Kinetoplastida; Trypanosomatidae; monoxenous kientoplastids; Leishmaniinae; molecular taxonomy; phylogeny

Four new species of monoxenous kinetoplastid parasites are described from Brachycera flies, namely Wallaceina raviniae Votypka et Lukes, 2014 and Crithidia otongatchiensis Votypka et Lukes, 2014 from Ecuador, Leptomonas moramango Votypka et Luke, 2014 from Madagascar, and Crithidia pragensis Votypka, Klepetkova et Lukes, 2014 from the Czech Republic. The new species are described here based on sequence analysis of their spliced leader (SL) RNA, glycosomal glyceraldehyde 3-phosphate dehydrogenase (gGAPDH) and small subunit (SSU) rRNA genes, as well as their morphology and ultrastructure. High-pressure freezing and Bernhard's EDTA regressive staining, used for the first time for monoxenous (one host) trypanosomatids, revealed the presence of viral particles with cytosolic localization in one and unique mitochondrial localization in another species. In accordance with previous observations, our results emphasize a discrepancy between morphology and molecular taxonomy of the family Trypanosomatidae?