Detection of Leishmania infantum and a Novel Phlebovirus (Balkan Virus) from Sand Flies in Albania.

OBJECTIVE: To organize entomological campaigns to trap sand flies in selected regions of Albania and to test them for the presence of existing or new phleboviruses and for leishmania DNA.

METHODS: Sand flies were collected in 14 locations from May to October 2014 using three different types of traps. Pools with a maximum of 30 individuals were prepared according to gender, trapping site, and trapping date; they were tested for the presence of (1) phlebovirus RNA with three different PCR systems (2) and Leishmania DNA using two different real-time PCR assays.

RESULTS: A total of 972 sand flies (568 females, 404 males) were aliquoted to 55 pools. Three pools (in two different regions) were positive for Leishmania infantum. Two pools (Kruje region) were positive for phlebovirus RNA and a 575-nucleotide (nt) colinearized sequence of a novel virus most closely related to but clearly distinct from Tehran virus (16% and 3% divergence at nt and amino acid levels). Next generation sequencing analysis indicated that this virus might be transmitted by either Phlebotomus neglectus, Phlebotomus tobbi, or both vectors.

CONCLUSIONS: Visceral leishmaniasis has been clinically recognized in Albania for at least 80 years; however, this is the first time that L. infantum, detected by molecular means, has been reported in sand flies in Albania. At the outset of this study, only Adria virus (Salehabad species) was recognized in Albania. A novel virus, Balkan virus, was identified and genetic analysis revealed that it belongs to the Sandfly fever Naples virus group containing human pathogens.