title: Molecular identification of field-collected Culicoides larvae in the southern part of Japan.
abstract: Although Culicoides biting midges act as a vector of important human and domestic animal diseases, their ecology is poorly understood. The lack of proper identification systems of Culicoides larvae is one of the main obstacles to progress in research. Based on mitochondrial sequences of 19 Japanese Culicoides species, we designed a universal primer set to amplify the partial sequence of the mitochondrial cytochrome c oxidase I (cox 1). The polymerase chain reaction product amplified from extracted DNA of Culicoides larvae using the primer set was directly sequenced, and species identification based on the variation at cox1 was conducted. Using the molecular identification system, we sorted 243 specimens of field-collected larvae from the southern part of Japan into 10 species including Culicoides arakawae (Arakawa), Culicoides oxystoma Kieffer, and Culicoides brevitarsis Kieffer, which are regarded as vectors of important livestock animal diseases. Eight species of Culicoides larvae, including C. arakawae and C. oxystoma, were recovered from active paddy fields and an abandoned paddy field. The result suggests that paddy fields contribute to breeding a variety of Culicoides species and maintenance and spread of Culicoides-borne pathogens. In contrast, larvae of C. brevitarsis were collected from cattle dung in pastures. The molecular identification system described herein using nucleotide sequences successfully achieved larval identification and will be useful for a better understanding of larval habitats of Culicoides biting midges.
authors.affiliation: Kyushu Research Station, National Institute of Animal Health, NARO, 2702, Chuzan, Kagoshima 891-0105, Japan. tyanase@affrc.go.jp
authors.forename: Tohru
authors.initials: T
authors.lastname: Yanase
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Matsumoto
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Matsumori
authors.forename: Maki
authors.initials: M
authors.lastname: Aizawa
authors.forename: Miki
authors.initials: M
authors.lastname: Hirata
authors.forename: Tomoko
authors.initials: T
authors.lastname: Kato
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Shirafuji
authors.forename: Makoto
authors.initials: M
authors.lastname: Yamakawa
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Tsuda
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Noda
mesh_terms: D000818:Animals; D002519:Ceratopogonidae / Q000145:classification* / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D017753:Ecosystem; D003576:Electron Transport Complex IV / Q000235:genetics / Q000378:metabolism; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism; D007564:Japan; D007814:Larva / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D013045:Species Specificity
chemical_list: D019476:Insect Proteins; D003576:Electron Transport Complex IV