A phylogeny of sand flies (Diptera: Psychodidae: Phlebotominae), using recent Ethiopian collections and a broad selection of publicly available DNA sequence data

Abstract

Sand flies in the psychodid subfamily Phlebotominae carry important human pathogens in the trypanosomatid protozoan genus Leishmania (Cupolillo). Despite the fact that hundreds of sequences for this group are now publicly available, they constitute different sets of taxa and genetic markers. Integrating these data to construct a molecular phylogeny of the family is a significant bioinformatics challenge. We used sequences of eight markers obtained from freshly collected sand flies from Ethiopia and combined them with over 1300 publicly available sequences, performing a combined analysis after generating single terminal sequences from ancestral reconstructions for some individual markers. The resulting phylogeny had 113 terminals and recovered Phlebotominae and certain species groups as monopheletic. Although the 20 outgroups in Psychodinae were recovered as a well-resolved clade with bootstrap support for many internal clades, Phlebotominae was recovered as several lineages with unclear relationships among them. However, phlebotomines clustered by geographic region, the most notable being all the New World species except Brumptomyia (Galati), which were recovered as monophyletic. Our phylogeny suggests a Sub-Saharan African or South Asian origin for the subfamily, which subsequently expanded to the north and west, and eventually to the New World. Supported species groups are often composed of widespread species with overlapping ranges. This result highlights the need for a large increase in the amount and diversity of molecular sequence data, and a broad selection of terminals, to test taxonomic hypotheses and examine speciation processes in this important group of flies.

Sergentomyia schwetzi: Salivary gland transcriptome, proteome and enzymatic activities in two lineages adapted to different blood sources

“…The phylogenetic and sequence analysis showed that most of the S. schwetzi salivary protein sequences are more divergent, compared to the Lutzomyia and Phlebotomus proteins. However, the results of phylogenetic comparison of S. schwetzi salivary protein families (antigen 5-related proteins, lufaxins, YRPs, and apyrases) were in accordance with the established sand fly phylogeny [70,71]. The S. schwetzi OBPs showed high sequence divergence, which is together with high gene duplication rates, typical for the sequences belonging to D7-related and PpSP15-like families [60].…”

“…Third protein, SschwYRP3, possessed 2 putative O-glycosylation sites and predicted Mw of 43.8 kDa (secreted variant). All sand fly YRPs sequences share similar Cys motif (CX 67-72 CX 122-123 CX [71][72][73][74][75][76] C modified according to [60]). Interestingly this sequence design was modified for all three SschwYRPs-SschwYRP1: CX 64 CX 120 CX 69 C; SschwYRP2: CX 57 CX 114 CX 73 C; SschwYRP3: CX 66 CX 138 CX 73 C (S7 Fig).…”

Engineering a vector-based pan-Leishmania vaccine for humans: proof of principle

“…However, using a sand fly saliva-based vaccine presents its own challenges. The natural evolutionary pressure that conditioned the speciation of the different Phlebotominae that transmit leishmaniasis 47 , brought as a logical consequence the lower potential for cross-reactivity (due to the known—although not strict—relation between T cell epitopes’ conservation and immunogenicity 48 ). Therefore defined sand fly salivary proteins will have a limited anti- Leishmania vaccine spectrum: effectiveness will probably be limited to the geographical region where the vector species is prevalent, and consequently to the parasite species and disease manifestation associated to that specific vector 21 , 22 .…”

Evolution, systematics and historical biogeography of sand flies of the subgenus Paraphlebotomus (Diptera, Psychodidae, Phlebotomus) inferred using restriction-site associated DNA markers

“…bergeroti) with poor support. Finally, Grace-Lema et al [ 20 ] used a patchy matrix of 7 genes (65% missing data, ca 45% of the species with more than 85% missing data) to infer a phylogeny of phlebotomine sand flies (113 terminals). However, as for the rest of the tree, relationships between the four species of Paraphlebotomus were poorly resolved.…”