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A novel method for the isolation of gastro-intestinal nematode eggs that allows automated analysis of digital images of egg preparations and high throughput screening

Published online by Cambridge University Press:  28 November 2001

T. H. M. MES
Affiliation:
Department of Infectious Diseases and Immunology, Division of Parasitology and Tropical Veterinary Medicine, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3508 TD, The Netherlands
H. W. PLOEGER
Affiliation:
Department of Infectious Diseases and Immunology, Division of Parasitology and Tropical Veterinary Medicine, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3508 TD, The Netherlands
M. TERLOU
Affiliation:
Faculty of Biology, Utrecht University, Padualaan 8, 3584 CH, Utrecht, The Netherlands
F. N. J. KOOYMAN
Affiliation:
Department of Infectious Diseases and Immunology, Division of Parasitology and Tropical Veterinary Medicine, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3508 TD, The Netherlands
M. P. J. VAN DER PLOEG
Affiliation:
Department of Infectious Diseases and Immunology, Division of Parasitology and Tropical Veterinary Medicine, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3508 TD, The Netherlands
M. EYSKER
Affiliation:
Department of Infectious Diseases and Immunology, Division of Parasitology and Tropical Veterinary Medicine, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3508 TD, The Netherlands

Abstract

A simple and robust method for the isolation of gastro-intestinal nematode eggs from faeces is described that uses both salt- and sugar solutions for flotation. Application of this ‘salt–sugar’; isolation method to large numbers of faecal samples of adult dairy cows indicates a 3- or 4-fold reduction in the proportion of e.p.g.-negative cows relative to studies that used other techniques for egg isolation. The procedure detects more eggs than the Wisconsin flotation method in replicate samples and in spiked egg-free faeces. The number of recovered eggs in spiked faecal samples is linear over a range of egg concentrations, and the transparent faecal preparations that result from the protocol can be stored as digital images which can be used as input for an efficient automated egg-counting procedure. The increased rate of processing of faeces combined with the large reduction of the percentage of e.p.g.-negative cows allows more accurate analysis of large numbers of adult or resistant animals for studies of nematode parasitism.

Type
Research Article
Copyright
© 2002 Cambridge University Press

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References

AGNEESSENS, J., CLAEREBOUT, E., DORNY, P., BORGSTEEDE, F. H. M. & VERCRUYSSE, J. (2000). Nematode parasitism in adult dairy cows in Belgium. Veterinary Parasitology 90, 8392.CrossRefGoogle Scholar
BORGSTEEDE, F. H. M., TIBBEN, J., CORNELISSEN, J. B. W. J., AGNEESSENS, J. & GAASENBEEK, C. P. H. (2000). Nematode parasites of adult dairy cattle in The Netherlands. Veterinary Parasitology 89, 287296.CrossRefGoogle Scholar
BURROWS, R. O., BEST, P. J. & PRESTON, J. M. (1980). Trichostrongylid egg output of dairy cows. Veterinary Record 107, 399401.CrossRefGoogle Scholar
COX, D. D. & TODD, A. C. (1962). Survey of gastro-intestinal parasitism in Wisconsin dairy cattle. Journal of the American Veterinary Medical Association 141, 706709.Google Scholar
GUTTIERES, V., TODD, A. C. & CROWLEY, J. W. (1979). Natural populations of helminths in Wisconsin dairy cows. Veterinary Medicine 74, 369374.Google Scholar
LEVINE, N. D., MEHRA, K. N., CLARK, D. T. & AVES, I. J. (1960). A comparison of nematode egg counting techniques for cattle and sheep faeces. American Journal of Veterinary Research 21, 511515.Google Scholar
MAFF, . (1977). Manual of Veterinary Parasitological Laboratory Techniques. Technical Bulletin 18. Her Majesty's Stationery Office, London, UK.
SOKAL, R. R. & ROHLF, F. J. (1981). Biometry. Freeman and Company, New York.
SMOTHERS, C. D., SUN, F. & DAYTON, A. D. (1999). Comparison of arithmetic and geometric means as measures of a central tendency in cattle nematode populations. Veterinary Parasitology 81, 211224.CrossRefGoogle Scholar
STOCKL, D., DEWITTE, K. & THIENPONT, L. M. (1998). Validity of linear regression in method comparison studies: is it limited by the statistical model or the quality of the analytical input data? Clinical Chemistry 44, 23402346.Google Scholar
THIENPONT, D., ROCHETTE, F. & VAN PARIJS, O. F. J. (1979). Diagnosing Helminthiasis by Coprological Examination. Janssen Research Foundation, Beerse, Belgium.