Hostname: page-component-586b7cd67f-2plfb Total loading time: 0 Render date: 2024-11-25T15:08:32.035Z Has data issue: false hasContentIssue false
  • English
  • Français

An improved technique for the cryopreservation of Onchocerca microfilariae

Published online by Cambridge University Press:  06 April 2009

P. J. Ham ,
S. Townson ,
E. R. James  and
A. E. Bianco
P. J. Ham
Affiliation:
Department of Medical Helmintnology, London School of Hygiene and Tropical Medicine, Keppel Street (Gower Street), London WC1E 7HT
S. Townson
Affiliation:
Department of Medical Helmintnology, London School of Hygiene and Tropical Medicine, Keppel Street (Gower Street), London WC1E 7HT
E. R. James
Affiliation:
Department of Medical Helmintnology, London School of Hygiene and Tropical Medicine, Keppel Street (Gower Street), London WC1E 7HT
A. E. Bianco
Affiliation:
Department of Medical Helmintnology, London School of Hygiene and Tropical Medicine, Keppel Street (Gower Street), London WC1E 7HT
Get access Rights & Permissions [Opens in a new window]

Summary

Experiments describe the use of ethanediol to store Onchocerca microfilariae in liquid nitrogen. The technique involves a 2-stage incubation of the parasites at 37 °C and 0 °C in ethanediol, before rapid cooling to − 196 °C. Viability has been assessed by motility, by migration in a proxy host and by development to the infective stage in the insect vector. A total of 71–79% of the cryopreserved microfilariae was shown to be viable compared to unfrozen controls. The technique is simple, inexpensive and very effective when compared to previous cryopreservation procedures for microfilariae and should be particularly suited for use under field conditions.

Type
Research Article
Information
Parasitology , Volume 83 , Issue 1 , August 1981 , pp. 139 - 146
Copyright
Copyright © Cambridge University Press 1981

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Bianco, A. E., Ham, P., El Sinnary, K. & Nelson, G. S. (1980). Large-scale recovery of Onchocerca microfilariae from naturally infected cattle and horses. Transactions of the Royal Society of Tropical Medicine and Hygiene 74, 109–10.Google Scholar
Eichler, D. A. (1973). Studies on Onchocerca gutturosa (Neumann, 1910) and its development in Simulium ornatum (Meigen, 1818). 3. Factors affecting the development of the parasite in its vector. Journal of Helminthology 47, 73–88.CrossRefGoogle ScholarPubMed
Farrant, J. (1977). Water transport and cell survival in cryobiological procedures. Philosophical Transactions of the Royal Society of London, B 278, 191–205.Google ScholarPubMed
Gehenio, P. M. & Luyet, B. J. (1953). The survival of myxamoebae after freezing in liquid nitrogen. Biodynamica 7, 175–80.Google ScholarPubMed
Ham, P. J., James, E. R. & Bianco, A. E. (1978). Preliminary studies on the cryopreservation of Onchocerca microfilariae using a proxy host model to assess viability. Transactions of the Royal Society of Tropical Medicine and Hygiene 72, 434.Google Scholar
Ham, P. J., James, E. R. & Bianco, A. E. (1979). Onchocerca spp: Cryopreservation of microfilariae and subsequent development in the insect host. Experimental Parasitology 47, 384–91.CrossRefGoogle ScholarPubMed
James, E. R. (1981 a). Schistosoma mansoni: Cryopreservation of schistosomula by 2-step addition of ethanediol and rapid cooling. Experimental Parasitology (in the Press).CrossRefGoogle Scholar
James, E. R. (1981 b). High cryoprotectant concentrations and the interaction of permeation, dehydration and time in cryopreservation of Schistosoma mansoni schistosomula. Cryobiology (in the Press).Google Scholar
Luyet, B. J. & Keane, J. F. (1952). Comparative efficiency of ethylene glycol, glucose and sodium chloride in protecting tissues against freezing injury. Biodynamica 7, 139 and 119–31.Google ScholarPubMed
Minjas, J. N. & Townson, H. (1980). The successful cryopreservation of microfilariae with hydroxyethyl starch as cryoprotectant. Annals of Tropical Medicine and Parasitology 74, 571–3.CrossRefGoogle ScholarPubMed
Miyata, A. (1975). On the cryo-biological study of the parasitic protozoa (3). Effects of temperature and time of equilibration with glycerol or DMSO on survival of Trichomonas vaginalis. Tropical Medicine 17, 55–64.Google Scholar
Nelson, G. S., Amin, M. A., Blackie, E. J. & Robson, N. (1966). The maintenance of Onchocerca gutturosa microfilariae in vitro and in vivo. Transactions of the Royal Society of Tropical Medicine and Hygiene 60, 17.CrossRefGoogle Scholar
Schiller, E. L., Turner, V. M., Marroqúin, H. F. & D'antonio, R. (1979). The cryopreservation and in vitro cultivation of larval Onchocerca volvulus. American Journal of Tropical Medicine and Hygiene 28, 997–1009.CrossRefGoogle ScholarPubMed
Taylor, R., Adams, G. D. J., Boardman, C. F. B. & Wallis, R. G. (1974). Cryoprotection – permeant vs nonpermeant additives. Cryobiology 11, 430–8.CrossRefGoogle ScholarPubMed