Resumo
Background: Snakebites are the main responsible for envenoming in dogs and the bothropic venom remains the mostcommon in Brazil, which can induce a necrotic skin wound. Hyperbaric oxygen therapy (HBOT) use 100% oxygen underhigh pressure and used to treat different wounds in human patients. To the authors knowledge, no reports regarding to usethe HBOT in skin wound caused by snakebite (Bothrops jararaca) are present in the literature. The present clinical caseaimed to describe the use of HBOT for the treatment of an extensive necrotic wound caused by jararaca snakebite in a dog.Case: A neutered 8-year-old mixed-breed dog, weighing 12 kg, was admitted with a 7-day history of extensive necroticwound was identified in the face and neck causing by a snakebite, and no sign of pain. The procedure of HBOT (singlesessions of 1.5 ATM, 45 min, repeated every 48 h, up to 12 sessions) was decided, and the complete blood cells, alanineaminotransferase, creatinine, creatine kinase, prothrombin time, activated partial thromboplastin time, wound clinicalevaluation were measured at the following time-points: 2nd, 5th, 10th, and 12th sessions. At the 5th session was identifiedleukopenia, neutropenia and lymphopenia. Wound re-epithelialization was initiated after the 5th session, and the completeepithelialization was identified at the 12th session of HBOT. During the HBOT no side effects were identified. Threemonths after the HBOT finished, the animal returned to the clinic and the clinical status evolved positively, and the woundwas completed healed.Discussion: This report described the treatment of an extensive necrotic skin wound caused by snakebite (Bothrops jararaca)in an 8-year-old, neutered, mixed-breed dog using the HBOT. The wound healing...
Assuntos
Animais , Cães , Cicatrização , Necrose/veterinária , Oxigenoterapia Hiperbárica/métodos , Oxigenoterapia Hiperbárica/veterinária , Venenos de Crotalídeos/antagonistas & inibidores , BothropsResumo
Background: Snakebites are the main responsible for envenoming in dogs and the bothropic venom remains the mostcommon in Brazil, which can induce a necrotic skin wound. Hyperbaric oxygen therapy (HBOT) use 100% oxygen underhigh pressure and used to treat different wounds in human patients. To the authors knowledge, no reports regarding to usethe HBOT in skin wound caused by snakebite (Bothrops jararaca) are present in the literature. The present clinical caseaimed to describe the use of HBOT for the treatment of an extensive necrotic wound caused by jararaca snakebite in a dog.Case: A neutered 8-year-old mixed-breed dog, weighing 12 kg, was admitted with a 7-day history of extensive necroticwound was identified in the face and neck causing by a snakebite, and no sign of pain. The procedure of HBOT (singlesessions of 1.5 ATM, 45 min, repeated every 48 h, up to 12 sessions) was decided, and the complete blood cells, alanineaminotransferase, creatinine, creatine kinase, prothrombin time, activated partial thromboplastin time, wound clinicalevaluation were measured at the following time-points: 2nd, 5th, 10th, and 12th sessions. At the 5th session was identifiedleukopenia, neutropenia and lymphopenia. Wound re-epithelialization was initiated after the 5th session, and the completeepithelialization was identified at the 12th session of HBOT. During the HBOT no side effects were identified. Threemonths after the HBOT finished, the animal returned to the clinic and the clinical status evolved positively, and the woundwas completed healed.Discussion: This report described the treatment of an extensive necrotic skin wound caused by snakebite (Bothrops jararaca)in an 8-year-old, neutered, mixed-breed dog using the HBOT. The wound healing...(AU)
Assuntos
Animais , Cães , Venenos de Crotalídeos/antagonistas & inibidores , Necrose/veterinária , Oxigenoterapia Hiperbárica/métodos , Oxigenoterapia Hiperbárica/veterinária , Cicatrização , BothropsResumo
Background: Snakebite envenoming is a condition that affects humans and domestic animals worldwide. Identification of the snake species involved in the envenomation is infrequent. Bothrops envenomation presents typical clinicopathological features. This report describes epidemiological, clinical, and pathological data of 2 cases of Bothrops envenomation in dogs, including the first case of Bothrops moojeni snake striking a domestic animal in Brazil. Cases: Case 1. A dog was witnessed to have a Bothrops moojeni snakebite on a farm. In the first 24 h, acute lameness, pain, diffuse swelling, focal bleeding at the left forelimb, and increased whole-blood clotting time were observed in the envenomed dog. Polyvalent antivenom was administered in addition to fluid therapy, analgesics, corticosteroids, and antibiotics. On the 5th day, the animal presented spontaneous bleeding at the wound site, thrombocytopenia, and increased whole-blood clotting time. An additional dose of polyvalent antivenom was administered, and local treatment at the snakebite site was initiated. After 13 days, the dog showed no clinical or laboratory changes and recovered entirely. Case 2. A mongrel dog was taken for a necropsy to determine the cause of death. Grossly, major findings included swelling in the nasal plane that extended to the neck and dissecting hemorrhage in the subcutaneous tissue and adjacent musculature. Hemorrhages were observed in the heart, parietal pleura, left forelimb, lumbar region, and perirenal tissue. Marked necrosis and disruption of small blood vessels and lymphatics within the deep dermis and subcutaneous tissue were the main microscopic findings close to the snakebite site. Additionally, degeneration and necrosis of muscle fibers and dissecting hemorrhage were observed in the head and neck tissues surrounding the snakebite site. Kidneys showed marked interstitial hemorrhage and acute tubular nephrosis. Discussion: Bothrops envenoming is characterized by local (hemorrhage, dermonecrosis, and myonecrosis) and systemic (coagulative disorders, systemic hemorrhage, and acute kidney injury) changes due to the effect of the main venom components such as phospholipase A2 and metalloproteinases. These changes are hallmarks for the bothropic envenomation, supporting the diagnosis in cases 1 and 2. In case 1, the dog developed a Bothrops moojeni snakebite envenomation, but the immediate treatment with antivenom allowed a favorable outcome. In case 2, gross and microscopic findings supported the presumptive diagnosis of fatal bothropic envenomation. A marked local reaction such as swelling, pain, bleeding, bruising, and tissue necrosis was observed in case 1. In case 2, the most significant local changes were swelling and edema at the head and neck, hemorrhage in the subcutaneous tissue, and adjacent musculature. Systemic effects were observed clinically as spontaneous bleeding, thrombocytopenia, increased whole-blood clotting time (Case 1), systemic hemorrhages, and acute tubular nephrosis (Case 2). A proper treatment probably prevented the development of acute renal failure in Case 1. Herein, we show the first case of accidental snakebite envenomation by B. moojeni in a dog in Brazil. Information is scarce on the identification of venomous snake species striking domestic animals. Fast detection of well-determined clinical and pathological findings of Bothrops envenomation is essential for a correct diagnosis, therapeutics, and a good prognosis, even in cases with an unknown history.
Assuntos
Animais , Cães , Mordeduras de Serpentes/fisiopatologia , Mordeduras de Serpentes/veterinária , Inibidores dos Fatores de Coagulação Sanguínea/análise , Venenos de Crotalídeos/toxicidade , BothropsResumo
Background: Several studies have been published on the characterization of Trimeresurus venoms. However, there is still limited information concerning the venom composition of Trimeresurus species distributed throughout Indonesia, which contributes to significant snakebite envenomation cases. The present study describes a comparative on the composition of T. albolabris, T. insularis, T. puniceus, and T. purpureomaculatus venoms originated from Indonesia. Methods: Protein content in the venom of four Trimeresurus species was determined using Bradford assay, and the venom proteome was elucidated using one-dimension SDS PAGE nano-ESI- LCMS/MS shotgun proteomics. Results: The venom of T. albolabris contained the highest protein content of 11.1 mg/mL, followed by T. puniceus, T. insularis and T. purpureomaculatus venom with 10.7 mg/mL, 8.9 mg/mL and 5.54 mg/mL protein, respectively. In total, our venomic analysis identified 65 proteins belonging to 16 protein families in T. purpureomaculatus; 64 proteins belonging to 18 protein families in T. albolabris; 58 different proteins belonging to 14 protein families in T. puniceus; and 48 different proteins belonging to 14 protein familiesin T. insularis. Four major proteins identified in all venoms belonged to snake venom metalloproteinase, C-type lectin, snake venom serine protease, and phospholipase A2. There were 11 common proteins in all venoms, and T. puniceus venom has the highest number of unique proteins compared to the other three venoms. Cluster analysis of the proteins and venoms showed that T. puniceus venom has the most distinct venom composition. Conclusions: Overall, the results highlighted venom compositional variation of four Trimeresurus spp. from Indonesia. The venoms appear to be highly similar, comprising at least four protein families that correlate with venom's toxin properties and function. This study adds more information on venom variability among Trimeresurus species within the close geographic origin and may contribute to the development of optimum heterologous antivenom.(AU)
Assuntos
Trimeresurus/fisiologia , Proteoma/análise , Venenos de Crotalídeos/química , IndonésiaResumo
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 , RuminantesResumo
In Central and South America, snakebite envenomation is mainly caused by Bothrops spp. snakes, whose venoms feature significant biochemical richness, including serine proteases. The available bothropic antivenoms are efficient in avoiding fatalities, but do not completely neutralize venom serine proteases, which are co-responsible for some disorders observed during envenomation. Methods: In order to search for tools to improve the antivenom's, 6-mer peptides were designed based on a specific substrate for Bothrops jararaca venom serine proteases, and then synthesized, with the intention to selectively inhibit these enzymes. Results: Using batroxobin as a snake venom serine protease model, two structurally similar inhibitor peptides were identified. When tested on B. jararaca venom, one of the new inhibitors displayed a good potential to inhibit the activity of the venom serine proteases. These inhibitors do not affect human serine proteases as human factor Xa and thrombin, due to their selectivity. Conclusion: Our study identified two small peptides able to inhibit bothropic serine proteases, but not human ones, can be used as tools to enhance knowledge of the venom composition and function. Moreover, one promising peptide (pepC) was identified that can be explored in the search for improving Bothrops spp. envenomation treatment.(AU)
Assuntos
Animais , Venenos de Serpentes , Antivenenos , Bothrops , Serina Proteases , PeptídeosResumo
In Central and South America, snakebite envenomation is mainly caused by Bothrops spp. snakes, whose venoms feature significant biochemical richness, including serine proteases. The available bothropic antivenoms are efficient in avoiding fatalities, but do not completely neutralize venom serine proteases, which are co-responsible for some disorders observed during envenomation. Methods: In order to search for tools to improve the antivenom's, 6-mer peptides were designed based on a specific substrate for Bothrops jararaca venom serine proteases, and then synthesized, with the intention to selectively inhibit these enzymes. Results: Using batroxobin as a snake venom serine protease model, two structurally similar inhibitor peptides were identified. When tested on B. jararaca venom, one of the new inhibitors displayed a good potential to inhibit the activity of the venom serine proteases. These inhibitors do not affect human serine proteases as human factor Xa and thrombin, due to their selectivity. Conclusion: Our study identified two small peptides able to inhibit bothropic serine proteases, but not human ones, can be used as tools to enhance knowledge of the venom composition and function. Moreover, one promising peptide (pepC) was identified that can be explored in the search for improving Bothrops spp. envenomation treatment.(AU)
Assuntos
Animais , Venenos de Serpentes , Antivenenos , Bothrops , Serina Proteases , PeptídeosResumo
Snake venoms are composed of pharmacologically active proteins that are evolutionarily diverse, stable and specific to targets. Hence, venoms have been explored as a source of bioactive molecules in treating numerous diseases. Recent evidences suggest that snake venom proteins may affect the formation of new blood vessels. Excessive angiogenesis has been implicated in several pathologies including tumours, diabetic retinopathy, arthritis, inter alia. In the present study, we have examined the effects of P-I metalloproteinases isolated from Bothrops moojeni (BmMP-1) and Bothrops atrox (BaMP-1) and L-amino acid oxidases (LAAO) isolated from B. moojeni (BmLAAO) and B. atrox (BaLAAO) on biochemical and functional aspects of angiogenesis. Methods: P-I metalloproteinases and LAAO were purified from venom by molecular size exclusion and ion-exchange chromatography and subsequently confirmed using mass spectrometry. The P-I metalloproteinases were characterized by azocaseinolytic, fibrinogenolytic and gelatinase activity and LAAO activity was assessed by enzyme activity on L-amino acids. Influence of these proteins on apoptosis and cell cycle in endothelial cells was analysed by flow cytometry. The angiogenic activity was determined by in vitro 3D spheroid assay, Matrigel tube forming assay, and in vivo agarose plug transformation in mice. Results: P-I metalloproteinases exhibited azocaseinolytic activity, cleaved α and partially β chain of fibrinogen, and displayed catalytic activity on gelatin. LAAO showed differential activity on L-amino acids. Flow cytometry analysis indicated that both P-I metalloproteinases and LAAO arrested the cells in G0/G1 phase and further induced both necrosis and apoptosis in endothelial cells. In vitro, P-I metalloproteinases and LAAO exhibited significant anti-angiogenic properties in 3D spheroid and Matrigel models by reducing sprout outgrowth and tube formation. Using agarose plug transplants in mice harbouring P-I metalloproteinases and LAAO we demonstrated a marked disruption of vasculature at the periphery. Conclusion: Our research suggests that P-I metalloproteinases and LAAO exhibit anti-angiogenic properties in vitro and in vivo.(AU)
Assuntos
Animais , Oxirredutases , Bothrops/fisiologia , Inibidores da Angiogênese , Venenos de Crotalídeos , MetaloproteasesResumo
Snake venoms are composed of pharmacologically active proteins that are evolutionarily diverse, stable and specific to targets. Hence, venoms have been explored as a source of bioactive molecules in treating numerous diseases. Recent evidences suggest that snake venom proteins may affect the formation of new blood vessels. Excessive angiogenesis has been implicated in several pathologies including tumours, diabetic retinopathy, arthritis, inter alia. In the present study, we have examined the effects of P-I metalloproteinases isolated from Bothrops moojeni (BmMP-1) and Bothrops atrox (BaMP-1) and L-amino acid oxidases (LAAO) isolated from B. moojeni (BmLAAO) and B. atrox (BaLAAO) on biochemical and functional aspects of angiogenesis. Methods: P-I metalloproteinases and LAAO were purified from venom by molecular size exclusion and ion-exchange chromatography and subsequently confirmed using mass spectrometry. The P-I metalloproteinases were characterized by azocaseinolytic, fibrinogenolytic and gelatinase activity and LAAO activity was assessed by enzyme activity on L-amino acids. Influence of these proteins on apoptosis and cell cycle in endothelial cells was analysed by flow cytometry. The angiogenic activity was determined by in vitro 3D spheroid assay, Matrigel tube forming assay, and in vivo agarose plug transformation in mice. Results: P-I metalloproteinases exhibited azocaseinolytic activity, cleaved α and partially β chain of fibrinogen, and displayed catalytic activity on gelatin. LAAO showed differential activity on L-amino acids. Flow cytometry analysis indicated that both P-I metalloproteinases and LAAO arrested the cells in G0/G1 phase and further induced both necrosis and apoptosis in endothelial cells. In vitro, P-I metalloproteinases and LAAO exhibited significant anti-angiogenic properties in 3D spheroid and Matrigel models by reducing sprout outgrowth and tube formation. Using agarose plug transplants in mice harbouring P-I metalloproteinases and LAAO we demonstrated a marked disruption of vasculature at the periphery. Conclusion: Our research suggests that P-I metalloproteinases and LAAO exhibit anti-angiogenic properties in vitro and in vivo.(AU)
Assuntos
Animais , Oxirredutases , Bothrops/fisiologia , Inibidores da Angiogênese , Venenos de Crotalídeos , MetaloproteasesResumo
In Brazil, snakes from the Bothrops genus are responsible for thousands of accidents, and their venoms are mainly composed of proteolytic enzymes. Although the antibothropic serum produced by the Brazilian Institutes is remarkably efficient, more studies are necessary, especially in veterinary medicine. The venom contain enzymes and non-enzymatic proteins that interfere with hemostasis leading to hemorrhage or even thrombosis. Possible treatment associations with known bothropic antivenom were the reason for the development of the present study. The aim of this study was to evaluate hemostasis alterations caused by Bothrops alternatus venom in rabbits followed by treatments with anti-bothropic serum, tranexamic acid and desmopressin. Twenty New Zealand rabbits were distributed into five groups (n=4) that were experimentally envenomed with 150mcg/kg of B. alternatus venom via intramuscular injection and treated as follow: Group 1 (G1) was the positive control and received venom and PBS/BSA; Group 2 (G2) was treated with tranexamic acid; Group 3 (G3) with desmopressin; Group 4 (G4) with tranexamic acid and anti-bothropic serum; and Group 5 (G5) with anti-bothropic serum and desmopressin. Blood samples were collected before venom administration, and one, four, eight and 12 hours after, for Partial activated partial thromboplastin time, Prothrombin Time, Thrombin Time and fibrinogen evaluation. Thrombin generation (TG) test was carried out with a pool of samples from final times (8 and 12h). At the end of 12h, all animals were euthanized and necropsy was conducted. Samples from muscle tissue, heart, lungs and kidney were analyzed. Classic coagulation tests showed no significant differences amongst groups and times. However, TG indicated that the venom causes a hypocoagulability state, which was not reversed by proposed treatments. Histology showed muscle inflammation, hemorrhage and necrosis, as well as hemorrhage in other tissues with no differences amongst groups. B. alternatus envenomation causes hypocoagulability detected by TG assay, but not through classical coagulation tests. The use of tranexamic acid and desmopressin for hemostasis stabilization after inoculation of the venom did not show advantage in coagulation restoration.(AU)
No Brasil, as serpentes do gênero Bothrops são responsáveis por milhares de acidentes, e seus venenos são compostos principalmente de enzimas proteolíticas. Embora o soro antiofídico produzido pelos institutos brasileiros seja notavelmente eficiente, mais estudos são necessários, especialmente na medicina veterinária. O veneno contem enzimas e proteínas não-enzimáticas que interferem com a hemostasia levando a hemorragias ou trombose. A associação de outros tratamentos ao soro antibotrópico foi a razão para o desenvolvimento do presente estudo. O objetivo deste estudo foi avaliar as alterações da hemostasia causadas pelo veneno de Bothrops alternatus em coelhos, após tratamento com soro antibotrópico, ácido tranexâmico e desmopressina. Vinte coelhos da Nova Zelândia foram distribuídos em cinco grupos (n = 4) que foram submetidos a experimentos com 150mcg/kg de veneno de B. alternatus por injeção intramuscular. O Grupo 1 (G1) foi o controle positivo e recebeu veneno e PBS / BSA, enquanto o Grupo 2 (G2) foi tratado com ácido tranexâmico, o Grupo 3 (G3) com desmopressina, o Grupo 4 (G4) com ácido tranexâmico e soro antibotrópico, e o Grupo 5 (G5) com soro antibotrópico e desmopressina. As amostras de sangue foram coletadas antes da administração do veneno, e uma, quatro, oito e 12 horas após os tratamentos para realização de tempo de tromboplastina parcial ativada parcial (TTPa), tempo de protrombina (TP), tempo de trombina (TT) e mensuração de fibrinogênio. Para o ensaio de geração de trombina (TG) foi realizado com um pool de amostras nos tempos finais (8 e 12h). Ao final das 12h, todos os animais foram sacrificados e a necropsia foi realizada. Amostras de tecido muscular, coração, pulmões e rins foram analisadas. Os testes TTPa, TP, TT e fibrinogênio não mostraram diferenças significativas entre os grupos e os tempos. No entanto, o TG indicou que o veneno causa um estado de hipocoagulabilidade, que não foi revertido pelos tratamentos propostos. Na histologia, foram observadas inflamação muscular, hemorragia e necrose, além de hemorragia em outros tecidos, sem diferenças entre os grupos. O envenenamento por B. alternatus causa hipocoagulabilidade detectada mais precocemente pelo teste de geração de trombina. O uso de ácido tranexâmico e desmopressina para estabilização da hemostasia após a inoculação do veneno não mostrou vantagem na restauração da coagulação.(AU)
Assuntos
Animais , Coelhos , Serpentes , Bothrops , Hemostasia , Técnicas HemostáticasResumo
In Brazil, snakes from the Bothrops genus are responsible for thousands of accidents, and their venoms are mainly composed of proteolytic enzymes. Although the antibothropic serum produced by the Brazilian Institutes is remarkably efficient, more studies are necessary, especially in veterinary medicine. The venom contain enzymes and non-enzymatic proteins that interfere with hemostasis leading to hemorrhage or even thrombosis. Possible treatment associations with known bothropic antivenom were the reason for the development of the present study. The aim of this study was to evaluate hemostasis alterations caused by Bothrops alternatus venom in rabbits followed by treatments with anti-bothropic serum, tranexamic acid and desmopressin. Twenty New Zealand rabbits were distributed into five groups (n=4) that were experimentally envenomed with 150mcg/kg of B. alternatus venom via intramuscular injection and treated as follow: Group 1 (G1) was the positive control and received venom and PBS/BSA; Group 2 (G2) was treated with tranexamic acid; Group 3 (G3) with desmopressin; Group 4 (G4) with tranexamic acid and anti-bothropic serum; and Group 5 (G5) with anti-bothropic serum and desmopressin. Blood samples were collected before venom administration, and one, four, eight and 12 hours after, for Partial activated partial thromboplastin time, Prothrombin Time, Thrombin Time and fibrinogen evaluation. Thrombin generation (TG) test was carried out with a pool of samples from final times (8 and 12h). At the end of 12h, all animals were euthanized and necropsy was conducted. Samples from muscle tissue, heart, lungs and kidney were analyzed. Classic coagulation tests showed no significant differences amongst groups and times. However, TG indicated that the venom causes a hypocoagulability state, which was not reversed by proposed treatments. Histology showed muscle inflammation, hemorrhage and necrosis, as well as hemorrhage in other tissues with no differences amongst groups. B. alternatus envenomation causes hypocoagulability detected by TG assay, but not through classical coagulation tests. The use of tranexamic acid and desmopressin for hemostasis stabilization after inoculation of the venom did not show advantage in coagulation restoration.(AU)
No Brasil, as serpentes do gênero Bothrops são responsáveis por milhares de acidentes, e seus venenos são compostos principalmente de enzimas proteolíticas. Embora o soro antiofídico produzido pelos institutos brasileiros seja notavelmente eficiente, mais estudos são necessários, especialmente na medicina veterinária. O veneno contem enzimas e proteínas não-enzimáticas que interferem com a hemostasia levando a hemorragias ou trombose. A associação de outros tratamentos ao soro antibotrópico foi a razão para o desenvolvimento do presente estudo. O objetivo deste estudo foi avaliar as alterações da hemostasia causadas pelo veneno de Bothrops alternatus em coelhos, após tratamento com soro antibotrópico, ácido tranexâmico e desmopressina. Vinte coelhos da Nova Zelândia foram distribuídos em cinco grupos (n = 4) que foram submetidos a experimentos com 150mcg/kg de veneno de B. alternatus por injeção intramuscular. O Grupo 1 (G1) foi o controle positivo e recebeu veneno e PBS / BSA, enquanto o Grupo 2 (G2) foi tratado com ácido tranexâmico, o Grupo 3 (G3) com desmopressina, o Grupo 4 (G4) com ácido tranexâmico e soro antibotrópico, e o Grupo 5 (G5) com soro antibotrópico e desmopressina. As amostras de sangue foram coletadas antes da administração do veneno, e uma, quatro, oito e 12 horas após os tratamentos para realização de tempo de tromboplastina parcial ativada parcial (TTPa), tempo de protrombina (TP), tempo de trombina (TT) e mensuração de fibrinogênio. Para o ensaio de geração de trombina (TG) foi realizado com um pool de amostras nos tempos finais (8 e 12h). Ao final das 12h, todos os animais foram sacrificados e a necropsia foi realizada. Amostras de tecido muscular, coração, pulmões e rins foram analisadas. Os testes TTPa, TP, TT e fibrinogênio não mostraram diferenças significativas entre os grupos e os tempos. No entanto, o TG indicou que o veneno causa um estado de hipocoagulabilidade, que não foi revertido pelos tratamentos propostos. Na histologia, foram observadas inflamação muscular, hemorragia e necrose, além de hemorragia em outros tecidos, sem diferenças entre os grupos. O envenenamento por B. alternatus causa hipocoagulabilidade detectada mais precocemente pelo teste de geração de trombina. O uso de ácido tranexâmico e desmopressina para estabilização da hemostasia após a inoculação do veneno não mostrou vantagem na restauração da coagulação.(AU)
Assuntos
Animais , Coelhos , Serpentes , Bothrops , Hemostasia , Técnicas HemostáticasResumo
The morphology of the male reproductive tract of Tomodon dorsatus was described in the austral seasons of the year considering macroscopic and microscopic variables. For this purpose, 56 specimens from the herpetological collection of the "Instituto Butantan" were used. Fragments of the testes, kidneys and ductus deferens were collected and submitted to histological routine. The peak of the testicular volume was observed in the summer and the epithelium of the seminiferous tubules had higher height in the summer (p=0.001). The testes were active throughout the year, however, the spermiogenesis peaked in the summer. There were spermatozoa in the lumen of the ductus deferens in all seasons of the year. Renal length was higher in autumn (p=0.027), and renal width did not show a significant increase (p=0.237). The diameter and epithelial height of the sexual segment of the kidney (SSK) showed hypertrophy in winter and spring, coinciding with the mating period. Based on findings of this study, we can suggest that, at the population level, the reproductive cycle of T. dorsatus can be considered seasonal semi-synchronous, due to the peak of spermiogenic activity in the hot season, and discontinuous at the individual level.(AU)
A morfologia do trato reprodutivo do macho de Tomodon dorsatus foi descrita nas estações climáticas do ano com base em variáveis macroscópicas e microscópicas. Para isto, foram usados 56 espécimes oriundos da coleção herpetológica do Instituto Butantan. Fragmentos dos testículos, rins e ductos deferentes foram coletados e submetidos à rotina histológica. O volume testicular foi maior no verão e o epitélio dos túbulos seminíferos mostrou uma maior altura no verão (p=0.001). Os testículos estavam ativos durante todo o ano, contudo, a espermiogênese foi maior no verão. Espermatozoides foram encontrados no lúmen do ducto deferente em todas as estações do ano. O comprimento renal foi maior no outono (p=0.027), e a largura renal não mostrou um aumento significativo (p=0.237). O diâmetro e a altura epitelial do segmento sexual do rim (SSR) mostrou hipertrofia nas estações inverno e primavera, coincidindo com o período reprodutivo. Com base nestes resultados, pode-se sugerir que, em nível populacional, o ciclo reprodutivo de T. dorsatus possa ser considerado semi-sincrônico sazonal, devido à atividade espermiogênica na estação quente, e descontínuo em nível individual.(AU)
Assuntos
Animais , Masculino , Estações do Ano , Testículo , Ducto Deferente , Hipertrofia , Espermatogênese , Bothrops , Padrões de ReferênciaResumo
The morphology of the male reproductive tract of Tomodon dorsatus was described in the austral seasons of the year considering macroscopic and microscopic variables. For this purpose, 56 specimens from the herpetological collection of the "Instituto Butantan" were used. Fragments of the testes, kidneys and ductus deferens were collected and submitted to histological routine. The peak of the testicular volume was observed in the summer and the epithelium of the seminiferous tubules had higher height in the summer (p=0.001). The testes were active throughout the year, however, the spermiogenesis peaked in the summer. There were spermatozoa in the lumen of the ductus deferens in all seasons of the year. Renal length was higher in autumn (p=0.027), and renal width did not show a significant increase (p=0.237). The diameter and epithelial height of the sexual segment of the kidney (SSK) showed hypertrophy in winter and spring, coinciding with the mating period. Based on findings of this study, we can suggest that, at the population level, the reproductive cycle of T. dorsatus can be considered seasonal semi-synchronous, due to the peak of spermiogenic activity in the hot season, and discontinuous at the individual level.(AU)
A morfologia do trato reprodutivo do macho de Tomodon dorsatus foi descrita nas estações climáticas do ano com base em variáveis macroscópicas e microscópicas. Para isto, foram usados 56 espécimes oriundos da coleção herpetológica do Instituto Butantan. Fragmentos dos testículos, rins e ductos deferentes foram coletados e submetidos à rotina histológica. O volume testicular foi maior no verão e o epitélio dos túbulos seminíferos mostrou uma maior altura no verão (p=0.001). Os testículos estavam ativos durante todo o ano, contudo, a espermiogênese foi maior no verão. Espermatozoides foram encontrados no lúmen do ducto deferente em todas as estações do ano. O comprimento renal foi maior no outono (p=0.027), e a largura renal não mostrou um aumento significativo (p=0.237). O diâmetro e a altura epitelial do segmento sexual do rim (SSR) mostrou hipertrofia nas estações inverno e primavera, coincidindo com o período reprodutivo. Com base nestes resultados, pode-se sugerir que, em nível populacional, o ciclo reprodutivo de T. dorsatus possa ser considerado semi-sincrônico sazonal, devido à atividade espermiogênica na estação quente, e descontínuo em nível individual.(AU)
Assuntos
Animais , Masculino , Estações do Ano , Testículo , Ducto Deferente , Hipertrofia , Espermatogênese , Bothrops , Padrões de ReferênciaResumo
The research and development of alternative treatments for snakebites (e.g., medicinal plants) is necessary due to the high costs of the existing ones. The effects of the aqueous extracts from Jacaranda decurrens leaves, roots, and xylopodium were analyzed upon the venom-induced (Bothrops spp. and Crotalus spp.) systemic and local toxicity. The extracts were able to partially inhibit the phospholipase activity of the venoms from Bothrops jararacussu and Crotalus durissus terrificus. The myotoxic, edema-inducing, coagulant, and hemorrhagic activities were also inhibited. The SDS-PAGE showed that the venom proteins were intact after their incubation with the extracts. This suggests that the possible mechanism of inhibition is not related to the degradation of the protein but rather to their binding to specific sites of the enzymes. The extracts significantly prolonged the survival time of animals in the lethality assay performed with Crotalus durissus terrificus venom and its toxin (crotoxin). The anti-ophidic activity of medicinal plants may aid in the management of snakebites in distant locations by reducing the victims local effects and time to heal.
Assuntos
Bignoniaceae/toxicidade , Plantas Medicinais/toxicidade , Técnicas In Vitro , Venenos de CrotalídeosResumo
The research and development of alternative treatments for snakebites (e.g., medicinal plants) is necessary due to the high costs of the existing ones. The effects of the aqueous extracts from Jacaranda decurrens leaves, roots, and xylopodium were analyzed upon the venom-induced (Bothrops spp. and Crotalus spp.) systemic and local toxicity. The extracts were able to partially inhibit the phospholipase activity of the venoms from Bothrops jararacussu and Crotalus durissus terrificus. The myotoxic, edema-inducing, coagulant, and hemorrhagic activities were also inhibited. The SDS-PAGE showed that the venom proteins were intact after their incubation with the extracts. This suggests that the possible mechanism of inhibition is not related to the degradation of the protein but rather to their binding to specific sites of the enzymes. The extracts significantly prolonged the survival time of animals in the lethality assay performed with Crotalus durissus terrificus venom and its toxin (crotoxin). The anti-ophidic activity of medicinal plants may aid in the management of snakebites in distant locations by reducing the victims local effects and time to heal.(AU)
Assuntos
Técnicas In Vitro , Bignoniaceae/toxicidade , Plantas Medicinais/toxicidade , Venenos de CrotalídeosResumo
Lack of complete genomic data of Bothrops jararaca impedes molecular biology research focusing on biotechnological applications of venom gland components. Identification of full-length coding regions of genes is crucial for the correct molecular cloning design. Methods: RNA was extracted from the venom gland of one adult female specimen of Bothrops jararaca. Deep sequencing of the mRNA library was performed using Illumina NextSeq 500 platform. De novo assembly of B. jararaca transcriptome was done using Trinity. Annotation was performed using Blast2GO. All predicted proteins after clustering step were blasted against non-redundant protein database of NCBI using BLASTP. Metabolic pathways present in the transcriptome were annotated using the KAAS-KEGG Automatic Annotation Server. Toxins were identified in the B. jararaca predicted proteome using BLASTP against all protein sequences obtained from Animal Toxin Annotation Project from Uniprot KB/Swiss-Pro database. Figures and data visualization were performed using ggplot2 package in R language environment. Results: We described the in-depth transcriptome analysis of B. jararaca venom gland, in which 76,765 de novo assembled isoforms, 96,044 transcribed genes and 41,196 unique proteins were identified. The most abundant transcript was the zinc metalloproteinase-disintegrin-like jararhagin. Moreover, we identified 78 distinct functional classes of proteins, including toxins, inhibitors and tumor suppressors. Other venom proteins identified were the hemolytic lethal factors stonustoxin and verrucotoxin. Conclusion: It is believed that the application of deep sequencing to the analysis of snake venom transcriptomes may represent invaluable insight on their biotechnological potential focusing on candidate molecules.(AU)
Assuntos
Animais , Bothrops , Bothrops/fisiologia , Proteoma , Venenos de Crotalídeos , Perfilação da Expressão Gênica , Metaloproteases , Transcriptoma , Biologia Molecular , Análise por Conglomerados , Sequenciamento de Nucleotídeos em Larga EscalaResumo
Background: The Brazils lancehead, Bothrops brazili, is a poorly studied pit viper distributed in lowlands of the equatorial rainforests of southern Colombia, northeastern Peru, eastern Ecuador, southern and southeastern Venezuela, Guyana, Suriname, French Guiana, Brazil, and northern Bolivia. Few studies have been reported on toxins isolated from venom of Ecuadorian and Brazilian B. brazili. The aim of the present study was to elucidate the qualitative and quantitative protein composition of B. brazili venom from Pará (Brazil), and to carry out a comparative antivenomics assessment of the immunoreactivity of the Brazilian antibothropic pentavalent antivenom [soro antibotrópico (SAB) in Portuguese] against the venoms of B. brazili and reference species, B. jararaca. Methods: We have applied a quantitative snake venomics approach, including reverse-phase and two-dimensional electrophoretic decomplexation of the venom toxin arsenal, LC-ESI-MS mass profiling and peptide-centric MS/MS proteomic analysis, to unveil the overall protein composition of B. brazili venom from Pará (Brazil). Using third-generation antivenomics, the specific and paraspecific immunoreactivity of the Brazilian SAB against homologous (B. jararaca) and heterologous (B. brazili) venoms was investigated. Results: The venom proteome of the Brazils lancehead (Pará) is predominantly composed of two major and three minor acidic (19%) and two major and five minor basic (14%) phospholipase A2 molecules; 7-11 snake venom metalloproteinases of classes PI (21%) and PIII (6%); 10-12 serine proteinases (14%), and 1-2 L-amino acid oxidases (6%). Other toxins, including two cysteine-rich secretory proteins, one C-type lectin-like molecule, one nerve growth factor, one 5'-nucleotidase, one phosphodiesterase, one phospholipase B, and one glutaminyl cyclase molecule, represent together less than 2.7% of the venom proteome...(AU)
Assuntos
Animais , Bothrops , Venenos de Crotalídeos/análise , Venenos de Crotalídeos/química , Venenos de Crotalídeos/antagonistas & inibidores , AntivenenosResumo
Bothrops atrox is known to be the pit viper responsible for most snakebites and human fatalities in the Amazon region. It can be found in a wide geographical area including northern South America, the east of Andes and the Amazon basin. Possibly, due to its wide distribution and generalist feeding, intraspecific venom variation was reported by previous proteomics studies. Sex-based and ontogenetic variations on venom compositions of Bothrops snakes were also subject of proteomic and peptidomic analysis. However, the venom peptidome of B. atrox remains unknown. Methods: We conducted a mass spectrometry-based analysis of the venom peptides of individual male and female specimens combining bottom-up and top-down approaches. Results: We identified in B. atrox a total of 105 native peptides in the mass range of 0.4 to 13.9 kDa. Quantitative analysis showed that phospholipase A2 and bradykinin potentiating peptides were the most abundant peptide families in both genders, whereas disintegrin levels were significantly increased in the venoms of females. Known peptides processed at non-canonical sites and new peptides as the Ba1a, which contains the SVMP BATXSVMPII1 catalytic site, were also revealed in this work. Conclusion: The venom peptidomes of male and female specimens of B. atrox were analyzed by mass spectrometry-based approaches in this work. The study points to differences in disintegrin levels in the venoms of females that may result in distinct pathophysiology of envenomation. Further research is required to explore the potential biological implications of this finding.(AU)
Assuntos
Animais , Peptídeos , Bothrops , Venenos de Crotalídeos/biossíntese , Caracteres Sexuais , Ecossistema Amazônico , PeptidomiméticosResumo
Pathogenesis of Bothrops envenomations is complex and despite numerous studies on the effects of this snake venom on various biological systems, relatively little is known about such effects on the male reproductive system. In the present study, the toxicological outcomes of the low molecular weight fraction (LMWF) of B. jararaca snake venom - containing a range of bioactive peptides - were investigated on the dynamics and structure of the seminiferous epithelium and 15P-1 Sertoli cells viability. Methods: LMWF (5 µg/dose per testis) venom was administered in male Swiss mice by intratesticular (i.t.) injection. Seven days after this procedure, the testes were collected for morphological and morphometric evaluation, distribution of claudin-1 in the seminiferous epithelium by immunohistochemical analyses of testes, and the nitric oxide (NO) levels were evaluated in the total extract of the testis protein. In addition, the toxicological effects of LMWF and crude venom (CV) were analyzed on the 15P-1 Sertoli cell culture. Results: LMWF induced changes in the structure and function of the seminiferous epithelium without altering claudin-1 distribution. LMWF effects were characterized especially by lost cells in the adluminal compartment of epithelium (spermatocytes in pachytene, preleptotene spermatocytes, zygotene spermatocytes, and round spermatid) and different stages of the seminiferous epithelium cycle. LMWF also increased the NO levels in the total extract of the testis protein and was not cytotoxic in concentrations and time tested in the present study. However, CV showed cytotoxicity at 10 μg/mL from 6 to 48 h of treatment. Conclusions: The major finding of the present study was that the LMWF inhibited spermatozoa production; principally in the spermiogenesis stage without altering claudin-1 distribution in the basal compartment. Moreover, NO increased by LMWF induce open of complexes junctions and release the germ cells of the adluminal compartment to the seminiferous tubule.(AU)
Assuntos
Animais , Masculino , Peptídeos , Epitélio Seminífero , Venenos de Serpentes , Espermatogênese , Bothrops , Produtos BiológicosResumo
Snakebite envenoming can be a life-threatening condition, for which emergency care is essential. The Bothrops (lancehead) genus is responsible for most snakebite-related deaths and permanent loss of function in human victims in Latin America. Bothrops spp. venom is a complex mixture of different proteins that are known to cause local necrosis, coagulopathy, and acute kidney injury. However, the long-term effects of these viper envenomings have remained largely understudied. Case presentation: Here, we present a case report of a 46-years old female patient from Las Claritas, Venezuela, who was envenomed by a snake from the Bothrops genus. The patient was followed for a 10-year period, during which she presented oliguric renal failure, culminating in kidney failure 60 months after the envenoming. Conclusion: In Latin America, especially in Brazil, where there is a high prevalence of Bothrops envenoming, it may be relevant to establish long-term outpatient programs. This would reduce late adverse events, such as chronic kidney disease, and optimize public financial resources by avoiding hemodialysis and consequently kidney transplantation.(AU)