ABSTRACT
Various factors, such as geographical origin, climate, sex, age and diet can influence the composition and pathophysiological activities of snake venoms. In this study, we examined the sexual and ontogenetic variations in the venom of Bothrops leucurus, a pitviper responsible for more than 80% of the snakebites in the state of Bahia, northeastern Brazilian. The venoms of 31 snakes were pooled according to sex and age (young, adult and old) and screened by SDS-PAGE (in reducing and non-reducing conditions), reverse-phase high performance liquid chromatography (RP-HPLC), gelatin zymography, and immunoblotting with therapeutic bothropic antivenom (BAV) from the Instituto Butantan. The electrophoretic and chromatographic profiles showed intraspecific ontogenetic variation, whereas sexual variations were less evident. All venoms showed gelatinolytic activity associated with 50-75 kDa protein bands. In addition, all venoms, regardless of the snakes' sex and age, cross-reacted to similar extents with BAV. Our findings show that B. leucurus venom changes during ontogenetic development and demonstrate sexual differences in its composition, indicating differences in biological activity.
Subject(s)
Bothrops , Crotalid Venoms , Animals , Brazil , Chromatography, High Pressure Liquid , Sex FactorsABSTRACT
Various factors, such as geographical origin, climate, sex, age and diet can influence the composition and pathophysiological activities of snake venoms. In this study, we examined the sexual and ontogenetic variations in the venom of Bothrops leucurus, a pitviper responsible for more than 80% of the snakebites in the state of Bahia, northeastern Brazilian. The venoms of 31 snakes were pooled according to sex and age (young, adult and old) and screened by SDS-PAGE (in reducing and non-reducing conditions), reverse-phase high performance liquid chromatography (RP-HPLC), gelatin zymography, and immunoblotting with therapeutic bothropic antivenom (BAV) from the Instituto Butantan. The electrophoretic and chromatographic profiles showed intraspecific ontogenetic variation, whereas sexual variations were less evident. All venoms showed gelatinolytic activity associated with 50–75 kDa protein bands. In addition, all venoms, regardless of the snakes' sex and age, cross-reacted to similar extents with BAV. Our findings show that B. leucurus venom changes during ontogenetic development and demonstrate sexual differences in its composition, indicating differences in biological activity.
ABSTRACT
Os venenos de serpentes são misturas complexas constituídas, em grande parte, de proteínas enzimáticas ou não; somando aproximadamente 90% do peso seco do veneno. O estudo da composição dos venenos colabora com o contínuo avanço biotecnológico, com o desenvolvimento de fármacos e na compreensão dos processos bioquímicos e fisiopatológicos dos organismos. O “grupo Bothrops atrox” corresponde a um conjunto de serpentes peçonhentas amplamente distribuídas na região neotropical e que apresenta um complexo padrão de variação morfológica interespecífico e intraespecífico. Por meio de experimentos in vitro e in vivo, e contribuindo para o melhor entendimento das variações interespecíficas, bem como da filogenia dentro do complexo, foi proposto um modelo de estudo composto por análises biológicas e bioquímicas de 4 amostras de “Pools” de venenos (1 amostra por espécie) de exemplares machos e fêmeas, de diversas idades e procedências das espécies Bothrops atrox, B. leucurus, B. marajoensis e B. moojeni, visando uma análise comparativa por meio da quantificação proteica, análise dos perfis eletroforéticos e de zimografia dos venenos bem como determinação da Dose Mínima Coagulante, atividade hemorrágica, letalidade e a soroneutralização destas atividades pelo antiveneno botrópico. Os venenos das espécies estudadas apresentaram semelhanças em relação à algumas atividades bioquímicas, porém mostrando maiores particularidades nos testes de atividade biológica, sendo as atividades hemorragica e de letalidade, assim como suas respectivas soroneutralizações.
ABSTRACT
Various factors, such as geographical origin, climate, sex, age and diet can influence the composition and pathophysiological activities of snake venoms. In this study, we examined the sexual and ontogenetic variations in the venom of Bothrops leucurus, a pitviper responsible for more than 80% of the snakebites in the state of Bahia, northeastern Brazilian. The venoms of 31 snakes were pooled according to sex and age (young, adult and old) and screened by SDS-PAGE (in reducing and non-reducing conditions), reverse-phase high performance liquid chromatography (RP-HPLC), gelatin zymography, and immunoblotting with therapeutic bothropic antivenom (BAV) from the Instituto Butantan. The electrophoretic and chromatographic profiles showed intraspecific ontogenetic variation, whereas sexual variations were less evident. All venoms showed gelatinolytic activity associated with 50–75 kDa protein bands. In addition, all venoms, regardless of the snakes' sex and age, cross-reacted to similar extents with BAV. Our findings show that B. leucurus venom changes during ontogenetic development and demonstrate sexual differences in its composition, indicating differences in biological activity.
