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The metabolic profile of adult Fasciola hepatica obtained from rafoxanide-treated sheep

Published online by Cambridge University Press:  06 April 2009

R. K. Prichard
Affiliation:
Division of Animal Health, CSIRO, McMaster Laboratory, Private Bag No. 1, Glebe, N.S.W. 2037, Australia

Summary

Sheep infected with adult Fasciola hepatica were drenched with rafoxanide. At 4, 8, 16 and 24 h after drenching the sheep were killed and the flukes removed, washed and rapidly frozen in liquid nitrogen. The content of key metabolites in the fermentation pathway were determined and compared with those in control F. hepatica, whose hosts were not treated with rafoxanide.

Rafoxanide decreased glycogen, malate, NADH and ATP levels. The level of other metabolites in the pathway increased for the first 8–16 h after rafoxanide treatment. The marked decrease in ATP and glycogen, and the increase in total [NAD+]/[NADH] and [oxaloacetate]/[malate], together with the changed content of other metabolites, led to the conclusion that the mode of action of rafoxanide against F. hepatica in vivo is by uncoupling oxidative phosphorylation.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1978

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References

REFERENCES

Barman, T. E. (1969). Enzyme Handbook. Berlin: Springer.Google Scholar
Barrett, J. (1975). The occurrence and intracellular distribution of nucleosidediphosphate kinase in parasitic helminths. Journal of Parasitology 61, 545–6.CrossRefGoogle Scholar
Barrett, J. & Beis, I. (1973 a). The redox state of the free nicotinamideadenine dinucleotide couple in the cytoplasm and mitochondria of muscle tissue from Ascaris lumbricoides (Nematoda). Comparative Biochemistry and Physiology 44A, 331–40.CrossRefGoogle ScholarPubMed
Barrett, J. & Beis, I. (1973 b). Studies on glycolysis in the muscle tissue of Ascaris lumbricoides (Nematoda). Comparative Biochemistry and Physiology 44B, 751–61.Google ScholarPubMed
Coles, G. C. & East, J. M. (1974). Unpublished observations cited in Coles, G. C. (1975). Fluke biochemistry – Fasciola and Schistosoma. Helminthological Abstracts 44, Ser. A, 147–62.Google Scholar
Corbett, J. R. (1974). The Biochemical Mode of Action of Pesticides. London and New York: Academic Press.Google Scholar
Corbett, J. R. & Goose, J. (1971). A possible biochemical mode of action of the fasciolicides nitroxynil, hexachlorophene and oxyclozanide. Pesticide Science 2, 119–21.CrossRefGoogle Scholar
Cornish, R. A. & Bryant, C. (1976). Changes in energy metabolism due to anthelmintics in Fasciola hepatica maintained in vitro. International Journal for Parasitology 6, 393–8.CrossRefGoogle ScholarPubMed
Czok, R. (1974). D-glycerate-3-phosphate. In Methods of Enzymatic Analysis, vol. 3 (ed. Bergmeyer, H. U.), pp. 1424–8. Weinheim, New York: Verlag Chemie, Academic Press.Google Scholar
Czok, R. & Lamprecht, W. (1974). Pyruvate, phosphoenolpyruvate and D-glycerate-2-phosphate. In Methods of Enzymatic Analysis, vol. 3 (ed. Bergmeyer, H. U.), pp. 1446–51. Weinheim, New York: Verlag Chemie, Academic Press.Google Scholar
de Zoeten, L. W. & Tipker, J. (1969). Intermediary metabolism of the liver fluke Fasciola hepatica. H. Hydrogen transport and phosphorylation. Hoppe-Seyler's Zeitschrift für physiologische Chemie 350, 691–5.Google Scholar
Düwel, D. & Metzger, H. (1973). 2,6-Dihydroxybenzoic acid anilides as fasciolicides. Journal of Medicinal Chemistry 16, 433–6.Google Scholar
Jaworek, D., Gruber, W. & Bergmeyer, H. U. (1974). Adenosine-5′-triphosphate. Determination with 3-phosphoglycerate kinase. In Methods of Enzymatic Analysis, vol. 4 (ed. Bergmeyer, H. U.), pp. 2097–101. Weinheim, New York: Verlag Chemie, Academic Press.Google Scholar
Klingenberg, M. (1974). Nicotinamide-adenine dinucleotides (NAD, NADP, NADH, NADPH). Spectrophotometric and fluorimetric methods. In Methods of Enzymatic Analysis, vol. 4 (ed. Bergmeyer, H. U.), pp. 2045–53. Weinheim, New York: Verlag Chemie, Academic Press.CrossRefGoogle Scholar
Mansour, T. E. (1959). Studies on the carbohydrate metabolism of the liver fluke, Fasciola hepatica. Biochimica et Biophysica Acta 34, 456–64.Google Scholar
Martin, J. B. & Doty, D. M. (1949). Determination of inorganic phosphate. Modification of isobutyl alcohol procedure. Analytical Chemistry 21, 965–7.CrossRefGoogle Scholar
Metzger, H. & Düwel, D. (1973). Investigations of metabolism in the liver fluke (Fasciola hepatica) as an aid to the development of new anthelmintics. International Journal of Biochemistry 4, 133–43.Google Scholar
Michal, G. & Beutler, H-O. (1974). D-fructose-1,6-diphosphate, dihydroxyacetone phosphate and D-glyceraldehyde-3-phosphate. In Methods of Enzymatic Analysis, vol. 3 (ed. Bergmeyer, H. U.), pp. 1314–19. Weinheim, New York: Verlag Chemie, Academic Press.Google Scholar
Möllering, H. (1974). L-( − )-malate. Determination with malate dehydrogenase and glutamate-oxaloacetate transaminase. In Methods of Enzymatic Analysis, vol. 3 (ed. Bergmeyer, H. U.), pp. 1589–93. Weinheim, New York: Verlag Chemie, Academic Press.Google Scholar
Mrozik, H., Jones, H., Friedman, J., Schwartzkopf, G., Schardt, R. A., Patchett, A. A., Hoff, D. R., Yakstis, J. J., Riek, R. F., Ostlind, D. A., Plishker, G. A., Butler, R. W., Cuckler, A. C. & Campbell, W. C. (1969). A new agent for the treatment of liver fluke infection (fascioliasis). Experientia 25, 883.Google Scholar
Noll, F. (1974). L-( + )-lactate. Determination with LDH, GPT and NAD. In Methods of Enzymatic Analysis, vol. 3 (ed. Bergmeyer, H. U.), pp. 1475–9. Weinheim, New York: Verlag Chemie, Academic Press.Google Scholar
Pfleiderer, G. (1963). Glycogen. Determination as D-glucose with hexokinase, pyruvic kinase and lactic dehydrogenase. In Methods of Enzymatic Analysis, (ed. Bergmeyer, H. U.), pp. 5962. Weinheim, New York: Verlag Chemie, Academic Press.Google Scholar
Prichard, R. K. (1976). Regulation of pyruvate kinase and phosphoenolpyruvate carboxykinase activity in adult Fasciola hepatica (Trematoda). International Journal for Parasitology 6, 227–33.Google Scholar
Prichard, R. K. & Schofield, P. J. (1968 a). A comparative study of the tricarboxylic acid cycle enzymes in Fasciola hepatica and rat liver. Comparative Biochemistry and Physiology 25, 1005–19.CrossRefGoogle ScholarPubMed
Prichard, R. K. & Schofield, P. J. (1968 b). Phosphoenolpyruvate carboxykinase in the adult liver fluke, Fasciola hepatica. Comparative Biochemistry and Physiology 24, 773–85.Google Scholar
Rolleston, F. S. (1972). A theoretical background to the use of measured concentrations of intermediates in study of the control of intermediary metabolism. Current Topics in Cellular Regulation 5, 4775.Google Scholar
Umbreit, W. W., Burris, R. H. & Stauffer, J. F. (1964). Manometric Techniques. Minnesota: Burgess.Google Scholar
Utter, M. F. & Kurahashi, K. (1954). Mechanism of action of oxalacetic carboxylase. The Journal of Biological Chemistry 207, 821–41.CrossRefGoogle ScholarPubMed
Van den Bossche, H. (1972). Studies on the phosphorylation in Ascaris mitochondria. In Comparative Biochemistry of Parasites, (ed. van den Bossche, H.), pp. 455–68. New York and London: Academic Press.Google Scholar
Veech, R. L., Eggleston, L. V. & Krebs, H. A. (1969). The redox state of free nicotinamide-adenine dinucleotide phosphate in the cytoplasm of rat liver. The Biochemical Journal 115, 609–19.Google Scholar
Veech, R. L., Raijman, L. & Krebs, H. A. (1970). Equilibrium relations between the cytoplasmic adenine nucleotide system and nicotinamide-adenine nucleotide system in rat liver. The Biochemical Journal 117, 499503.Google Scholar
Wahlefeld, A. W. (1974). Oxaloacetate. UV spectrophotometric determination. In Methods of Enzymatic Analysis, vol. 3 (ed. Bergmeyer, H. U.), pp. 1604–8. Weinheim, New York: Verlag Chemie, Academic Press.Google Scholar
Williamson, D. H., Lund, P. & Krebs, H. A. (1967). The redox state of free nicotinamide-adenine dinucleotide in the cytoplasm and mitochondria of rat liver. The Biochemical Journal 103, 514–27.Google Scholar
Williamson, J. R. (1965). Glycolytic control mechanisms. I. Inhibition of glycolysis by acetate and pyruvate in the isolated, perfused rat heart. The Journal of Biological Chemistry 240, 2308–21.CrossRefGoogle ScholarPubMed
Williamson, J. R. (1974). Succinate. In Methods of Enzymatic Analysis, vol. 3 (ed. Bergmeyer, H. U.), pp. 1616–21. Weinheim, New York: Verlag Chemie, Academic Press.Google Scholar
Williamson, R. L. & Metcalf, R. L. (1967). Salicylanilides: a new group of active uncouplers of oxidative phosphorylation. Science 158, 1694–5.Google Scholar