title: Description of microsporidia in simulids: molecular and morphological characterization of microsporidia in the larvae of Simulium pertinax Kollar (Diptera: Simuliidae).
abstract: BACKGROUND
  Microsporidia constitute the most common black fly pathogens, although the species' diversity, seasonal occurrence and transmission mechanisms remain poorly understood. Infections by this agent are often chronic and non-lethal, but they can cause reduced fecundity and decreased longevity. The objective of this study was to identify microsporidia infecting Simulium (Chirostilbia) pertinax (Kollar, 1832) larvae from Caraguatatuba, State of São Paulo, Brazil, by molecular and morphological characterization.
  
  
  METHODS
  Larvae were collected at a single point in a stream in a rural area of the city and were kept under artificial aeration until analysis. Polydispyrenia spp. infection was characterized by the presence of at least 32 mononuclear spores measuring 6.9 ± 1.0 × 5.0 ± 0.7 µm in persistent sporophorous vesicles. Similarly, Amblyospora spp. were characterized by the presence of eight uninucleate spores measuring 4.5 × 3.5 µm in sporophorous vesicles.
  
  
  RESULTS
  The molecular analysis confirmed the presence of microsporidian DNA in the 8 samples (prevalence of 0.51%). Six samples (Brazilian larvae) were related to Polydispyrenia simulii and Caudospora palustris reference sequences but in separate clusters. One sample was clustered with Amblyospora spp. Edhazardia aedis was the positive control taxon.
  
  
  CONCLUSIONS
  Samples identified as Polydispyrenia spp. and Amblyospora spp. were grouped with P. simulii and Amblyospora spp., respectively, corroborating previous results. However, the 16S gene tree showed a considerable distance between the black fly-infecting Amblyospora spp. and the mosquito-infecting spp. This distance suggests that these two groups are not congeneric. Additional genomic region evaluation is necessary to obtain a coherent phylogeny for this group.
authors.affiliation: Laboratório de Parasitologia, Instituto Butantan, São Paulo, SP, Brazil.
authors.forename: Isabel Maria Vicente Guedes de
authors.initials: IM
authors.lastname: Carvalho
authors.affiliation: Laboratório de Imunoparasitologia, Centro de Pesquisa Gonçalo Moniz, Fundação Oswaldo Cruz, Salvador, BA, Brasil.
authors.forename: Artur Trancoso Lopo de
authors.initials: AT
authors.lastname: Queiroz
authors.affiliation: Laboratório de Entomologia Médica, Superintendência de Controle de Endemias, São Paulo, SP, Brazil.
authors.forename: Rosiane Brito de
authors.initials: RB
authors.lastname: Moraes
authors.affiliation: Disciplina de Infectologia, Universidade Federal de São Paulo, São Paulo, SP, Brazil.
authors.forename: Helio Benites
authors.initials: HB
authors.lastname: Gil
authors.affiliation: Departamento de Medicina, Disciplina de Gastroenterologia, Universidade Federal de São Paulo, São Paulo, SP, Brazil.
authors.forename: Rafael
authors.initials: R
authors.lastname: Alves
authors.affiliation: Laboratório de Simulídeos, Superintendência de Controle de Endemias, Caraguatatuba, SP, Brasil.
authors.forename: Andréa de Barros Pinto
authors.initials: Ade B
authors.lastname: Viviani
authors.affiliation: United States Department of Agriculture, Mosquito and Fly Research Unit, Gainesville, Florida, USA.
authors.forename: James John
authors.initials: JJ
authors.lastname: Becnel
authors.affiliation: Laboratório de Entomologia Médica, Superintendência de Controle de Endemias, São Paulo, SP, Brazil.
authors.forename: Carlos José Pereira da Cunha de
authors.initials: CJ
authors.lastname: Araujo-Coutinho
mesh_terms: D000818:Animals; D007814:Larva / Q000382:microbiology; D016814:Microsporidia / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012621:Seasons; D012843:Simuliidae / Q000145:classification / Q000382:microbiology*