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Molecular and cellular probes1996; 10(5); 371-378; doi: 10.1006/mcpr.1996.0050

Species markers for equine strongyles detected in intergenic rDNA by PCR-RFLP.

Abstract: Five species of equine strongyle belonging to the subfamily Strongylinae (Strongylus edentatus, S. equinus, S. vulgaris, Oesophagodontus robustus and Triodontophorus serratus) and 11 species belonging to the subfamily Cyathostominae (Poteriostomum imparidentatum, P. ratzii, Cylicocyclus insignis, Cc. leptostomus, Cc. nassatus, Cylicostephanus calicatus, Cs. longibursatus, Cs. goldi, Cyathostomum catinatum, Cy. labiatum and Cy. pateratum) were characterized using a polymerase chain reaction-linked restriction fragment length polymorphism technique (PCR-RFLP). Internal transcribed spacer ribosomal DNA was amplified from genomic DNA by polymerase chain reaction (PCR) using conserved primers, digested separately with six restriction endonucleases (AluI, BfaI, CfoI, Hae III, VSpI and XbaI) and the fragments separated by agarose gel electrophoresis. The PCR products of the three Strongylus species were approx. 90-100 bp smaller in size compared with those of the other 13 species. The PCR-RFLP analysis of the rDNA region spanning the first and second internal transcribed spacers plus the 5.85 rDNA gene (ITS+) produced characteristic patterns for each of the 16 species examined, and no variation in RFLP patterns was detected within the species Cy. catinatum, where multiple isolates were analysed. The study demonstrates that the internal transcribed spacer sequences provide genetic markers for the species identification of a range of equine strongyles. These markers will be of use for the identification of egg and larval stages, where morphological characters alone are unreliable. The results also indicate that the spacer sequences will be of use to study the systematics of equine strongyles.
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Publication Date: 1996-10-01 PubMed ID: 8910892DOI: 10.1006/mcpr.1996.0050Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article studies the use of a scientific technique to identify 16 different species of equine strongyles, a type of parasitic worm in horses, using genetic markers. The aim is to enable easier identification of the worms and contribute to understanding their systematics.

Methodology

  • The study focused on certain species of equine strongyle from the subfamilies Strongylinae and Cyathostominae.
  • The researchers used a technique known as the polymerase chain reaction-linked restriction fragment length polymorphism (PCR-RFLP) to classify the different species.
  • This technique involves amplifying the internal transcribed spacer (ITS) ribosomal DNA from the worm’s genomic DNA using conserved primers.
  • The amplified DNA was then digested with six different restriction enzymes and the resulting fragments separated using agarose gel electrophoresis.

Findings

  • The research revealed that the PCR products of three Strongylus species were noticeably smaller in size compared with the PCR products of the other 13 species.
  • The PCR-RFLP analysis of the region of ribosomal DNA that included the first and second internal transcribed spacers and the 5.85 rDNA gene provided specific patterns for each of the 16 species.
  • The researchers did not observe any variation in RFLP patterns within the species Cy. catinatum, where multiple isolates were analyzed.

Significance and Application

  • This study demonstrates that the ITS sequences provide genetic markers for identifying different species of equine strongyles.
  • These genetic markers will be particularly useful for identifying egg and larval stages of the worms, where morphological characteristics may not be distinct enough for accurate identification.
  • The research also suggests that these spacer sequences could be useful in studying the systematics of equine strongyles, potentially contributing to improved management and treatment of these parasites in horses.

Cite This Article

APA
Gasser RB, Stevenson LA, Chilton NB, Nansen P, Bucknell DG, Beveridge I. (1996). Species markers for equine strongyles detected in intergenic rDNA by PCR-RFLP. Mol Cell Probes, 10(5), 371-378. https://doi.org/10.1006/mcpr.1996.0050

Publication

Molecular and cellular probes
ISSN: 0890-8508
NlmUniqueID: 8709751
Country: England
Language: English
Volume: 10
Issue: 5
Pages: 371-378

Researcher Affiliations

Gasser, R B
  • Department of Veterinary Science, University of Melbourne, Victoria, Australia.
Stevenson, L A
    Chilton, N B
      Nansen, P
        Bucknell, D G
          Beveridge, I

            MeSH Terms

            • Animals
            • DNA, Helminth / genetics
            • DNA, Ribosomal / genetics
            • Genetic Markers
            • Horses
            • Polymerase Chain Reaction / methods
            • Polymorphism, Restriction Fragment Length
            • Species Specificity
            • Strongylida / genetics
            • Victoria

            Citations

            This article has been cited 12 times.
            1. Ma X, Bai X, Li H, Ding J, Zhang H, Qiu Y, Wang J, Liu X, Liu M, Tang B, Xu N. A rapid and visual detection assay for Clonorchis sinensis based on recombinase polymerase amplification and lateral flow dipstick. Parasit Vectors 2023 May 19;16(1):165.
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              pubmed: 32489382
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              doi: 10.1016/j.ijppaw.2018.07.005pubmed: 30094177google scholar: lookup
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            8. Kharchenko V, Kuzmina T, Trawford A, Getachew M, Feseha G. Morphology and diagnosis of some fourth-stage larvae of cyathostomines (Nematoda: Strongyloidea) in donkeys Equus asinus L. from Ethiopia. Syst Parasitol 2009 Jan;72(1):1-13.
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            9. Posedi J, Drögemüller M, Schnieder T, Höglund J, Lichtenfels JR, von Samson-Himmelstjerna G. Microchip capillary electrophoresis-based genetic comparison of closely related cyathostomin nematode parasites of horses using randomly amplified polymorphic DNA polymerase chain reaction. Parasitol Res 2004 Mar;92(5):421-9.
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            10. Wang T, Chen X, Yan X, Su Y, Gao W, Liu C, Wang W. Progress in serology and molecular biology of equine parasite diagnosis: sustainable control strategies. Front Vet Sci 2025;12:1663577.
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              doi: 10.1111/evj.14134pubmed: 39012065google scholar: lookup
            12. Buono F, Veneziano V, Veronesi F, Molento MB. Horse and donkey parasitology: differences and analogies for a correct diagnostic and management of major helminth infections. Parasitology 2023 Oct;150(12):1119-1138.
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