In vitro hemolytic activity of Bothrops lanceolatus (fer-de-lance) venom

Abstract

Bothrops lanceolatus venom contains a variety of enzymatic and biological activities. The present work investigated the hemolytic activity of this venom and its phospholipase A 2 (PLA 2 ). Bothrops lanceolatus venom (6.7 µg/mL) caused indirect hemolysis of cow, horse, rat and sheep erythrocytes, with horse erythrocytes being the most sensitive; no direct hemolysis was observed. Hemolysis in sheep erythrocytes was concentration-dependent (5-11.7 µg/mL) and markedly attenuated by heating the venom for 30 minutes at ≥ 40°C and by the PLA 2 inhibitor p-bromophenacyl bromide. An acidic PLA 2 (5 µg/mL) purified from B. lanceolatus venom also caused hemolysis. This PLA 2 showed immunoprecipitin lines with antivenom against B. lanceolatus, which suggests that the enzymatic and hemolytic activities of this enzyme may be neutralized during antivenom therapy. These results indicate that B. lanceolatus venom and its PLA 2 can cause hemolysis in vitro.

New Insights into Immunopathology Associated to Bothrops lanceolatus Snake Envenomation: Focus on PLA2 Toxin

“…It is important to mention that we have selected the lowest concentration of PLA 2 , able to promote direct hemolysis (Figure 3A: 5 mg), to perform the next experiments of the present study. This PLA 2 hemolytic dose is also in line with data from the literature [31].…”

“…It is important to mention that we have selected the lowest concentration of PLA2, able to promote direct hemolysis (Figure 3A: 5 mg), to perform the next experiments of the present study. This PLA2 hemolytic dose is also in line with data from the literature [31]. A,B) Hemoglobin release after incubation with increasing concentrations of PLA2 (5, 10, 15, 20 and 25 µg/mL), crude venom (25 µg/mL) or saline (negative control) for 30 min at 37 °C.…”