Abstract
Autochthonous transmissions of mosquito-borne diseases were described in close proximity to Slovenia where the knowledge of mosquito species and the viruses they transmit remains very scarce. Therefore we aimed to assess the burden mosquitoes bring to the local community and to test mosquitoes for the presence of the selected zoonotic arboviruses, i. e. Dengue (DENV), West Nile (WNV) and Chikungunya (CHIKV). A pilot mosquito monitoring was conducted in 2012 within two municipalities based on adult mosquito trapping. Altogether 2007 adult mosquitoes were caught. Females of Aedes albopictus and Culex pipiens represented the majority of the total mosquito catch. In sites with more households and nearby greenery, catches of Ae. albopictus were more abundant. Highest numbers of Cx. pipiens females were caught close to a nature reserve, where many bird species stop during their migrations. Trapping results are consistent with both species’ feeding behaviour. Female mosquitoes were grouped into 58 pools according to species, location and date of catch. Real time RT-PCRs for WNV, DENV and CHIKV were performed, but no arboviral genomes were detected in any of the tested mosquito pools. However, amplicons were obtained in three pools when using generic RT-PCR markers for flaviviruses. The viruses detected were propagated and isolated in mosquito cell line C3/36 and identified with sequencing as mosquito-only flaviviruses (MOFs). The important contributions of this study are a successful first isolation of MOFs and confirmation of the presence of the invasive mosquito Ae. albopictus in Slovenia.
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Abbreviations
- IMS:
-
Invasive mosquito species
- MOFs:
-
Mosquito-only flaviviruses
- DENV:
-
Dengue virus
- WNV:
-
West Nile virus
- CHIKV:
-
Chikungunya virus
- UTM :
-
Universal transport medium
- CPE:
-
Cytopathic effect
References
Alm E, Lesko B, Lindegren G, Ahlm C, Söderholm S, Falk KI, Lagerqvist N (2014) Universal single-probe RT-PCR assay for diagnosis of dengue virus infections. PLoS Negl Trop Dis 8(12):e3416. https://doi.org/10.1371/journal.pntd.0003416
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410. https://doi.org/10.1016/S0022-2836(05)80360-2
Angelini R, Finarelli AC, Angelini P, Po C, Petropulacos K, Macini P, Fiorentini C, Fortuna C, Venturi G, Romi R, Majori G, Nicoletti L, Rezza G, Cassone A (2007) An outbreak of chikungunya fever in the province of Ravenna, Italy. Euro Surveill 12(9). https://doi.org/10.2807/esw.12.36.03260-en
Bakonyi T, Hubálek Z, Rudolf I, Nowotny N (2005) Novel Flavivirus or new lineage of West Nile virus, Central Europe. Emerg Inf Dis 11(2):225–231. https://doi.org/10.3201/eid1102.041028
Bakonyi T, Ivanics É, Erdélyi K, Ursu K, Ferenczi E, Weissenböck H, Nowotny N (2006) Lineage 1 and 2 strains of encephalitic West Nile virus, Central Europe. Emerg Infect Dis 12(4):618. https://doi.org/10.3201/eid1204.051379
Bakonyi T, Ferenczi E, Erdélyi K, Kutasi O, Csörgő T, Seidel B, Weissenböck H, Brugger K, Bán E, Nowotny N (2013) Explosive spread of a neuroinvasive lineage 2 West Nile virus in Central Europe, 2008/2009. Vet Microbiol 165(1–2):61–70. https://doi.org/10.1016/j.vetmic.2013.03.005
Becker N, Petrić D, Zgomba M, Boase C, Dahl C, Lane J, Kaisera A (2003) Mosquitoes and their control, vol 2. Springer, New York. https://doi.org/10.1007/978-3-540-92874-4
Bolling BG, Eisen L, Moore CG, Blair CD (2011) Insect-specific flaviviruses from Culex mosquitoes in Colorado, with evidence of vertical transmission. Am J Trop Med Hygiene 851:169–177. https://doi.org/10.4269/ajtmh.2011.10-0474
Bustamante DM, Lord CC (2010) Sources of error in the estimation of mosquito infection rates used to assess risk of arbovirus transmission. Am J Trop Med Hyg 82(6):1172–1184. https://doi.org/10.4269/ajtmh.2010.09-0323
Calzolari M, Ze-Ze L, Ruzek D, Vázquez A, Jeffries C, Defilippo F, Costa Osório H, Kilian P, Ruíz S, Fooks RA, Maioli G, Amaro F, Tlustý M, Figuerola J, Medlock JM, Bonilauri P, João Alves M, Šebesta O, Tenorio A, Vaux AGC, Bellini R, Gelbič I, Sánchez-Seco MP, Johnson N, Dottori M (2012) Detection of mosquito-only flaviviruses in Europe. J Gen Virol 93:1215–1225. https://doi.org/10.1099/vir.0.040485-0
Carrieri M, Bacchi M, Bellini R, Maini S (2003) On the competition occurring between Aedes albopictus and Culex pipiens (Diptera: Culicidae) in Italy. Environ Entomol 326:1313–1321. https://doi.org/10.1603/0046-225X-32.6.1313
ECDC European Centre for Disease Control (2017) West Nile Fever. In: Historical data by year https://ecdc.europa.eu/en/west-nile-fever/surveillance-and-disease-data/historical
Farajollahi A, Fonseca DM, Kramer LD, Kilpatrick AM (2011) “Birds biting” mosquitoes and human disease: a review of the role of Culex pipiens mosquitoes in epidemiology. Infect Genet Evol 11(7):1577–1585. https://doi.org/10.1016/j.meegid.2011.08.013
Gjenero-Margan I, Aleraj B, Krajcar D, Lesnikar V, Klobucar A, Pem-Novosel I, Kurečić-Filipović S, Komparak S, Martić R, Đuričić S, Betica-Radić L, Okmadžić J, Vilibić-Čavlek T, Babić-Erceg A, Turković B, Avšić-Županc T, Radić I, Ljubić M, Šarac K, Benić N , Mlinarić-Galinović G (2011) Autochthonous dengue fever in Croatia, August-September 2010. Euro Surveill 169:19805. https://doi.org/10.2807/ese.16.09.19805-en
Grisenti M, Vaszquez A, Herrero L Cuevas L, Perez-Pastrana E, Arnoldi D, Rosà R, Capelli G, Tenorio A, Sánchez-Seco MP, Rizzoli A (2015) Wide detection of Aedes flavivirus in North-Eastern Italy – a European hotspot of emerging mosquito borne diseases. J Gen Virol 96:420–430. https://doi.org/10.1099/vir.0.069625-0
Gu W, Novak RJ (2004) Short report: detection probability of arbovirus infection in mosquito populations. Am J Trop Med Hyg 71(5):636–638. https://doi.org/10.4269/ajtmh.2004.71.636
Gutsevich AV, Monchadskii AS , Shtakelberg AA (1974) Fauna of the USSR Diptera 3 4. Mosquitoes, family Culicidae
Halstead SB (2007) Dengue. Lancet 370(9599):1644–1652. https://doi.org/10.1016/S0140-6736(07)61687-0
Hawley WA (1988) The biology of Aedes albopictus. J Am Mosq Control Assoc, Supplement 1:1–39
Hlad B, Skoberne P (2001) Characteristics of Biological and Landscape Diversity in Slovenia. Biological and Landscape Diversity in Slovenia: An Overview PDF. Ljubljana: Environmental Agency of the Republic of Slovenia, Ministry of the Environment and Spatial Planning
Hoshino K, Isawa H, Tsuda Y, Sawabe K, Kobayashi M (2009) Isolation and characterization of a new insect flavivirus from Aedes albopictus and Aedes flavopictus mosquitoes in Japan. Virology 3911:119–129. https://doi.org/10.1016/j.virol.2009.06.025
Hubalek Z, Halouzka J (1999) West Nile fever-a reemerging mosquito-borne viral disease in Europe. Emerg Infect Dis 55:643. https://doi.org/10.3201/eid0505.990505
Kalan K, Ivović V, Glasnović P, Bužan E (2017) Presence and potential distribution of Aedes albopictus and Aedes japonicus japonicus (Diptera: Culicidae) in Slovenia. J Med Entomol 1–9. https://doi.org/10.1093/jme/tjx150
Kim DY, Guzman H, Bueno R, Dennett JA, Auguste AJ, Carrington CVF, Popov VL, Weaver SC, Beasley DWC, Robert B, Tesh RB (2009) Characterization of Culex Flavivirus (Flaviviridae) strains isolated from mosquitoes in the United States and Trinidad. Virology 3861:154–159. https://doi.org/10.1016/j.virol.2008.12.034
Panning M, Grywna K, van Esbroeck M, Emmerich P, Drosten C (2006) Chikungunya fever in travelers returning to Europe from the Indian Ocean region. Emerg Infect Dis 14(3): 416–422. https://doi.org/10.3201/eid1403.070906
Paz S, Semenza JC (2013) Environmental drivers of West Nile fever epidemiology in Europe and Western Asia – a review. Int J Environ Res Public Health 10:3543–3562. https://doi.org/10.3390/ijerph10083543
Di Sabatino D, Bruno R, Sauro F, Danzetta ML, Cito F, Iannetti S, Narcisi V, De Massis F, Calistri P (2014) Epidemiology of West Nile disease in Europe and in the Mediterranean Basin from 2009 to 2013. Biomed Res Int. https://doi.org/10.1155/2014/907852
Scaramozzino N, Crance JM, Jouan A, DeBriel DA, Stoll F, Garin D (2001) 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 39(5):1922–1927. https://doi.org/10.1128/JCM.39.5.1922-1927.2001
Schaffner F, Angel G, Geoffroy B, Hervy JP, Rhaiem A, Brunhes J (2001) The mosquitoes of Europe – an identification and training programme. CD-rom., IRDEditions. In: EID Mediterranee. Montpellier, France
Schwartz O, Albert ML (2010) Biology and pathogenesis of chikungunya virus. Nat Rev Microbiol 8:491–500. https://doi.org/10.1038/nrmicro2368
Tomasello D, Schlagenhauf P (2013) Chikungunya and dengue autochthonous cases in Europe, 2007–2012. Travel Med Infect Dis 115:274–284. https://doi.org/10.1016/j.tmaid.2013.07.006
Tovornik D (1983) Review of some bio-ecological research data on the Culicidae fauna in Slovenia Yugoslavia. Acta entomol Yugoslavica 19:19–26
Tovornik D (1990) O ekologiji larvalnih komarjev Diptera: Culicidae. v manjši mirujoči mlaki na Ljubljanskem barju. Biol Vestn 4:47–68
Trpiš M, Tovornik D (1958) Faunistische, okologische und zoogeographische bemerkungen zu den stechmücken Jugoslawien. Bilogia, Bratislava 13:721–793
White DJ, Kramer LD, Backenson PB, Lukacik G, Johnson G, Oliver JA, Howard JJ, Means RG, Eidson M, Gotham I, Kulasekera V, Campbell S (2001) Mosquito surveillance and polymerase chain reaction detection of West Nile virus, New York state. Emerg Infect Dis 7(4):643–649. https://doi.org/10.3201/eid0704.010407
Acknowledgments
The authors acknowledge the financial support from the Institute of Public Health Koper, Municipality of Nova Gorica and Slovenian Research Agency (research core funding No. (100-12-1988)). We are grateful to dr. Heidi Christine Hauffe and dr. Matthew Schwarzkopf for English corrections. We also wish to thank Tjaša Zagoršek, Eva Praprotnik and Jana Zoratti for their help with field and laboratory work.
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Kalan, K., Avšič-Županc, T., Rus, K.R. et al. Mosquito-only flaviviruses, isolated from Aedes albopictus in Slovenia: results of a pilot mosquito monitoring program. Biologia 73, 1277–1282 (2018). https://doi.org/10.2478/s11756-018-0135-1
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DOI: https://doi.org/10.2478/s11756-018-0135-1