Current work was carried out to assess the possible genotoxic effect of the natural sweetener Stevia in causing DNA damage, in order to achieve such a purpose, the genetic material of mice (Mus musculus, 2n=40) and human (Homo sapiens, 2n = 46) were chosen and used employing the following short-term genotoxic bioassays recommended by Environmental Protection Agency (EPA-USA): 1- Analysis of chromosomal abnormalities in mice bone-marrow cells; 2-Analysis of mice primary spermatocytes 3-Analysis of micronucleated polychromatic erythrocytes in mice; and 4- Chromosomal aberrations in human lymphocyte culture. The obtained results clearly showed that Stevia extractwas proven to be negative in causing DNA damage, which gave a strong evidence, at the level of the present study, that Stevia extract is recommended to be suitable for human usage