Temporal Detection Limits of Remnant Larval Bloodmeals in Nymphal Ixodes scapularis (Say, Ixodida: Ixodidae) Using Two Next-Generation Sequencing DNA Barcoding Assays
| Autor: | Sarah Dolynskyj, Genevieve Lumsden, L. Robbin Lindsay, Evgeny Zakharov, J. Scott Weese, Claire M. Jardine |
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| Rok vydání: | 2020 |
| Předmět: |
Nymph
030231 tropical medicine Zoology DNA barcoding DNA sequencing Host-Parasite Interactions Electron Transport Complex IV 03 medical and health sciences 0302 clinical medicine Limit of Detection parasitic diseases Animals DNA Barcoding Taxonomic 030304 developmental biology 0303 health sciences Larva Ixodes General Veterinary biology Cytochrome c oxidase subunit I High-Throughput Nucleotide Sequencing Feeding Behavior biology.organism_classification Blood Infectious Diseases Genetic Techniques Ixodes scapularis Insect Science Vertebrates Parasitology Rabbits Primer (molecular biology) Ixodidae |
| Zdroj: | Journal of Medical Entomology. 58:821-829 |
| ISSN: | 1938-2928 0022-2585 |
| Popis: | Using next-generation sequencing DNA barcoding, we aimed to determine: 1) if the larval bloodmeal can be detected in Ixodes scapularis nymphs and 2) the post-moult temporal window for detection of the larval bloodmeal. Subsets of 30 nymphs fed on a domestic rabbit (Oryctolagus cuniculus Linnaeus, Lagomorphia: Leporidae) as larvae were reared and frozen at 11 time points post-moult, up to 150 d. Vertebrate DNA was amplified using novel universal (UP) and species-specific primers (SSP) and sequenced for comparison against cytochrome c oxidase subunit I barcodes to infer host identification. Detectable bloodmeals decreased as time since moult increased for both assays. For the SSP assay, detection of bloodmeals decreased from 96.7% (n = 29/30) in day 0 nymphs to 3.3% (n = 1/30) and 6.7% (n = 2/30) at 4- and 5-mo post-moult, respectively. A shorter temporal detection period was achieved with the UP assay, declining from 16.7% (n = 5/30) in day 0 nymphs to 0/30 in 3-d-old nymphs. Bloodmeal detection was nonexistent for the remaining cohorts, with the exception of 1/30 nymphs at 2-mo post-moult. Host detection was significantly more likely using the SSP assay compared to the UP assay in the first three time cohorts (day 0: χ 2 = 39.1, P < 0.005; day 2: χ 2 = 19.2, P < 0.005; day 3: χ 2 = 23.3, P < 0.005). Regardless of the primer set used, the next-generation sequencing DNA barcoding assay was able to detect host DNA from a larval bloodmeal in the nymphal life stage; however, a short window with a high proportion of detection post-moult was achieved. |
| Databáze: | OpenAIRE |
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