Aim Phlebotominae sandflies are primary vectors of phleboviruses, causing the sandfly fever disease. The aim of this study was to detect and report the presence of flaviviruses in Phlebotominae sandflies captured in Bosnia and Herzegovina. Methods After a microscopic and morphometric analysis, the final identification of collected Phlebotomus specimens was confirmed by PCR, using a hemi-nested polymerase chain reaction on extracted and reversely transcribed RNA. Results We obtained a 155 nt long fragment of the viral non-structural protein 5 (NS5) gene (GenBank accession no. MN090154). The acquired nucleotide sequence, provisionally named as Drežnica, showed a maximum of 70-80% identity in 70-88% (110-137 nucleotides) of the query coverage with several Anopheles, Sabethes, Calbertado and Culex flaviviruses. Maximum likelihood phylogenetic analysis showed that the new flavivirus Drežnica clusters together with the flavivirus isolated from Culiseta annulata mosquitos. Conclusion We report the presence of flavivirus in Phlebotominae sandflies, captured in Drežnica, Herzegovina for the first time. The next phase of research will be directed towards virus cultivation, obtaining a longer or complete virus sequence and clarifying the medical and epidemiological importance of the Drežnica virus.
Hall T, Biosciences I, Carlsbad C. BioEdit: an important software for molecular biology. GERF Bull Biosci 2011:60–1.
2
Özkale Y, Özkale M, Kiper P, Cetinkaya B, Erol I. Sadfly fever: two case reports. Turk Pediatri Ars 2016:110.
3
Moureau G, Ninove L, Izri A, Cook S, Lamballerie X, Charrel R. Flavivirus RNA in phlebotomine sandflies. Vector Borne Zoonotic Dis 2010:195–7.
4
Genbank. 15AD.
5
Pick A. Zur Pathologie und Therapie einer eigenthümlichen endemischen Krankheitsform. Wien Med Wochenschr 1886:1141–5.
6
Terzin A, Matuka S, Fornazaric M, Hlaca D. Antibodies against some Arbor-Viruses and against the Bedsonia Antigen in Sera of Men, Sheep and Cattle in Bosnia and Herzegovina. Acta Med Iugosl 1962:301–17.
7
Vesenjak-Hirjan J, Porterfield J, Arslanagić E. Arboviruses in the Mediterranean countries. Stuttgart 1980.
8
Ayhan N, Alten B, Ivovic V, Dvořák V, Martinkovic F, Omeragic J, et al. Direct evidence for an expanded circulation area of the recently identified Balkan virus (Sandfly fever Naples virus species) in several countries of the Balkan archipelago. Parasit Vectors 2017:402.
9
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. Dis 2016:802–6.
10
Sánchez-Secso M, Echevarría J, Hernández L, Estévez D, Navarro-Marí J, Tenorio A. Detection and identification of Toscana and other phleboviruses by RT-nested-PCR assays with degenerated primers. J Med Virol 2003:140–9.
11
Scaramozzino N, Crance J, Jouan A, Debriel D, Stoll F, Garin D. Comparison of flavivirus universal primer pairs and development of a rapid, highly sensitive heminested reverse transcription-PCR assay for detection of flaviviruses targeted to a conserved region of the NS5 gene sequences. J Clin Microbiol 2001:1922–7.
12
Cusi M, Savellini G, Zanelli G. Toscana virus epidemiology: from Italy to beyond. Open Virol J 2010:22.
13
Altschul S, Madden T, Schäffer A, Zhang J, Zhang Z, Miller W, et al. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 1997:3389–402.
14
Kumar S, Stecher G, Li M, Knyaz C, Tamura K. MEGA X: molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol 2018:1547–9.
15
Tamura K, Nei M. Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. Mol Biol Evol 1993:512–26.
16
Tamura K, Nei M, Kumar S. Prospects for inferring very large phylogenies by using the neighbor-joining method. Proc Natl Acad Sci 2004:11030–5.
17
Genbank. 15AD.
18
Alkan C, Zapata S, Bichaud L, Moureau G, Lemey P, Firth A, et al. Ecuador Paraiso Escondido virus, a new flavivirus isolated from New World sand flies in Ecuador, is the first representative of a novel clade in the genus Flavivirus. J Virol 2015:11773–85.
19
Tesh R, Saidi S, Javadian E, Loh P, Nadim A. Isfahan virus, a new vesiculovirus infecting humans, gerbils, and sandflies in Iran. Am J Trop Med Hyg 1977:299–306.
20
Tesh R, Modi G. Growth and transovarial transmission of Chandipura virus (Rhabdoviridae: Vesiculovirus) in Phlebotomus papatasi. Am J Trop Med Hyg 1983:621–3.
21
Vasilakis N, Widen S, Mayer S, Seymour R, Wood T, Popov V, et al. Niakha virus: a novel member of the family Rhabdoviridae isolated from phlebotomine sandflies in Senegal. Virology 2013:80–9.
22
Guler S, Guler E, Caglayik D, Kokoglu O, Ucmak H, Bayrakdar F, et al. A sandfly fever virus outbreak in the East Mediterranean region of Turkey. Int J Infect Dis 2012:244–6.
23
Weaver S, Reisen W. Present and future arboviral threats. Antiviral Res 2010:328–45.
24
Dash A, Bhatia R, Sunyoto T, Mourya D. Emerging and re-emerging arboviral diseases in Southeast Asia. J Vector Borne Dis 2013:77.
25
Go Y, Balasuriya U, Lee C. Zoonotic encephalitides caused by arboviruses: transmission and epidemiology of alphaviruses and flaviviruses. Clin Exp Vaccine Res 2014:58–77.
26
Competition for financing / co-financing scientific / scientific-research and research-development projects in the Federation of Bosnia and Herzegovina in 2017 n.d.
27
Blitvich B. Arboviruses: Molecular Biology, Evolution and Control. Nikos Vasilakis and Duane J. Gubler. Am J Trop Med Hyg 2016:488–9.
28
Alatoom A, Payne D. An overview of arboviruses and bunyaviruses. Lab Med 2009:237–40.
29
Hollidge B, González-Scarano F, Soldan S. Arboviral encephalitides: transmission, emergence, and pathogenesis. J Neuroimmune Pharmacol 2010:428–42.
30
Chastel C. Asymptomatic infections in man: A Trojan horse for the introduction and spread of mosquito-borne arboviruses in non-endemic areas? Bull Soc Pathol Exot 1990:213–9.
31
Olivia W, Obanda L, Bucht V, Mosomtai G, Otieno G, Ahlm V, et al. Global emergence of Alphaviruses that cause arthritis in humans. Infect Ecol Epidemiol 2015:29853.
32
Karabatsos N. International catalogue of arboviruses: including certain other viruses of vertebrates. 3rd ed. San Antonio, Tex: Published for the Subcommittee on Information Exchange of the American Committee on Arthropod-borne Viruses by the 1985.
33
Sabin A, Philip C, Paul J. Phlebotomus (Pappataci or Sandfly) fever: a disease of military importance summary of existing knowledge and preliminary report of original investigations. J Am Med Assoc 1944:693–9.
34
Tufan Z, Weidmann M, Bulut C, Kinikli S, Hufert F, Dobler G, et al. Clinical and laboratory findings of a sandfly fever Turkey Virus outbreak in Ankara. J Infect 2011:375–81.
35
Palacios G, Tesh R, Savji N, Da Rosa A, Guzman H, Bussetti A, et al. Characterization of the Sandfly fever Naples species complex and description of a new Karimabad species complex (genus Phlebovirus, family Bunyaviridae). J Gen Virol 2014:292.
36
Kocak Tufan Z, Tasyaran M, Sandfly G, Fever. A Mini Review Virol Mycol 2013:2161–0517.
37
Ozbel Y. The infections transmitted by sand flies in Turkey. Vet Fak Derg 2013:225–8.
38
Nicoletti L, Ciufolini M, Verani P. Imported Virus Infections 1996:41–7.
39
Nicoletti L, Verani P, Caciolli S, Ciufolini M, Renzi A, Bartolozzi D, et al. Central nervous system involvement during infection by Phlebovirus toscana of residents in natural foci in central Italy (1977-1988). Am J Trop Med Hyg 1991:429–34.
40
Dionisio D, Valassina M, Ciufolini M, Vivarelli A, Esperti F, Cusi M, et al. Encephalitis without meningitis due to sandfly fever virus serotype Toscana. Clin Infect Dis 2001:1241–3.
41
Hukić M, Saumović-Bešić I. Sandfly-Pappataci fever in Bosnia and Herzegovina: the new-old disease. Bosn J Basic Med Sci 2009:39.
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