Introduction: Snakebite envenoming is considered by the World Health Organization (WHO) as a neglected tropical disease. Currently, Bothrops snake venoms are being studied intensively, but there is little knowledge about Bothrops roedingeri venom. Objectives: To biochemically characterize B. roedingeri total venom and evaluate its myotoxic, edematogenic, and hemorrhagic activity. Materials and methods: We characterized B. roedingeri venom enzymatic activity by determining the phospholipase A2 and the proteolytic and fibrinogenolytic action using SDS-PAGE electrophoresis while we characterized its venom toxicity by determining the minimum hemorrhagic dose, the minimum edema dose, and the local and systemic myotoxic effects. Results: Bothrops roedingeri venom showed a PLA2 activity of 3.45 ± 0.11 nmoles/min, proteolytic activity of 0.145 ± 0.009 nmoles/min, and a fibrinogen coagulation index of 6.67 ± 1.33 seconds. On the other hand, it produced an minimum hemorrhagic dose of 24.5 jg, an minimum edema dose of 15.6 jg, and a pronounced local myotoxic effect evidenced by the elevation of plasma creatine kinase levels after intramuscular inoculation. The venom showed no systemic myotoxicity. Conclusions: Bothrops roedingeri venom has local hemorrhagic, edematogenic, and myotoxic activity. Enzymatically, it has high PLA2 activity, which would be responsible for the myotoxic and edematogenic effects. It also has proteolytic activity, which could affect coagulation given its ability to degrade fibrinogen, and it causes bleeding through the metalloproteases.