Vector soup: high-throughput identification of Neotropical phlebotomine sand flies using metabarcoding.
Phlebotomine sand flies are haematophagous dipterans of primary medical importance. They represent the only proven vectors of leishmaniasis worldwide and are involved in the transmission of various other pathogens. Studying the ecology of sand flies is crucial to understand the epidemiology of leishmaniasis and further control this disease. A major limitation in this regard is that traditional morphological-based methods for sand fly species identifications are time-consuming and require taxonomic expertise. DNA metabarcoding holds great promise in overcoming this issue by allowing the identification of multiple species from a single bulk sample. Here, we assessed the reliability of a short insect metabarcode located in the mitochondrial 16S rRNA for the identification of Neotropical sand flies, and constructed a reference database for 40 species found in French Guiana. Then, we conducted a metabarcoding experiment on sand flies mixtures of known content and showed that the method allows an accurate identification of specimens in pools. Finally, we applied metabarcoding to field samples caught in a 1-ha forest plot in French Guiana. Besides providing reliable molecular data for species-level assignations of phlebotomine sand flies, our study proves the efficiency of metabarcoding based on the mitochondrial 16S rRNA for studying sand fly diversity from bulk samples. The application of this high-throughput identification procedure to field samples can provide great opportunities for vector monitoring and eco-epidemiological studies.
CNRS, Université Toulouse III Paul Sabatier, ENFA, UMR5174 EDB (Laboratoire Evolution et Diversité Biologique), Toulouse, France.|UMR MIVEGEC (IRD 224 - CNRS 5290 - Université de Montpellier), 911 Avenue Agropolis, F34394, Montpellier, France.
Arthur
A
Kocher
Laboratoire des Identifications Fongiques et Entomo-parasitologiques, Mennecy, France.
Jean-Charles
JC
Gantier
Medical Entomology Unit, Institut Pasteur de la Guyane, 23 Avenue Pasteur, BP 6010, 97306, Cayenne Cedex, French Guiana.
Pascal
P
Gaborit
CNRS, Université Toulouse III Paul Sabatier, ENFA, UMR5174 EDB (Laboratoire Evolution et Diversité Biologique), Toulouse, France.
Lucie
L
Zinger
CNRS, Université Toulouse III Paul Sabatier, ENFA, UMR5174 EDB (Laboratoire Evolution et Diversité Biologique), Toulouse, France.
Helene
H
Holota
GeT-PlaGe, Genotoul, INRA Auzeville, 31326, Castanet-Tolosan, France.
Sophie
S
Valiere
Medical Entomology Unit, Institut Pasteur de la Guyane, 23 Avenue Pasteur, BP 6010, 97306, Cayenne Cedex, French Guiana.
Isabelle
I
Dusfour
Medical Entomology Unit, Institut Pasteur de la Guyane, 23 Avenue Pasteur, BP 6010, 97306, Cayenne Cedex, French Guiana.
Romain
R
Girod
UMR MIVEGEC (IRD 224 - CNRS 5290 - Université de Montpellier), 911 Avenue Agropolis, F34394, Montpellier, France.
Anne-Laure
AL
Bañuls
CNRS, Université Toulouse III Paul Sabatier, ENFA, UMR5174 EDB (Laboratoire Evolution et Diversité Biologique), Toulouse, France.
Jerome
J
Murienne
D000818:Animals; D016000:Cluster Analysis; D058893:DNA Barcoding, Taxonomic / Q000379:methods*; D004272:DNA, Mitochondrial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D005616:French Guiana; D007303:Insect Vectors; D056186:Metagenomics / Q000379:methods*; D010802:Phylogeny; D011576:Psychodidae / Q000145:classification* / Q000235:genetics* / Q000254:growth & development; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA
D004272:DNA, Mitochondrial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S
DNA barcode; French Guiana; Psychodidae; leishmaniasis; mitochondrial 16S rRNA gene; vector monitoring