Parasitology Research

, Volume 95, Issue 4, pp 287–292

Identification of strongyle eggs from anthelmintic-treated horses using a PCR-ELISA based on intergenic DNA sequences

  • J. E. Hodgkinson
  • K. L. Freeman
  • J. R. Lichtenfels
  • S. Palfreman
  • S. Love
  • J. B. Matthews
Original Paper

Abstract

The efficacy of five daily fenbendazole (FBZ) treatments was tested against benzimidazole-resistant cyathostomins in naturally infected horses (n=13). Horses were treated with pyrantel embonate (PYR) to remove adult strongyles followed, 7 days later, by a 5-day course of FBZ. The PYR treatment produced an average faecal egg count reduction of 98%. All samples were negative by faecal egg count 7 days after the start of the FBZ treatment. Positive egg counts were observed from 28 days after the start of FBZ treatment and all horses displayed positive faecal egg counts by 77 days after treatment. Strongyle eggs were harvested from the faeces of the horses prior to treatment and then weekly from 42 to 70 days post-treatment. DNA was obtained from eggs in groups of ten. A PCR-ELISA, based on species-specific differences in intergenic DNA sequences, was used to identify the presence of six cyathostomin species. In pre-treatment samples, Cyathostomum catinatum was detected in nine out of the 13 horses and Cylicostephanus longibursatus, Cylicostephanus goldi and Cylicocyclus nassatus, were found in samples from eight animals. Cylicocyclus ashworthi and Cylicocyclus insigne were not detected pre-treatment. After anthelmintic treatment, C. catinatum and C. longibursatus were most frequently detected, followed by C. nassatus, C. goldi and C. ashworthi. C. insigne was detected at only one time point in a sample from a single horse.

References

  1. Barger IA (1986) The statistical distribution of trichostrongylid nematodes in grazing lambs. Int J Parasitol 15:645–649CrossRefGoogle Scholar
  2. Boersema JH, Borgsteede FHM, Eysker M, Saedt I (1995) The reappearance of strongyle eggs in faeces of horses treated with pyrantel embonate. Vet Q 17:18–20Google Scholar
  3. Bucknell DG, Gasser RB, Beveridge I (1995) The prevalence and epidemiology of gastrointestinal parasites of horses in Victoria, Australia. Int J Parasitol 25:711–724CrossRefGoogle Scholar
  4. Chandler KJ, Collins MC, Love S (2001) Efficacy of a five-day course of fenbendazole in benzimidazole resistant cyathostomes. Vet Rec 147:661–662Google Scholar
  5. Craven J, Bjorn H, Henriksen SA, Nansen P, Larsen M, Lendal S (1998) Survey of anthelmintic resistance on Danish horse farms using 5 different methods of calculating faecal egg count reduction. Equine Vet J 30:289–293Google Scholar
  6. Crofton HD (1971) A model of host-parasite relationships. Parasitology 63:343–364Google Scholar
  7. Dowdall SM, Proudman CJ, Klei TR, Mair T, Matthews (2004) Characterisation of IgG(T) serum antibody responses to two larval antigen complexes in horses naturally- or experimentally-infected with cyathostomins. Int J Parasitol 34:101–108CrossRefGoogle Scholar
  8. Duncan JL, Bairden KB, Abbot EM (1998) Elimination of mucosal cyathostome larvae by five daily treatments with fenbendazole. Vet Rec 142:268–271Google Scholar
  9. Eysker M, Boersema JH, Kooman FNL (1992) The effect of ivermectin against inhibited early 3rd stage, late 3rd stage and 4th stage larvae and adult stages of the cyathostomes in Shetland ponies and expulsion of these helminths. Vet Parasitol 42:295–302CrossRefGoogle Scholar
  10. Gawor JJ (1995) The prevalence and abundance of internal parasites in working horses autopsied in Poland. Vet Parasitol 58:99–108CrossRefGoogle Scholar
  11. Giles CJ, Urquhart KA, Longstaffe JA (1985) Larval cyathostomiasis (immature trichoneme induced enteropathy), a report of 15 clinical cases. Equine Vet J 17:196–201Google Scholar
  12. Hodgkinson JE, Love S, Lichtenfels JR, Palfreman S, Ramsey YH, Matthews JB (2001) Evaluation of the specificity of five oligoprobes for identification of cyathostomin species from horses. Int J Parasitol 31:197–204CrossRefGoogle Scholar
  13. Hodgkinson JE, Lictenfels JR, Mair TS, Cripps P, Freeman KL, Ramsey YH, Love S, Matthews JB (2003) A PCR-ELISA for the identification of cyathostomin fourth-stage larvae from clinical cases of larval cyathostominosis. Int J Parasitol 33:1427–1435CrossRefGoogle Scholar
  14. Hung GC, Chilton NB, Beveridge I, McDonnell A, Lichtenfels JR, Gasser RB (1997) Molecular delineation of Cylicocyclus nassatus and C. ashworthi (Nematoda: Strongylidae). Int J Parasitol 27:601–607CrossRefGoogle Scholar
  15. Kaplan RM (2002) Anthelmintic resistance in nematodes of horses. Vet Res 33:491–507CrossRefGoogle Scholar
  16. Kaye JN, Love S, Lichtenfels JR, Mckeand JB (1998) Comparative sequence analysis of the intergenic spacer (IGS) region of cyathostome species. Int J Parasitol 28:831–836CrossRefGoogle Scholar
  17. Klei TR, Chapman MR, French DD, Taylor HW (1993) Evaluation of ivermectin at an elevated dose against encysted equine cyathostome larvae. Vet Parasitol 47:99–106CrossRefGoogle Scholar
  18. Lichtenfels JR, Kharchenko VA, Sommer C, Ito M, (1997) Key characters for the microscopical identification of Cylicocyclus nassatus and Cylicocyclus ashworthi (Nematoda, Cyathostominae) of the horse, Equus caballus. J Helminthol Soc Wash 64:120–127Google Scholar
  19. Love S, Mckeand JB (1997) Cyathostomosis: practical issues of treatment and control. Equine Vet Ed 9:253–256Google Scholar
  20. Lyons E (2003) Population-S benzimidazole- and tetrahydropyrimidine-resistant small strongyles in a pony herd in Kentucky(1977–1999): effects of anthelmintic treatment on the parasites as determined in critical tests. Parasitol Res 91:407–411CrossRefGoogle Scholar
  21. Lyons ET, Tolliver SC (2003) Field test data on small strongyles in evaluation of activity of fenbendazole given once a day for 5 consecutive days to thoroughbred yearlings on two farms in Kentucky in 2002 and 2003. Parasitol Res 91:312–315CrossRefGoogle Scholar
  22. Mair TS, Pearson GR (1995) Multifocal non-strangulating intestinal infarction associated with larval cyathostomiasis in a pony. Equine Vet J 27:154–155Google Scholar
  23. Murphy D, Love S (1997) The pathogenic effects of experimental cyathostome infections in ponies. Vet Parasitol 70:99–110CrossRefGoogle Scholar
  24. Ogbourne CP (1976) The prevalence, relative abundance and site distribution of nematodes of the subfamily Cyathostominae in horses killed in Britain. J Helminthol 50:203–214Google Scholar
  25. Patterson P, Coles GC (2000) Winter transmission of equine nematodes. Vet Rec 147:251–252Google Scholar
  26. Proudman C, Matthews JB (2000) Control of intestinal parasites in horses. Equine Prac 22:90–97Google Scholar
  27. Reinemeyer CR, Smith SA, Gabel AA, Herd RP (1984) The prevalence and intensity of internal parasites of horses in the USA. Vet Parasitol 15:75–83CrossRefGoogle Scholar
  28. Silva AV, Costa HM, Santos HA, Carvalho RO (1999) Cyathostominae (Nematoda) parasites of Equus caballus in some Brazilian states. Vet Parasitol 86:15–21CrossRefGoogle Scholar
  29. Torbert BJ, Klei TR, Lichtenfels JR, Chapman MR (1986) A survey in Louisiana of intestinal helminths of ponies with little exposure to anthelmintics. J Parasitol 72:926–930Google Scholar
  30. Uhlingher CA (1990) Effects of three anthelmintic schedules on the incidence of colic in horses. Equine Vet J 22:251–254Google Scholar
  31. Urquhart GM, Armour J, Duncan JL, Dunn AM, Jennings FW (1996) Veterinary Parasitology, 2nd edn. Blackwell, Oxford, pp 276–277Google Scholar
  32. Vanloon G, Deprez P, Muylle E, Sustronk B (1995) Larval cyathostomiasis as a cause of death in two regularly dewormed horses. J Vet Med 42:301–306Google Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • J. E. Hodgkinson
    • 1
  • K. L. Freeman
    • 2
  • J. R. Lichtenfels
    • 3
  • S. Palfreman
    • 2
  • S. Love
    • 4
  • J. B. Matthews
    • 5
  1. 1.Department of Veterinary Parasitology, Faculty of Veterinary Science, Liverpool School of Tropical MedicineUniversity of LiverpoolLiverpool UK
  2. 2.Department of Veterinary Clinical Science, Faculty of Veterinary ScienceUniversity of LiverpoolLeahurstUK
  3. 3.Animal Parasitic Diseases Laboratory, Agricultural Research ServiceUS Department of AgricultureBeltsvilleUSA
  4. 4.Department of Veterinary Clinical Studies, Faculty of Veterinary MedicineUniversity of GlasgowGlasgowUK
  5. 5.Division of ParasitologyMoredun Research InstitutePentlands Science ParkUK

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