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Title: SPECIFICITY OF SIX OLIGOPROBES FOR CYATHOSTOMIN NEMATODE PARASITES OF HORSES

Author
item HODGKINSON, JANE - UNIV LIVERPOOL, UK
item LOVE, SANDY - UNIV GLASGOW, UK
item Lichtenfels, James
item RAMSEY, YVONNE - UNIV GLASGOW, UK
item MATTHEWS, JACQUELINE - UNIV LIVERPOOL, UK

Submitted to: Parasitology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/29/2000
Publication Date: N/A
Citation: N/A

Interpretive Summary: Strongyloid nematodes are a major cause of morbidity and mortality in equines in the United States. Resistance to antiparasitic drugs (currently the only means of controlling the nematodes) is common and alternative control methods for these parasites are needed. The 40 species in horses worldwide can be identified by a few authorities using comparative anatomy of adult stages, but larval stages are exceptionally difficult to identify and eggs are impossible to identify to even subfamily level. The development of diagnostic DNA probes that can be used to identify all stages of the nematodes including eggs in the feces is a high priority need. This report describes five oligoprobes for Cylicocyclus ashworthi, C. nassatus, Cyathostomum pateratum, Cylicostephanus longibursatus and C. goldi, five of the most commonly occurring species, and a sixth probe that detects all nematode species in the group. In combination these probes have the potential to identify any stage of the nematodes including eggs in the feces, and to provide an estimate of the intensity of the infection. The results will be used by researchers worldwide working to control these economically important nematodes of horses, especially in the pharmaceutical industry, the Food and Drug Administration, and scientists evaluating biological control agents and antiparasitic drugs for larval cyathostomiasis, an emerging disease of horses worldwide.

Technical Abstract: This study reports the investigation of five oligoprobes designed from short sequences within the IGS region for identification of individual cyathostomin species and a sixth probe designed from a conserved region of the IGS to detect all species. The species for which probes were designed included; Cylicocyclus ashworthi, C. nassatus, Cyathostomum pateratum, Cylicostephanus longibursatus and C. goldi. The amplification of intergenic spacer DNA from 16 cyathostomin species using primers designed to conserved regions of intergenic spacer sequence allowed sequence comparison and identification of these five species-specific probes. Subsequent Southern blotting of amplified products from 16 cyathostomin species with the oligonucleotide probes labeled with digoxigenin showed that four of these probes hybridised specifically to individual species with little or no cross- hybridisation to other closely related species. Unfortunately, the probe designed to detect C. pateratum cross hybridised with products generated from individuals of the closely related species, Cyathostomum catinatum. The sixth probe recognized all 16 cyathostomin species.