title: Detection of Culex flavivirus and Aedes flavivirus nucleotide sequences in mosquitoes from parks in the city of São Paulo, Brazil.
abstract: The dengue viruses are widespread in Brazil and are a major public health concern. Other flaviviruses also cause diseases in humans, although on a smaller scale. The city of São Paulo is in a highly urbanized area with few green spaces apart from its parks, which are used for recreation and where potential vertebrate hosts and mosquito vectors of pathogenic Flavivirus species can be found. Although this scenario can contribute to the transmission of Flavivirus to humans, little is known about the circulation of members of this genus in these areas. In light of this, the present study sought to identify Flavivirus infection in mosquitoes (Diptera: Culicidae) collected in parks in the city of São Paulo. Seven parks in different sectors of the city were selected. Monthly mosquito collections were carried out in each park from March 2011 to February 2012 using aspiration and traps (Shannon and CD C-CO2). Nucleic acids were extracted from the mosquitoes collected and used for reverse-transcriptase and real-time polymerase chain reactions with genus-specific primers targeting a 200-nucleotide region in the Flavivirus NS5 gene. Positive samples were sequenced, and phylogenetic analyses were performed. Culex and Aedes were the most frequent genera of Culicidae collected. Culex flavivirus (CxFV)-related and Aedes flavivirus (AEFV)- related nucleotide sequences were detected in 17 pools of Culex and two pools of Aedes mosquitoes, respectively, among the 818 pools of non-engorged females analyzed. To the best of our knowledge, this is the first report of CxFV and AEFV in the city of São Paulo and Latin America, respectively. Both viruses are insect- specific flaviviruses, a group known to replicate only in mosquito cells and induce a cytopathic effect in some situations. Hence, our data suggests that CxFV and AEFV are present in Culex and Aedes mosquitoes, respectively, in parks in the city of São Paulo. Even though Flavivirus species of medical importance were not detected, surveillance is recommended in the study areas because of the presence of vertebrates and mosquitoes that could act as amplifying hosts and vectors of flaviviruses, providing the required conditions for circulation of these viruses.
authors.affiliation: Laboratório de Protozoologia, Instituto de Medicina Tropical de São Paulo, Universidade de São Paulo, Av. Dr. Enéas de Carvalho Aguiar, 470, CEP 05403-000 São Paulo, SP, Brazil. Electronic address: licianatal@usp.br.
authors.forename: Licia Natal
authors.initials: LN
authors.lastname: Fernandes
authors.affiliation: Departamento de Epidemiologia, Faculdade de Saúde Pública, Universidade de São Paulo, Av. Dr. Arnaldo, 715, CEP 01246-904 São Paulo, SP, Brazil.
authors.forename: Marcia Bicudo
authors.initials: MB
authors.lastname: de Paula
authors.affiliation: Laboratório de Fauna Sinantrópica, Centro de Controle de Zoonoses, Prefeitura de São Paulo, Rua Santa Eulália, 86, CEP 02031-020 São Paulo, SP, Brazil.
authors.forename: Alessandra Bergamo
authors.initials: AB
authors.lastname: Araújo
authors.affiliation: Departamento de Parques e Áreas Verdes, Prefeitura de São Paulo, Avenida IV Centenário, CEP 04030-090 São Paulo, SP, Brazil.
authors.forename: Elisabeth Fernandes Bertoletti
authors.initials: EF
authors.lastname: Gonçalves
authors.affiliation: Laboratório de Virologia, Instituto de Medicina Tropical de São Paulo, Universidade de São Paulo, Av. Dr. Enéas de Carvalho Aguiar, 470, CEP 05403-000 São Paulo, SP, Brazil.
authors.forename: Camila Malta
authors.initials: CM
authors.lastname: Romano
authors.affiliation: Departamento de Epidemiologia, Faculdade de Saúde Pública, Universidade de São Paulo, Av. Dr. Arnaldo, 715, CEP 01246-904 São Paulo, SP, Brazil.
authors.forename: Delsio
authors.initials: D
authors.lastname: Natal
authors.affiliation: Laboratório de Protozoologia, Instituto de Medicina Tropical de São Paulo, Universidade de São Paulo, Av. Dr. Enéas de Carvalho Aguiar, 470, CEP 05403-000 São Paulo, SP, Brazil.
authors.forename: Rosely dos Santos
authors.initials: Rdos S
authors.lastname: Malafronte
authors.affiliation: Departamento de Epidemiologia, Faculdade de Saúde Pública, Universidade de São Paulo, Av. Dr. Arnaldo, 715, CEP 01246-904 São Paulo, SP, Brazil.
authors.forename: Mauro Toledo
authors.initials: MT
authors.lastname: Marrelli
authors.affiliation: Laboratório de Virologia, Instituto de Medicina Tropical de São Paulo, Universidade de São Paulo, Av. Dr. Enéas de Carvalho Aguiar, 470, CEP 05403-000 São Paulo, SP, Brazil; Fundação Pró-Sangue, Hemocentro de São Paulo, Departamento de Biologia molecular, Av. Dr. Enéas de Carvalho Aguiar, 155, CEP 05403-000 São Paulo, SP, Brazil.
authors.forename: José Eduardo
authors.initials: JE
authors.lastname: Levi
mesh_terms: D000330:Aedes / Q000821:virology*; D000818:Animals; D001483:Base Sequence; D001938:Brazil; D002947:Cities; D003465:Culex / Q000821:virology*; D005260:Female; D005416:Flavivirus / Q000235:genetics* / Q000302:isolation & purification*; D006801:Humans; D000068316:Parks, Recreational; D010802:Phylogeny
keywords: Flavivirus; Mosquitoes; Parks; Polymerase chain reaction