engleski

Can crickets replace mice in the venom toxicity test?

The assay of lethal toxicity of venom (determination of median lethal dose of venom LD50) of snake venoms appears to be an important step to assess (and compare) venom toxic activity. This assay causes animal suffering, pain and death, but also employing large number of mice, were identified as assays requiring urgent replacement with alternative methods. Several species of snake include arthropods in their food and venom toxicity test on mice is not suitable for evaluate venom toxicity. There are 16 species of snake in Croatia, only three of them are venomous: long nosed viper (Vipera ammodytes), European adder (Vipera berus) and meadow viper (Vipera ursinii). Besides being fed to mammals, Vipera ursinii spp include insects in their diets. Crickets (Gryllus assimilis) were used as a model which mimics the natural insect prey. Here we investigate for the first time the composition and biological activity of the Vum venom composition and biological activity. The Vum venom is less lethally toxic in mice than the V. a. ammodytes (Vaa) venom, however the pattern of mice dying indicates the presence of a strong neurotoxic component. Interestingly, a two-dimensional gel electrophoresis revealed a lack of basic phospholipases, which are known neurotoxic components of Vipera venoms. Western blot of non-reduced Vum venom with anti- ammodytoxin antibodies gave no signal. Mass spectrometric identification of SDS-PAGE bands showed the lack of ammodytoxins, as well. This was finally proven by ELISA, in which Vum venom coated wells were not recognized by anti-Atx antibodies. Taken all together, VuCro venom might be a good starting material for the discovery of a novel neurotoxic component in Vipera venoms with potentially insecticidal activity, which we would not detect by venom toxicity test on mice.

venom ; lethal toxicity test ; crickets ; mice

nije evidentirano