title: Phlebovirus and Leishmania detection in sandflies from eastern Thrace and northern Cyprus.
abstract: BACKGROUND
  Phlebotomine sandflies are vectors of several pathogens with significant impact for public health. This study was conducted to investigate and characterize phlebovirus and Leishmania infections in vector sandflies collected in the eastern Thrace region in Turkey and Northern Cyprus, where previous data indicate activity of these agents.
  
  
  METHODS
  Field sampling of sandflies was performed at 4 locations in Edirne and Tekirdag provinces of eastern Thrace and at 17 locations in Lefkosa, Girne, Magosa and Guzelyurt provinces of northern Cyprus. In sandfly pools, phlebovirus RNA and Leishmania DNA were screened via a generic polymerase chain reaction (PCR) and kinetoplast minicircle PCR, respectively. Selected sandfly specimens unsuitable for pathogen detection were identified to species level. Cytochrome oxidase 1 gene region was used for DNA barcoding of selected specimens and pathogen positive pools. Positive amplicons were cloned and characterized by sequencing.
  
  
  RESULTS
  A total of 2690 sandflies, collected from Eastern Thrace (15.4%) and Northern Cyprus (84.6%) were evaluated. Morphological examination of 780 specimens from Cyprus exhibited Phlebotomus perfiliewi sensu lato (72.6%), Phlebotomus tobbi (19.7%), Phlebotomus papatasi (2.8%), Laroussius sp. (1.6%) and Sergentomyia azizi (1.6%), Sergentomyia sp. (0.9%), Sergentomyia minuta (0.5%) and Phleobotomus jacusieli (0.1%) species. Pathogen screening was performed in 1910 specimens distributed in 195 pools. In eight pools of P.tobbi sandflies collected in Cyprus, Leishmania infantum DNA was demonstrated. Toscana virus (TOSV) genotype A sequences were identified in two pools of P. perfiliewi s.l. and one pool of P.tobbi sandflies from Cyprus. Co-infection of TOSV and Leishmania infantum was characterized in a P.tobbi pool. Sequences belonging to novel phleboviruses are revealed in three P. perfiliewi s.l. pools. One sequence, provisionally named Edirne virus, identified in Edirne province in eastern Thrace, demonstrated the highest rate of genomic similarity to Adria and Salehabad viruses. Furthermore, Girne 1 and Girne 2 viruses, identified in Girne province, revealed similarities to TOSV and Sandfly Fever Sicilian virus and related strains, respectively.
  
  
  CONCLUSIONS
  Activity of TOSV genotype A strains in Cyprus and co-infection of sandfly vectors with L. infantum was documented for the first time. Novel phlebovirus strains of unknown medical significance was identified in sampling regions.
authors.affiliation: Faculty of Medicine, Department of Medical Microbiology, Virology Unit, Hacettepe University, Morphology Building 3rd Floor, 06100, Sihhiye, Ankara, Turkey. ekoray@hacettepe.edu.tr.
authors.forename: Koray
authors.initials: K
authors.lastname: Ergunay
authors.affiliation: Faculty of Sciences, Department of Biology, Division of Ecology, Hacettepe University, Ankara, Turkey. ozgeerisoz@yahoo.com.
authors.forename: Ozge Erisoz
authors.initials: OE
authors.lastname: Kasap
authors.affiliation: Faculty of Medicine, Department of Medical Microbiology, Virology Unit, Hacettepe University, Morphology Building 3rd Floor, 06100, Sihhiye, Ankara, Turkey. serraorsten@gmail.com.
authors.forename: Serra
authors.initials: S
authors.lastname: Orsten
authors.affiliation: Faculty of Veterinary Medicine, Department of Parasitology, Istanbul University, Istanbul, Turkey. keremoter@hotmail.com.
authors.forename: Kerem
authors.initials: K
authors.lastname: Oter
authors.affiliation: Faculty of Sciences, Department of Biology, Division of Ecology, Hacettepe University, Ankara, Turkey. gunayf@gmail.com.
authors.forename: Filiz
authors.initials: F
authors.lastname: Gunay
authors.affiliation: Faculty of Veterinary Medicine, Department of Virology, Ankara University, Ankara, Turkey. zeynepakkutay@gmail.com.
authors.forename: Ayse Zeynep Akkutay
authors.initials: AZ
authors.lastname: Yoldar
authors.affiliation: Advanced Technology Education, Research and Application Center, Mersin University, Mersin, Turkey. enderdin@gmail.com.
authors.forename: Ender
authors.initials: E
authors.lastname: Dincer
authors.affiliation: Faculty of Sciences, Department of Biology, Division of Ecology, Hacettepe University, Ankara, Turkey. kaynas@hacettepe.edu.tr.
authors.forename: Bulent
authors.initials: B
authors.lastname: Alten
authors.affiliation: Faculty of Veterinary Medicine, Department of Virology, Ankara University, Ankara, Turkey. ozkul@ankara.edu.tr.
authors.forename: Aykut
authors.initials: A
authors.lastname: Ozkul
mesh_terms: D000818:Animals; D003535:Cyprus; D007303:Insect Vectors / Q000469:parasitology* / Q000821:virology*; D007891:Leishmania / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D010691:Phlebotomus / Q000469:parasitology* / Q000821:virology*; D016856:Phlebovirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D010802:Phylogeny; D014421:Turkey