The Wallacean island of Timor is of particular biological interest due to its relatively large size and transitional location between the Indo-Malayan and Australasian biogeographic realms. However, the origins and levels of endemism of its invertebrate fauna are poorly known. A recent study of Timorese ants revealed a diverse fauna with predominantly Indo-Malayan affinities, but species-level taxonomy was considered to be too poorly understood for an analysis of levels of endemism. The highly diverse Old World tropical genus Polyrhachis represents a notable exception, and here we analyse levels of endemism in the Polyrhachis fauna of Timor and surrounding islands. We supplement the species listed in the previous study with additional collections to record a total of 35 species of Polyrhachis from Timor and surrounding islands. Only 14 (40%) of the 35 species could be named (P. constricta, P. costulata, P. gab, P. sokolova, P. hera, P. illaudata, P. rixosa, P. acantha chrysophanes, P. saevissima, P. bicolor, P. cryptoceroides, P. dives, P. longipes and P. olybria), and the large majority of the remaining species have not previously been collected. These are very likely to be endemic to Timor and surrounding islands, and point to remarkably high levels (>50%) of endemism in the regional ant fauna.