Use of a reverse line blot assay to survey small strongyle (Strongylida: Cyathostominae) populations in horses before and after treatment with ivermectin

Mariana Ionita, Daniel K. Howe, Eugene T. Lyons, Sharon C. Tolliver, Ray M. Kaplan, Ioan Liviu Mitrea, Michelle Yeargan

Research output: Contribution to journalArticlepeer-review

27 Scopus citations


A sensitive and specific PCR hybridization assay was applied for species-specific monitoring of the small strongyle (Strongylida: Cyathostominae) populations in horses in a herd before and after treatment with the anthelmintic drug ivermectin. Fecal samples were collected pre- and post-treatment weekly from eight individual horses (four foals and four yearlings) for 6 weeks to determine counts of strongyle eggs per gram of feces (EPGs). Additionally, one foal and one yearling were nontreated controls. Also, one horse, from another herd known to be infected with Strongylus spp., was a positive control for these parasites. Genomic DNA was obtained from eggs in groups of approximately 6000-7000 eggs except for two samples containing low EPGs in which 450 eggs were used. Amplification of the intergenic spacers (IGSs) of ribosomal DNA (rDNA) of small and large strongyles followed by reverse line blot (RLB) assay were performed to identify the presence of the 12 most common equine small strongyle species and to discriminate them from Strongylus spp. Overall, 11 small strongyle species were identified in pretreatment samples. In the samples collected at 4 weeks after ivermectin treatment, eight small strongyle species were identified and four of them were predominant (Cylicocyclus nassatus, Cylicostephanus longibursatus, Cylicostephanus calicatus and Cylicostephanus minutus). At 5 and 6 weeks post-treatment, the RLB assay analysis showed almost the same composition in the small strongyle population as before treatment. Strongylus spp. were identified only in samples collected from the positive control horse for these parasites. These data confirm the ability of the PCR-RLB technique for simultaneous species-specific differentiation of equine strongyle eggs, indicating a valuable way of furthering drug-resistance studies.

Original languageEnglish
Pages (from-to)332-337
Number of pages6
JournalVeterinary Parasitology
Issue number3-4
StatePublished - Mar 25 2010

Bibliographical note

Funding Information:
This work was supported by the Albert and Lorraine Clay Research Fellowship awarded by Department of Veterinary Science, Gluck Equine Research Center, University of Kentucky, and Grant Id_729/2007 awarded by CNCSIS , Romania.


  • Cyathostomins
  • Horse
  • Ivermectin
  • PCR
  • Reverse line blot (RLB)
  • Small strongyles

ASJC Scopus subject areas

  • Parasitology
  • General Veterinary


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