Phlebotomine sand flies (Diptera: Phlebotominae) are the principal vectors of Leishmania spp. (Kinetoplastida: Trypanosomatidae). In Central Europe, Phlebotomus mascittii is the predominant species, but largely understudied. To better understand factors driving its current distribution, we infer patterns of genetic diversity by testing for signals of population expansion based on two mitochondrial genes and model current and past climate and habitat suitability for seven post-glacial maximum periods, taking 19 climatic variables into account. Consequently, we elucidate their connections by environmental-geographical network analysis. Most analyzed populations share a main haplotype tracing back to a single glacial maximum refuge area on the Mediterranean coasts of South France, which is supported by network analysis. The rapid range expansion of Ph. mascittii likely started in the early mid-Holocene epoch until today and its spread possibly followed two routes. The first one was through northern France to Germany and then Belgium, and the second across the Ligurian coast through present-day Slovenia to Austria, toward the northern Balkans. Here we present a combined approach to reveal glacial refugia and post-glacial spread of Ph. mascittii and observed discrepancies between the modelled and the current known distribution might reveal yet overlooked populations and potential further spread.
- MeSH
- hmyz - vektory genetika MeSH
- Leishmania * MeSH
- Phlebotomus * genetika MeSH
- Psychodidae * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Evropa MeSH
The subgenus Transphlebotomus comprises sand fly species with distribution markedly restricted to the Mediterranean basin and suspected of Leishmania transmission. Only three species, Phlebotomus mascittii, Phlebotomus canaaniticus and Phlebotomus economidesi, have been described up to the present. Due to their similar morphology, proper identification remains difficult and relies mainly on molecular markers. We studied sand fly species of this subgenus from Crete and south-western coast of Anatolia. Based on the sequencing analysis of mitochondrial genes (cytochrome b, NADH dehydrogenase subunit 4, cytochrome oxidase I), two new Transphlebotomus species were identified and subsequently distinguished also by morphological characters: Phlebotomus anatolicus n. sp. and Phlebotomus killicki n. sp. Moreover, Ph. economidesi, previously only recorded from Cyprus, was found in Turkey sympatrically with these two new species. Based on the divergence time estimates, the first split has occurred in the subgenus Transphlebotomus ∼10 million years ago and the paleogeographic events took place around the Aegean and Mediterranean regions were suggested as the main drivers of the diversification of the subgenus. Our findings indicate that for Transphlebotomus species, morphological identification should be confirmed by molecular approaches, especially for investigations concerning their possible vectorial role in Leishmania transmission.
- MeSH
- cytochromy b genetika MeSH
- druhová specificita MeSH
- fylogeneze MeSH
- fylogeografie MeSH
- hmyz - vektory anatomie a histologie klasifikace genetika MeSH
- hmyzí proteiny genetika MeSH
- leishmanióza přenos MeSH
- lidé MeSH
- multilokusová sekvenční typizace MeSH
- Phlebotomus anatomie a histologie klasifikace genetika MeSH
- vznik druhů (genetika) MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Evropa MeSH
BACKGROUND: Rapid, accurate and high-throughput identification of vector arthropods is of paramount importance in surveillance programmes that are becoming more common due to the changing geographic occurrence and extent of many arthropod-borne diseases. Protein profiling by MALDI-TOF mass spectrometry fulfils these requirements for identification, and reference databases have recently been established for several vector taxa, mostly with specimens from laboratory colonies. METHODS: We established and validated a reference database containing 20 phlebotomine sand fly (Diptera: Psychodidae, Phlebotominae) species by using specimens from colonies or field-collections that had been stored for various periods of time. RESULTS: Identical biomarker mass patterns ('superspectra') were obtained with colony- or field-derived specimens of the same species. In the validation study, high quality spectra (i.e. more than 30 evaluable masses) were obtained with all fresh insects from colonies, and with 55/59 insects deep-frozen (liquid nitrogen/-80 °C) for up to 25 years. In contrast, only 36/52 specimens stored in ethanol could be identified. This resulted in an overall sensitivity of 87 % (140/161); specificity was 100 %. Duration of storage impaired data counts in the high mass range, and thus cluster analyses of closely related specimens might reflect their storage conditions rather than phenotypic distinctness. A major drawback of MALDI-TOF MS is the restricted availability of in-house databases and the fact that mass spectrometers from 2 companies (Bruker, Shimadzu) are widely being used. We have analysed fingerprints of phlebotomine sand flies obtained by automatic routine procedure on a Bruker instrument by using our database and the software established on a Shimadzu system. The sensitivity with 312 specimens from 8 sand fly species from laboratory colonies when evaluating only high quality spectra was 98.3 %; the specificity was 100 %. The corresponding diagnostic values with 55 field-collected specimens from 4 species were 94.7 % and 97.4 %, respectively. CONCLUSIONS: A centralized high-quality database (created by expert taxonomists and experienced users of mass spectrometers) that is easily amenable to customer-oriented identification services is a highly desirable resource. As shown in the present work, spectra obtained from different specimens with different instruments can be analysed using a centralized database, which should be available in the near future via an online platform in a cost-efficient manner.
- MeSH
- entomologie metody MeSH
- hmyzí proteiny analýza MeSH
- molekulární sekvence - údaje MeSH
- Psychodidae chemie klasifikace MeSH
- respirační komplex IV genetika MeSH
- sekvenční analýza DNA MeSH
- senzitivita a specificita MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice metody MeSH
- teplota MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- validační studie MeSH
- MeSH
- feces parazitologie MeSH
- Gorilla gorilla parazitologie MeSH
- Leishmania major izolace a purifikace MeSH
- zdroje nemoci * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- dopisy MeSH
- komentáře MeSH
