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Viability of Onchocerca volvulus in vitro

Published online by Cambridge University Press:  06 April 2009

G. Strotel ,
A. Bokhof  and
J. C. W. Comley
G. Strotel
Affiliation:
Bernhard Nocht Institute for Tropical Medicine, Department of Helminthology and Entomology, Bernhard Nocht Strasse 74, D-2000 Hamburg 36, Germany
A. Bokhof
Affiliation:
Bernhard Nocht Institute for Tropical Medicine, Department of Helminthology and Entomology, Bernhard Nocht Strasse 74, D-2000 Hamburg 36, Germany
J. C. W. Comley
Affiliation:
The Wellcome Research Labs, Beckenham, Kent, UK
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Summary

The use of a selective schedule of tests to identify a viable population of isolated adult Onchocerca volvulus (Nematoda: Filarioidea) has been investigated in a large worm population. The study was initiated to develop methodology appropriate to test new candidate macrofilaricides for their in vitro activity against O. volvulus. After removal from the host the viability of isolated intact parasites was estimated by assessing the motility indices of male worms, and the colorimetric quantification of the reduction of the bioreducible tetrazolium reagent XTT and lactate output by female worms. Additionally the motility of whole females and the movement of inner organs of female worms were scored quantitatively. These response parameters were used to sort the adult worms into viability groups at the start of the in vitro culture. The adult worms were then observed for 6 days and viability was assessed regularly during the culture period. At the end of the culture period, the reduction of the water-insoluble tetrazolium reagent MTT was used to determine the formazan formed by the entire male and female worms. The response parameters used at the start of the culture proved to be highly predictive for detecting viable and non-viable adult worms. In the group of worms selected as ‘viable’ around 70% kept their motility and metabolic activity at a high level until the end of the culture compared to the initial level. In contrast, none of the female worms and only 13% of the male worms categorized as ‘poorly viable’ demonstrated a motility index or metabolic level at the end of the culture period that was comparable to that of the worms in the ‘viable’ groups. For female worms the lactate output correlated significantly with weight whereas no correlation was seen between MTT reduction and weight.

Keywords

Onchocerca volvulusin vitro cultureviabilityselective schedule
Type
Research Article
Information
Parasitology , Volume 107 , Issue 2 , August 1993 , pp. 175 - 182
Copyright
Copyright © Cambridge University Press 1993

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References

REFERENCES

Bennett, J. L. & Pax, R. A. (1986). Micromotility meter: an instrument designated to evaluate the action of drugs on motility of larval and adult nematodes. Parasitology 93, 341–6.CrossRefGoogle Scholar
Comley, J. C. W., Szopa, T., Strote, G., BÜttner, M., Darge, K. & Büttner, D. W. (1989 a). A preliminary assessment of the feasibility of selectively evaluating promising antifilarials in vitro against adult Onchocerca volvulus. Parasitology 99, 417–25.Google Scholar
Comley, J. C. W., Townson, S., Rees, M. J. & Dobinson, A. (1989 b). The further application of MTT–formazan colorimetry to studies on filarial worm viability. Tropical Medicine and Parasitology 40, 311–16.Google Scholar
Comley, J. C. W., Rees, M. J., Turner, C. H. & Jenkins, D. C. (1989 c). Colorimetric quantification of filarial viability. International Journal for Parasitology 19, 77–83.Google Scholar
Comley, J. C. W. & Turner, C. H. (1990). Potential of a soluble tetrazolium/formazan assay for the evaluation of filarial viability. International Journal for Parasitology 20, 251–5.Google Scholar
Kozek, W. J. & Figueroa-Marroquin, H. (1982). Attempts to establish Onchocerca volvulus infection in primates and small laboratory animals. Acta Tropica 39, 317–24.Google ScholarPubMed
MacKenzie, N. E., Vande Waa, E. A., Gooley, P. R., Williams, J. F., Bennett, J. L., Bjorge, S. M., Baille, T. A. & Geary, T. G. (1989). Comparison of glycolysis and glutaminolysis in Onchocerca volvulus and Brugia pahangi by 13C nuclear magnetic resonance spectroscopy. Parasitology 99, 427–35.Google Scholar
Mosmann, T. (1988). Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. Journal of Immunological Methods 65, 55–63.Google Scholar
Mössinger, J. (1991) In vitro cultivation of adult Litomosoides carinii: evaluation of basic culture media, gas phases and supplements. Parasitology 103, 85–95.CrossRefGoogle ScholarPubMed
Nowak, M., Hutchinson, G. W. & Copeman, D. B. (1987). In vitro drug screening in isolated male Onchocerca gibsoni using motility suppression. Tropical Medicine and Parasitology 38, 128–30.Google Scholar
Paull, K. D., Shoemaker, R. H., Body, M. R., Parsons, J. L., Risbood, P. A., Barbera, W. A., Sharma, M. N., Baker, D. C., Hand, E., Scudiero, D. A., Monks, A., Alley, M. C. & Grote, M. (1988). The synthesis of XTT: a new tetrazolium reagent that is bioreducible to a water-soluble formazan. Journal of Heterocyclic Chemistry 25, 911–14.CrossRefGoogle Scholar
Pax, R. A., Williams, J. F. & Guderian, R. H. (1988). In vitro motility of isolated adults and segments of Onchocerca volvulus, Brugia pahangi and Acanthocheilonema viteae. Tropical Medicine and Parasitology 39, 450–5.Google Scholar
Satti, M. Z., Vande Waa, E. A., Bennett, J. L., Williams, J. F., Conder, G. A. & McCall, J. W. (1988). Comparative effects of anthelmintics on motility in vitro of Onchocerca gutturosa, Brugia pahangi and Acanthocheilonema viteae. Tropical Medicine and Parasitology 39, 480–3.Google ScholarPubMed
Schulz-Key, H., Albiez, E. J. & Büttner, D. W. (1977). Isolation of living adult Onchocerca volvulus from nodules. Tropical Medicine and Parasitology 28, 428–30.Google ScholarPubMed
Scudiero, D. A., Shoemaker, R. H., Paull, K. D., Monks, A., Tierney, S., Nofziger, T. H., Currens, M. J., Seniff, D. & Boyd, M. R. (1988). Evaluation of a soluble tetrazolium/formazan assay for cell growth and drug sensitivity in culture using human and other tumour cell lines. Cancer Research 48, 4827–33.Google Scholar
Slater, T. F., Sawyer, B. & StrÄuli, U. (1963). Studies on succinate—tetrazolium reductase systems. III. Points of coupling of four different tetrazolium salts. Biochimica et Biophysica Acta 77, 383–93.CrossRefGoogle ScholarPubMed
Strote, G., Wieland, S., Darge, K. & Comley, J. C. W. (1990 a). In vitro assessment of the activity of anthelmintic compounds on adults of Onchocerca volvulus. Acta Leidensia 59, 285–96.Google Scholar
Strote, G., Darge, K. & Bonow, I. (1990 b). Morphological alterations of male Onchocerca volvulus after in vitro exposure to mel w and milbemycin a confirming the results of viability tests. Tropical Medicine and Parasitology 41, 429–36.Google Scholar
Strote, G., Bokhof, A. & Darge, K. (1992). The viability of adult Onchocerca volvulus after long-distance transportation. Transactions of the Royal Society of Tropical Medicine and Hygiene 86, 66.CrossRefGoogle ScholarPubMed
Townson, S., Connelly, C., Dobinson, A. & Muller, R. (1987). Drug activity against Onchocerca gutturosa males in vitro: a model for chemotherapeutic research on onchocerciasis. Journal of Helminthology 61, 271–81.CrossRefGoogle Scholar
Townson, S., Shay, K. E., Dobinson, A. R., Connelly, C., Comley, J. C. W. & Zea-Flores, G. (1989). Onchocerca gutturosa and O. volvulus: studies on the viability and drug responses of cryopreserved adult worms in vitro. Transactions of the Royal Society of Tropical Medicine and Hygiene 83, 664–9.Google Scholar
Walter, R. D. (1988). Preparation and shipping of Onchocerca volvulus material. Tropical medicine and Parasitology 39, 448–9.Google ScholarPubMed