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Here we described a new trypanosomatid species, Phytomonas lipae, parasitizing the dock bug Coreus marginatus based on axenic culture and in vivo material. Using light and electron microscopy we characterized the development of this flagellate in the intestine, hemolymph and salivary glands of its insect host. The intestinal promastigotes of Phytomonas lipae do not divide and occur only in the anterior part of the midgut. From there they pass into hemolymph, increasing in size, and then to salivary glands, where they actively proliferate without attachment to the host's epithelium and form infective endomastigotes. We conducted molecular phylogenetic analyses based on 18s rRNA, gGAPDH and HSP83 gene sequences, of which the third marker performed the best in terms of resolving phylogenetic relationships within the genus Phytomonas. Our inference demonstrated rather early origin of the lineage comprising the new species, right after that of P. oxycareni, which represents the earliest known branch within the Phytomonas clade. This allowed us to compare the development of P. lipae and three other Phytomonas spp. in their insect hosts and reconstruct the vectorial part of the life cycle of their common ancestor.
- MeSH
- fylogeneze MeSH
- Heteroptera parazitologie MeSH
- Kinetoplastida MeSH
- pravděpodobnostní funkce MeSH
- proteiny tepelného šoku genetika MeSH
- protozoální proteiny genetika MeSH
- RNA ribozomální 18S genetika MeSH
- slinné žlázy parazitologie MeSH
- stadia vývoje * MeSH
- střeva parazitologie MeSH
- Trypanosomatina klasifikace genetika fyziologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
To further investigate the diversity of Trypanosomatidae we have examined the species present within the flea (Siphonaptera) population in the Czech Republic. Out of 1549 fleas, 239 were found to be infected by trypanosomatids. Axenic cultures were established from 90 infected specimens and 29 of them were further characterized. Molecular phylogenetic analysis of the SL RNA, gGAPDH, and SSU rRNA genes revealed a striking diversity within this group and analyzed isolates were classified into 16 Typing units (TUs) of which 15 typified new species. In addition to one Trypanosoma species, two TUs grouped within the sub-family Leishmaniinae, two clustered together with Herpetomonas, wheras 11 TUs formed a novel clade branching off between Trypanosoma spp. and remaining trypanosomatids. We propose to recognize this clade as a new genus Blechomonas and a new subfamily Blechomonadinae, and provide molecular and morphological description of 11 TUs representing this genus. Our finding of such an ancient host-specific group sheds new light at the origin of Trypanosomatidae and the roots of dixenous parasitism. The strict host restriction of Blechomonas to Siphonaptera with adult fleas' dependence on blood meal may reflect passing of parasites from larvae through pupae to adults and implies potential transmission to the warm-blooded vertebrates.
- MeSH
- biodiverzita * MeSH
- fylogeneze MeSH
- hlodavci MeSH
- molekulární sekvence - údaje MeSH
- Siphonaptera klasifikace genetika parazitologie MeSH
- Trypanosomatina klasifikace genetika izolace a purifikace fyziologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
Four new species of monoxenous kinetoplastid parasites are described from Brachycera flies, namely Wallaceina raviniae Votýpka et Lukes, 2014 and Crithidia otongatchiensis Votýpka et Lukes, 2014 from Ecuador, Leptomonas moramango Votypka et Lukes, 2014 from Madagascar, and Crithidia pragensis Votýpka, Klepetková 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. All four newly described species are represented by typical morphotypes (mainly choano- and promastigotes) and are virtually indistinguishable from other monoxenous trypanosomatids by morphology. Nevertheless, they all differ in their phylogenetic affinities. Whereas three of them grouped within the recently defined subfamily Leishmaniinae, which includes numerous representatives of the genera Leishmania Ross, 1903, Crithidia Léger, 1902 and Leptomonas Kent, 1880, the fourth species clusters together with the 'collosoma' clade (named after 'Leptomonas' collosoma Wallace, Clark, Dyer et Collins, 1960). Here we demonstrate that the 'collosoma' group represents the elusive genus Wallaceina Podlipaev, Frolov et Kolesnikov, 1999. We redefine this genus in molecular terms based on similarities of the respective molecular markers and propose to use this taxon name for the group of species of the 'collosoma' clade.