title: Sources of blood meals of sylvatic Triatoma guasayana near Zurima, Bolivia, assayed with qPCR and 12S cloning.
abstract: BACKGROUND
  In this study we compared the utility of two molecular biology techniques, cloning of the mitochondrial 12S ribosomal RNA gene and hydrolysis probe-based qPCR, to identify blood meal sources of sylvatic Chagas disease insect vectors collected with live-bait mouse traps (also known as Noireau traps). Fourteen T. guasayana were collected from six georeferenced trap locations in the Andean highlands of the department of Chuquisaca, Bolivia.
  
  
  RESULTS
  We detected four blood meals sources with the cloning assay: seven samples were positive for human (Homo sapiens), five for chicken (Gallus gallus) and unicolored blackbird (Agelasticus cyanopus), and one for opossum (Monodelphis domestica). Using the qPCR assay we detected chicken (13 vectors), and human (14 vectors) blood meals as well as an additional blood meal source, Canis sp. (4 vectors).
  
  
  CONCLUSIONS
  We show that cloning of 12S PCR products, which avoids bias associated with developing primers based on a priori knowledge, detected blood meal sources not previously considered and that species-specific qPCR is more sensitive. All samples identified as positive for a specific blood meal source by the cloning assay were also positive by qPCR. However, not all samples positive by qPCR were positive by cloning. We show the power of combining the cloning assay with the highly sensitive hydrolysis probe-based qPCR assay provides a more complete picture of blood meal sources for insect disease vectors.
authors.affiliation: Department of Biology, University of Vermont, Burlington, Vermont, United States of America; Vector-borne Diseases Section, Tennessee Department of Health, Nashville, Tennessee, United States of America.
authors.forename: David E
authors.initials: DE
authors.lastname: Lucero
authors.affiliation: Facultad de Bioquímica, Universidad de San Francisco Xavier de Chuquisaca, Sucre, Bolivia.
authors.forename: Wilma
authors.initials: W
authors.lastname: Ribera
authors.affiliation: Facultad de Bioquímica, Universidad de San Francisco Xavier de Chuquisaca, Sucre, Bolivia.
authors.forename: Juan Carlos
authors.initials: JC
authors.lastname: Pizarro
authors.affiliation: Departamento de Entomología, Servicio Departamental de Salud, Sucre, Bolivia.
authors.forename: Carlos
authors.initials: C
authors.lastname: Plaza
authors.affiliation: Department of Biology, University of Vermont, Burlington, Vermont, United States of America.
authors.forename: Levi W
authors.initials: LW
authors.lastname: Gordon
authors.affiliation: Department of Biology, University of Vermont, Burlington, Vermont, United States of America.
authors.forename: Reynaldo
authors.initials: R
authors.lastname: Peña
authors.affiliation: Department of Biology, University of Vermont, Burlington, Vermont, United States of America.
authors.forename: Leslie A
authors.initials: LA
authors.lastname: Morrissey
authors.affiliation: Department of Biology, University of Vermont, Burlington, Vermont, United States of America.
authors.forename: Donna M
authors.initials: DM
authors.lastname: Rizzo
authors.affiliation: Department of Biology, University of Vermont, Burlington, Vermont, United States of America.
authors.forename: Lori
authors.initials: L
authors.lastname: Stevens
mesh_terms: D000818:Animals; D001838:Bolivia; D014355:Chagas Disease / Q000635:transmission; D002645:Chickens; D003001:Cloning, Molecular; D004032:Diet / Q000145:classification; D005247:Feeding Behavior / Q000502:physiology*; D006801:Humans; D007303:Insect Vectors / Q000502:physiology*; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods; D014224:Triatoma / Q000502:physiology*