A karyological and morphological analysis of 97 specimens from eastern Australia belonging to the Gyratrix hermaphroditus species complex was performed. Based on karyotype and on details of sclerotized structures of the copulatory organ, the existence of at least eight sibling species in eastern Australia could be recognised. Some of the siblings have a wide distribution across eastern and northern Australia. Populations of wide-ranging species often showed degrees of karyological and morphological differentiation. The diversity of the group is particularly high in tropical Australia. Distribution of siblings appears to be affected by ecological and physical barriers, and determined by sediment texture.