A repeatable and quantitative DNA metabarcoding assay to characterize mixed strongyle infections in horses |
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Authors: | Jocelyn Poissant Stefan Gavriliuc Jennifer Bellaw Elizabeth M Redman Russell W Avramenko David Robinson Matthew L Workentine Todd K Shury Emily J Jenkins Philip D McLoughlin Martin K Nielsen John S Gilleard |
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Institution: | 1. Department of Ecosystem and Public Health, University of Calgary, 3280 Hospital Drive, Calgary, AB T2N 4Z6, Canada;2. M.H. Gluck Equine Research Center, Department of Veterinary Science, University of Kentucky, Lexington, KY, USA;3. Department of Comparative Biology and Experimental Medicine, Host-Parasite Interactions (HPI) Program, University of Calgary, 3280 Hospital Drive, Calgary, AB T2N 4Z6, Canada;4. Faculty of Veterinary Medicine, University of Calgary, 3280 Hospital Drive, Calgary, AB T2N 4Z6, Canada;5. Parks Canada Agency, 52 Campus Drive, Saskatoon, SK S7N 5B4, Canada;6. Department of Veterinary Microbiology, University of Saskatchewan, 52 Campus drive, Saskatoon, SK S7N 5B4, Canada;7. Department of Biology, University of Saskatchewan, 112 Science Place, Saskatoon, SK S7N 5E2, Canada |
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Abstract: | Horses are ubiquitously infected by a diversity of gastro-intestinal parasitic helminths. Of particular importance are nematodes of the family Strongylidae, which can significantly impact horse health and performance. However, knowledge about equine strongyles remains limited due to our inability to identify most species non-invasively using traditional morphological techniques. We developed a new internal transcribed spacer 2 (ITS2) DNA metabarcoding ‘nemabiome’ assay to characterise mixed strongyle infections in horses and assessed its performance by applying it to pools of infective larvae from fecal samples from an experimental herd in Kentucky, USA and two feral horse populations from Sable Island and Alberta, Canada. In addition to reporting the detection of 33 different species with high confidence, we illustrate the assay’s repeatability by comparing results generated from aliquots from the same fecal samples and from individual horses sampled repeatedly over multiple days or months. We also validate the quantitative potential of the assay by demonstrating that the proportion of amplicon reads assigned to different species scales linearly with the number of larvae present. This new tool significantly improves equine strongyle diagnostics, presenting opportunities for research on species-specific anthelmintic resistance and the causes and consequences of variation in mixed infections. |
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Keywords: | Cyathostomin Diagnostic Host-parasite interaction Nematode Nemabiome Parasitology |
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