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.
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
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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
Researcher Affiliations
- Department of Veterinary Science, University of Melbourne, Victoria, Australia.
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 9 times.- 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.
- Jürgenschellert L, Krücken J, Bousquet E, Bartz J, Heyer N, Nielsen MK, von Samson-Himmelstjerna G. Occurrence of Strongylid Nematode Parasites on Horse Farms in Berlin and Brandenburg, Germany, With High Seroprevalence of Strongylus vulgaris Infection.. Front Vet Sci 2022;9:892920.
- Meigouni M, Makki M, Haniloo A, Askari Z, Mobedi I, Naddaf SR, Boenke N, Stollner T, Aali A, Heidari Z, Mowlavi G. Herbivores Coprolites from Chehrabad Salt Mine of Zanjan, Iran (Sassanid Era, 224-651 AD) Reveals Eggs of Strongylidae and Anoplocephalidae Helminths.. Iran J Parasitol 2020 Jan-Mar;15(1):109-114.
- Saeed MA, Beveridge I, Abbas G, Beasley A, Bauquier J, Wilkes E, Jacobson C, Hughes KJ, El-Hage C, O'Handley R, Hurley J, Cudmore L, Carrigan P, Walter L, Tennent-Brown B, Nielsen MK, Jabbar A. Systematic review of gastrointestinal nematodes of horses from Australia.. Parasit Vectors 2019 Apr 29;12(1):188.
- Camp LE, Radke MR, Shihabi DM, Pagan C, Yang G, Nadler SA. Molecular phylogenetics and species-level systematics of Baylisascaris.. Int J Parasitol Parasites Wildl 2018 Dec;7(3):450-462.
- Hoberg EP, Burek-Huntington K, Beckmen K, Camp LE, Nadler SA. Transuterine infection by Baylisascaris transfuga: Neurological migration and fatal debilitation in sibling moose calves (Alces alces gigas) from Alaska.. Int J Parasitol Parasites Wildl 2018 Dec;7(3):280-288.
- Poppert S, Heideking M, Agostini H, Fritzenwanker M, Wüppenhorst N, Muntau B, Henneke P, Kern W, Krücken J, Junker B, Hufnagel M. Diffuse Unilateral Subacute Neuroretinitis Caused by Ancylostoma Hookworm.. Emerg Infect Dis 2017 Feb;23(2):343-344.
- 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.
- 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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