Análise da atividade fosfolipásica do veneno de serpentes do gênero Bothrops: um estudo comparativo

Abstract

Snake envenomation was reintroduced as a neglected tropical disease by the World Health Organization (WHO) in 2017 and represents a serious public health problem for tropical and subtropical regions of developing countries. Species of the genus Bothrops are considered Category 1 of medical importance by the WHO, accounting for approximately 85% of snakebites that occur throughout the year in Brazil. Variations in the venom composition of snakes of the same species can be determined according to genetics, diet, gender, geographicaldistribution, age or even seasonality. The objective is to investigate the phospholipase activity comparatively in adult snakes of the species: Bothrops alternatus, Bothrops atrox, Bothrops jararaca, Bothrops jararacussu, Bothrops leucurus, Bothrops moojeni, Bothrops neuwiedi and Bothrops pauloensis, as well as the identification of variations in the enzymatic activity of L - amino acid oxidase and immunoenzymatic variations of the toxins present in these venoms. Methods of characterization of biochemical

activity and immunorecognition were used: proteindosage by the Bradford method; SDS- containing polyacrylamide gel electrophoresis; High Performance Liquid Chromatography –

Reverse Phase; phospholipase activity with NOBA substrate; L-amino acid oxidase activity on L-methionine and Western Blotting. The results show interspecific and intraspecific differences. In the electrophoresis gel it was possible to observe differences involving absence, presence and intensity of bands. PLA2 and LAAO activities were notably higher in males although Balt1 and Bju1 were exceptions, with their phospholipase activity lower than that of females (Balt2 and Bju2). The commercial Bothrops antivenom produced by the Butantan Institute presents a good pattern of band recognition forming a profile similar to the gels, regardless of gender and only in the regions between 37 and 25 kDa there was a lack of recognition in bands. Although all the chromatograms presenteda very similar profile, it is noticeable that there was a notable gain in variability, some quantitative differences were observed in the chromatographic profiles of the venoms studied.