Utilising a novel surveillance system to investigate species of () (Diptera: Ceratopogonidae) as the suspected vectors of (Kinetoplastida: Trypanosomatidae) in the Darwin region of Australia.

Up until recently, Australia was considered free of due to the absence of phlebotomine sandfly species (Diptera: Phlebotominae) known to transmit parasites in other parts of the world. The discovery of () (Kinetoplastida: Trypanosomatidae) in Northern Australia sparked questions as to the existence of alternative vectors of . This has added to the complexity of fully understanding the parasite's interaction with its vector, which is known to be very specific. Previous findings demonstrated infection beyond the blood meal stage in the day-biting midges () Kieffer (Diptera: Ceratopogonidae) implicating them in the parasite's life cycle. Currently, there is no conclusive evidence demonstrating this suspected vector to transmit . to a naïve host. Therefore, this research aimed to investigate the vector competency of day-biting midge () to transmit . utilising a novel technology that preserves nucleic acids. Honey-soaked Flinders Technology Associates (FTA®) filter-paper cards were used to obtain saliva expectorated from biting midges while sugar-feeding. . () were aspirated directly off macropods from a known -transmission site and were kept in a waxed-paper container holding a honey-coated FTA® card for feeding. Insect identification and Taqman quantitative real-time PCR (qPCR) screening assays revealed DNA in () up to 7 days post field-collection, and in an unidentified biting midge, previously known as ) sp.1. Moreover, 7/145 (4.83%) of FTA® cards were confirmed positive with . DNA after exposure to field-collected ). Additionally, FTA® cards were found to be a valuable surveillance tool, given the ease of use in the field and laboratory. Overall, our findings support previous reports on . transmission by an alternative vector to phlebotomine sandflies. Further studies identifying and isolating infective promastigotes is necessary to resolve questions on the vector.