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

Published online by Cambridge University Press:  13 April 2012

K. CWIKLINSKI ,
F. N. J. KOOYMAN ,
D. C. K. VAN DOORN ,
J. B. MATTHEWS  and
J. E. HODGKINSON
K. CWIKLINSKI*
Affiliation:
Veterinary Parasitology, Department of Infection Biology, Institute of Infection and Global Health, University of Liverpool, Liverpool L69 7ZJ, UK
F. N. J. KOOYMAN
Affiliation:
Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3584 CL Utrecht, The Netherlands
D. C. K. VAN DOORN
Affiliation:
Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3584 CL Utrecht, The Netherlands
J. B. MATTHEWS
Affiliation:
Moredun Research Institute, Penicuik, Midlothian EH26 0PZ, Scotland
J. E. HODGKINSON
Affiliation:
Veterinary Parasitology, Department of Infection Biology, Institute of Infection and Global Health, University of Liverpool, Liverpool L69 7ZJ, UK
*
*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
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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†.

Keywords

cyathostominsIGSintra-specificinter-specificRLB hybridization

Information

Type
Research Article
Information
Parasitology , Volume 139 , Issue 8 , July 2012 , pp. 1063 - 1073
Copyright
Copyright © Cambridge University Press 2012

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