Leeches have long been considered potential vectors for the aquatic lineage of trypanosomes, while bloodsucking insects are generally considered as the vectors for the terrestrial lineage of trypanosomes. cyclops based on genetic similarity and biogeography considerations. cyclops we propose Trypanosoma cyclops australiensis as a subspecies of T. bilineata is a host for at least five lineages of Trypanosoma sp. Blood-meal analysis of leeches showed that leeches primarily contained blood from swamp wallaby (Wallabia bicolour), human (Homo sapiens) or horse (Equus sp.). Phylogeny of 18S rDNA/GAPDH/ND5 DNA sequences from primary cultures and subclones showed the trypanosomes were monophyletic, with T. Novy–MacNeal–Nicolle slopes with liquid overlay were used to isolate trypanosomes into culture that proved similar in morphology to Trypanosoma cyclops in that they contained a large numbers of acidocalcisomes. bilineata contained trypanosomes diversity profiling by deep amplicon sequencing of 18S rDNA indicated the presence of four different clusters related to the Trypanosoma (Megatrypanum) theileri. PCR targeting the 28S rDNA demonstrated that over 95% of C. Phylogenetic analysis ensued to further define the populations present. Consequently, Chtonobdella bilineata leeches were investigated for the presence of trypanosome species by polymerase chain reaction (PCR), DNA sequencing and in vitro isolation. Previously, it was suggested that haemadipsid leeches represent an important vector of trypanosomes amongst native animals in Australia.
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