A new species of the spider genus Loxosceles, L. mrazig sp. n., found in Tunisia is described and illustrated. The male bulb shows a high degree of morphological similarity to that of L. gaucho from Brazil, but the pro- portions of the palpal segments and the general colouration of the body reveal significant differences between the two species. A distance analysis of the sequences of the mitochondrial gene cox1 reveals that the specimen from Tunisia shows high genetic distance from L. gaucho (more than 20%). The American species L. gaucho and L. laeta form a sister group to the Mediterranean representatives (L. rufescens and the Tunisian specimen). Taxonomy, Araneae, Loxosceles, new species, Tunisia.