RESUMO
Variability in snake venom composition is well-documented and crucial for understanding snake ecology and predicting snakebites. In this study, we characterize the venom composition and biological activities of newborn female and male Bothrops moojeni and their mother. Our results reveal significant differences between the venom of newborn females and males, demonstrating a broad and diverse range of proteins. The venoms of newborn females showed higher serine protease effects, increased hemorrhagic activity, and greater lethality compared to the venom of newborn males. However, no differences were observed in phospholipase A2 and coagulant activity. The differences in protein composition and toxic activities between maternal and neonatal venom, as well as between the venoms of newborn females and males, contribute to understanding the diverse outcomes of snakebites. These results underscore the importance of considering sex and ontogeny in understanding venom composition in snakes.
Assuntos
Animais Recém-Nascidos , Bothrops , Venenos de Crotalídeos , Animais , Bothrops/classificação , Bothrops/fisiologia , Feminino , Masculino , Fatores SexuaisRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Lauraceae family includes Nectandra angustifolia a species widely used in the folk medicine of South America against various maladies. It is commonly used to treat different types of processes like inflammation, pain, and snakebites. Snakes of the Bothrops genus are responsible for about 97% of the ophidic accidents in northeastern Argentina. AIM OF THE STUDY: To evaluate the anti-snake activity of the phytochemicals present in N. angustifolia extracts, identify the compounds, and evaluate their inhibitory effect on phospholipase A2 (PLA2) with in vitro and in silico assays. METHODS: Seasonal variations in the alexiteric potential of aqueous, ethanolic and hexanic extracts were evaluated by inhibition of coagulant, haemolytic, and cytotoxic effects of B. diporus venom. The chemical identity of an enriched fraction obtained by bio-guided fractioning was established by UPLC-MS/MS analysis. Molecular docking studies were carried out to investigate the binding mechanisms of the identified compounds to PLA2 enzyme from snake venom. RESULTS: All the extracts inhibited venom coagulant activity. However, spring ethanolic extract achieved 100% inhibition of haemolytic activity. Bio-guide fractioning led to an enriched fraction (F4) with the highest haemolytic inhibition. Five flavonoids were identified in this fraction; molecular docking and Molecular Dynamics (MD) simulations indicated the binding mechanisms of the identified compounds. The carbohydrates present in some of the compounds had a critical effect on the interaction with PLA2. CONCLUSION: This study shows, for the first time, which compounds are responsible for the anti-snake activity in Nectandra angustifolia based on in vitro and in silico assays. The results obtained in this work support the traditional use of this species as anti-snake in folk medicine.
Assuntos
Bothrops , Venenos de Crotalídeos , Lauraceae , Animais , Flavonoides/farmacologia , Simulação de Acoplamento Molecular , Cromatografia Líquida , Extratos Vegetais/uso terapêutico , Espectrometria de Massas em Tandem , Bothrops/fisiologia , Fosfolipases A2/metabolismoRESUMO
Disintegrins are a group of cysteine-rich proteins found in a wide variety of snake venoms. These proteins selectively bind to integrins, which play a fundamental role in the regulation of many physiological and pathological processes. Here, we report the NMR chemical shift assignments for 1H, 15N, and 13C nuclei in the backbone and side chains of recombinant disintegrin Jarastatin (rJast), which was further validated by secondary structure prediction using the TALOS-N server. Taken together, these data are essential to perform NMR-based experiments, including structure determination, backbone dynamics, mapping ligand sites and enabling a deeper understanding of the effect of hydrophobic surface clusters, which are important elements to stabilize some 3D proteins structure/folding.
Assuntos
Bothrops , Venenos de Crotalídeos , Sequência de Aminoácidos , Animais , Bothrops/fisiologia , Venenos de Crotalídeos/química , Desintegrinas/química , Ressonância Magnética Nuclear BiomolecularRESUMO
Snake envenomation is considered a public health problem in tropical countries, where they occur in a high incidence. The present study reports the snake envenomation that occurred in Mato Grosso do Sul state (Brazil) between 2007 and 2017. Epidemiological data were obtained from the online platform of the Notification Disease Information System and were analyzed according to biome. A total of 5568 cases of snake envenomations were recorded during the study period, where the highest frequency was registered between October and April. The majority of envenomations occurred in working-age males (20 to 39 years), caused mainly by Bothrops snakes, and the duration of care after the envenomation in most cases took three hours. The municipalities that showed the highest snake envenomations case per 100,000 inhabitants presents low population density, and have their economy based on agricultural activity, which is a risk factor to snake envenomations. To the Mato Grosso do Sul state, the total number of snake envenomations had a positive relationship with the size of the municipality. Since this, larger areas usually have a mosaic of environments, which may harbor higher richness and abundance of snakes, and can cause more snake encounters with the population, resulting in more snake envenomations.
Assuntos
Mordeduras de Serpentes/epidemiologia , Adulto , Animais , Bothrops/fisiologia , Brasil/epidemiologia , Feminino , Humanos , Masculino , Estações do Ano , Mordeduras de Serpentes/parasitologia , Adulto JovemRESUMO
The emergence of the diagonal of open/dry vegetations, including Chaco, Cerrado and Caatinga, is suggested to have acted as a dispersal barrier for terrestrial organisms by fragmenting a single large forest that existed in South America into the present Atlantic and Amazon forests. Here we tested the hypothesis that the expansion of the South American diagonal of open/dry landscapes acted as a vicariant process for forest lanceheads of the genus Bothrops, by analyzing the temporal range dynamics of those snakes. We estimated ancestral geographic ranges of the focal lancehead clade and its sister clade using a Bayesian dated phylogeny and the BioGeoBEARS package. We compared nine Maximum Likelihood models to infer ancestral range probabilities and their related biogeographic processes. The best fitting models (DECTS and DIVALIKETS) recovered the ancestor of our focal clade in the Amazon biogeographic region of northwestern South America. Vicariant processes in two different subclades resulted in disjunct geographic distributions in the Amazon and the Atlantic Forest. Dispersal processes must have occurred mostly within the Amazon and the Atlantic Forest and not between them. Our results suggest the fragmentation of a single ancient large forest into the Atlantic and Amazon forests acting as a driver of vicariant processes for the snake lineage studied, highlighting the importance of the diagonal of open/dry landscapes in shaping distribution patterns of terrestrial biota in South America.
Assuntos
Bothrops/fisiologia , Animais , Teorema de Bayes , Bothrops/classificação , Florestas , Filogenia , Filogeografia , Dinâmica Populacional , América do Sul , Clima TropicalRESUMO
Assisted reproductive technologies such as cooling, freezing, and artificial insemination are essential in conserving genetic diversity of critically endangered reptiles like the golden lancehead pitviper (Bothrops insularis). Therefore, we examined viability of semen samples from captive golden lanceheads diluted in coconut-water based extender over a 48-h period of cooling (5°C). Semen evaluations were performed immediately after the dilution and at 6, 24, and 48 h, using computer-assisted sperm analysis and stains to assess plasma membrane and acrosomal status. Our findings showed that the extender and protocol employed here were effective in preserving golden lancehead pitviper spermatozoa for a short periods of time (48 h), allowing semen samples to be transported to distant locations for immediate use without the setbacks of cryopreservation.
Assuntos
Bothrops/fisiologia , Temperatura Baixa , Preservação do Sêmen/veterinária , Sêmen/fisiologia , Animais , Masculino , Análise do Sêmen , Preservação do Sêmen/métodos , Fatores de TempoRESUMO
The two-striped forest-pitviper (Bothrops bilineatus) is an arboreal snake that is currently represented by two subspecies (B. b. bilineatus and B. b. smaragdinus) that comprise a species complex, and its distribution is in the Amazon and the Atlantic Forest. The rarity of encounters with this snake is reflected in the low occurrence of cases of snakebites throughout its geographic distribution and the resulting low number of published clinical reports. However, in some areas, B. bilineatus proves to be more frequent and causes envenomations in a greater proportion. Herein, we review the main aspects of the species complex B. bilineatus, including its biology, ecology, taxonomy, morphology, genetic and molecular studies, geographic distribution, conservation status, venom, pathophysiology and clinical aspects, and epidemiology. In addition, the different antivenoms available for the treatment of envenomations caused by B. bilineatus are presented along with suggestions for future studies that are needed for a better understanding of the snakebites caused by this snake.
Assuntos
Bothrops , Adulto , Animais , Antivenenos/uso terapêutico , Bothrops/anatomia & histologia , Bothrops/genética , Bothrops/fisiologia , Brasil , Conservação dos Recursos Naturais , Venenos de Crotalídeos/toxicidade , Florestas , Humanos , Masculino , Mordeduras de Serpentes/epidemiologia , Mordeduras de Serpentes/terapiaRESUMO
Ectotherms depend on temperature to maintain their physiological functions and through behavioral changes, they can avoid overheating in their habitats. The voluntary thermal maximum (VTMax ) represents the maximum temperature tolerated by individuals before actively moving to a colder place. However, if and how VTMax might change after capture and in captivity remains understudied. We investigate if measurements taken in captivity are a good proxy for thermal tolerance of wild individuals. As thermal history has been shown to affect behavioral response and physiological parameters, herein we hypothesized that VTMax of the neotropical viper Bothrops pauloensis varies throughout the captivity period. We measured the VTMax of individuals immediately after capture and in three trials during a short-term period in captivity. Measurements were done by recording their body temperature at which they exited a heating box experimental setup. In contrast to our hypothesis, the VTMax was not significantly affected by time in captivity but there was interindividual variation. There were also no significant differences between field and captivity measurements, in spite of the small effect size. Our results indicate that the VTMax of this snake population is not affected by a short-term captivity period. Furthermore, an invariant VTMax might indicate low phenotypic plasticity, as individuals do not appear to adjust their tolerance to short-term exposure to higher temperatures and potential vulnerability to threats such as global warming. We expect that our results can contribute to understanding the effect of captivity on thermal tolerance in neotropical squamates, allowing for insights into their thermal physiology and ecology.
Assuntos
Comportamento Animal/fisiologia , Regulação da Temperatura Corporal/fisiologia , Bothrops/fisiologia , Criação de Animais Domésticos , Animais , Feminino , MasculinoRESUMO
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)