title: Bacterial diversity of the American sand fly Lutzomyia intermedia using high-throughput metagenomic sequencing.
abstract: Parasites of the genus Leishmania cause a broad spectrum of diseases, collectively known as leishmaniasis, in humans worldwide. American cutaneous leishmaniasis is a neglected disease transmitted by sand fly vectors including Lutzomyia intermedia, a proven vector. The female sand fly can acquire or deliver Leishmania spp. parasites while feeding on a blood meal, which is required for nutrition, egg development and survival. The microbiota composition and abundance varies by food source, life stages and physiological conditions. The sand fly microbiota can affect parasite life-cycle in the vector.
  
  
  
  We performed a metagenomic analysis for microbiota composition and abundance in Lu. intermedia, from an endemic area in Brazil. The adult insects were collected using CDC light traps, morphologically identified, carefully sterilized, dissected under a microscope and the females separated into groups according to their physiological condition: (i) absence of blood meal (unfed = UN); (ii) presence of blood meal (blood-fed = BF); and (iii) presence of developed ovaries (gravid = GR). Then, they were processed for metagenomics with Illumina Hiseq Sequencing in order to be sequence analyzed and to obtain the taxonomic profiles of the microbiota.
  
  
  
  Bacterial metagenomic analysis revealed differences in microbiota composition based upon the distinct physiological stages of the adult insect. Sequence identification revealed two phyla (Proteobacteria and Actinobacteria), 11 families and 15 genera; 87 % of the bacteria were Gram-negative, while only one family and two genera were identified as Gram-positive. The genera Ochrobactrum, Bradyrhizobium and Pseudomonas were found across all of the groups.
  
  
  
  The metagenomic analysis revealed that the microbiota of the Lu. intermedia female sand flies are distinct under specific physiological conditions and consist of 15 bacterial genera. The Ochrobactrum, Bradyrhizobium and Pseudomonas were the common genera. Our results detailing the constituents of Lu. intermedia native microbiota contribute to the knowledge regarding the bacterial community in an important sand fly vector and allow for further studies to better understand how the microbiota interacts with vectors of human parasites and to develop tools for biological control.
authors: Laboratory of Medical Entomology, René Rachou Research Centre (FIOCRUZ-MG), Belo Horizonte, Minas Gerais, Brazil.
authors: Carolina Cunha
authors: CC
authors: Monteiro
authors: Laboratory of Medical Entomology, René Rachou Research Centre (FIOCRUZ-MG), Belo Horizonte, Minas Gerais, Brazil.
authors: Luis Eduardo Martinez
authors: LE
authors: Villegas
authors: Laboratory of Medical Entomology, René Rachou Research Centre (FIOCRUZ-MG), Belo Horizonte, Minas Gerais, Brazil.
authors: Thais Bonifácio
authors: TB
authors: Campolina
authors: Laboratory of Medical Entomology, René Rachou Research Centre (FIOCRUZ-MG), Belo Horizonte, Minas Gerais, Brazil.
authors: Ana Clara Machado Araújo
authors: AC
authors: Pires
authors: Centro de Pesquisas Gonçalo Moniz (CPqGM)-Fundação Oswaldo Cruz (FIOCRUZ), Salvador, Bahia, Brazil.
authors: Jose Carlos
authors: JC
authors: Miranda
authors: Laboratory of Medical Entomology, René Rachou Research Centre (FIOCRUZ-MG), Belo Horizonte, Minas Gerais, Brazil.
authors: Paulo Filemon Paolucci
authors: PF
authors: Pimenta
authors: Laboratory of Medical Entomology, René Rachou Research Centre (FIOCRUZ-MG), Belo Horizonte, Minas Gerais, Brazil. nagila@cpqrr.fiocruz.br.
authors: Nagila Francinete Costa
authors: NF
authors: Secundino
mesh_terms: D000818:Animals; D001419:Bacteria / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D005260:Female; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D056186:Metagenomics / Q000379:methods*; D011576:Psychodidae / Q000382:microbiology*
keywords: Lutzomyia intermedia; Metagenomics; Native microbiota