We present the first karyological study on Kloss's mole Euroscaptor klossi from Chiang Rai Province of Thailand. We used differential chromosome staining methods and determined the diploid and fundamental autosomal numbers to be 36 and 54, respectively. Compared with the G- and C-banded karyotypes of the Japanese mountain mole E. mizura, the karyotype of E. klossi involved a reciprocal translocation followed by a pericentric inversion of chromosomes 1 and 16 of the ancestral karyotype of E. mizura. Some minor C-band duplications supplemented the karyological differences between the two species. The karyotype of E. klossi is distinct from that of the Malaysian mole E. micrura malayana examined previously and these species have distinct scenarios in diverging from the ancestral karyotype of the genus Euroscaptor.