Molecular detection of Eastern Equine Encephalitis virus in mosquitoes from La Pintada (Antioquia)

Molecular detection of Eastern Equine Encephalitis virus in mosquitoes from La Pintada (Antioquia)

ABSTRACT

Objective. The detection of emerging and re-emerging arboviruses in mosquitoes from urban and rural areas, is fundamental for predict possible epidemic outbreaks in human populations. The Municipality of La Pintada (Antioquia), is characterized by the presence of dry tropical forest relicts, fishing, tourism, farms and mining. An entomological research was performed for explore the possible circulation of arboviruses of public health importance. Materials and methods. Mosquitoes were captured in urban and rural sites in February-April of 2012. The specimens were stored in liquid nitrogens tanks and were grouped using taxonomic keys for genera. RNA extraction from pools and generic/nested RT-PCR was performed for Flavivirus, Alphavirus, Orthobunyavirus (Group Bunyamwera) and Phlebovirus. Results. 1274 mosquitoes were collected, mainly belonging to Culex and Aedes genera. RNA extracts of 64 pools were tested by RT-PCR and one pool was positive for Alphavirus. Sequencing of the RT- PCR product and the analysis with sequences storage in GenBank designate the presence of Eastern equine encephalitis virus (EEEV). Conclusions. This is the first record of natural infection from EEEV in mosquitoes from La Pintada (Antioquia), an area with ecological elements that favor the emergence of emerging and re-emerging arboviruses of medical and veterinarian importance.

1. Jones K. Global trends in emerging infectious diseases. Nat 2008; 451:990-993. http://dx.doi.org/10.1038/nature06536
3. Weaver S, Reisen W. Present and Future arboviral threats. Antiviral Res 2010; 85(2):328-345. http://dx.doi.org/10.1016/j.antiviral.2009.10.008
5. Groot H, Morales A, Romero M, Ferro C, Prías E, Vidales H, et al. Estudios de arbovirosis en Colombia en la década de 1970. Biomédica 1996; 16:331-344. http://dx.doi.org/10.7705/biomedica.v16i4.915
7. Mesa FA, Cardenas JA, Villamil LC. Las Encefalitis Equinas en Colombia. Bogotá D.C, Editorial Universidad Nacional de Colombia; 2005.
9. Osorio J, Ciuoderis K, Lopera J, Piedrahita L, Murphy D, Levasseur J et al. Characterization of west nile viruses isolated from captive American Flamingoes (Phoenicopterous ruber) in Medellin, Colombia. Am J Trop Med Hyg 2012; 87(3):565-572.
10. http://dx.doi.org/10.4269/ajtmh.2012.11-0655
12. Ferro C, Boshell J, Moncayo A, Gonzalez M, Ahumada M, Kang W, et al. Natural Enzootic Vectors of Venezuelan equine encephalitis virus in the Magdalena Valley, Colombia. Emerg Infect Dis 2003; 9(1):49-54. http://dx.doi.org/10.3201/eid0901.020136
14. Mattar S, Komar N, Young G, Alvarez J, Gonzalez M. Seroconversion for West Nile and St. Louis encephalitis viruses among sentinel horses in Colombia. Mem Inst Oswaldo Cruz 2011; 106(8):976-979. http://dx.doi.org/10.1590/S0074-02762011000800012
16. Sánchez-Seco M, Rosario D, Quiroz E, Guzmán G, Tenorio A. A generic nested-RT-PCR followed by sequencing for detection and identification of members of the alphavirus genus. J Virol Methods 2001; 95:153-161. http://dx.doi.org/10.1016/S0166-0934(01)00306-8
18. Sánchez-Seco M, Rosario D, Domingo C, Hernández L, Valdés K, Guzmán M, Tenorio A. Generic RT-nested-PCR for detection of flaviviruses using degenerated primers and internal control followed by sequencing for specific identification. J Virol Methods 2005; 126:101-109. http://dx.doi.org/10.1016/j.jviromet.2005.01.025
20. Sánchez-Seco M, Echevarría J, Hernández L, Estévez D, Navarro-Mari J, 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.
21. http://dx.doi.org/10.1002/jmv.10465
23. Kuno G, Mitchell C, Chang G, Smith G. Detecting bunyaviruses of the Bunyamwera and California serogroups by a PCR technique. J Clin Microbiol 1996; 34(5):1184-1188.
25. Gu W, Unnasch T, Katholi C, Lampman R, Novak R. Fundamental issues in mosquito surveillance for arboviral transmission. Trans R Soc Trop Med Hyg 2008; 102(8):817-822.
26. http://dx.doi.org/10.1016/j.trstmh.2008.03.019
28. Calisher C, Maness K, Lord R, Coleman P. Identification of two South American strains of Eastern equine encephalomyelitis virus from migrant birds captured on the Mississippi delta. Am J Epidemiol 1971; 94(2):172-178.
30. Carrera J, Forrester N, Wang E, Vittor A, Haddow A, López-Vergés S et al. Eastern equine encephalitis in Latin America. N Engl J Med 2013; 369(8):732-744. http://dx.doi.org/10.1056/NEJMoa1212628
32. Arrigo N, Paige A, Weaver S. Evolutionary Patterns of Eastern Equine Encephalitis Virus in North versus South America Suggest Ecological Differences and Taxonomic Revision. J Virol 2010; 84(2):1014-1025. http://dx.doi.org/10.1128/JVI.01586-09