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Sources of blood meals of sylvatic Triatoma guasayana near Zurima, Bolivia, assayed with qPCR and 12S cloning.

Lucero DE, Ribera W, Pizarro JC, Plaza C, Gordon LW, Peña R, Morrissey LA, Rizzo DM, Stevens L - PLoS Negl Trop Dis (2014)

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

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.

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.

Methodology/principal findings: 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/significance: 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.

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Related in: MedlinePlus

Noireau sylvatic insect vector trap.Traps consisted of plastic containers with a live mouse and wire mesh tops. Double sided sticky tape was placed on the outside of the traps.
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pntd-0003365-g002: Noireau sylvatic insect vector trap.Traps consisted of plastic containers with a live mouse and wire mesh tops. Double sided sticky tape was placed on the outside of the traps.

Mentions: Trapping many insect vectors is challenging due to the low success rates of traps [27]. We baited traps with live mice (also known as Noireau traps) because a review of the literature suggested these traps attract T. infestans, the principal insect vector in the region, more successfully than traps without mice [28] in laboratory studies, and that they have been successful in previous field studies [27], [29]. Traps consisted of opaque bottles (15×7 cm) covered with double-sided tape. A mouse was placed inside with a small piece of apple; and the opening was sealed with a metal screening mesh to prevent adult vectors and large nymphs from entering the trap (Figure 2). One to four traps were placed at six georeferenced sampling locations for a total of 17 traps (Figure 1). Traps west of the riverbed were placed in sylvan areas, while the eastern traps were between the village of Zurima and agricultural areas.


Sources of blood meals of sylvatic Triatoma guasayana near Zurima, Bolivia, assayed with qPCR and 12S cloning.

Lucero DE, Ribera W, Pizarro JC, Plaza C, Gordon LW, Peña R, Morrissey LA, Rizzo DM, Stevens L - PLoS Negl Trop Dis (2014)

Noireau sylvatic insect vector trap.Traps consisted of plastic containers with a live mouse and wire mesh tops. Double sided sticky tape was placed on the outside of the traps.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4256209&req=5

pntd-0003365-g002: Noireau sylvatic insect vector trap.Traps consisted of plastic containers with a live mouse and wire mesh tops. Double sided sticky tape was placed on the outside of the traps.
Mentions: Trapping many insect vectors is challenging due to the low success rates of traps [27]. We baited traps with live mice (also known as Noireau traps) because a review of the literature suggested these traps attract T. infestans, the principal insect vector in the region, more successfully than traps without mice [28] in laboratory studies, and that they have been successful in previous field studies [27], [29]. Traps consisted of opaque bottles (15×7 cm) covered with double-sided tape. A mouse was placed inside with a small piece of apple; and the opening was sealed with a metal screening mesh to prevent adult vectors and large nymphs from entering the trap (Figure 2). One to four traps were placed at six georeferenced sampling locations for a total of 17 traps (Figure 1). Traps west of the riverbed were placed in sylvan areas, while the eastern traps were between the village of Zurima and agricultural areas.

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

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.

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.

Methodology/principal findings: 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/significance: 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.

Show MeSH
Related in: MedlinePlus