BACKGROUND: Recently, Balkan virus (BALKV, family Phenuiviridae, genus Phlebovirus) was discovered in sand flies collected in Albania and genetically characterised as a member of the Sandfly fever Naples species complex. To gain knowledge concerning the geographical area where exposure to BALKV exists, entomological surveys were conducted in 2014 and 2015, in Croatia, Bosnia and Herzegovina (BH), Kosovo, Republic of Macedonia and Serbia. RESULTS: A total of 2830 sand flies were trapped during 2014 and 2015 campaigns, and organised as 263 pools. BALKV RNA was detected in four pools from Croatia and in one pool from BH. Phylogenetic relationships were examined using sequences in the S and L RNA segments. Study of the diversity between BALKV sequences from Albania, Croatia and BH showed that Albanian sequences were the most divergent (9-11% [NP]) from the others and that Croatian and BH sequences were grouped (0.9-5.4% [NP]; 0.7-5% [L]). The sand fly infection rate of BALKV was 0.26% in BH and 0.27% in Croatia. Identification of the species content of pools using cox1 and cytb partial regions showed that the five BALKV positive pools contained Phlebotomus neglectus DNA; in four pools, P neglectus was the unique species, whereas P. tobbi DNA was also detected in one pool. CONCLUSIONS: We report here (i) the first direct evidence that the Balkan virus initially described in coastal Albania has a much wider dissemination area than originally believed, (ii) two real-time RT-PCR assays that may be useful for further screening of patients presenting with fever of unknown origin that may be caused by Balkan virus infection, (iii) entomological results suggesting that Balkan virus is likely transmitted by Phlebotomus neglectus, and possibly other sand fly species of the subgenus Larroussius. So far, BALKV has been detected only in sand flies. Whether BALKV can cause disease in humans is unknown and remains to be investigated.

Department of Parasitology and Parasitic Diseases Faculty of Veterinary Medicine Ss Cyril and Methodius University Skopje Republic of Macedonia

Department of Parasitology Veterinary Faculty of Sarajevo Zmaja od Bosne 90 71000 Sarajevo Bosnia and Herzegovina

Faculty of Agriculture Laboratory for Medical and Veterinary Entomology University of Novi Sad Novi Sad Serbia

Faculty of Science Department of Biology Ecology Division VERG Labs Hacettepe University Beytepe Ankara Turkey

Faculty of Science Department of Parasitology Charles University Prague Czech Republic

Faculty of Veterinary Medicine Department of Parasitology and Parasitic Diseases with Clinics University of Zagreb Zagreb Croatia

National Institute of Public Health Pristina Kosovo

UMR Emergence des Pathologies Viralesˮ Marseille France

University of Primorska FAMNIT Koper Slovenia

Zobrazit více v PubMed

Depaquit J, Grandadam M, Fouque F, Andry PE, Peyrefitte C. Arthropod-borne viruses transmitted by phlebotomine sand flies in Europe: a review. Euro Surveill. 2010;15(10):19507. PubMed

Alkan C, Bichaud L, de Lamballerie X, Alten B, Gould EA, Charrel RN. Sandfly-borne phleboviruses of Eurasia and Africa: epidemiology, genetic diversity, geographic range, control measures. Antivir Res. 2013;100(1):54–74. doi: 10.1016/j.antiviral.2013.07.005. PubMed DOI

Charrel RN, Moureau G, Temmam S, Izri A, Marty P, Parola P, et al. Massilia virus, a novel Phlebovirus (Bunyaviridae) isolated from sandflies in the Mediterranean. Vector Borne Zoonotic Dis. 2009;9(5):519–530. doi: 10.1089/vbz.2008.0131. PubMed DOI PMC

Zhioua E, Moureau G, Chelbi I, Ninove L, Bichaud L, Derbali M, et al. Punique virus, a novel phlebovirus, related to sandfly fever Naples virus, isolated from sandflies collected in Tunisia. J Gen Virol. 2010;91(5):1275–1283. doi: 10.1099/vir.0.019240-0. PubMed DOI PMC

Papa A, Velo E, Bino S. A novel phlebovirus in Albanian sandflies. Clin Microbiol Infect. 2011;17(4):585–587. doi: 10.1111/j.1469-0691.2010.03371.x. PubMed DOI

Remoli ME, Fortuna C, Marchi A, Bucci P, Argentini C, Bongiorno G, et al. Viral isolates of a novel putative phlebovirus in the Marche Region of Italy. Am J Trop Med Hyg. 2014;90(4):760–763. doi: 10.4269/ajtmh.13-0457. PubMed DOI PMC

Alkan C, Alwassouf S, Piorkowski G, Bichaud L, Tezcan S, Dincer E, et al. Isolation, genetic characterization, and seroprevalence of Adana virus, a novel phlebovirus belonging to the Salehabad virus complex, in Turkey. J Virol. 2015;89(8):4080–4091. doi: 10.1128/JVI.03027-14. PubMed DOI PMC

Amaro F, Hanke D, Zé-Zé L, Alves MJ, Becker SC, Höper D. Genetic characterization of Arrabida virus, a novel phlebovirus isolated in South Portugal. Virus Res. 2016;214:19–25. doi: 10.1016/j.virusres.2016.01.004. PubMed DOI

Alkan C, Erisoz Kasap O, Alten B, de Lamballerie X, Charrel RN. Sand fly-borne phlebovirus isolations from Turkey: new insight into the sandfly fever Sicilian and sandfly fever Naples species. PLoS Negl Trop Dis. 2016;10(3):e0004519. doi: 10.1371/journal.pntd.0004519. PubMed DOI PMC

Anagnostou V, Pardalos G, Athanasiou-Metaxa M, Papa A. Novel phlebovirus in febrile child, Greece. Emerg Infect Dis. 2011;17(5):940–1. PubMed PMC

Ayhan N, Velo E, de Lamballerie X, Kota M, Kadriaj P, Ozbel Y, et al. Detection of Leishmania infantum and a novel phlebovirus (Balkan Virus) from sand flies in Albania. Vector Borne Zoonotic Dis. 2016;16(12):802–806. doi: 10.1089/vbz.2016.2002. PubMed DOI

Sánchez-Seco MP, Echevarría JM, Hernández L, Estévez D, Navarro-Marí JM, Tenorio A. Detection and identification of Toscana and other phleboviruses by RT-nested-PCR assays with degenerated primers. J Med Virol. 2003;71(1):140–149. doi: 10.1002/jmv.10465. PubMed DOI

Charrel RN, Izri A, Temmam S, Delaunay P, Toga I, Dumon H, et al. Cocirculation of 2 genotypes of Toscana virus, southeastern France. Emerg Infect Dis. 2007;13(3):465–468. doi: 10.3201/eid1303.061086. PubMed DOI PMC

Lambert AJ, Lanciotti RS. Consensus amplification and novel multiplex sequencing method for S segment species identification of 47 viruses of the Orthobunyavirus, Phlebovirus, and Nairovirus genera of the family Bunyaviridae. J Clin Microbiol. 2009;47(8):2398–2404. doi: 10.1128/JCM.00182-09. PubMed DOI PMC

Pick A. Zur Pathologie und Therapie einer eigenthümlichen endemischen Krankheitsform. Wien Med Wschr. 1886;33:1141–1145.

Pick A. Beiträge zur Pathologie und Therapie einer eigenthümlichen Krankheitsform (Gastro-enteritis climatica) Prager Med Wschr. 1887;12:364.

Terzin AL, Matuka S, Fornazarić MR, Hlača DM. Antibodies against some arboviruses and against the Bedsonia antigen in sera of men, sheep and cattle in Bosnia and Herzegovina. Acta Medica Yugoslavica. 1962;16(3–4):301–317.

Vesenjak-Hirjan J. Arboviruses in Yugoslavia. In: Vesenjak-Hirjan J, editor. Arboviruses in the Mediterranean countries. Stuttgart-New York: Gustav Fischer Verlag; 1980. pp. 165–177.

Gligić A, Mišcević Z, Tesh RB, Travassos da Rosa A, Zivković V. First isolations of Naples sandfly fever virus in Yugoslavia. Mikrobiologija. 1982;19:167–175.

Hukić M, Salimović-Besić I. Sandfly-Pappataci fever in Bosnia and Herzegovina: the new-old disease. Bosn J Basic Med Sci. 2009;9(1):39–43. PubMed PMC

Hukić M, Numanović F, Sisirak M, Moro A, Dervović E, Jakovec S, Besić IS. Surveillance of wildlife zoonotic diseases in the Balkans Region. Med Glas (Zenica) 2010;7(2):96–105. PubMed

Tesh RB, Saidi S, Gajdamovic SJ, Rodhain F, Vesenjak-Hirjan J. Serological studies on the epidemiology of sandfly fever in the old world. Bull World Health Organ. 1976;54(6):663–674. PubMed PMC

Vesenjak-Hirjan J, Punda-Polić V, Dobe M. Geographical distribution of arboviruses in Yugoslavia. J Hyg Epidemiol Microbiol Immunol. 1991;35(2):129–140. PubMed

Borcić B, Punda V. Sandfly fever epidemiology in Croatia. Acta Med Iugosl. 1987;41(2):89–97. PubMed

Punda-Polić V, Calisher CH, Vesenjak-Hirjan J. Neutralizing antibodies for sandfly fever Naples virus in human sera on the island of Mljet. Acta Med Iugosl. 1990;44(1):15–20. PubMed

Punda-Polić V, Mohar B, Duh D, Bradarić N, Korva M, Fajs L, et al. Evidence of an autochthonous Toscana virus strain in Croatia. J Clin Virol. 2012;55(1):4–7. doi: 10.1016/j.jcv.2012.06.006. PubMed DOI

Punda-Polić V, Jerončić A, Mohar B, Šiško KK. Prevalence of Toscana virus antibodies in residents of Croatia. Clin Microbiol Infect. 2012;18(6):E200–E203. doi: 10.1111/j.1469-0691.2012.03840.x. PubMed DOI

Karabatsos N. Supplement to International Catalogue of Arboviruses including certain other viruses of vertebrates. Am J Trop Med Hyg. 1978;27:372. doi: 10.4269/ajtmh.1978.27.372. PubMed DOI

First Detection and Molecular Analysis of Leishmania infantum DNA in Sand Flies of Kosovo

Sand flies (Diptera: Psychodidae) in eight Balkan countries: historical review and region-wide entomological survey