Skip to main content
×
Home
    • Aa
    • Aa

New insights into sequence variation in the IGS region of 21 cyathostomin species and the implication for molecular identification

  • K. CWIKLINSKI (a1), F. N. J. KOOYMAN (a2), D. C. K. VAN DOORN (a2), J. B. MATTHEWS (a3) and J. E. HODGKINSON (a1)...
Summary
SUMMARY

Cyathostomins comprise a group of 50 species of parasitic nematodes that infect equids. Ribosomal DNA sequences, in particular the intergenic spacer (IGS) region, have been utilized via several methodologies to identify pre-parasitic stages of the commonest species that affect horses. These methods rely on the availability of accurate sequence information for each species, as well as detailed knowledge of the levels of intra- and inter-specific variation. Here, the IGS DNA region was amplified and sequenced from 10 cyathostomin species for which sequence was not previously available. Also, additional IGS DNA sequences were generated from individual worms of 8 species already studied. Comparative analysis of these sequences revealed a greater range of intra-specific variation than previously reported (up to 23%); whilst the level of inter-specific variation (3–62%) was similar to that identified in earlier studies. The reverse line blot (RLB) method has been used to exploit the cyathostomin IGS DNA region for species identification. Here, we report validation of novel and existing DNA probes for identification of cyathostomins using this method and highlight their application in differentiating life-cycle stages such as third-stage larvae that cannot be identified to species by morphological means.

Copyright
Corresponding author
*Corresponding author: Veterinary Parasitology, Department of Infection Biology, Institute of Infection and Global Health, University of Liverpool, Liverpool L69 7ZJ, UK. Tel: +44 (0)151 7941586. Fax: +44 (0)151 7941519. E-mail: K.Cwiklinski@liverpool.ac.uk
Linked references
Hide All

This list contains references from the content that can be linked to their source. For a full set of references and notes please see the PDF or HTML where available.

R. Appels and J. Dvorak (1982). Relative rates of divergence of spacer and gene sequences within the rDNA region of species in the Triticeae: implications for the maintenance of homogeneity of a repeated gene family. Theoretical and Applied Genetics 63, 361365. doi: 10.1007/BF00303907.

S. M. Baker and T. Platt (1986). Pol I transcription: which comes first, the end or the beginning? Cell 47, 839840. doi: 10.1016/0092-8674(86)90795-6.

D. G. Bucknell , R. B. Gasser and I. Beveridge (1995). The prevalence and epidemiology of gastrointestinal parasites of horses in Victoria, Australia. International Journal for Parasitology 25, 711724. doi: 10.1016/0020-7519(94)00214-9.

D. Cernanská , B. Paoletti , I. Kralova-Hromadova , R. Iorio , P. Cudekova , P. Milillo and D. Traversa (2009). Application of a Reverse Line Blot hybridisation assay for the species-specific identification of cyathostomins (Nematoda, Strongylida) from benzimidazole-treated horses in the Slovak Republic. Veterinary Parasitology 160, 171174. doi: 10.1016/j.vetpar.2008.10.078.

M. R. Chapman , D. D. French and T. R. Klei (2002). Gastrointestinal helminths of ponies in Louisiana: a comparison of species currently prevalent with those present 20 years ago. The Journal of Parasitology 88, 11301134. doi: 10.1645/0022-3395(2002)088[1130:GHOPIL]2.0.CO;2.

R. F. De Winter and T. Moss (1986). The ribosomal spacer in Xenopus laevis is transcribed as part of the primary ribosomal RNA. Nucleic Acids Research 14, 60416051. doi: 10.1093/nar/14.15.6041.

J. J. Elder and B. J. Turner (1995). Concerted evolution of repetitive DNA sequences in eukaryotes. The Quarterly Review of Biology 70, 297320.

M. Eysker and V. S. Pandey (1989). Small strongyle infections in donkeys from the Highveld in Zimbabwe. Veterinary Parasitology 30, 345349. doi: 10.1016/0304-4017(89)90104-0.

R. B. Flavell , M. O'Dell , W. F. Thompson , M. Vincentz , R. Sardana and R. F. Barker (1986). The differential expression of ribosomal RNA genes. Philosophical Transactions of the Royal Society of London, B 314, 385397. doi: 10.1098/rstb.1986.0060.

R. B. Gasser and S. E. Newton (2000). Genomic and genetic research on bursate nematodes: significance, implications and prospects. International Journal for Parasitology 30, 509534. doi: 10.1016/S0020-7519(00)00021-7.

C. J. Giles , K. A. Urquhart and J. A. Longstaffe (1985). Larval cyathostomiasis (immature trichonema-induced enteropathy): a report of 15 clinical cases. Equine Veterinary Journal 17, 196201. doi: 10.1111/j.2042-3306.1985.tb02469.x.

C. A. Harrington and D. M. Chikaraishi (1987). Transcription of spacer sequences flanking the rat 45S ribosomal DNA gene. Molecular and Cellular Biology 7, 314325. doi: 10.1128/MCB.7.1.314.

J. E. Hodgkinson , K. L. Freeman , J. R. Lichtenfels , S. Palfreman , S. Love and J. B. Matthews (2005). Identification of strongyle eggs from anthelmintic-treated horses using a PCR-ELISA based on intergenic DNA sequences. Parasitology Research 95, 287292. doi: 10.1007/s00436-004-1289-z.

J. E. Hodgkinson , J. R. Lichtenfels , T. S. Mair , P. Cripps , K. L. Freeman , Y. H. Ramsey , S. Love and J. B. Matthews (2003). A PCR-ELISA for the identification of cyathostomin fourth-stage larvae from clinical cases of larval cyathostominosis. International Journal for Parasitology 33, 14271435. doi: 10.1016/S0020-7519(03)00140-1.

J. E. Hodgkinson , S. Love , J. R. Lichtenfels , S. Palfreman , Y. H. Ramsey and J. B. Matthews (2001). Evaluation of the specificity of five oligoprobes for identification of cyathostomin species from horses. International Journal for Parasitology 31, 197204. doi: 10.1016/S0020-7519(00)00161-2.

G. C. Hung , R. B. Gasser , I. Beveridge , and N. B. Chilton (1999 b). Species-specific amplification by PCR of ribosomal DNA from some equine strongyles. Parasitology 119, 6980.

G. C. Hung , N. B. Chilton , I. Beveridge , X. Q. Zhu , J. R. Lichtenfels and R. B. Gasser (1999 a). Molecular evidence for cryptic species within Cylicostephanus minutus (Nematoda: Strongylidae). International Journal for Parasitology 29, 285291. doi: 10.1016/S0020-7519(98)00203-3.

M. Ionita , D. K. Howe , E. T. Lyons , S. C. Tolliver , R. M. Kaplan , I. L. Mitrea and M. Yeargan (2010). Use of a reverse line blot assay to survey small strongyle (Strongylida: Cyathostominae) populations in horses before and after treatment with ivermectin. Veterinary Parasitology 168, 332337. doi: 10.1016/j.vetpar.2009.11.021.

J. N. Kaye , S. Love , J. R. Lichtenfels and J. B. McKeand (1998). Comparative sequence analysis of the intergenic spacer region of cyathostome species. International Journal for Parasitology 28, 831836. doi: 10.1016/S0020-7519(98)00031-9.

V. Kharchenko , T. Kuzmina , A. Trawford , M. Getachew and G. Feseha (2009). Morphology and diagnosis of some fourth-stage larvae of cyathostomines (Nematoda: Strongyloidea) in donkeys Equus asinus L. from Ethiopia. Systematic Parasitology 72, 113. doi: 10.1007/s11230-008-9152-8.

S. Kornaś , J. Cabaret , M. Skalska and B. Nowosad (2010). Horse infection with intestinal helminths in relation to age, sex, access to grass and farm system. Veterinary Parasitology 174, 285291. doi: 10.1016/j.vetpar.2010.09.007.

P. Labhart and R. H. Reeder (1986). Characterization of three sites of RNA 3′ end formation in the Xenopus ribosomal gene spacer. Cell 45, 431443. doi: 10.1016/0092-8674(86)90329-6.

J. R. Lichtenfels , V. A. Kharchenko and G. M. Dvojnos (2008). Illustrated identification keys to strongylid parasites (Strongylidae: Nematoda) of horses, zebras and asses (Equidae). Veterinary Parasitology 156, 4161. doi: 10.1016/j.vetpar.2008.04.026.

J. R. Lichtenfels , V. A. Kharchenko , R. C. Krecek and L. M. Gibbons (1998). An annotated checklist by genus and species of 93 species level names for 51 recognized species of small strongyles (Nematoda: Strongyloidea: Cyathostominea) of horses, asses and zebras of the world. Veterinary Parasitology 79, 6579. doi: 10.1016/S0304-4017(98)00149-6.

S. Love , D. Murphy and D. Mellor (1999). Pathogenicity of cyathostome infection. Veterinary Parasitology 85, 113121; discussion 121–2, 215–25. doi: 10.1016/S0304-4017(99)00092-8.

E. T. Lyons , S. C. Tolliver and J. H. Drudge (1999). Historical perspective of cyathostomes: prevalence, treatment and control programs. Veterinary Parasitology 85, 97111; discussion 111–2, 215–25. doi: 10.1016/S0304-4017(99)00091-6.

E. T. Lyons , S. C. Tolliver , J. H. Drudge , S. Stamper , T. W. Swerczek and D. E. Granstrom (1996). A study (1977–1992) of population dynamics of endoparasites featuring benzimidazole-resistant small strongyles (population S) in Shetland ponies. Veterinary Parasitology 66, 7586. doi: 10.1016/S0304-4017(96)00998-3.

S. Matthee , R. C. Krecek and L. M. Gibbons (2002). Cylicocyclus asini n.sp. (Nematoda: Cyathostominae) from donkeys Equus asinus in South Africa. Systematic Parasitology 51, 2935. doi: 10.1023/A:1012989810087.

S. Matthee , R. C. Krecek and M. A. McGeoch (2004). A comparison of the intestinal helminth communities of Equidae in Southern Africa. Journal of Parasitology 90, 12631273. doi: 10.1645/GE-3353.

S. Matthee , R. C. Krecek and S. A. Milne (2000). Prevalence and biodiversity of helminth parasites in donkeys from South Africa. Journal of Parasitology 86, 756762. doi: 10.1645/0022-3395(2000)086[-756:PABOHP]2.0.CO;2.

T. Moss and V. Y. Stefanovsky (1995). Promotion and regulation of ribosomal transcription in eukaryotes by RNA polymerase I. Progress in Nucleic Acid Research and Molecular Biology 50, 2566. doi: 10.1016/S0079-6603(08)60810-7.

S. W. Reid , T. S. Mair , M. H. Hillyer and S. Love (1995). Epidemiological risk factors associated with a diagnosis of clinical cyathostomiasis in the horse. Equine Veterinary Journal 27, 127130. doi: 10.1111/j.2042-3306.1995.tb03048.x.

D. Traversa , R. Iorio , T. R. Klei , V. A. Kharchenko , J. Gawor , D. Otranto and O. A. Sparagano (2007). New method for simultaneous species-specific identification of equine strongyles (Nematoda, Strongylida) by reverse line blot hybridization. Journal of Clinical Microbiology 45, 29372942. doi: 10.1128/JCM.00714-07.

D. Traversa , R. Iorio , D. Otranto , A. Giangaspero , P. Milillo and T. R. Klei (2009). Species-specific identification of equine cyathostomes resistant to fenbendazole and susceptible to oxibendazole and moxidectin by macroarray probing. Experimental Parasitology 121, 9295. doi: 10.1016/j.exppara.2008.10.001.

D. Traversa , P. Milillo , H. Barnes , G. Von Samson-Himmelstjerna , S. Schurmann , J. Demeler , D. Otranto , R. P. Lia , S. Perrucci , A. Frangipane Di Regalbono , P. Beraldo , D. Amodie , K. Rohn , R. Cobb and A. Boeckh (2010). Distribution and species-specific occurrence of cyathostomins (Nematoda, Strongylida) in naturally infected horses from Italy, United Kingdom and Germany. Veterinary Parasitology 168, 8492. doi: 10.1016/j.vetpar.2009.10.006.

D. C. van Doorn , F. N. Kooyman , M. Eysker , J. E. Hodgkinson , J. A. Wagenaar and H. W. Ploeger (2010). In vitro selection and differentiation of ivermectin resistant cyathostomin larvae. Veterinary Parasitology 174, 292299. doi: 10.1016/j.vetpar.2010.08.021.

G. van Loon , P. Deprez , E. Muylle and B. Sustronck (1995). Larval cyathostomiasis as a cause of death in two regularly dewormed horses. Zentralblatt für Veterinarmedizin.Reihe A 42, 301306. doi: 10.1111/j.1439-0442.1995.tb00381.

Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

Parasitology
  • ISSN: 0031-1820
  • EISSN: 1469-8161
  • URL: /core/journals/parasitology
Please enter your name
Please enter a valid email address
Who would you like to send this to? *
×

Keywords:

Metrics

Altmetric attention score

Full text views

Total number of HTML views: 2
Total number of PDF views: 21 *
Loading metrics...

Abstract views

Total abstract views: 146 *
Loading metrics...

* Views captured on Cambridge Core between September 2016 - 22nd September 2017. This data will be updated every 24 hours.