The extreme biological diversity of Oceanian archipelagos has long stimulated research in ecology and evolution. However, parasitic protists in this geographic area remained neglected and no molecular analyses have been carried out to understand the evolutionary patterns and relationships with their hosts. Papua New Guinea (PNG) is a biodiversity hotspot containing over 5% of the world's biodiversity in less than 0.5% of the total land area. In the current work, we examined insect heteropteran hosts collected in PNG for the presence of trypanosomatid parasites. The diversity of insect flagellates was analysed, to our knowledge for the first time, east of Wallace's Line, one of the most distinct biogeographic boundaries of the world. Out of 907 investigated specimens from 138 species and 23 families of the true bugs collected in eight localities, 135 (15%) were infected by at least one trypanosomatid species. High species diversity of captured hosts correlated with high diversity of detected trypanosomatids. Of 46 trypanosomatid Typing Units documented in PNG, only eight were known from other geographic locations, while 38 TUs (~83%) have not been previously encountered. The widespread trypanosomatid TUs were found in both widely distributed and endemic/sub-endemic insects. Approximately one-third of the endemic trypanosomatid TUs were found in widely distributed hosts, while the remaining species were confined to endemic and sub-endemic insects. The TUs from PNG form clades with conspicuous host-parasite coevolutionary patterns, as well as those with a remarkable lack of this trait. In addition, our analysis revealed new members of the subfamilies Leishmaniinae and Strigomonadinae, potentially representing new genera of trypanosomatids.

Biology Centre Institute of Entomology Czech Academy of Sciences 370 05 České Budějovice Czechia; New Guinea Binatang Research Center Madang Papua New Guinea; University of South Bohemia Faculty of Sciences 370 05 České Budějovice Czechia

Biology Centre Institute of Parasitology Czech Academy of Sciences 370 05 České Budějovice Czechia; Department of Parasitology Faculty of Science Charles University 128 44 Prague Czechia

Biology Centre Institute of Parasitology Czech Academy of Sciences 370 05 České Budějovice Czechia; University of South Bohemia Faculty of Sciences 370 05 České Budějovice Czechia

Department of Entomology National Museum 193 00 Prague Czechia

Life Science Research Centre Faculty of Science University of Ostrava 710 00 Ostrava Czechia

Life Science Research Centre Faculty of Science University of Ostrava 710 00 Ostrava Czechia; Institute of Environmental Technologies Faculty of Science University of Ostrava 710 00 Ostrava Czechia; Martsinovsky Institute of Medical Parasitology Tropical and Vector Borne Diseases Sechenov University Moscow Russia

Life Science Research Centre Faculty of Science University of Ostrava 710 00 Ostrava Czechia; Zoological Institute of the Russian Academy of Sciences St Petersburg 199034 Russia

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