno de B. moojeni exibiu atividade antibacteriana quando testada sozinha ou em combinação com ciprofloxacina. Os dados obtidos poderão contribuir para a pesquisa de novos agentes antimicrobianos contra S. aureus.(AU)


Assuntos
Animais , Venenos de Serpentes/análise , Venenos de Serpentes/intoxicação , Viperidae/microbiologia , Antibacterianos/análise , Staphylococcus aureus/classificação
16.
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1484661

Resumo

The blood plasma of numerous snake species naturally comprises endogenous phospholipase A2 inhibitors, which primarily neutralize toxic phospholipases A2 that may eventually reach their circulation. This inhibitor type is generally known as snake blood phospholipase A2 inhibitors (sbPLIs). Most, if not all sbPLIs are oligomeric glycosylated proteins, although the carbohydrate moiety may not be essential for PLA2 inhibition in every case. The presently known sbPLIs belong to one of three structural classes namely sbPLI, sbPLI or sbPLI depending on the presence of characteristic C-type lectin-like domains, leucine-rich repeats or three-finger motifs, respectively. Currently, the most numerous inhibitors described in the literature are sbPLIs and sbPLIs, whereas sbPLIs are rare. When the target PLA2 is a Lys49 homolog or an Asp49 myotoxin, the sbPLI is denominated a myotoxin inhibitor protein (MIP). In this brief overview, the most relevant data on sbPLIs will be presented. Representative examples of sbPLIs and sbPLIs from two Old World Gloydius brevicaudus and Malayopython reticulatus and two New World Bothrops alternatus and Crotalus durissus terrificus snake species will be emphasized.


Assuntos
Animais , Viperidae/imunologia , Viperidae/metabolismo , Viperidae/sangue , /análise , /química
17.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 22: [1-7], Janeiro 19, 2016. tab
Artigo em Inglês | VETINDEX | ID: vti-15434

Resumo

The blood plasma of numerous snake species naturally comprises endogenous phospholipase A2 inhibitors, which primarily neutralize toxic phospholipases A2 that may eventually reach their circulation. This inhibitor type is generally known as snake blood phospholipase A2 inhibitors (sbPLIs). Most, if not all sbPLIs are oligomeric glycosylated proteins, although the carbohydrate moiety may not be essential for PLA2 inhibition in every case. The presently known sbPLIs belong to one of three structural classes namely sbPLI, sbPLI or sbPLI depending on the presence of characteristic C-type lectin-like domains, leucine-rich repeats or three-finger motifs, respectively. Currently, the most numerous inhibitors described in the literature are sbPLIs and sbPLIs, whereas sbPLIs are rare. When the target PLA2 is a Lys49 homolog or an Asp49 myotoxin, the sbPLI is denominated a myotoxin inhibitor protein (MIP). In this brief overview, the most relevant data on sbPLIs will be presented. Representative examples of sbPLIs and sbPLIs from two Old World Gloydius brevicaudus and Malayopython reticulatus and two New World Bothrops alternatus and Crotalus durissus terrificus snake species will be emphasized.(AU)


Assuntos
Animais , Viperidae/sangue , Viperidae/imunologia , Viperidae/metabolismo , Inibidores de Fosfolipase A2/análise , Inibidores de Fosfolipase A2/química
18.
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-954786

Resumo

Background In the Atlantic forest of the North and Northeast regions of Brazil, local population often uses the fruit juice and the aqueous extract of leaves of soursop (Annona muricata L.) to treat Lachesis muta rhombeata envenomation. Envenomation is a relevant health issue in these areas, especially due to its severity and because the production and distribution of antivenom is limited in these regions. The aim of the present study was to evaluate the relevance of the use of soursop leaf extract and its juice against envenomation by Lachesis muta rhombeata. Methods We evaluated the biochemical, hematological and hemostatic parameters, the blood pressure, the inflammation process and the lethality induced by Lachesis muta rhombeata snake venom. We also assessed the action of the aqueous extract of leaves (AmL) and juice (AmJ) from A. muricata on the animal organism injected with L. m. rhombeata venom (LmrV) in the laboratory environment. Results LmrV induced a decrease of total protein, albumin and glucose; and increase of creatine kinase, aspartate aminotransferase, and urea concentrations. It provoked hemoconcentration followed by reduction of hematocrit, an increase in prothrombin time and partial thromboplastin time and a decrease of the blood pressure. LmrV induced the release of interleukin-6, an increase in neutrophils and changes in the serum protein profile, characteristic of the acute inflammatory process. LD50 values were similar for the groups injected with LmrV and treated or untreated with AmJ and AmL. Both treatments play a role on the maintenance of blood glucose, urea and coagulation parameters and exert a protective action against the myotoxicity. However, they seem to worsen the hypotension caused by LmrV. Conclusion The treatments with AmJ and AmL present some beneficial actions, but they might intensify some effects of the venom. Therefore, additional studies on A. muricata are necessary to enable its use as natural antivenom for bushmaster snakebite.(AU)


Assuntos
Mordeduras de Serpentes , Venenos de Serpentes , Antivenenos , Lachesis muta , Viperidae , Creatina Quinase , Annona , Miotoxicidade
19.
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-954791

Resumo

Background Snakebites cause considerable death and injury throughout the globe, particularly in tropical regions, and pose an important yet neglected threat to public health. In 2008, the Centre Anti Poison et de Parmacovigilance du Maroc (CAPM) started to set up a specific strategy for the control of snakebites that was formalized in 2012. The aim of the present study is to describe and update the epidemiological characteristics of snakebites notified to CAPM between 2009 and 2013. Methods This retrospective five-year study included all cases of snakebites notified to CAPM by mail or phone. Results During the study period, 873 snakebite cases were reported to CAPM, an average incidence of 2.65 cases per 100,000 inhabitants with 218 cases each year. The highest incidence was found in Tangier-Tetouan region with 357 cases (40.9 %) followed by Souss Massa Draa region with 128 cases (14.6 %). The average age of patients was 26.8 ± 17.2 years. The male to female sex ratio was 1.67:1 and 77 % of cases occurred in rural areas. The bites occurred mainly in spring (44 %) followed by summer (42 %). Snake species was identified in 54 cases (6.2 %): colubrids represented 31 % (n = 18) and vipers 67 % (n = 36), mainly Daboia mauritanica, Bitis arietans and Cerastes cerastes. In 311 cases (35.6 %), the patients showed viper syndrome. Thrombocytopenia was observed in 23.5 % of viper syndrome cases, whereas, compartment syndrome was observed in 7.6 % patients. FAV-Afrique® was administered in 41 patients (5 %). In patients treated with antivenom, 38 patients recovered and three died. Twenty-seven deaths were reported (3.9 %). Conclusion Despite specific efforts to better understand the epidemiology of snakebites in Morocco (incidence, severity, snake species involved), it remains underestimated. Therefore, further work is still necessary to ensure accessibility of appropriate antivenom against venomous species and to improve the management of envenomation in Morocco.(AU)


Assuntos
Centros de Controle de Intoxicações , Mordeduras de Serpentes , Antivenenos , Epidemiologia , Viperidae
20.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 22: [1-12], Abril 29, 2016. tab, graf
Artigo em Inglês | VETINDEX | ID: vti-15796

Resumo

In the Atlantic forest of the North and Northeast regions of Brazil, local population often uses the fruit juice and the aqueous extract of leaves of soursop (Annona muricata L.) to treat Lachesis muta rhombeata envenomation. Envenomation is a relevant health issue in these areas, especially due to its severity and because the production and distribution of antivenom is limited in these regions. The aim of the present study was to evaluate the relevance of the use of soursop leaf extract and its juice against envenomation by Lachesis muta rhombeata. Methods We evaluated the biochemical, hematological and hemostatic parameters, the blood pressure, the inflammation process and the lethality induced by Lachesis muta rhombeata snake venom. We also assessed the action of the aqueous extract of leaves (AmL) and juice (AmJ) from A. muricata on the animal organism injected with L. m. rhombeata venom (LmrV) in the laboratory environment. Results LmrV induced a decrease of total protein, albumin and glucose; and increase of creatine kinase, aspartate aminotransferase, and urea concentrations. It provoked hemoconcentration followed by reduction of hematocrit, an increase in prothrombin time and partial thromboplastin time and a decrease of the blood pressure. LmrV induced the release of interleukin-6, an increase in neutrophils and changes in the serum protein profile, characteristic of the acute inflammatory process. LD50 values were similar for the groups injected with LmrV and treated or untreated with AmJ and AmL. Both treatments play a role on the maintenance of blood glucose, urea and coagulation parameters and exert a protective action against the myotoxicity. However, they seem to worsen the hypotension caused by LmrV. Conclusion The treatments with AmJ and AmL present some beneficial actions, but they might intensify some effects of the venom. Therefore, additional studies on A. muricata are necessary to enable its use as natural antivenom for bushmaster snakebite.(AU)


Assuntos
Annona/efeitos adversos , Annona/intoxicação , Antivenenos/análise , Antivenenos/química , Lachesis muta/administração & dosagem , Lachesis muta/análise
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