Abstract
The Brazilian lancehead (Bothrops moojeni) has a wide distribution in Brazil and represents a serious public health hazard. Previous works reported that the symptoms of snakebites caused by B. moojeni juveniles' bites were mainly related to coagulation, while those caused by adults' bites had a more prominent local damage. In this work, we analyzed the venoms of B. moojeni at different life stages to better understand the ontogeny shift in this species. Snakes were grouped by age and sex, and venom pools were formed accordingly. Compositional analyses by one-dimensional electrophoresis (1-DE), chromatography, and mass spectrometry revealed that ontogenetic changes might be mostly related to phospholipase A2 (PLA2) and metalloproteases. Regarding the venoms functional aspect, proteolytic, L-amino acid oxidase, PLA2, and coagulant in vitro activities were assayed, but only the first and the last ones showed age-related changes, with the venom of snakes up to 1 year-old displaying lower proteolytic and higher coagulant activities, while those from 2 years-old onward presented the opposite relation. The venoms of 3 years-old snakes were exceptions to the compositional and functional pattern of adults as both venoms presented profiles similar to neonates. Sex-related differences were observed in specific groups and were not age-related. In vivo experiments (median lethal dose and hemorrhagic activity) were statistically similar between neonates and adults, however we verified that the adult venom killed mice faster comparing to the neonates. All venoms were mostly recognized by the antibothropic serum and displayed similar profiles to 1-DE in western blotting. In conclusion, the Brazilian lancehead venom showed ontogenetic shift in its composition and activities. Furthermore, this change occurred in snakes from 1 to 2 years-old, and interestingly the venom pools from 3 years-old snakes had particular characteristics, which highlights the importance of comprehensive studies to better understand venom variability.
Highlights
Classified as a neglected tropical disease by the World Health Organization, snakebites are an important public health hazard, especially in tropical and subtropical areas [1]
Most of bothropic venoms are mainly composed by metalloproteases (SVMP), serine proteases (SVSP), phospholipases A2 (PLA2), L-amino acid oxidases (LAAO), and C-type lectins (CTL) [1, 10, 11]
LAAOs are known to be quite labile when exposed to temperature, pH, and even to the proteases present in the venom [16,17,18] and their main effects are related to cytotoxicity by inducing the release of H2O2, induction of hemorrhage and apoptosis, and inhibition and induction of platelet aggregation [19]
Summary
Classified as a neglected tropical disease by the World Health Organization (in Category A), snakebites are an important public health hazard, especially in tropical and subtropical areas [1]. In the case of the Bothrops venom, the main effects reported are involved in coagulation disorders, hemorrhage and local tissue damage [7, 8], due to single toxins and/or the synergistic action of the venom’s compounds [9]. LAAOs are known to be quite labile when exposed to temperature, pH, and even to the proteases present in the venom [16,17,18] and their main effects are related to cytotoxicity by inducing the release of H2O2, induction of hemorrhage and apoptosis, and inhibition and induction of platelet aggregation [19]. CTLs disrupts blood homeostasis by inducing/inhibiting platelet aggregation or activating/consuming coagulation factors [20]
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