Determination of in vivo toxicity and in vitro cytotoxicity of venom from the Cypriot blunt-nosed viper Macrovipera lebetina lebetina and antivenom production

A Nalbantsoy,Mz Yildiz,F Sayim,I Deliloglu-Gurhan,H Arikan,B Gocmen,Nu Karabay-Yavasoglu

doi:10.1590/s1678-91992012000200011

A Nalbantsoy, Mz Yildiz + Show 5 more

Open Access

https://doi.org/10.1590/s1678-91992012000200011

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Abstract

The venomous Levantine viper, Macrovipera lebetina lebetina is endemic to Cyprus. The objective of this study was to investigate in vitro cytotoxicity, in vivo lethality, and antivenom production followed by a re-immunization schedule in mice against Macrovipera lebetina lebetina venom. The LD50 value was estimated as 7.58 mg/kg within 24 hours by different venom doses administrated intraperitoneally in mice. Freund's complete and incomplete adjuvants were used for first and second immunization of mice in antivenom production. A cell-based assay was performed to determine the effects of Macrovipera lebetina lebetina venom and antivenom neutralizing potency on L929 cell viability. The snake venom toxicity and cytotoxicity were examined and comparison of results showed good correlation, the LD50 value was tenfold higher than the IC50 value. The IC50 value was 0.62 ± 0.18 mL after 48 hours treatment while the calculated value was 1.62 ± 0.25 mL for the culture media totally refreshed after two hours treatment with venom. The in vitro efficacy of antivenom against Macrovipera lebetina lebetina venom was found to be low. This is the first report that describes the in vivo and in vitro toxic effects of Macrovipera lebetina lebetina venom and antivenom production against this species.

Highlights

Snake envenomation is a major public health issue that provokes thousands of deaths throughout the globe
Our interpretation of the results suggests that the LD50 value is approximately ten times greater than the IC50 value for crude venom from Macrovipera lebetina lebetina
This study indicated that Macrovipera lebetina lebetina antivenom production was achieved in a mouse model

Summary

Introduction

Snake envenomation is a major public health issue that provokes thousands of deaths throughout the globe. There are nearly 3,000 different species of snakes found in the world of which approximately 300 are venomous [13]. The Levantine viper, Macrovipera lebetina, is a venomous viper species found, in Cyprus, the Middle East, northern Africa, Cyclades, Turkey and central Asia [4,5,6,7,8]. The subspecies lebetina seems to be endemic to Cyprus. It is the only snake in Cyprus which can be dangerous to humans [9]. As reported by Cesaretli and Ozkan [10], the National Poison Information Center recorded 550 snakebite cases between 1995 and

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