Archives of Virology

, Volume 161, Issue 8, pp 2183–2188 | Cite as

Insect-specific flaviviruses in Aedes mosquitoes in Greece

  • Anna Papa
  • Elpida Papadopoulou
  • Ravish Paliwal
  • Stella Kalaitzopoulou
  • Spiros Mourelatos
  • Matthias Niedrig
Original Article

Abstract

Mosquitoes of the genus Aedes are known vectors of pathogenic flaviviruses, and insect-specific flaviviruses (ISFs) have been detected in members of this genus in numerous parts of the world. In order to gain insight into whether Aedes mosquitoes in Greece are infected by flaviviruses, 1173 Aedes spp. mosquitoes collected in 2010 and 2012 were grouped in 53 pools and tested by RT nested PCR using flavivirus generic primers. Eight pools (15.09 %) were found to be PCR positive: five pools (5/53, 9.4 %) contained RNA sequences related to Ochlerotatus caspius flavivirus (OCFV), an ISF previously detected in the Iberian peninsula, two pools (2/53, 3.8 %) contained sequences related to a mosquito flavivirus detected in Aedes vexans (AeveV) in Italy and the Czech Republic, and one pool contained a DNA sequence that was too short to identify accurately. The highest OCFV prevalence (12.9 %) was observed in August 2010 in the regional unit of Thessaloniki. Similar sequences were later obtained from two Culex spp. pools collected in 2013 in the same regions. A genetic difference of 0.2-1.4 % was seen among the Greek OCFV strains, which differed by 2.2-4.1 % from the Iberian strains and by 6.2-11.1 % from the Finnish Hanko virus. The genetic distances among strains varied depending on the genome region (genes for E, NS3 and NS5 proteins), with NS3 being the most variable. The present study shows no evidence of infection of Aedes mosquitoes with known pathogenic flaviviruses, but it expands the geographic distribution of OCFV in the eastern Mediterranean area. Any implication of ISFs for public health (either directly or through interactions with other flaviviruses in the mosquitoes) remains to be elucidated.

Notes

Compliance with ethical standards

Funding

The study was funded by the Development Agency of Thessaloniki, SA. The collaboration between Aristotle University of Thessaloniki and Robert Koch Institute was supported by IKYDA 2013 (granted by the State Scholarships Foundation in Greece and German Academic Exchange Service-DAAD).

Conflict of interest

All authors declare that they have no conflict of interest.

Ethical approval

Not needed.

References

  1. 1.
    Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410CrossRefPubMedGoogle Scholar
  2. 2.
    Aranda C, Sanchez-Seco MP, Caceres F, Escosa R, Galvez JC, Masia M, Marques E, Ruiz S, Alba A, Busquets N, Vazquez A, Castella J, Tenorio A (2009) Detection and monitoring of mosquito flaviviruses in Spain between 2001 and 2005. Vector Borne Zoonotic Dis 9:171–178CrossRefPubMedGoogle Scholar
  3. 3.
    Barrera R, MacKay A, Amador M, Vasquez J, Smith J, Diaz A, Acevedo V, Caban B, Hunsperger EA, Munoz-Jordan JL (2010) Mosquito vectors of West Nile virus during an epizootic outbreak in Puerto Rico. J Med Entomol 47:1185–1195CrossRefPubMedGoogle Scholar
  4. 4.
    Blitvich BJ, Firth AE (2015) Insect-specific flaviviruses: a systematic review of their discovery, host range, mode of transmission, superinfection exclusion potential and genomic organization. Viruses 7:1927–1959CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Bolling BG, Olea-Popelka FJ, Eisen L, Moore CG, Blair CD (2012) Transmission dynamics of an insect-specific flavivirus in a naturally infected Culex pipiens laboratory colony and effects of co-infection on vector competence for West Nile virus. Virology 427:90–97CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Calzolari M, Bonilauri P, Bellini R, Caimi M, Defilippo F, Maioli G, Albieri A, Medici A, Veronesi R, Pilani R, Gelati A, Angelini P, Parco V, Fabbi M, Barbieri I, Lelli D, Lavazza A, Cordioli P, Dottori M (2010) Arboviral survey of mosquitoes in two northern Italian regions in 2007 and 2008. Vector Borne Zoonotic Dis 10:875–884CrossRefPubMedGoogle Scholar
  7. 7.
    Calzolari M, Ze-Ze L, Ruzek D, Vazquez A, Jeffries C, Defilippo F, Osorio HC, Kilian P, Ruiz S, Fooks AR, Maioli G, Amaro F, Tlusty M, Figuerola J, Medlock JM, Bonilauri P, Alves MJ, Sebesta O, Tenorio A, Vaux AG, Bellini R, Gelbic I, Sanchez-Seco MP, Johnson N, Dottori M (2012) Detection of mosquito-only flaviviruses in Europe. J Gen Virol 93:1215–1225CrossRefPubMedGoogle Scholar
  8. 8.
    Cammisa-Parks H, Cisar LA, Kane A, Stollar V (1992) The complete nucleotide sequence of cell fusing agent (CFA): homology between the nonstructural proteins encoded by CFA and the nonstructural proteins encoded by arthropod-borne flaviviruses. Virology 189:511–524CrossRefPubMedGoogle Scholar
  9. 9.
    Crochu S, Cook S, Attoui H, Charrel RN, De Chesse R, Belhouchet M, Lemasson JJ, de Micco P, de Lamballerie X (2004) Sequences of flavivirus-related RNA viruses persist in DNA form integrated in the genome of Aedes spp. mosquitoes. J Gen Virol 85:1971–1980CrossRefPubMedGoogle Scholar
  10. 10.
    Darsie RF, Samanidou-Voyadjoglou A (1997) Keys for the identification of the mosquitoes of Greece. J Am Mosq Control Assoc 13:247–254PubMedGoogle Scholar
  11. 11.
    Engler O, Savini G, Papa A, Figuerola J, Groschup MH, Kampen H, Medlock J, Vaux A, Wilson AJ, Werner D, Jost H, Goffredo M, Capelli G, Federici V, Tonolla M, Patocchi N, Flacio E, Portmann J, Rossi-Pedruzzi A, Mourelatos S, Ruiz S, Vazquez A, Calzolari M, Bonilauri P, Dottori M, Schaffner F, Mathis A, Johnson N (2013) European surveillance for West Nile virus in mosquito populations. Int J Environ Res Public Health 10:4869–4895CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Fall AG, Diaite A, Etter E, Bouyer J, Ndiaye TD, Konate L (2012) The mosquito Aedes (Aedimorphus) vexans arabiensis as a probable vector bridging the West Nile virus between birds and horses in Barkedji (Ferlo, Senegal). Med Vet Entomol 26:106–111CrossRefPubMedGoogle Scholar
  13. 13.
    Farfan-Ale JA, Lorono-Pino MA, Garcia-Rejon JE, Hovav E, Powers AM, Lin M, Dorman KS, Platt KB, Bartholomay LC, Soto V, Beaty BJ, Lanciotti RS, Blitvich BJ (2009) Detection of RNA from a novel West Nile-like virus and high prevalence of an insect-specific flavivirus in mosquitoes in the Yucatan Peninsula of Mexico. Am J Trop Med Hyg 80:85–95PubMedPubMedCentralGoogle Scholar
  14. 14.
    Ferreira DD, Cook S, Lopes A, de Matos AP, Esteves A, Abecasis A, de Almeida AP, Piedade J, Parreira R (2013) Characterization of an insect-specific flavivirus (OCFVPT) co-isolated from Ochlerotatus caspius collected in southern Portugal along with a putative new Negev-like virus. Virus Genes 47:532–545CrossRefPubMedGoogle Scholar
  15. 15.
    Hobson-Peters J, Yam AW, Lu JW, Setoh YX, May FJ, Kurucz N, Walsh S, Prow NA, Davis SS, Weir R, Melville L, Hunt N, Webb RI, Blitvich BJ, Whelan P, Hall RA (2013) A new insect-specific flavivirus from northern Australia suppresses replication of West Nile virus and Murray Valley encephalitis virus in co-infected mosquito cells. PLoS One 8:e56534CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Huhtamo E, Moureau G, Cook S, Julkunen O, Putkuri N, Kurkela S, Uzcategui NY, Harbach RE, Gould EA, Vapalahti O, de Lamballerie X (2012) Novel insect-specific flavivirus isolated from northern Europe. Virol 433:471–478CrossRefGoogle Scholar
  17. 17.
    Kuno G, Chang GJ, Tsuchiya KR, Karabatsos N, Cropp CB (1998) Phylogeny of the genus Flavivirus. J Virol 72:73–83PubMedPubMedCentralGoogle Scholar
  18. 18.
    Papa A, Bakonyi T, Xanthopoulou K, Vazquez A, Tenorio A, Nowotny N (2011) Genetic characterization of West Nile virus lineage 2, Greece, 2010. Emerg Infect Dis 17:920–922CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Papa A, Xanthopoulou K, Gewehr S, Mourelatos S (2011) Detection of West Nile virus lineage 2 in mosquitoes during a human outbreak in Greece. Clin Microbiol Infect 17:1176–1180CrossRefPubMedGoogle Scholar
  20. 20.
    Papa A (2013) West Nile virus infections in humans-Focus on Greece. J Clin Virol 58:351–353CrossRefPubMedGoogle Scholar
  21. 21.
    Papa A, Papadopoulou E, Gavana E, Kalaitzopoulou S, Mourelatos S (2013) Detection of West Nile virus lineage 2 in Culex mosquitoes, Greece, 2012. Vector Borne Zoonotic Dis 13:682–684CrossRefPubMedGoogle Scholar
  22. 22.
    Papa A, Xanthopoulou K, Tsioka A, Kalaitzopoulou S, Mourelatos S (2013) West Nile virus in mosquitoes in Greece. Parasitol Res 112:1551–1555CrossRefPubMedGoogle Scholar
  23. 23.
    Papa A, Papadopoulou E, Kalaitzopoulou S, Tsioka K, Mourelatos S (2014) Detection of West Nile virus and insect-specific flavivirus RNA in Culex mosquitoes, central Macedonia, Greece. Trans R Soc Trop Med Hyg 108:555–559CrossRefPubMedGoogle Scholar
  24. 24.
    Roiz D, Vazquez A, Rosso F, Arnoldi D, Girardi M, Cuevas L, Perez-Pastrana E, Sanchez-Seco MP, Tenorio A, Rizzoli A (2012) Detection of a new insect flavivirus and isolation of Aedes flavivirus in Northern Italy. Parasit Vectors 5:223CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Roiz D, Vazquez A, Seco MP, Tenorio A, Rizzoli A (2009) Detection of novel insect flavivirus sequences integrated in Aedes albopictus (Diptera: Culicidae) in Northern Italy. Virol J 6:93CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Sanchez-Seco MP, Rosario D, Domingo C, Hernandez L, Valdes K, Guzman MG, Tenorio A (2005) Generic RT-nested-PCR for detection of flaviviruses using degenerated primers and internal control followed by sequencing for specific identification. J Virol Methods 126:101–109CrossRefPubMedGoogle Scholar
  27. 27.
    Schlesinger RW (1971) New opportunities in biological research offered by arthropod cell cultures. I. some speculations on the possible role of arthropods in the evolution of arboviruses. Curr Top Microbiol Immunol 55:241–245PubMedGoogle Scholar
  28. 28.
    Stollar V, Thomas VL (1975) An agent in the Aedes aegypti cell line (Peleg) which causes fusion of Aedes albopictus cells. Virol 64:367–377CrossRefGoogle Scholar
  29. 29.
    Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Tiawsirisup S, Kinley JR, Tucker BJ, Evans RB, Rowley WA, Platt KB (2008) Vector competence of Aedes vexans (Diptera: Culicidae) for West Nile virus and potential as an enzootic vector. J Med Entomol 45:452–457CrossRefPubMedGoogle Scholar
  31. 31.
    Vazquez A, Sanchez-Seco MP, Palacios G, Molero F, Reyes N, Ruiz S, Aranda C, Marques E, Escosa R, Moreno J, Figuerola J, Tenorio A (2012) Novel flaviviruses detected in different species of mosquitoes in Spain. Vector Borne Zoonotic Dis 12:223–229CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag Wien 2016

Authors and Affiliations

  • Anna Papa
    • 1
  • Elpida Papadopoulou
    • 1
  • Ravish Paliwal
    • 2
  • Stella Kalaitzopoulou
    • 3
  • Spiros Mourelatos
    • 3
  • Matthias Niedrig
    • 2
  1. 1.Department of Microbiology, Medical SchoolAristotle University of ThessalonikiThessalonikiGreece
  2. 2.Robert Koch InstituteBerlinGermany
  3. 3.EcoDevelopment SAThessalonikiGreece

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