Sandfly-Borne Phlebovirus Isolations from Turkey: New Insight into the Sandfly fever Sicilian and Sandfly fever Naples Species

Southern Anatolia in Turkey at the border with Syria, where many refugee camps are settled, is endemic for sandfly-borne leishmaniasis. Sandfly-borne phleboviruses are also known to circulate in this region, although their relevance in terms of medical implications is virtually unknown. Therefore, the specific objectives of our study were firstly to identify isolate and characterise potentially pathogenic phleboviruses in sandflies; secondly to determine the complete genomic sequence of any viruses that we were able to isolate; and thirdly, to further our understanding of the potential medical importance and epidemiological significance of these viruses. To achieve these objectives, we organised field campaigns in 2012 and 2013. Two new phleboviruses (Toros and Zerdali viruses) were isolated and characterized by complete genome sequencing and phylogenetic analyses. Toros virus was genetically most closely related to Corfou virus within the Sandfly fever Sicilian group. Zerdali virus was most closely related to Tehran virus within the Sandfly fever Naples species. Although these new viruses belong to genetic groups that include several human pathogens, it is not yet clear if Toros and Zerdali viruses can infect humans and cause disease such as sandfly fever. Consequently, the availability of these genetically characterized infectious viruses will enable seroprevalence studies to establish their medical importance in this region and to assist the health agencies to develop appropriate and effective disease control strategies. Background Many studies have presented virus sequences which suggest the existence of a variety of putative new phleboviruses transmitted by sandflies in the Old World. However, in most of these studies, only partial sequences in the polymerase or the nucleoprotein genes were characterised. Therefore to further our understand of the presence and potential medical importance of sandfly-borne phleboviruses that circulate in southern Anatolia, we initiated field campaigns in 2012 and 2013 designed to identify, isolate and characterise phleboviruses in sandflies in this region Methodology/Principal Findings An entomological investigation encompassing 8 villages in Adana, Mediterranean Turkey was performed in August and September 2012 and 2013. A total of 11,302 sandflies were collected and grouped into 797 pools which were tested for the presence of phleboviruses using specific primers for RT-PCR analysis and also cell culture methods for virus isolation. Seven pools were PCR positive, and viruses were isolated from three pools of sandflies, resulting in the identification of two new viruses that we named Zerdali virus and Toros virus. Phylogenetic analysis based on full-length genomic sequence showed that Zerdali virus was most closely related with Tehran virus (and belongs to the Sandfly fever Naples species), whereas Toros virus was closest to Corfou virus. Conclusions/Significance The results indicate that a variety of phleboviruses are co-circulating in this region of southern Anatolia. Based on our studies, these new viruses clearly belong to genetic groups that include several human pathogens. However, whether or not Toros and Zerdali viruses can infect humans and cause diseases such as sandfly fever remains to be investigated.