Evaluation of a larval development assay (DrenchRite®) for the detection of anthelmintic resistance in cyathostomin nematodes of horses
Abstract
A larval development assay (LDA, DrenchRite) was evaluated to determine the effectiveness of this method in detecting anthelmintic resistance in cyathostomin nematodes of horses. A total of 15 horse farms from Georgia and South Carolina (USA) and Population S ponies from the University of Kentucky (USA) were included in this study. Nematode eggs were extracted from pooled fecal samples and placed into the wells of a DrenchRite plate for testing against thiabendazole (TBZ), levamisole (LEV) and 2 ivermectin (IVM) analogs (IVM-1, IVM-2). After a 7-day incubation larvae in each well were counted and data were analyzed by logistic regression. Resistance status of each farm for different drugs was determined in a separate study using a fecal egg count reduction test. LDA were performed on the 15 farms once, however, the Population S cyathostomins were assayed on 3 separate occasions to estimate the consistency of results between assays. Mean TBZ LC50 for oxibendazole resistant, suspected resistant and sensitive farms were 0.2015, 0.1625, and 0.1355 microM, respectively. For LEV, mean LC50 for PYR resistant, suspected resistant and sensitive farms were 1.590, 1.8018 and 1.4219 microM, respectively. All 15 farms had worms susceptible to IVM; mean LC50 for IVM-1 and for IVM-2 were 7.5727 and 87.9718 nM, respectively. A linear mixed model was fitted to the data to determine the relationship between LC50 and LC95 and resistance status for each farm. No meaningful relations were found. Consistency of assays varied between drugs, being best for TBZ and worst for LEV and IVM-1. All farms in this study had benzimidazole-resistant nematodes; therefore usefulness of DrenchRite for discriminating susceptibility versus resistance to this drug class could not be accurately assessed. Moreover, since all farms tested were sensitive to IVM and resistance to this drug class has not yet been reported in cyathostomins, it is not possible to assess accurately the usefulness of DrenchRite LDA for detecting IVM resistance at this time. Assay results for LEV suggest that LEV in a LDA does not yield data that is useful in estimating PYR efficacy in vivo. Based on results for PYR/LEV, the current high prevalence of benzimidazole resistance, no known cases of IVM resistance, and the sometimes extreme variation in results seen in many of the assays, DrenchRite LDA cannot be considered a useful tool for the diagnosis of resistance in cyathostomins of horses at present.
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In vitro bioassays used for detection of anthelmintic resistance in parasitic nematodes are highly species-specific and drug class-specific in their ability to provide useful levels of discrimination between susceptible and resistant isolates.
“This finding builds on previous works demonstrating that in vitro bioassays used for detection of anthelmintic resistance in parasitic nematodes are highly species-specific and drug class-specific in their ability to provide useful levels of discrimination between susceptible and resistant isolates [43, 57, 63].”
From Multiple drug resistance in the canine hookworm Ancylostoma caninum: an emerging threat? (2019) · cites this paper as [43]
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- Authors
- Tandon, R., & Kaplan, R. M.
- Affiliations
- University of Georgia (US)
- Corresponding author
- Kaplan, R.M.
- Venue
- Veterinary Parasitology
- Publisher
- Elsevier BV
- Published
- 2004-05
- Volume
- 121
- Issue
- 1-2
- Pages
- 125-142
- ISSN
- 0304-4017 (ISSN-L)
- OpenAlex
- W2048164308
- Cited by
- 49
- References
- 46
- Citations by year
- 2025: 1 · 2023: 2 · 2022: 1 · 2020: 2 · 2019: 3 · 2018: 3 · 2017: 5 · 2016: 3 · 2015: 5 · 2014: 1 · 2013: 1 · 2012: 3
- FWCI
- 4.61
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Veterinary Parasitology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- University of Kentucky
- University of Georgia
- Topics
- Health Sciences › Veterinary › Small Animals › Helminth infection and control
- Health Sciences › Veterinary › Small Animals › Veterinary Pharmacology and Anesthesia
- Health Sciences › Veterinary › Equine › Veterinary Equine Medical Research
- Keywords
- anthelmintic resistance, cyathostomins, horses, larval development assay, thiabendazole, levamisole, oxibendazole, pyrantel pamoate, benzimidazole resistance, faecal egg count reduction test
- MeSH terms
- Animals, Antinematodal Agents, Drug Resistance, Feces, Horse Diseases, Horses, Ivermectin, Larva, Levamisole, Nematoda, Nematode Infections, Parasite Egg Count, Reagent Kits, Diagnostic, Regression Analysis, Thiabendazole, Reproducibility of Results, Linear Models
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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