Caracterização da peçonha de Bothrops pubescens através de técnicas proteômicas e avaliação da sua toxicidade em músculo biventer cervicis de pintainho.

Abstract

The Bothrops genus comprises several species of snakes that occur in the American continent, from Mexico to Argentina. Envenomation by snakes of the Bothrops genus is characterized by local effects, including hemorrhage, swelling, pain, and myonecrosis, as well as systemic effects such as coagulopathies and renal failure. The venom composition of some Bothrops species is already known through venomics or indirectly through transcriptomic studies. Bothrops pubescens is a medium-sized venomous snake that occurs exclusively in Rio Grande do Sul and Uruguay. It was observed that its diet consists of centipedes, amphibians, lizards, snakes, mammals and birds and there is ontogenetic variation in the diet. Considering the great potential of the molecules found in Bothrops venoms, the occurrence of B. pubescens exclusively in the Pampa Biome and the lack of information about its venom, this study aimed to elucidate the composition of the venom of B. pubescens through proteomic techniques, as well as to analyze its effect on the chick biventer cervicis neuromuscular junction. For this, we performed BUP, TDP and nTDP analyses. For toxinological analysis, chick biverter cervicis was exposed to B. pubescens venom at different concentrations. With the BUP technique, 89 protein groups belonging to 13 families of toxins were identified, while with the TDP technique, 40 unique proteoforms belonging to 6 families of toxins were identified. We also identified a multi-proteoform complex of a LAAO using nTDP. In the analysis of the venom’s toxicity, we demonstrated that the venom of B. pubescens in high concentrations is capable of causing total and irreversible neuromuscular blockade against isolated preparations of birds, but in reduced concentrations, it produces a partial and reversible blockade, demonstrating its toxicity for the somatic nervous system