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1.
Pesqui. vet. bras ; 42: e07105, 2022. ilus
Artigo em Inglês | VETINDEX | ID: biblio-1386822

Resumo

In Brazil, snakebites are often cited as a cause of mortality in ruminants, but there are discrepancies in the literature regarding its actual prevalence, either by lack of diagnosis or by mistakes in the differential diagnosis. Among the factors that hinder the diagnosis are included the inconsistencies to distinguish between accidents caused by Bothrops and Crotalus, responsible for over 90% of the cases. For the diagnosis of accidents involving Lachesis muta, both the neurotropic and the proteolytic/hemolytic effects must be considered, similar to what is described in Crotalus scutulatus. This article describes the main clinical, pathological, and laboratory findings observed in envenoming by the aforementioned snakes and suggests procedures for establishing the diagnosis and differential diagnosis starting from a logical sequence, based on epidemiological evidence, clinical, laboratory, and pathological findings.


No Brasil, acidentes ofídicos são frequentemente citados como causa de mortalidade em ruminantes, mas existem discrepâncias em relação a sua atual prevalência, seja por falta de diagnóstico ou por erros no diagnóstico diferencial. Entre os fatores que dificultam o diagnóstico estão as inconsistências para distinguir entre os acidentes causados por Bothrops e Crotalus, responsáveis por mais de 90% dos casos. Para o diagnóstico de envenenamentos por Lachesis muta, devem ser considerados os efeitos neurotrópico e proteolítico/hemolíticos concomitantes, a exemplo do que ocorre com algumas cascavéis norte-americanas (Crotalus scutulatus, entre elas). Este artigo descreve os principais achados clinicopatológicos e laboratoriais observados em casos de envenenamento pelas serpentes citadas e sugere um roteiro simplificado para o estabelecimento do diagnóstico e diagnóstico diferencial, a partir de uma sequência lógica, baseada em evidências epidemiológicas e achados clínicos, laboratoriais e patológicos.


Assuntos
Animais , Mordeduras de Serpentes/diagnóstico , Mordeduras de Serpentes/mortalidade , Crotalus , Bothrops , Mordeduras de Serpentes/veterinária , Ruminantes
2.
J. venom. anim. toxins incl. trop. dis ; 26: e20200016, 2020. graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1135158

Resumo

South American rattlesnakes are represented in Brazil by a single species, Crotalus durissus, which has public health importance due to the severity of its envenomation and to its wide geographical distribution. The species is subdivided into several subspecies, but the current classification is controversial. In Brazil, the venoms of C. d. terrificus and C. d. collilineatus are used for hyperimmunization of horses for antivenom production, even though the distinction of these two subspecies are mostly by their geographical distribution. In this context, we described a comparative compositional and functional characterization of individual C. d. collilineatus and C. d. terrificus venoms from three Brazilian states. Methods: We compared the compositional patterns of C. d. terrificus and C. d. collilineatus individual venoms by 1-DE and RP-HPLC. For functional analyzes, the enzymatic activities of PLA2, LAAO, and coagulant activity were evaluated. Finally, the immunorecognition of venom toxins by the crotalic antivenom produced at Butantan Institute was evaluated using Western blotting. Results: The protein profile of individual venoms from C. d. collilineatus and C. d. terrificus showed a comparable overall composition, despite some intraspecific variation, especially regarding crotamine and LAAO. Interestingly, HPLC analysis showed a geographic pattern concerning PLA2. In addition, a remarkable intraspecific variation was also observed in PLA2, LAAO and coagulant activities. The immunorecognition pattern of individual venoms from C. d. collilineatus and C. d. terrificus by crotalic antivenom produced at Butantan Institute was similar. Conclusions: The results highlighted the individual variability among the venoms of C. durissus ssp. specimens. Importantly, our data point to a geographical variation of C. durissus ssp. venom profile, regardless of the subspecies, as evidenced by PLA2 isoforms complexity, which may explain the increase in venom neurotoxicity from Northeastern through Southern Brazil reported for the species.(AU)


Assuntos
Animais , Crotalus , Venenos Elapídicos , Fosfolipases A2 , Localizações Geográficas
3.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 26: e20200016, 2020. graf
Artigo em Inglês | VETINDEX | ID: vti-32213

Resumo

South American rattlesnakes are represented in Brazil by a single species, Crotalus durissus, which has public health importance due to the severity of its envenomation and to its wide geographical distribution. The species is subdivided into several subspecies, but the current classification is controversial. In Brazil, the venoms of C. d. terrificus and C. d. collilineatus are used for hyperimmunization of horses for antivenom production, even though the distinction of these two subspecies are mostly by their geographical distribution. In this context, we described a comparative compositional and functional characterization of individual C. d. collilineatus and C. d. terrificus venoms from three Brazilian states. Methods: We compared the compositional patterns of C. d. terrificus and C. d. collilineatus individual venoms by 1-DE and RP-HPLC. For functional analyzes, the enzymatic activities of PLA2, LAAO, and coagulant activity were evaluated. Finally, the immunorecognition of venom toxins by the crotalic antivenom produced at Butantan Institute was evaluated using Western blotting. Results: The protein profile of individual venoms from C. d. collilineatus and C. d. terrificus showed a comparable overall composition, despite some intraspecific variation, especially regarding crotamine and LAAO. Interestingly, HPLC analysis showed a geographic pattern concerning PLA2. In addition, a remarkable intraspecific variation was also observed in PLA2, LAAO and coagulant activities. The immunorecognition pattern of individual venoms from C. d. collilineatus and C. d. terrificus by crotalic antivenom produced at Butantan Institute was similar. Conclusions: The results highlighted the individual variability among the venoms of C. durissus ssp. specimens. Importantly, our data point to a geographical variation of C. durissus ssp. venom profile, regardless of the subspecies, as evidenced by PLA2 isoforms complexity, which may explain the increase in venom neurotoxicity from Northeastern through Southern Brazil reported for the species.(AU)


Assuntos
Animais , Venenos de Serpentes/análise , Venenos de Serpentes/classificação , Características de Residência , Crotalus
4.
Acta sci. vet. (Impr.) ; 48(suppl.1): Pub.521-4 jan. 2020. ilus, tab
Artigo em Inglês | VETINDEX | ID: biblio-1458348

Resumo

Background: South American rattlesnake (Crotalus durissus spp.) envenomation is rarely reported in small animals andlivestock in Brazil. Minor swelling at the snakebite site, skeletal muscle, and renal damage, and severe neurological signscharacterize the crotalic envenomation. This case report aims to present epidemiological, clinical, and pathological dataof two cases of Crotalus durissus spp envenomation in dogs in the Northeast of Brazil.Cases: Envenomation by Crotalus durissus spp. was recorded in two dogs in Patos, State of Paraíba, Brazil. In Case 1,the dog presented flaccid paralysis, hyporeflexia, a deficit of cranial nerves, epistaxis, and gingival hemorrhages. Laboratory assay showed proteinuria, myoglobinuria, regenerative thrombocytopenia, and increased serum activities of creatinekinase (CK), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP). The dogwas medicated with crotalic antivenom and wholly recovered from local and systemic clinical signs. In Case 2, the dogdied and was detected fang marks at the ventral region of the left mandible (two small parallel perforations spaced 2.0 cmapart) at the snakebite site. Cyanosis of the oral cavity, congestion, and hemorrhages in several organs were observed atnecropsy. Tubular nephrosis, muscular necrosis, hepatocytes swelling were observed. The owners witnessed snakebites,and the rattlesnakes (Crotalus durissus spp.) identified by the rattle at the end portion of the tail in both cases.Discussion: Natural South American rattlesnake envenomation presents complex clinical signs that makes diagnosis achallenge for veterinary practitioners. The criteria for the correct diagnosis and observed in the two dogs include witness ofthe snakebite, identification of the snake, detection of fang marks, clinical-pathological findings, and therapeutic responseto treatment with specific anti-venom....


Assuntos
Animais , Cães , Doenças Musculares/veterinária , Mordeduras de Serpentes/epidemiologia , Mordeduras de Serpentes/patologia , Venenos de Crotalídeos , Brasil , Crotalus
5.
Acta sci. vet. (Online) ; 48(suppl.1): Pub. 521, July 19, 2020. ilus, tab
Artigo em Inglês | VETINDEX | ID: vti-31873

Resumo

Background: South American rattlesnake (Crotalus durissus spp.) envenomation is rarely reported in small animals andlivestock in Brazil. Minor swelling at the snakebite site, skeletal muscle, and renal damage, and severe neurological signscharacterize the crotalic envenomation. This case report aims to present epidemiological, clinical, and pathological dataof two cases of Crotalus durissus spp envenomation in dogs in the Northeast of Brazil.Cases: Envenomation by Crotalus durissus spp. was recorded in two dogs in Patos, State of Paraíba, Brazil. In Case 1,the dog presented flaccid paralysis, hyporeflexia, a deficit of cranial nerves, epistaxis, and gingival hemorrhages. Laboratory assay showed proteinuria, myoglobinuria, regenerative thrombocytopenia, and increased serum activities of creatinekinase (CK), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP). The dogwas medicated with crotalic antivenom and wholly recovered from local and systemic clinical signs. In Case 2, the dogdied and was detected fang marks at the ventral region of the left mandible (two small parallel perforations spaced 2.0 cmapart) at the snakebite site. Cyanosis of the oral cavity, congestion, and hemorrhages in several organs were observed atnecropsy. Tubular nephrosis, muscular necrosis, hepatocytes swelling were observed. The owners witnessed snakebites,and the rattlesnakes (Crotalus durissus spp.) identified by the rattle at the end portion of the tail in both cases.Discussion: Natural South American rattlesnake envenomation presents complex clinical signs that makes diagnosis achallenge for veterinary practitioners. The criteria for the correct diagnosis and observed in the two dogs include witness ofthe snakebite, identification of the snake, detection of fang marks, clinical-pathological findings, and therapeutic responseto treatment with specific anti-venom....(AU)


Assuntos
Animais , Cães , Venenos de Crotalídeos , Mordeduras de Serpentes/epidemiologia , Mordeduras de Serpentes/patologia , Doenças Musculares/veterinária , Brasil , Crotalus
6.
Acta sci. vet. (Online) ; 47: Pub. 1662, May 30, 2019. ilus, tab, graf
Artigo em Inglês | VETINDEX | ID: vti-19610

Resumo

Background: Accidents caused by venom of Crotalus durissus snakes, popularly known in Brazil as rattlesnake, aresecond in relation to the occurrence and first place in deaths in humans and animals, mainly due to the great neurotoxic,myotoxic, coagulant, nephrotoxic and hepatotoxic potential of their venom. The effects observed are due to the action ofthe main poison fractions and among them we can mention crotoxin (representing 50% of the total poison), crotamine,gyroxine and conxulxin. The present study aimed to analyze the liver of experimentally poisoned Wistar rats with venomof Crotalus durissus terrificus by means of histological and fractal analysis. The hypothesis is that the venom of Crotalusdurissus terrificus is can induce hepatic damage at the dose recommended in this study, that its alterations can be quantifiedby the fractal dimension and that the antiofidic serum botropic crotalic be able to minimize the hepatic lesions inducedby the venom.Materials, Methods & Results: Ninety rats were distributed into different groups and treated with: control group (GC, n= 30) 0.9% sodium chloride solution; venom group (GV, n = 30) crotalic venom at the dose of 1 mg/kg; (GVS, n = 30)crotalic venom at the dose of 1 mg/Kg and antiofidic serum 6 h after the application of the venom at the dose recommendedby the manufacturer. Liver samples were collected at 2 h (M1), 8 h (M2) and 24 h (M3) after venom administration andsubmitted to histological analysis and fractal dimension (DF) using the ImageJ® software and box-counting method. Procedures for collecting, processing and analyzing samples were standardized. For statistical analyzes, after the normalitywas verified by the Shapiro-Wilk test and homogeneity by the Bartlett test, the data were submitted to analysis of variance(ANOVA) with Duncan test contrast with a significance level of 5%. No significant lesions were observed in GC, howevernecrosis...(AU)


Assuntos
Animais , Ratos , Ratos Wistar , Fígado/anatomia & histologia , Venenos de Crotalídeos/análise , Crotalus , Fractais , Análise de Variância
7.
Acta sci. vet. (Impr.) ; 47: Pub.1662-2019. ilus, tab, graf
Artigo em Inglês | VETINDEX | ID: biblio-1458060

Resumo

Background: Accidents caused by venom of Crotalus durissus snakes, popularly known in Brazil as rattlesnake, aresecond in relation to the occurrence and first place in deaths in humans and animals, mainly due to the great neurotoxic,myotoxic, coagulant, nephrotoxic and hepatotoxic potential of their venom. The effects observed are due to the action ofthe main poison fractions and among them we can mention crotoxin (representing 50% of the total poison), crotamine,gyroxine and conxulxin. The present study aimed to analyze the liver of experimentally poisoned Wistar rats with venomof Crotalus durissus terrificus by means of histological and fractal analysis. The hypothesis is that the venom of Crotalusdurissus terrificus is can induce hepatic damage at the dose recommended in this study, that its alterations can be quantifiedby the fractal dimension and that the antiofidic serum botropic crotalic be able to minimize the hepatic lesions inducedby the venom.Materials, Methods & Results: Ninety rats were distributed into different groups and treated with: control group (GC, n= 30) 0.9% sodium chloride solution; venom group (GV, n = 30) crotalic venom at the dose of 1 mg/kg; (GVS, n = 30)crotalic venom at the dose of 1 mg/Kg and antiofidic serum 6 h after the application of the venom at the dose recommendedby the manufacturer. Liver samples were collected at 2 h (M1), 8 h (M2) and 24 h (M3) after venom administration andsubmitted to histological analysis and fractal dimension (DF) using the ImageJ® software and box-counting method. Procedures for collecting, processing and analyzing samples were standardized. For statistical analyzes, after the normalitywas verified by the Shapiro-Wilk test and homogeneity by the Bartlett test, the data were submitted to analysis of variance(ANOVA) with Duncan test contrast with a significance level of 5%. No significant lesions were observed in GC, howevernecrosis...


Assuntos
Animais , Ratos , Crotalus , Fígado/anatomia & histologia , Ratos Wistar , Venenos de Crotalídeos/análise , Análise de Variância , Fractais
8.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 24: 34, Dec. 17, 2018. graf
Artigo em Inglês | VETINDEX | ID: vti-19379

Resumo

Background: Our group has previously performed a proteomic study verifying that individual variations can occur among Crotalus durissus collilineatus venoms. These variations may lead to differences in venom toxicity and may result in lack of neutralization of some components by antivenom. In this way, this study aimed to evaluate the Brazilian anticrotalic serum capacity in recognizing twenty-two Crotalus durissus collilineatus venoms, as well as their fractions. Methods: The indirect enzyme-linked immunosorbent assay (ELISA) was chosen to evaluate the efficacy of heterologous anticrotalic serum produced by Instituto Butantan (Brazil) in recognizing the twenty-two Crotalus durissus collilineatus venoms and the pool of them. Moreover, the venom pool was fractionated using reversed-phase fast protein liquid chromatography (RP-FPLC) and the obtained fractions were analyzed concerning antivenom recognition. Results: Evaluation of venom variability by ELISA showed that all venom samples were recognized by the Brazilian anticrotalic antivenom. However, some particular venom fractions were poorly recognized. Conclusion: This study demonstrated that the Brazilian anticrotalic serum recognizes all the different twenty-two venoms of C. d. collilineatus and their fractions, although in a quantitatively different way, which may impact the effectiveness of the antivenom therapy. These results confirm the need to use a pool of venoms with the greatest possible variability in the preparation of antivenoms, in order to improve their effectiveness.(AU)


Assuntos
Animais , Crotalus , Venenos de Crotalídeos/análise , Venenos de Crotalídeos/química , Antivenenos/análise , Ensaio de Imunoadsorção Enzimática
9.
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1040379

Resumo

Our group has previously performed a proteomic study verifying that individual variations can occur among Crotalus durissus collilineatus venoms. These variations may lead to differences in venom toxicity and may result in lack of neutralization of some components by antivenom. In this way, this study aimed to evaluate the Brazilian anticrotalic serum capacity in recognizing twenty-two Crotalus durissus collilineatus venoms, as well as their fractions. Methods: The indirect enzyme-linked immunosorbent assay (ELISA) was chosen to evaluate the efficacy of heterologous anticrotalic serum produced by Instituto Butantan (Brazil) in recognizing the twenty-two Crotalus durissus collilineatus venoms and the pool of them. Moreover, the venom pool was fractionated using reversed-phase fast protein liquid chromatography (RP-FPLC) and the obtained fractions were analyzed concerning antivenom recognition. Results: Evaluation of venom variability by ELISA showed that all venom samples were recognized by the Brazilian anticrotalic antivenom. However, some particular venom fractions were poorly recognized. Conclusion: This study demonstrated that the Brazilian anticrotalic serum recognizes all the different twenty-two venoms of C. d. collilineatus and their fractions, although in a quantitatively different way, which may impact the effectiveness of the antivenom therapy. These results confirm the need to use a pool of venoms with the greatest possible variability in the preparation of antivenoms, in order to improve their effectiveness.(AU)


Assuntos
Antivenenos/toxicidade , Crotalus , Venenos de Crotalídeos , Ensaio de Imunoadsorção Enzimática
10.
J. venom. anim. toxins incl. trop. dis ; 24: 39, 2018. graf, ilus
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-984689

Resumo

For the past 80 years, Crotoxin has become one of the most investigated isolated toxins from snake venoms, partially due to its major role as the main toxic component in the venom of the South American rattlesnake Crotalus durissus terrificus. However, in the past decades, progressive studies have led researchers to shift their focus on Crotoxin, opening novel perspectives and applications as a therapeutic approach. Although this toxin acts on a wide variety of biological events, the modulation of immune responses is considered as one of its most relevant behaviors. Therefore, the present review describes the scientific investigations on the capacity of Crotoxin to modulate anti-inflammatory and immunosuppressive responses, and its application as a medicinal immunopharmacological approach. In addition, this review will also discuss its mechanisms, involving cellular and molecular pathways, capable of improving pathological alterations related to immune-associated disorders.(AU)


Assuntos
Venenos de Serpentes , Produtos Biológicos , Antivenenos , Crotalus , Crotoxina/imunologia , Imunidade , Imunossupressores
11.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 24: 39, Jan. 24, 2018. ilus, graf
Artigo em Inglês | VETINDEX | ID: vti-18999

Resumo

For the past 80 years, Crotoxin has become one of the most investigated isolated toxins from snake venoms, partially due to its major role as the main toxic component in the venom of the South American rattlesnake Crotalus durissus terrificus. However, in the past decades, progressive studies have led researchers to shift their focus on Crotoxin, opening novel perspectives and applications as a therapeutic approach. Although this toxin acts on a wide variety of biological events, the modulation of immune responses is considered as one of its most relevant behaviors. Therefore, the present review describes the scientific investigations on the capacity of Crotoxin to modulate anti-inflammatory and immunosuppressive responses, and its application as a medicinal immunopharmacological approach. In addition, this review will also discuss its mechanisms, involving cellular and molecular pathways, capable of improving pathological alterations related to immune-associated disorders.(AU)


Assuntos
Animais , Crotalus cascavella , Crotoxina/imunologia , Crotoxina/uso terapêutico , Imunossupressores , Imunidade Inata , Imunidade Adaptativa , Anti-Inflamatórios , Inflamação/terapia
12.
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-976024

Resumo

In recent decades, snake venom disintegrins have received special attention due to their potential use in anticancer therapy. Disintegrins are small and cysteine-rich proteins present in snake venoms and can interact with specific integrins to inhibit their activities in cell-cell and cell-ECM interactions. These molecules, known to inhibit platelet aggregation, are also capable of interacting with certain cancer-related integrins, and may interfere in important processes involved in carcinogenesis. Therefore, disintegrin from Crotalus durissus collilineatus venom was isolated, structurally characterized and evaluated for its toxicity and ability to interfere with cell proliferation and migration in MDA-MB-231, a human breast cancer cell line. Methods: Based on previous studies, disintegrin was isolated by FPLC, through two chromatographic steps, both on reversed phase C-18 columns. The isolated disintegrin was structurally characterized by Tris-TricineSDS-PAGE, mass spectrometry and N-terminal sequencing. For the functional assays, MTT and wound-healing assays were performed in order to investigate cytotoxicity and effect on cell migration in vitro, respectively. Results: Disintegrin presented a molecular mass of 7287.4 Da and its amino acid sequence shared similarity with the disintegrin domain of P-II metalloproteases. Using functional assays, the disintegrin showed low cytotoxicity (15% and 17%, at 3 and 6 µg/mL, respectively) after 24 h of incubation and in the wound-healing assay, the disintegrin (3 µg/mL) was able to significantly inhibit cell migration (24%, p < 0.05), compared to negative control. Conclusion: Thus, our results demonstrate that non-RGD disintegrin from C. d. collilineatus induces low cytotoxicity and inhibits migration of human breast cancer cells. Therefore, it may be a very useful molecular tool for understanding ECM-cell interaction cancer-related mechanisms involved in an important integrin family that highlights molecular aspects of tumorigenesis. Also, non-RGD disintegrin has potential to serve as an agent in anticancer therapy or adjuvant component combined with other anticancer drugs.(AU)


Assuntos
Venenos de Serpentes , Crotalus , Desintegrinas , Neoplasias da Mama
13.
J. Venom. Anim. Toxins incl. Trop. Dis. ; 24: 28, Nov. 29, 2018. tab, graf
Artigo em Inglês | VETINDEX | ID: vti-18439

Resumo

Background:In recent decades, snake venom disintegrins have received special attention due to their potential use in anticancer therapy. Disintegrins are small and cysteine-rich proteins present in snake venoms and can interact with specific integrins to inhibit their activities in cell-cell and cell-ECM interactions. These molecules, known to inhibit platelet aggregation, are also capable of interacting with certain cancer-related integrins, and may interfere in important processes involved in carcinogenesis. Therefore, disintegrin from Crotalus durissus collilineatus venom was isolated, structurally characterized and evaluated for its toxicity and ability to interfere with cell proliferation and migration in MDA-MB-231, a human breast cancer cell line.Methods:Based on previous studies, disintegrin was isolated by FPLC, through two chromatographic steps, both on reversed phase C-18 columns. The isolated disintegrin was structurally characterized by Tris-Tricine-SDS-PAGE, mass spectrometry and N-terminal sequencing. For the functional assays, MTT and wound-healing assays were performed in order to investigate cytotoxicity and effect on cell migration in vitro, respectively.Results:Disintegrin presented a molecular mass of 7287.4 Da and its amino acid sequence shared similarity with the disintegrin domain of P-II metalloproteases. Using functional assays, the disintegrin showed low cytotoxicity (15% and 17%, at 3 and 6 μg/mL, respectively) after 24 h of incubation and in the wound-healing assay, the disintegrin (3 μg/mL) was able to significantly inhibit cell migration (24%, p < 0.05), compared to negative control.Conclusion:Thus, our results demonstrate that non-RGD disintegrin from C. d. collilineatus induces low cytotoxicity and inhibits migration of human breast cancer cells. Therefore, it may be a very useful molecular tool for understanding ECM-cell interaction cancer-related mechanisms involved in...(AU)


Assuntos
Humanos , Animais , Venenos de Crotalídeos/análise , Desintegrinas/análise , Movimento Celular , Adesão Celular , Neoplasias da Mama/tratamento farmacológico , Crotalus
14.
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-954844

Resumo

Background: Classically, Crotalus durissus terrificus (Cdt) venom can be described, according to chromatographic criteria, as a simple venom, composed of four major toxins, namely: gyroxin, crotamine, crotoxin and convulxin. Crotoxin is a non-covalent heterodimeric neurotoxin constituted of two subunits: an active phospholipase A2 and a chaperone protein, termed crotapotin. This molecule is composed of three peptide chains connected by seven disulfide bridges. Naturally occurring variants/isoforms of either crotoxin or crotapotin itself have already been reported. Methods: The crude Cdt venom was separated by using RP-HPLC and the toxins were identified by mass spectrometry (MS). Crotapotin was purified, reduced and alkylated in order to separate the peptide chains that were further analyzed by mass spectrometry and de novo peptide sequencing. Results: The RP-HPLC profile of the isolated crotapotin chains already indicated that the α chain would present isoforms, which was corroborated by the MS and tandem mass spectrometry analyses. Conclusion: It was possible to observe that the Cdt crotapotin displays a preferred amino acid substitution pattern present in the α chain, at positions 31 and 40. Moreover, substitutions could also be observed in ß and γ chains (one for each). The combinations of these four different peptides, with the already described chains, would produce ten different crotapotins, which is compatible to our previous observations for the Cdt venom.(AU)


Assuntos
Animais , Espectrometria de Massas , Isoformas de Proteínas , Venenos de Crotalídeos , Crotoxina , Fosfolipases A2 , Neurotoxinas
15.
Artigo em Inglês | VETINDEX | ID: vti-31521

Resumo

Background: Classically, Crotalus durissus terrificus (Cdt) venom can be described, according to chromatographic criteria, as a simple venom, composed of four major toxins, namely: gyroxin, crotamine, crotoxin and convulxin. Crotoxin is a non-covalent heterodimeric neurotoxin constituted of two subunits: an active phospholipase A2 and a chaperone protein, termed crotapotin. This molecule is composed of three peptide chains connected by seven disulfide bridges. Naturally occurring variants/isoforms of either crotoxin or crotapotin itself have already been reported. Methods: The crude Cdt venom was separated by using RP-HPLC and the toxins were identified by mass spectrometry (MS). Crotapotin was purified, reduced and alkylated in order to separate the peptide chains that were further analyzed by mass spectrometry and de novo peptide sequencing. Results: The RP-HPLC profile of the isolated crotapotin chains already indicated that the α chain would present isoforms, which was corroborated by the MS and tandem mass spectrometry analyses. Conclusion: It was possible to observe that the Cdt crotapotin displays a preferred amino acid substitution pattern present in the α chain, at positions 31 and 40. Moreover, substitutions could also be observed in ß and γ chains (one for each). The combinations of these four different peptides, with the already described chains, would produce ten different crotapotins, which is compatible to our previous observations for the Cdt venom.(AU)


Assuntos
Animais , Espectrometria de Massas , Isoformas de Proteínas , Venenos de Crotalídeos , Crotoxina , Fosfolipases A2 , Neurotoxinas
16.
Artigo em Inglês | LILACS-Express | LILACS, VETINDEX | ID: biblio-1484721

Resumo

Abstract Background Classically, Crotalus durissus terrificus (Cdt) venom can be described, according to chromatographic criteria, as a simple venom, composed of four major toxins, namely: gyroxin, crotamine, crotoxin and convulxin. Crotoxin is a non-covalent heterodimeric neurotoxin constituted of two subunits: an active phospholipase A2 and a chaperone protein, termed crotapotin. This molecule is composed of three peptide chains connected by seven disulfide bridges. Naturally occurring variants/isoforms of either crotoxin or crotapotin itself have already been reported. Methods The crude Cdt venom was separated by using RP-HPLC and the toxins were identified by mass spectrometry (MS). Crotapotin was purified, reduced and alkylated in order to separate the peptide chains that were further analyzed by mass spectrometry and de novo peptide sequencing. Results The RP-HPLC profile of the isolated crotapotin chains already indicated that the chain would present isoforms, which was corroborated by the MS and tandem mass spectrometry analyses. Conclusion It was possible to observe that the Cdt crotapotin displays a preferred amino acid substitution pattern present in the chain, at positions 31 and 40. Moreover, substitutions could also be observed in and chains (one for each). The combinations of these four different peptides, with the already described chains, would produce ten different crotapotins, which is compatible to our previous observations for the Cdt venom.

17.
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1040378

Resumo

Background Snakebite treatment requires administration of an appropriate antivenom that should contain antibodies capable of neutralizing the venom. To achieve this goal, antivenom production must start from a suitable immunization protocol and proper venom mixtures. In Brazil, antivenom against South American rattlesnake (Crotalus durissus terrificus) bites is produced by public institutions based on the guidelines defined by the regulatory agency of the Brazilian Ministry of Health, ANVISA. However, each institution uses its own mixture of rattlesnake venom antigens. Previous works have shown that crotamine, a toxin found in Crolatus durissus venom, shows marked individual and populational variation. In addition, serum produced from crotamine-negative venoms fails to recognize this molecule. Methods In this work, we used an antivenomics approach to assess the cross-reactivity of crotalic antivenom manufactured by IVB towards crotamine-negative venom and a mixture of crotamine-negative/crotamine-positive venoms. Results We show that the venom mixture containing 20% crotamine and 57% crotoxin produced a strong immunogenic response in horses. Antivenom raised against this venom mixture reacted with most venom components including crotamine and crotoxin, in contrast to the antivenom raised against crotamine-negative venom. Conclusions These results indicate that venomic databases and antivenomics analysis provide a useful approach for choosing the better venom mixture for antibody production and for the subsequent screening of antivenom cross-reactivity with relevant snake venom components.(AU)


Assuntos
Mordeduras e Picadas , Antivenenos , Crotalus cascavella , Venenos de Crotalídeos , Formação de Anticorpos
18.
Artigo em Inglês | VETINDEX | ID: vti-32806

Resumo

Background Snakebite treatment requires administration of an appropriate antivenom that should contain antibodies capable of neutralizing the venom. To achieve this goal, antivenom production must start from a suitable immunization protocol and proper venom mixtures. In Brazil, antivenom against South American rattlesnake (Crotalus durissus terrificus) bites is produced by public institutions based on the guidelines defined by the regulatory agency of the Brazilian Ministry of Health, ANVISA. However, each institution uses its own mixture of rattlesnake venom antigens. Previous works have shown that crotamine, a toxin found in Crolatus durissus venom, shows marked individual and populational variation. In addition, serum produced from crotamine-negative venoms fails to recognize this molecule. Methods In this work, we used an antivenomics approach to assess the cross-reactivity of crotalic antivenom manufactured by IVB towards crotamine-negative venom and a mixture of crotamine-negative/crotamine-positive venoms. Results We show that the venom mixture containing 20% crotamine and 57% crotoxin produced a strong immunogenic response in horses. Antivenom raised against this venom mixture reacted with most venom components including crotamine and crotoxin, in contrast to the antivenom raised against crotamine-negative venom. Conclusions These results indicate that venomic databases and antivenomics analysis provide a useful approach for choosing the better venom mixture for antibody production and for the subsequent screening of antivenom cross-reactivity with relevant snake venom components.(AU)


Assuntos
Mordeduras e Picadas , Antivenenos , Crotalus cascavella , Venenos de Crotalídeos , Formação de Anticorpos
19.
Artigo em Inglês | LILACS-Express | LILACS, VETINDEX | ID: biblio-1484733

Resumo

Abstract Background Snakebite treatment requires administration of an appropriate antivenom that should contain antibodies capable of neutralizing the venom. To achieve this goal, antivenom production must start from a suitable immunization protocol and proper venom mixtures. In Brazil, antivenom against South American rattlesnake (Crotalus durissus terrificus) bites is produced by public institutions based on the guidelines defined by the regulatory agency of the Brazilian Ministry of Health, ANVISA. However, each institution uses its own mixture of rattlesnake venom antigens. Previous works have shown that crotamine, a toxin found in Crolatus durissus venom, shows marked individual and populational variation. In addition, serum produced from crotamine-negative venoms fails to recognize this molecule. Methods In this work, we used an antivenomics approach to assess the cross-reactivity of crotalic antivenom manufactured by IVB towards crotamine-negative venom and a mixture of crotamine-negative/crotamine-positive venoms. Results We show that the venom mixture containing 20% crotamine and 57% crotoxin produced a strong immunogenic response in horses. Antivenom raised against this venom mixture reacted with most venom components including crotamine and crotoxin, in contrast to the antivenom raised against crotamine-negative venom. Conclusions These results indicate that venomic databases and antivenomics analysis provide a useful approach for choosing the better venom mixture for antibody production and for the subsequent screening of antivenom cross-reactivity with relevant snake venom components.

20.
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-954818

Resumo

Background Globally, snake envenomation is a well-known cause of death and morbidity. In many cases of snakebite, myonecrosis, dermonecrosis, hemorrhage and neurotoxicity are present. Some of these symptoms may be provoked by the envenomation itself, but others are secondary effects of the produced oxidative stress that enhances the damage produced by the venom toxins. The only oxidative stress effect known in blood is the change in oxidation number of Fe (from ferrous to ferric) in hemoglobin, generating methemoglobin but not in other macromolecules. Currently, the effects of the overproduction of methemoglobin derived from snake venom are not extensively recorded. Therefore, the present study aims to describe the oxidative stress induced by Crotalus molossus nigrescens venom using erythrocytes. Methods Human erythrocytes were washed and incubated with different Crotalus molossus nigrescens venom concentrations (0-640 μg/mL). After 24 h, the hemolytic activity was measured followed by attenuated total reflectance-Fourier transform infrared spectroscopy, non-denaturing PAGE, conjugated diene and thiobarbituric acid reactive substances determination. Results Low concentrations of venom (<10 μg/mL) generates oxyhemoglobin release by hemolysis, whereas higher concentrations produced a hemoglobin shift of valence, producing methemoglobin (>40 μg/mL). This substance is not degraded by proteases present in the venom. By infrared spectroscopy, starting in 80 μg/mL, we observed changes in bands that are associated with protein damage (1660 and 1540 cm−1) and lipid peroxidation (2960, 2920 and 1740 cm−1). Lipid peroxidation was confirmed by conjugated diene and thiobarbituric acid reactive substance determination, in which differences were observed between the control and erythrocytes treated with venom. Conclusions Crotalus molossus nigrescens venom provokes hemolysis and oxidative stress, which induces methemoglobin formation, loss of protein structure and lipid peroxidation.(AU)


Assuntos
Animais , Venenos de Serpentes , Análise Espectral , Metemoglobina , Oxiemoglobinas , Crotalus , Estresse Oxidativo , Eritrócitos , Espectroscopia de Infravermelho com Transformada de Fourier
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