Determination of Toxic Activities in Bothrops spp. Snake Venoms Using Animal-Free Approaches: Correlation Between In Vitro Versus In Vivo Assays

Toxicol Sci. 2015 Oct;147(2):458-65. doi: 10.1093/toxsci/kfv140. Epub 2015 Jul 8.

Abstract

The main purpose of this study is to investigate the in vitro toxic effects of 5 Bothrops spp. snake venoms, which are part of the antigenic mixture used for the production of Brazilian antivenom, and evaluate their correlation with the in vivo toxic activities of Bothrops spp. venoms. The correlation analysis could be helpful for the replacement of living animals experimentation for in vitro bioassay. Cytotoxicity, L-amino acid oxidase (LAAO), proteolitic (serine and metalloproteinase), hyaluronidase (Hyal), and phospholipase A2 (PLA2) activities were estimated and the correlation coefficient was determined for each activity in relation to lethality, edema, hemorrhage and necrosis induced in live animals by B. jararaca, B. alternatus, B. jararacussu, B. neuwiedi, and B. moojeni venoms. The lethal activity in mice was highly related to Hyal activity (r = 0.94, p < .05), edema related to PLA2 activity (r = 0.94, p < .05), whereas the necrotizing activity showed high correlation with LAAO activity (r = 0.83, p < .05). A very significant correlation between in vitro cytotoxicity and LAAO activities was also observed (r = 0.97, p < .05).

Keywords: Bothrops venoms; cytotoxic activity; hyaluronidase activity; in vitro assays; lethal activity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animal Testing Alternatives / methods*
  • Animals
  • Antigens, Human Platelet / drug effects
  • Antigens, Human Platelet / metabolism
  • Antivenins / pharmacology
  • Bothrops*
  • Cell Line / drug effects
  • Cell Survival / drug effects
  • Crotalid Venoms / toxicity*
  • Humans
  • Hyaluronoglucosaminidase / drug effects
  • Hyaluronoglucosaminidase / metabolism
  • In Vitro Techniques
  • L-Amino Acid Oxidase / drug effects
  • L-Amino Acid Oxidase / metabolism
  • Mice
  • Proteolysis / drug effects

Substances

  • Antigens, Human Platelet
  • Antivenins
  • Crotalid Venoms
  • L-Amino Acid Oxidase
  • Hyaluronoglucosaminidase