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The order Piroplasmida encompasses two main families:  Babesiidae  and  Theileriidae , containing tick-borne pathogens of veterinary and medical importance worldwide. While only three genera ( Babesia ,  Cytauxzoon  and  Theileria ) comprising piroplasm parasites are currently recognised, phylogenetic studies at the 18S rRNA (18S) gene suggest that these organisms represent at least ten lineages, one of which comprises the relatively unique and highly diverse  Theileria  spp. from Australian marsupials and ticks. As an alternative to analysing 18S sequences alone, sequencing of mitochondrial genes has proven to be useful for the elucidation of evolutionary relationships amongst some groups of piroplasms. This research aimed to characterise piroplasms from Australian native mammals and ticks using multiple genetic markers (18S, cytochrome c, oxidase subunit III ( cox3)  and cytochrome B (c ytB )) and microscopy. For this, nearly complete piroplasm-18S sequences were obtained from 32 animals belonging to six marsupial species: eastern bettong ( Bettongia gaimardi ), eastern quoll ( Dasyurus viverrinus ), eastern grey kangaroo ( Macropus giganteus ), swamp wallaby ( Wallabia bicolor ), quokka ( Setonix brachyurus ) and Gilbert’s potoroo ( Potorous gilbertii ). The organisms detected represented eight novel  Theileria  genotypes, which formed five sub-clades within the main marsupial clade containing previously reported Australian marsupial and tick-derived  Theileria  spp. A selection of both novel and previously described Australian piroplasms at the 18S were also successfully characterised, for the first time, at the  cox3  and  cytB  loci, and corroborated the position of Australian native theilerias in a separate, well-supported clade. Analyses of the  cox3  and  cytB  genes also aided in the taxonomic resolution within the clade of Australian Piroplasmida. Importantly, microscopy and molecular analysis at multiple loci led to the discovery of a unique piroplasm species that clustered with the Australian marsupial theilerias, for which we propose the name  Theileria lupei  n. sp.

plos one

Sequence analyses at mitochondrial and nuclear loci reveal a novel Theileria sp. and aid in the phylogenetic resolution of piroplasms from Australian marsupials and ticks