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. 2014 Dec 12:7:575.
doi: 10.1186/s13071-014-0575-6.

Phlebovirus and Leishmania detection in sandflies from eastern Thrace and northern Cyprus

Koray Ergunay  1 , Ozge Erisoz Kasap  2 , Serra Orsten  3 , Kerem Oter  4 , Filiz Gunay  5 , Ayse Zeynep Akkutay Yoldar  6 , Ender Dincer  7 , Bulent Alten  8 , Aykut Ozkul  9
Affiliations

Phlebovirus and Leishmania detection in sandflies from eastern Thrace and northern Cyprus

Koray Ergunay et al. Parasit Vectors. .

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.

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Figures

Figure 1
Figure 1
Illustrative map of locations targeted for sampling in the study.
Figure 2
Figure 2
Neighbour-joining analysis of the partial cytochrome c oxidase I gene sequences obtained from Phlebovirus and Leishmania positive sandfly pools. Kimura two-parameter distance model was used for taxon identity tree construction. Sequences characterized in this study are marked (coloured triangles) whereas standard sequences are indicated as organism and GenBank accession number (if available).
Figure 3
Figure 3
Neighbour-joining analysis of the Phlebovirus partial nucleotide (A) and aminoacid (B) sequences. Jukes-Cantor and p-distance models are employed for nucleotide and aminoacid data, respectively. Viruses included in the analysis are indicated with name, isolate identifier and GenBank accession number. Uukuniemi virus is included as an outlier. Sequences characterized in this study are marked (coloured circles). Toscana virus isolates Thr2012 and Ank2012 demonstrate sequences previously characterized in Eastern Thrace and Central Anatolia, Turkey.
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