On the integrative taxonomy of Trichophoromyia Barretto, 1962 and its relationship with Nyssomyia Barretto, 1962 (Diptera, Psychodidae, Phlebotominae): species delimitation, phylogeny, genus and subgenus description.

BACKGROUND: The sand fly genus Trichophoromyia Barretto, 1962 is one of the most diverse inthe subfamily Phlebotominae. The taxonomy and systematics of this group is complex due toboth a high similarity among species and unclear relationships among other sand fly groupswithin the subtribe Psychodopygina Galati, 1995. Despite their great relevance as vectors of Leishmania spp., few studies have explored the usefulness of molecular markers in studyingthe diversity of this group. METHODS: Here, we evaluated the use of barcode sequences of the cytochrome coxidasesubunit I gene (COI) for identifying several Trichophoromyia spp., by inferring intra- andinterspecific genetic distances, in addition to performing a set of several single-locus speciesdelimitation approaches using discovery methods. Moreover, we employed a multilocusdataset of four independent molecular markers (COI , ITS2 , 28S and PARA) to infer thephylogenetic species tree, estimate divergence times and delimit species under a validationmodel. RESULTS: The phylogenetic inferences confirmed the paraphyly of Trichophoromyia and Nyssomy ia Barretto, 1962. Thus, two new genera, named Reburrus gen. nov. and Shawmyia gen. nov., were proposed to accommodate sand flyspecies that did not fit in the aforementioned groups. Additionally, a new subgenus wasproposed: Trichophoromyia (Dilermandomyia) subg. nov., containing most speciesof Trichophoromyia . A recent speciation history was also estimated, with most of the speciesstudied diversifying during the Pleistocene. However, our dataset was insufficient to fullyresolve relationships within Trichophoromyia (Dilermandomyia) subg. nov. Many speciesshowed paraphyletic patterns in the gene trees, and some could not be reliably identified anddelimited using both COI barcodes and multilocus tools. CONCLUSIONS: The sand fly genus Trichophoromyia exhibits a complex diversification history.Our phylogenetic inference and morphological observations of Nyssomyia and Trichophoromy ia, allowed us to propose new groups for the Psychodopygina subtribe. However, theprevalence of species-level paraphyletic patterns for Trichophoromyia (Dilermandomyia) subg.nov., showed that further assessment of this group requires a broader locus sampling combined with detailed morphological analysis.