title: Simultaneous identification of host, ectoparasite and pathogen DNA via in-solution capture.
abstract: Ectoparasites frequently vector pathogens from often unknown pathogen reservoirs to both human and animal populations. Simultaneous identification of the ectoparasite species, the wildlife host that provided their most recent blood meal(s), and their pathogen load would greatly facilitate the understanding of the complex transmission dynamics of vector-borne diseases. Currently, these identifications are principally performed using multiple polymerase chain reaction (PCR) assays. We developed an assay (EctoBaits) based on in-solution capture paired with high-throughput sequencing to simultaneously identify ectoparasites, host blood meals and pathogens. We validated our in-solution capture results using double-blind PCR assays, morphology and collection data. The EctoBaits assay effectively and efficiently identifies ectoparasites, blood meals, and pathogens in a single capture experiment, allowing for high-resolution taxonomic identification while preserving the DNA sample for future analyses.
authors.affiliation: Center for Conservation and Evolutionary Genetics, Smithsonian Conservation Biology Institute, 3001 Connecticut Avenue NW, Washington, DC, 20008, USA.
authors.forename: Michael G
authors.initials: MG
authors.lastname: Campana
authors.affiliation: Center for Conservation and Evolutionary Genetics, Smithsonian Conservation Biology Institute, 3001 Connecticut Avenue NW, Washington, DC, 20008, USA.|Division of Mammals, National Museum of Natural History, Smithsonian Institution, MRC 108, P.O. Box 37012, Washington, DC, 20013-7012, USA.
authors.forename: Melissa T R
authors.initials: MT
authors.lastname: Hawkins
authors.affiliation: Center for Conservation and Evolutionary Genetics, Smithsonian Conservation Biology Institute, 3001 Connecticut Avenue NW, Washington, DC, 20008, USA.
authors.forename: Lauren H
authors.initials: LH
authors.lastname: Henson
authors.affiliation: Center for Conservation and Evolutionary Genetics, Smithsonian Conservation Biology Institute, 3001 Connecticut Avenue NW, Washington, DC, 20008, USA.
authors.forename: Kristin
authors.initials: K
authors.lastname: Stewardson
authors.affiliation: Department of Ecology, Evolution and Marine Biology, University of California Santa Barbara, Santa Barbara, CA, 93106, USA.
authors.forename: Hillary S
authors.initials: HS
authors.lastname: Young
authors.affiliation: Smithsonian Conservation Biology Institute, National Zoological Park, 1500 Remount Rd., Front Royal, VA, 22630, USA.
authors.forename: Leah R
authors.initials: LR
authors.lastname: Card
authors.affiliation: Center for Conservation and Evolutionary Genetics, Smithsonian Conservation Biology Institute, 3001 Connecticut Avenue NW, Washington, DC, 20008, USA.
authors.forename: Justin
authors.initials: J
authors.lastname: Lock
authors.affiliation: National Museums of Kenya, Box 40658-00100, Nairobi, Kenya.
authors.forename: Bernard
authors.initials: B
authors.lastname: Agwanda
authors.affiliation: Department of Biology, B322 Gottwald Center for the Sciences, University of Richmond, 28 Westhampton Way, Richmond, VA, 23173, USA.
authors.forename: Jory
authors.initials: J
authors.lastname: Brinkerhoff
authors.affiliation: Department of Biological Sciences, Old Dominion University, Norfolk, VA, 23529, USA.
authors.forename: Holly D
authors.initials: HD
authors.lastname: Gaff
authors.affiliation: Division of Mammals, National Museum of Natural History, Smithsonian Institution, MRC 108, P.O. Box 37012, Washington, DC, 20013-7012, USA.
authors.forename: Kristofer M
authors.initials: KM
authors.lastname: Helgen
authors.affiliation: Center for Conservation and Evolutionary Genetics, Smithsonian Conservation Biology Institute, 3001 Connecticut Avenue NW, Washington, DC, 20008, USA.|Division of Mammals, National Museum of Natural History, Smithsonian Institution, MRC 108, P.O. Box 37012, Washington, DC, 20013-7012, USA.
authors.forename: Jesús E
authors.initials: JE
authors.lastname: Maldonado
authors.affiliation: Smithsonian Conservation Biology Institute, National Zoological Park, 1500 Remount Rd., Front Royal, VA, 22630, USA.
authors.forename: William J
authors.initials: WJ
authors.lastname: McShea
authors.affiliation: Center for Conservation and Evolutionary Genetics, Smithsonian Conservation Biology Institute, 3001 Connecticut Avenue NW, Washington, DC, 20008, USA.
authors.forename: Robert C
authors.initials: RC
authors.lastname: Fleischer
mesh_terms: D000818:Animals; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D017753:Ecosystem; D060005:Genotyping Techniques / Q000379:methods*; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008967:Molecular Biology / Q000379:methods*; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
chemical_list: D004247:DNA
keywords: DNA; blood meal; ectoparasite; high-throughput sequencing; in-solution capture; pathogen