Sand flies (Diptera: Psychodidae: Phlebotominae) are blood-feeding insects that transmit the protozoan parasites Leishmania spp. and various arthropod-borne (arbo) viruses. While in Mediterranean parts of Europe the sand fly fauna is diverse, in Central European countries including Austria mainly Phlebotomus mascittii is found, an assumed but unproven vector of Leishmania infantum. To update the currently understudied sand fly distribution in Austria, a sand fly survey was performed and other entomological catches were screened for sand flies. Seven new trapping locations of Ph. mascittii are reported including the first record in Vienna, representing also one of the first findings of this species in a city. Morphological identification, supported by fluorescence microscopy, was confirmed by two molecular approaches, including sequencing and matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) protein profiling. Sand fly occurrence and activity were evaluated based on surveyed locations, habitat requirements and climatic parameters. Moreover, a first comparison of European Ph. mascittii populations was made by two marker genes, cytochrome c oxidase subunit 1 (COI), and cytochrome b (cytb), as well as MALDI-TOF mass spectra. Our study provides new important records of Ph. mascittii in Austria and valuable data for prospective entomological surveys. MALDI-TOF MS protein profiling was shown to be a reliable tool for differentiation between sand fly species. Rising temperatures and globalization demand for regular entomological surveys to monitor changes in species distribution and composition. This is also important with respect to the possible vector competence of Ph. mascittii.

BioCeV Institute of Microbiology of the Czech Academy of Sciences 252 50 Vestec Czech Republic

Department of Biodiversity FAMNIT University of Primorska 6000 Koper Capodistria Slovenia

Department of Dermatology and Tropical Medicine Military Medical Cluster East Austrian Armed Forces 1210 Vienna Austria

Department of Integrative Biology and Biodiversity Research Institute of Zoology University of Natural Resources and Life Sciences Vienna 1180 Vienna Austria

Department of Parasitology Faculty of Science Charles University Prague 128 43 Prague Czech Republic

Department of Pathobiology Institute of Parasitology University of Veterinary Medicine Vienna 1210 Vienna Austria

Division Microsystems Engineering Technical University of Applied Sciences Wildau 15745 Wildau Germany

Division Molecular Biotechnology and Functional Genomics Technical University of Applied Sciences Wildau 15745 Wildau Germany

Federal Ministry of Defence Division of Science Research and Development 1090 Vienna Austria

Institute of Specific Prophylaxis and Tropical Medicine Center for Pathophysiology Infectiology and Immunology Medical University of Vienna 1090 Vienna Austria

Research Platform Models and Simulation Leibniz Centre for Agricultural Landscape Research 15374 Müncheberg Germany

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Akhoundi M., Kuhls K., Cannet A., Votýpka J., Marty P., Delaunay P., Sereno D. A Historical Overview of the Classification, Evolution, and Dispersion of Leishmania Parasites and Sandflies. PLoS Negl. Trop. Dis. 2016;10:e0004349. doi: 10.1371/journal.pntd.0004349. PubMed DOI PMC

Ready P.D. Biology of Phlebotomine Sand Flies as Vectors of Disease Agents. Annu. Rev. Entomol. 2013;58:227–250. doi: 10.1146/annurev-ento-120811-153557. PubMed DOI

Naucke T.J., Pesson B. Presence of Phlebotomus (Transphlebotomus) mascittii Grassi, 1908 (Diptera: Psychodidae) in Germany. Parasitol. Res. 2000;86:335–336. doi: 10.1007/s004360050053. PubMed DOI

Naucke T.J., Schmitt C. Is leishmaniasis becoming endemic in Germany? Int. J. Med. Microbiol. 2004;293:179–181. doi: 10.1016/S1433-1128(04)80036-6. PubMed DOI

Naucke T.J., Lorentz S., Rauchenwald F., Aspöck H. Phlebotomus (Transphlebotomus) mascittii Grassi, 1908, in Carinthia: First record of the occurrence of sandflies in Austria (Diptera: Psychodidae: Phlebotominae) Parasitol. Res. 2011;109:1161–1164. doi: 10.1007/s00436-011-2361-0. PubMed DOI

Poeppl W., Obwaller A.G., Weiler M., Burgmann H., Mooseder G., Lorentz S., Rauchenwald F., Aspöck H., Walochnik J., Naucke T.J. Emergence of sandflies (Phlebotominae) in Austria, a Central European country. Parasitol. Res. 2013;112:4231–4237. doi: 10.1007/s00436-013-3615-9. PubMed DOI PMC

Obwaller A.G., Poeppl W., Naucke T.J., Luksch U., Mooseder G., Aspöck H., Walochnik J. Stable populations of sandflies ( Phlebotominae ) in Eastern Austria: A comparison of the trapping seasons 2012 and 2013. Trends Entomol. 2014;2:1–5.

Dvořák V., Hlavackova K., Kocisova A., Volf P. First record of Phlebotomus (Transphlebotomus) mascittii in Slovakia. Parasite. 2016;23:48. doi: 10.1051/parasite/2016061. PubMed DOI PMC

Kniha E., Dvořák V., Milchram M., Obwaller A.G., Koehsler M., Poeppl W., Antoniou M., Chaskopoulou A., Paronyan L., Stefanovska J., et al. Phlebotomus (Adlerius) simici Nitzulescu, 1931: First record in Austria and phylogenetic relationship with other Adlerius species. Parasit. Vectors. 2020 (in press) PubMed PMC

Aransay A.M., Testa J.M., Morillas-Márquez F., Lucientes J., Ready P.D. Distribution of sandfly species in relation to canine leishmaniasis from the Ebro Valley to Valencia, northeastern Spain. Parasitol. Res. 2004;94:416–420. doi: 10.1007/s00436-004-1231-4. PubMed DOI

Depaquit J., Naucke T.J., Schmitt C., Ferté H., Léger N. A molecular analysis of the subgenus Transphlebotomus Artemiev, 1984 (Phlebotomus, Diptera, Psychodidae) inferred from ND4 mtDNA with new northern records of Phlebotomus mascittii Grassi, 1908. Parasitol. Res. 2005;95:113–116. doi: 10.1007/s00436-004-1254-x. PubMed DOI

Veronesi E., Pilani R., Carrieri M., Bellini R. Trapping sand flies (Diptera: Psychodidae) in the Emilia-Romagna region of northern Italy. J. Vector Ecol. 2007;32:313–318. doi: 10.3376/1081-1710(2007)32[313:TSFDPI]2.0.CO;2. PubMed DOI

Naucke T.J., Menn B., Massberg D., Lorentz S. Winter activity of Phlebotomus (Transphlebotomus) mascittii, Grassi 1908 (Diptera: Psychodidae) on the island of Corsica. Parasitol. Res. 2008;103:477–479. doi: 10.1007/s00436-008-1002-8. PubMed DOI

Bosnić S., Gradoni L., Khoury C., Maroli M. A review of leishmaniasis in Dalmatia (Croatia) and results from recent surveys on phlebotomine sandflies in three southern counties. Acta Trop. 2006;99:42–49. doi: 10.1016/j.actatropica.2006.06.009. PubMed DOI

Grimm F., Gessler M., Jenni L. Aspects of sandfly biology in southern Switzerland. Med. Vet. Entomol. 1993;7:170–176. doi: 10.1111/j.1365-2915.1993.tb00671.x. PubMed DOI

Praprotnik E., Zupan S., Ivović V. Morphological and Molecular Identification of Phlebotomus mascittii Grassi, 1908 Populations From Slovenia. J. Med. Entomol. 2019;56:565–568. doi: 10.1093/jme/tjy176. PubMed DOI

Vaselek S., Dvořák V., Hlavackova K., Ayhan N., Halada P., Oguz G., Ivović V., Ozbel Y., Charrel R.N., Alten B., et al. A survey of sand flies (Diptera, Phlebotominae) along recurrent transit routes in Serbia. Acta Trop. 2019;197:105063. doi: 10.1016/j.actatropica.2019.105063. PubMed DOI

Farkas R., Tánczos B., Bongiorno G., Maroli M., Dereure J., Ready P.D. First Surveys to Investigate the Presence of Canine Leishmaniasis and Its Phlebotomine Vectors in Hungary. Vector-Borne Zoonotic Dis. 2011;11:823–834. doi: 10.1089/vbz.2010.0186. PubMed DOI

Dantas-Torres F., Tarallo V.D., Latrofa M.S., Falchi A., Lia R.P., Otranto D. Ecology of phlebotomine sand flies and Leishmania infantum infection in a rural area of southern Italy. Acta Trop. 2014;137:67–73. doi: 10.1016/j.actatropica.2014.04.034. PubMed DOI

Berdjane-Brouk Z., Charrel R.N., Bitam I., Hamrioui B., Izri A. Record of Phlebotomus (Transphlebotomus) mascittii Grassi, 1908 and Phlebotomus (Laroussius) chadli Rioux, Juminer & Gibily, 1966 female in Algeria. Parasite. 2011;18:337–339. PubMed PMC

Melaun C., Krüger A., Werblow A., Klimpel S. New record of the suspected leishmaniasis vector Phlebotomus (Transphlebotomus) mascittii Grassi, 1908 (Diptera: Psychodidae: Phlebotominae)—The northernmost phlebotomine sandfly occurrence in the Palearctic region. Parasitol. Res. 2014;113:2295–2301. doi: 10.1007/s00436-014-3884-y. PubMed DOI

Obwaller A.G., Karakus M., Poeppl W., Töz S., Özbel Y., Aspöck H., Walochnik J. Could Phlebotomus mascittii play a role as a natural vector for Leishmania infantum? New data. Parasit. Vectors. 2016;9:458. doi: 10.1186/s13071-016-1750-8. PubMed DOI PMC

Zanet S., Sposimo P., Trisciuoglio A., Giannini F., Strumia F., Ferroglio E. Epidemiology of Leishmania infantum, Toxoplasma gondii, and Neospora caninum in Rattus rattus in absence of domestic reservoir and definitive hosts. Vet. Parasitol. 2014;199:247–249. doi: 10.1016/j.vetpar.2013.10.023. PubMed DOI

Bogdan C., Schönian G., Bañuls A.L., Hide M., Pratlong F., Lorenz E., Röllinghoff M., Mertens R. Visceral leishmaniasis in a German child who had never entered a known endemic area: Case report and review of the literature. Clin. Infect. Dis. 2001;32:302–306. doi: 10.1086/318476. PubMed DOI

Koehler K., Stechele M., Hetzel U., Domingo M., Schönian G., Zahner H., Burkhardt E. Cutaneous leishmaniosis in a horse in southern Germany caused by Leishmania infantum. Vet. Parasitol. 2002;109:9–17. doi: 10.1016/S0304-4017(02)00246-7. PubMed DOI

Kollaritsch H., Emminger W., Zaunschirm A., Aspöck H. Suspected Autochthonous Kala-azar in Austria. Lancet. 1989;1:901–902. doi: 10.1016/S0140-6736(89)92895-X. PubMed DOI

Beyreder J. Ein Fall von Leishmaniose in Niederösterreich. Wien. Med. Wochenschr. 1962;115:900–901. PubMed

Kasap O.E., Dvořák V., Depaquit J., Alten B., Votypka J., Volf P. Phylogeography of the subgenus Transphlebotomus Artemiev with description of two new species, Phlebotomus anatolicus n. sp. and Phlebotomus killicki n. sp. Infect. Genet. Evol. 2015;34:467–479. doi: 10.1016/j.meegid.2015.05.025. PubMed DOI

Lafri I., Almeras L., Bitam I., Caputo A., Yssouf A., Forestier C.L., Izri A., Raoult D., Parola P. Identification of Algerian Field-Caught Phlebotomine Sand Fly Vectors by MALDI-TOF MS. PLoS Negl. Trop. Dis. 2016;10:e0004351. doi: 10.1371/journal.pntd.0004351. PubMed DOI PMC

Pareyn M., Dvořák V., Halada P., Van Houtte N., Medhin G., de Kesel W., Merdekios B., Massebo F., Leirs H., Volf P. An integrative approach to identify sand fly vectors of leishmaniases in Ethiopia by morphological and molecular techniques. Parasit. Vectors. 2020;13:580. doi: 10.1186/s13071-020-04450-2. PubMed DOI PMC

Oerther S., Jöst H., Heitmann A., Lühken R., Krüger A., Steinhausen I., Brinker C., Lorentz S., Marx M., Schmidt-Chanasit J., et al. Phlebotomine sand flies in Southwest Germany: An update with records in new locations. Parasit. Vectors. 2020;13:173. doi: 10.1186/s13071-020-04058-6. PubMed DOI PMC

Rossi E., Bongiorno G., Ciolli E., Di Muccio T., Scalone A., Gramiccia M., Gradoni L., Maroli M. Seasonal phenology, host-blood feeding preferences and natural Leishmania infection of Phlebotomus perniciosus (Diptera, Psychodidae) in a high-endemic focus of canine leishmaniasis in Rome province, Italy. Acta Trop. 2008;105:158–165. doi: 10.1016/j.actatropica.2007.10.005. PubMed DOI

Ivović V., Kalan K., Zupan S., Bužan E. Illegal waste sites as a potential micro foci of Mediterranean Leishmaniasis: First records of phlebotomine sand flies (Diptera: Psychodidae) from Slovenia. Acta Vet. Brno. 2015;65:348–357.

Trájer A.J., Sebestyén V. The changing distribution of Leishmania infantum Nicolle, 1908 and its Mediterranean sandfly vectors in the last 140 kys. Sci. Rep. 2019;9:11820. doi: 10.1038/s41598-019-48350-7. PubMed DOI PMC

Simsek F.M., Alten B., Caglar S.S., Ozbep Y., Aytekin A.M., Kaynas S., Belen A., Kasap O.E., Yaman M., Rastgeldi S. Distribution and altitudinal structuring of phlebotomine sand flies (Diptera: Psychodidae) in southern Anatolia, Turkey: Their relation to human cutaneous leishmaniasis. J. Vector Ecol. 2007;32:269–279. doi: 10.3376/1081-1710(2007)32[269:DAASOP]2.0.CO;2. PubMed DOI

Tichy H., Kallina W. Insect hygroreceptor responses to continuous changes in humidity and air pressure. J. Neurophysiol. 2010;103:3274–3286. doi: 10.1152/jn.01043.2009. PubMed DOI PMC

Leschnik M., Löwenstein M., Edelhofer R., Kirtz G. Imported non-endemic, arthropod-borne and parasitic infectious diseases in Austrian dogs. Wien. Klin. Wochenschr. 2008;120:59–62. doi: 10.1007/s00508-008-1077-3. PubMed DOI

Aspöck H., Walochnik J. When sandflies move north. Public Health. 2009;20:24–31.

Aspöck H. Postglacial formation and fluctuations of the biodiversity of Central Europe in the light of climate change. Parasitol. Res. 2008;103:10–13. doi: 10.1007/s00436-008-1057-6. PubMed DOI

Depaquit J. Molecular systematics applied to Phlebotomine sandflies: Review and perspectives. Infect. Genet. Evol. 2014;28:744–756. doi: 10.1016/j.meegid.2014.10.027. PubMed DOI

Depaquit J., Ferté H., Léger N., Lefranc F., Alves-Pires C., Hanafi H., Maroli M., Morillas-Márquez F., Rioux J.A., Svobodova M., et al. ITS 2 sequences heterogeneity in Phlebotomus sergenti and Phlebotomus similis (Diptera, Psychodidae): Possible consequences in their ability to transmit Leishmania tropica. Int. J. Parasitol. 2002;32:1123–1131. doi: 10.1016/S0020-7519(02)00088-7. PubMed DOI

QGIS Development Team QGIS Geographic Information System. Open Source Geospatial Foundation Project. [(accessed on 29 October 2019)]; Available online: http://qgis.osgeo.org.

Lewis D.J. A taxonomic review of the genus Phlebotomus (Diptera: Psychodidae) Bull. Br. Mus. Nat. Hist. 1982;45:121–209.

Kumar S., Stecher G., Li M., Knyaz C., Tamura K. MEGA X: Molecular evolutionary genetics analysis across computing platforms. Mol. Biol. Evol. 2018;35:1547–1549. doi: 10.1093/molbev/msy096. PubMed DOI PMC

Dvořák V., Halada P., Hlacvackiva K., Dokianakis E., Antoniou M., Volf P. Identification of phlebotomine sand flies (Diptera: Psychodidae) by matrix-assisted laser desorption/ionization time of flight mass spectrometry. Parasit. Vectors. 2014;7:21. doi: 10.1186/1756-3305-7-21. PubMed DOI PMC

Halada P., Hlavackova K., Risueño J., Berriatua E., Volf P., Dvořák V. Effect of trapping method on species identification of phlebotomine sandflies by MALDI-TOF MS protein profiling. Med. Vet. Entomol. 2018;32:388–392. doi: 10.1111/mve.12305. PubMed DOI

Dvořák V., Tsirigotakis N., Pavlou C., Dokianakis E., Akhoundi M., Halada P., Volf P., Depaquit J., Antoniou M. Sand fly fauna of Crete and the description of Phlebotomus (Adlerius) creticus n. sp. Parasit. Vectors. 2020;13:547. doi: 10.1186/s13071-020-04358-x. PubMed DOI PMC

El Tai N.O., Osman F.O., El Far M., Presber W., Schönian G. Genetic heterogeneity of ribosomal internal transcribed spacer in clinical samples of Leishmania donovani spotted on filter paper as revealed by single-strand conformation polymorphisms and sequencing. Trans. R. Soc. Trop. Med. Hyg. 2000;94:575–579. doi: 10.1016/S0035-9203(00)90093-2. PubMed DOI

Librado P., Rozas J. DnaSP v5: A software for comprehensive analysis of DNA polymorphism data. Bioinformatics. 2009;25:1451–1452. doi: 10.1093/bioinformatics/btp187. PubMed DOI

Bandelt H.-J., Forster P., Röhl A. Median-joining networks for inferring intraspecific phylogenies. Mol. Biol. Evol. 1999;16:37–48. doi: 10.1093/oxfordjournals.molbev.a026036. PubMed DOI

Leigh J.W., Bryant D. Popart: Full-feature software for haplotype network construction. Methods Ecol. Evol. 2015;6:1110–1116. doi: 10.1111/2041-210X.12410. DOI

R Foundation for Statistical Computing; Vienna, Austria: [(accessed on 5 May 2020)]. R Core Team R: A Language and Environment for Statistical Computing. Available online: https://www.R-project.org/

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