G A Lumsden1, E V Zakharov2, S Dolynskyj2, J S Weese3, L R Lindsay4, C M Jardine3,5. 1. Ontario Veterinary College, University of Guelph, 50 Stone Road E., Guelph, ON, N1G2W1, Canada. glumsden@uoguelph.ca. 2. Canadian Centre for DNA Barcoding, Biodiversity Institute of Ontario, University of Guelph, 579 Gordon St., Guelph, ON, N1G2W1, Canada. 3. Ontario Veterinary College, University of Guelph, 50 Stone Road E., Guelph, ON, N1G2W1, Canada. 4. Public Health Agency of Canada, National Microbiology Laboratory, 1015 Arlington St., Winnipeg, MB, R3E 3R2, Canada. 5. Canadian Wildlife Health Cooperative, Ontario Veterinary College, University of Guelph, 50 Stone Road E., Guelph, ON, N1G2W1, Canada.
Abstract
OBJECTIVE: Our objective was to apply next-generation sequence-based DNA barcoding to identify the remnant larval bloodmeals in wild-caught host-seeking (unengorged) Ixodes scapularis nymphs (n = 216). To infer host species identification, vertebrate DNA was amplified using universal primers for cytochrome c oxidase subunit I (COI) and sequenced using next-generation sequencing (NGS) for comparison against known barcode references. RESULTS: Bloodmeal identification was unsuccessful in most samples (99% of 216 specimens) demonstrating a very low detection rate of this assay. Sequences that surpassed quality thresholds were obtained for 41.7% of nymphs (n = 90) and of those, confident species identification was obtained for 15.6% of nymphs (n = 14). Wild host identifications were only obtained from 2 specimens, where DNA from the eastern grey squirrel (Sciurus carolinensis) was identified. Human and bovine DNA was identified in remaining nymphs and considered to be contaminants. Further optimization of the technique is required to improve detection of remnant bloodmeals in host-seeking nymphs.
OBJECTIVE: Our objective was to apply next-generation sequence-based DNA barcoding to identify the remnant larval bloodmeals in wild-caught host-seeking (unengorged) Ixodes scapularis nymphs (n = 216). To infer host species identification, vertebrate DNA was amplified using universal primers for cytochrome c oxidase subunit I (COI) and sequenced using next-generation sequencing (NGS) for comparison against known barcode references. RESULTS: Bloodmeal identification was unsuccessful in most samples (99% of 216 specimens) demonstrating a very low detection rate of this assay. Sequences that surpassed quality thresholds were obtained for 41.7% of nymphs (n = 90) and of those, confident species identification was obtained for 15.6% of nymphs (n = 14). Wild host identifications were only obtained from 2 specimens, where DNA from the eastern grey squirrel (Sciurus carolinensis) was identified. Human and bovine DNA was identified in remaining nymphs and considered to be contaminants. Further optimization of the technique is required to improve detection of remnant bloodmeals in host-seeking nymphs.
Authors: Katie M Clow; Nicholas H Ogden; L Robbin Lindsay; Pascal Michel; David L Pearl; Claire M Jardine Journal: Vector Borne Zoonotic Dis Date: 2016-02-12 Impact factor: 2.133
Authors: Pierre-François Humair; Véronique Douet; Francisca Morán Cadenas; Leo M Schouls; Ingrid Van De Pol; Lise Gern Journal: J Med Entomol Date: 2007-09 Impact factor: 2.278