Development of an innovative zebrafish (Danio rerio) smear/imprint in vitro for rapid preliminary inspection of the snake venom toxinological effects

Abstract

ABSTRACT The current assay describes the implementation of an innovative animal model to in vestigate toxin pathology in vivo, with the possibility of using zebrafish larvae (Danio rerio) (7dpf) smear/imprint technique for preliminary inspection of snake venom activities. Molecu lar exclusion chromatography was done using proteins less than 30 kDa for the larvae and thus be assured that we had neurotoxins in the inoculum, since neurotoxins are generally of low molecular weight. The cuaima (Lachesis muta muta) venom, in addition to being neuro toxic, it is also haemorrhagic and proteolytic. Evident toxins damages were caused by the snake venom on zebrafish larvae, such as epidermal, neurotoxic and myotoxic alterations that by direct testing of a larval smear/imprint procedure they were quickly and easily demonstrat ed. Here was established a number of alterations, such as an obvious curvature in the caudal segment, which was accompanied by the death of the population of epidermis cells (tested by acridine orange) at the end of the larval tail; cells with highly vacuolated cytoplasm; nerve cells with a high presence of vacuolated cell bodies and irregular band pattern in the sarco mere units of muscle tissue. This method will allow to have a quick preliminary vision of the injuries that the snake venom is capable of producing in several tissues, showing in terms of time an advantage on the classical histopathological methods.