Species-specific amplification by PCR of ribosomal DNA from some equine strongyles.
Abstract: The first and second internal transcribed spacer sequences of 28 morphologically-defined species of horse strongyle were characterized, and specific oligonucleotide primers were designed for some species based on the nucleotide differences. Utilizing these primers, a PCR approach was developed for the specific amplification of ribosomal DNA of Strongylus vulgaris, Cyathostomum catinatum, Cylicocyclus nassatus, Cylicostephanus longibursatus or Cylicostephanus goldi. The method allowed the species-specific amplification of parasite DNA derived from faecal samples and/or copro-cultures, demonstrating the potential of the approach for the diagnosis of equine strongyloidosis. The establishment of this PCR assay also has implications for studying the biology and epidemiology of equine strongyles and anthelmintic resistance using faecal egg count reduction tests.
Publication Date: 1999-08-14 PubMed ID: 10446706DOI: 10.1017/s0031182099004497Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Anthelmintic Resistance
- Diagnosis
- Diagnostic Technique
- Disease Diagnosis
- Disease Management
- DNA
- Epidemiology
- Equine Diseases
- Equine Health
- Fecal Egg Count
- Horses
- Infection
- Infectious Disease
- Laboratory Methods
- Molecular biology
- Parasites
- Polymerase Chain Reaction
- Species Comparison
- Strongyles
- Veterinary Medicine
- Veterinary Research
Summary
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The research article presents the development of a Polymerase Chain Reaction (PCR) method to specifically amplify the ribosomal DNA of certain species of horse parasites or strongyles, enhancing the potential for their diagnosis and further study.
Objective of the Study
The main objective of the study was:
- To characterize the internal transcribed spacer sequences of 28 horse strongyle species.
- To design specific oligonucleotide primers for some species based on the nucleotide differences.
- To develop a PCR method for the specific amplification of ribosomal DNA of Strongylus vulgaris, Cyathostomum catinatum, Cylicocyclus nassatus, Cylicostephanus longibursatus or Cylicostephanus goldi.
Methodology
Through this research, the researchers followed several steps:
- Initially, they characterized the first and second internal transcribed spacer sequences of 28 different species of horse strongyle.
- They utilized the differences in nucleotide sequences to design specific oligonucleotide primers for some species.
- Employing these primers, they developed a PCR approach to specifically amplify the ribosomal DNA of Strongylus vulgaris, Cyathostomum catinatum, Cylicocyclus nassatus, Cylicostephanus longibursatus, and Cylicostephanus goldi.
- They tested the method using parasite DNA derived from faecal samples and/or copro-cultures.
Results and Implications
The results showed that:
- The developed PCR method enabled the species-specific amplification of parasite DNA from faecal samples and/or copro-cultures.
- This result demonstrated the potential of the developed PCR approach for diagnosing equine strongyloidosis – a condition caused by strongyle infestation in horses.
- The establishment of this PCR assay will also aid in studying the biology and epidemiology of equine strongyles.
- Furthermore, it has strong implications for understanding anthelmintic resistance via faecal egg count reduction tests, potentially leading towards improved treatment strategies and prevention measures.
Cite This Article
APA
Hung GC, Gasser RB, Beveridge I, Chilton NB.
(1999).
Species-specific amplification by PCR of ribosomal DNA from some equine strongyles.
Parasitology, 119 ( Pt 1), 69-80.
https://doi.org/10.1017/s0031182099004497 Publication
Researcher Affiliations
- Department of Veterinary Science, University of Melbourne, Australia.
MeSH Terms
- Animals
- Base Composition
- DNA Primers
- DNA, Ribosomal / genetics
- DNA, Ribosomal / isolation & purification
- Feces / parasitology
- Horse Diseases / parasitology
- Horses
- Molecular Sequence Data
- Polymerase Chain Reaction / methods
- RNA, Ribosomal, 18S / genetics
- RNA, Ribosomal, 28S / genetics
- RNA, Ribosomal, 5.8S / genetics
- Sequence Analysis, DNA
- Species Specificity
- Strongylida / classification
- Strongylida / genetics
- Strongylida Infections / veterinary
Citations
This article has been cited 16 times.- 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.
- Martins AV, Coelho AL, Corrêa LL, Ribeiro MS, Lobão LF, Palmer JPS, Moura LC, Molento MB, Barbosa ADS. First microscopic and molecular parasitological survey of Strongylus vulgaris in Brazilian ponies. Rev Bras Parasitol Vet 2023;32(3):e006323.
- 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.
- 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.
- 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.
- 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.
- 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.
- Bredtmann CM, Krücken J, Murugaiyan J, Kuzmina T, von Samson-Himmelstjerna G. Nematode Species Identification-Current Status, Challenges and Future Perspectives for Cyathostomins. Front Cell Infect Microbiol 2017;7:283.
- Clough D, Råberg L. Contrasting patterns of structural host specificity of two species of Heligmosomoides nematodes in sympatric rodents. Parasitol Res 2014 Dec;113(12):4633-9.
- Rakhshandehroo E, Sharifiyazdi H, Shayegh H, Ahmadi A. Molecular and morphological comparison of two different types of Habronema muscae (Nematoda: Habronematidae) in horse. Parasitol Res 2014 Dec;113(12):4439-45.
- Roeber F, Jex AR, Gasser RB. Next-generation molecular-diagnostic tools for gastrointestinal nematodes of livestock, with an emphasis on small ruminants: a turning point?. Adv Parasitol 2013;83:267-333.
- Roeber F, Jex AR, Gasser RB. Advances in the diagnosis of key gastrointestinal nematode infections of livestock, with an emphasis on small ruminants. Biotechnol Adv 2013 Dec;31(8):1135-52.
- 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.
- Lin RQ, Ai L, Zou FC, Verweij JJ, Jiang Q, Li MW, Song HQ, Zhu XQ. A multiplex PCR tool for the specific identification of Oesophagostomum spp. from pigs. Parasitol Res 2008 Sep;103(4):993-7.
- Traversa D, Iorio R, Klei TR, Kharchenko VA, Gawor J, Otranto D, Sparagano OA. New method for simultaneous species-specific identification of equine strongyles (nematoda, strongylida) by reverse line blot hybridization. J Clin Microbiol 2007 Sep;45(9):2937-42.
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