Bothrops snake venoms have been proved toxic to a variety of cell types, in both in vivo and in vitro models. Studies on the pharmacological actions of Bothrops venoms from Argentina are relatively scarce and the direct action of the crude venoms has not been assessed using cell culture models. In this work, we investigated the cytotoxicity of crude venoms from B. alternatus and B. diporus in a skeletal muscle (C2C12) cell line, which is commonly used as a model for studying the myotoxic action of snake venom. Both venoms (1.25-50 µg/mL) induced an early and significant decrease in cell viability. The cytotoxic concentration 50 (CC50), determined three hours after exposure, revealed that B. diporus venom was significantly more cytotoxic (CC50: 2 µg/mL) than B. alternatus (CC50: 5.8 µg/mL). To investigate the cell death mechanism involved, myoblast cells were examined by phase contrast microscopy and after acridine orange and ethidium bromide fluorescence staining, respectively. Our data clearly demonstrated that an apoptotic mechanism mediated this cell line destruction. The current study aimed to provide new information on the cytotoxicity mechanisms of Argentine Bothrops snake venoms on a skeletal muscle cell line

Acid proteases from sábalo stomach mucosa were recovered using salting-out procedure. This single step produced an enzyme extract purified 1.8-fold over the crude extract with a recovery of 45.1% of its initial proteolytic activity. Sábalo proteases exhibited the highest activity at 45 °C-pH 2.0, showed pH stability between 2.0 and 5.0 and retained more than 70% of its activity after incubation at pH 7.0 for 2 h. Fish extract was unstable at temperatures greater than 45 °C. Its activity was inhibited by pepstatin A but not by PMSF, while EDTA and SDS showed partial inhibitory effects. Presence of CaCl2 and MgCl2 increased the proteolytic activity, while increasing concentrations of NaCl strongly decreased it. In addition, compared to the acid extraction method, the use of sábalo enzymatic extract increased 1.7 times the yield of collagen extraction.