This study examined genetic variation within and among species of Cloacina found in the common wallaroo (Macropus robustus) collected at different localities from mainland Australia, and evaluated geographical distance as a potential driver for genetic variation. The first and second internal transcribed spacers (ITS-1 and ITS-2=ITS) of nuclear ribosomal DNA were used to characterize individuals of 17 morphospecies of Cloacina that parasitize Macropus robustus and its sub-species. Results revealed intraspecific variation in ITS within some morphospecies of Cloacina. Phylogenetic analyses showed little correlation between host speciation patterns and geographical location for the majority of the nematode species, although it did suggest geographical distance was a driver for speciation within Cloacina communis, C. phaethon and C. parva. Our results suggest that nucleotide variation within Cloacina species is complex, and is likely to be propagated by factors such as geographical distance and host sub-species. Further studies determining factors involved in speciation, such as host-parasite relationships, are needed to improve our understanding of the diversity of populations of species of Cloacina.