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

Abstract

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 LD 50 value was estimated as 7.58mg/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 LD 50 value was tenfold higher than the IC 50 value. The IC 50 value was 0.62 ± 0.18 µg/mL after 48 hours treatment while the calculated value was 1.62 ± 0.25 µg/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.

Screening of cytotoxic and antimicrobial activity potential of AnatolianMacrovipera lebetina obtusa(Ophidia: Viperidae) crude venom

“…Snake venoms are a well-known natural source in drug development and discovery studies (Lewis & Garcia 2003;Koh et al 2006;Fox & Serrano 2007;Vyas et al 2013). Although there are published reports showing selected biological activities of crude venom or purified proteins of different subspecies of M. lebetina (Tõnismägi et al 2006;Son et al 2007;Bazaa et al 2009;Nalbantsoy et al 2012;Park et al 2012;Shebl et al 2012a;Morjen et al 2013), the combination of cancer cell lines and microorganisms included for screening in the present study has not been chosen in earlier studies, particularly on M. l. obtusa subspecies venom. The present study investigated cytotoxic effects on cancer and non-cancerous cells and the antimicrobial properties of Anatolian M. l. obtusa crude venom in order to assess its potential for further bioactivity-guided characterization studies.…”

“…Recent studies on the biological and proteomic characterization of various viper venoms in Turkey have been carried out (Igci & Demiralp 2012;Nalbantsoy et al 2012Nalbantsoy et al , 2013Topyıldız & Hayretdag 2012;Yalcin et al 2014). As a contribution to the authors' ongoing studies on Turkish venomous snakes, the aim of this study was to screen the cytotoxic and antimicrobial activities of Anatolian M. l. obtusa crude venom on various cancer cells and microorganisms, to assess its potential as a source of bioactive peptides/proteins which may have therapeutic value.…”

In Vitro Cytotoxic and Proapoptotic Activities of Anatolian Macrovipera Lebetina Obtusa (Dwigubski, 1832) Crude Venom on Cultured K562 Human Chronic Myelogenous Leukemia Cells.

“…24,25 Recently, snake venom studies aiming to make biological and proteomic characterization have been done. 8,[26][27][28][29][30] This study aimed to investigate the in vitro cytotoxicity and proapoptotic activity of crude venom extracted from Anatolian M. l. obtusa against K562 human chronic myelogenous leukemia cancer cell line.…”

Structure-based drug design and biological evaluation of 2-acetamidobenzothiazole derivative as EGFR kinase inhibitor

“…One of these groups served as control and received normal saline. The other groups were subjected to acute intraperitoneal toxicity study using the tested compound in different concentrations and mice were observed for toxic symptoms, signs of poisoning and mortality continuously after dosing 21 . Finally, the number of survivors in each group was noted after 24 h. The arithmetical method of Karber was used for the determination of LD 50 values (mg/kg) of the selected compounds 22 .…”

“…The LD 50 is defined as the amount of the substance required to kill 50% of a given test population, it is usually expressed as the amount of substance per kg of body weight. Compound 4 was screened for potential acute toxicity in mice 21 and the dose (mg/kg) required to kill 50% of animals (LD 50 ) within 24 h was calculated. Compound 4 showed LD 50 value of 365 mg/kg, while the LD 50 value for the reference antitumor agent, 5-fluorouracil was 115 mg/kg.…”