A repeatable and quantitative DNA metabarcoding assay to characterize mixed strongyle infections in horses.
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.
Copyright © 2020 Australian Society for Parasitology. All rights reserved.
Publication Date: 2020-11-23 PubMed ID: 33242465DOI: 10.1016/j.ijpara.2020.09.003Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The study developed a DNA metabarcoding assay to provide a detailed profile of mixed strongyle (a family of parasitic worms) infections in horses. The technique’s reliability and quantitative ability were verified through repeat testing of fecal samples, with results demonstrating a correlation between the number of larvae and the percentage of assigned amplicon reads. The findings pave the way for advanced research into species-specific drug resistance and dynamics of mixed infections.
Article Overview
- The research was centered on creating and validating a DNA metabarcoding assay capable of profiling mixed strongyle infections in horses. Strongyles are parasitic nematodes that can significantly impact horse health and performance.
- It aimed to overcome the limitations of traditional methods that rely on morphology and are unable to non-invasively identify most strongyle species.
Methodology
- The researchers designed an internal transcribed spacer 2 (ITS2) DNA metabarcoding ‘nemabiome’ assay for this purpose.
- The newly-developed assay was applied on pools of infective larvae obtained from fecal samples of an experimental herd in Kentucky, USA, and two wild horse populations from Sable Island and Alberta, Canada.
Findings
- The assay resulted in the detection of 33 different strongyle species with a high degree of confidence.
- Repeatability of the assay was verified by testing aliquots from the same fecal sample and from individual horses on multiple occasions.
- The research also confirmed the assay’s quantitative potential by showing a linear correlation between the proportion of amplicon reads assigned to different species and the number of larvae present.
Implications
- The successful development and validation of this assay contributes to significant improvement in the diagnostics of equine strongyle infections.
- It opens up new possibilities for research into species-specific anthelmintic (anti-parasitic) resistance and the factors influencing variation in mixed infections.
Cite This Article
APA
Poissant J, Gavriliuc S, Bellaw J, Redman EM, Avramenko RW, Robinson D, Workentine ML, Shury TK, Jenkins EJ, McLoughlin PD, Nielsen MK, Gilleard JS.
(2020).
A repeatable and quantitative DNA metabarcoding assay to characterize mixed strongyle infections in horses.
Int J Parasitol, 51(2-3), 183-192.
https://doi.org/10.1016/j.ijpara.2020.09.003 Publication
Researcher Affiliations
- Department of Ecosystem and Public Health, University of Calgary, 3280 Hospital Drive, Calgary, AB T2N 4Z6, Canada. Electronic address: jocelyn.poissant@ucalgary.ca.
- Department of Ecosystem and Public Health, University of Calgary, 3280 Hospital Drive, Calgary, AB T2N 4Z6, Canada.
- M.H. Gluck Equine Research Center, Department of Veterinary Science, University of Kentucky, Lexington, KY, USA.
- Department of Comparative Biology and Experimental Medicine, Host-Parasite Interactions (HPI) Program, University of Calgary, 3280 Hospital Drive, Calgary, AB T2N 4Z6, Canada.
- Department of Comparative Biology and Experimental Medicine, Host-Parasite Interactions (HPI) Program, University of Calgary, 3280 Hospital Drive, Calgary, AB T2N 4Z6, Canada.
- Faculty of Veterinary Medicine, University of Calgary, 3280 Hospital Drive, Calgary, AB T2N 4Z6, Canada.
- Faculty of Veterinary Medicine, University of Calgary, 3280 Hospital Drive, Calgary, AB T2N 4Z6, Canada.
- Parks Canada Agency, 52 Campus Drive, Saskatoon, SK S7N 5B4, Canada.
- Department of Veterinary Microbiology, University of Saskatchewan, 52 Campus drive, Saskatoon, SK S7N 5B4, Canada.
- Department of Biology, University of Saskatchewan, 112 Science Place, Saskatoon, SK S7N 5E2, Canada.
- M.H. Gluck Equine Research Center, Department of Veterinary Science, University of Kentucky, Lexington, KY, USA.
- Department of Comparative Biology and Experimental Medicine, Host-Parasite Interactions (HPI) Program, University of Calgary, 3280 Hospital Drive, Calgary, AB T2N 4Z6, Canada.
MeSH Terms
- Alberta
- Animals
- Anthelmintics / therapeutic use
- Coinfection
- DNA Barcoding, Taxonomic
- Feces
- Horse Diseases / diagnosis
- Horse Diseases / drug therapy
- Horses
- Parasite Egg Count / veterinary
- Strongyle Infections, Equine / diagnosis
Citations
This article has been cited 11 times.- Courtot É, Boisseau M, Dhorne-Pollet S, Serreau D, Gesbert A, Reigner F, Basiaga M, Kuzmina T, Lluch J, Annonay G, Kuchly C, Diekmann I, Krücken J, von Samson-Himmelstjerna G, Mach N, Sallé G. Comparison of two molecular barcodes for the study of equine strongylid communities with amplicon sequencing.. PeerJ 2023;11:e15124.
- Boisseau M, Dhorne-Pollet S, Bars-Cortina D, Courtot É, Serreau D, Annonay G, Lluch J, Gesbert A, Reigner F, Sallé G, Mach N. Species interactions, stability, and resilience of the gut microbiota - Helminth assemblage in horses.. iScience 2023 Feb 17;26(2):106044.
- Boisseau M, Mach N, Basiaga M, Kuzmina T, Laugier C, Sallé G. Patterns of variation in equine strongyle community structure across age groups and gut compartments.. Parasit Vectors 2023 Feb 11;16(1):64.
- Davey ML, Kamenova S, Fossøy F, Solberg EJ, Davidson R, Mysterud A, Rolandsen CM. Faecal metabarcoding provides improved detection and taxonomic resolution for non-invasive monitoring of gastrointestinal nematode parasites in wild moose populations.. Parasit Vectors 2023 Jan 18;16(1):19.
- Halvarsson P, Tydén E. The complete ITS2 barcoding region for Strongylus vulgaris and Strongylus edentatus.. Vet Res Commun 2023 Sep;47(3):1767-1771.
- Baltrušis P, Halvarsson P, Höglund J. Estimation of the impact of three different bioinformatic pipelines on sheep nemabiome analysis.. Parasit Vectors 2022 Aug 11;15(1):290.
- Malsa J, Courtot É, Boisseau M, Dumont B, Gombault P, Kuzmina TA, Basiaga M, Lluch J, Annonay G, Dhorne-Pollet S, Mach N, Sutra JF, Wimel L, Dubois C, Guégnard F, Serreau D, Lespine A, Sallé G, Fleurance G. Effect of sainfoin (Onobrychis viciifolia) on cyathostomin eggs excretion, larval development, larval community structure and efficacy of ivermectin treatment in horses.. Parasitology 2022 Sep;149(11):1439-1449.
- Titcomb GC, Pansu J, Hutchinson MC, Tombak KJ, Hansen CB, Baker CCM, Kartzinel TR, Young HS, Pringle RM. Large-herbivore nemabiomes: patterns of parasite diversity and sharing.. Proc Biol Sci 2022 May 11;289(1974):20212702.
- Papaiakovou M, Littlewood DTJ, Doyle SR, Gasser RB, Cantacessi C. Worms and bugs of the gut: the search for diagnostic signatures using barcoding, and metagenomics-metabolomics.. Parasit Vectors 2022 Apr 1;15(1):118.
- Tombak KJ, Hansen CB, Kinsella JM, Pansu J, Pringle RM, Rubenstein DI. The gastrointestinal nematodes of plains and Grevy's zebras: Phylogenetic relationships and host specificity.. Int J Parasitol Parasites Wildl 2021 Dec;16:228-235.
- Davey ML, Utaaker KS, Fossøy F. Characterizing parasitic nematode faunas in faeces and soil using DNA metabarcoding.. Parasit Vectors 2021 Aug 21;14(1):422.
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