title: Detection of a new insect flavivirus and isolation of Aedes flavivirus in Northern Italy.
abstract: BACKGROUND
  During recent years, numerous novel 'insect flaviviruses' have been discovered in natural mosquito populations. In a previous study we described the presence of flavivirus DNA sequences integrated in Aedes albopictus (Asian tiger mosquito) populations from Northern Italy in 2007.
  
  
  METHODS
  During 2008 we collected and tested Aedes females for flavivirus presence and developed phylogenetic analysis, virus isolation, electron microscopy studies and RNAse treatments.
  
  
  RESULTS
  We detected a high prevalence of flavivirus in Ae. albopictus (77.5%). The phylogenetic analysis identified the insect flavivirus sequences as Aedes flavivirus (AEFV) recently described in Japan, and that may have been introduced in Italy travelling with the tiger mosquito. Some of these pools grew in C6/36 cells, producing cytopathic effects, and the RNase treatment results showed the presence of the detected sequences in RNA forms. Furthermore, we detected a new insect flavivirus in one pool of Aedes cinereus/geminus mosquitoes. Phylogenetic analysis of this virus shows that it forms a distinct cluster within the clade of insect flavivirus.
  
  
  CONCLUSIONS
  This is the first study to report a high prevalence, to describe the seasonal activity and an isolation of the insect flavivirus Aedes flavivirus in Europe. Moreover we describe the detection of a new insect flavivirus detected from Ae. cinereus mosquitoes from Italy. These flavivirus may be common, ubiquitous and diverse in nature and we discuss the implications of the insect flavivirus group in virus evolution and transmission.
authors.affiliation: Wetland Ecology Department, Doñana Biological Station, CSIC, Sevilla, Spain. davidroiz@gmail.com
authors.forename: David
authors.initials: D
authors.lastname: Roiz
authors.forename: Ana
authors.initials: A
authors.lastname: Vázquez
authors.forename: Fausta
authors.initials: F
authors.lastname: Rosso
authors.forename: Daniele
authors.initials: D
authors.lastname: Arnoldi
authors.forename: Matteo
authors.initials: M
authors.lastname: Girardi
authors.forename: Laureano
authors.initials: L
authors.lastname: Cuevas
authors.forename: Esperanza
authors.initials: E
authors.lastname: Perez-Pastrana
authors.forename: Mari Paz
authors.initials: MP
authors.lastname: Sánchez-Seco
authors.forename: Antonio
authors.initials: A
authors.lastname: Tenorio
authors.forename: Annapaola
authors.initials: A
authors.lastname: Rizzoli
mesh_terms: D000330:Aedes / Q000821:virology*; D000818:Animals; D016000:Cluster Analysis; D005260:Female; D005416:Flavivirus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification / Q000648:ultrastructure; D007558:Italy; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D010802:Phylogeny; D012367:RNA, Viral / Q000235:genetics; D017422:Sequence Analysis, DNA; D014771:Virion / Q000648:ultrastructure; D014776:Virus Cultivation
chemical_list: D012367:RNA, Viral