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Studies on the cryopreservation of Dictyocaulus viviparus (Nematoda) third-stage larvae

Published online by Cambridge University Press:  18 November 2009

E. R. James  and
R. Peacock
E. R. James
Affiliation:
Department of Ophthalmology, Medical University of South Carolina, 171 Ashley Avenue, Charleston, South Carolina 29425, USA.
R. Peacock
Affiliation:
Glaxo Group Research Ltd., Ware, Hertfordshire SGI 2 ODJ, England.
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Abstract

Various cooling (01–5.100 Cmin−1) and warming (2O–6,8OO½Cmin−1) rates, stepped cooling schedules and four cryoprotective additives (dimethyl sulphoxide, methanol, ethanediol and glycerol) were investigated in cryopreservation studies with Dictyocaulus viviparus third-stage larvae. Exsheathment with sodium hypochlorite was essential to achieve significant survival. With uninterrupted cooling, highest survival (30% normally motile) was achieved with rates of 10–70½C min−1. Survival was higher (50–75%) using 1 ½C min−1 to — 10½C followed by plunging into liquid nitrogen. The optimum warming rate was 6,8OO½C min−1. The use of cryoprotectants led to marginally lower survival while varying the suspending media had no significant effect on survival. X-irradiated, exsheathed third-stage larvae cryopreserved by the optimum protocol yielded 38–3 ±4–2% survival. Two calves each infected with 45,000 (15,000 viable) exsheathed, unirradiated, C“ryopreserved third-stage larvae harboured 494 worms (11% infectivity) and 355 worms (0-8%) at necropsy. Numbers of first-stage larvae in the faeces reached 420/G and 105/G respectively 27 days after infection.

Keywords

CryopreservationDictyocaulusCooling rateWarming rateCryoprotectants.
Type
Research Papers
Information
Journal of Helminthology , Volume 60 , Issue 1 , March 1986 , pp. 65 - 73
Copyright
Copyright © Cambridge University Press 1986

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References

Anderson, F. L. & Levine, N. D. (1968) Effect of Desiccation on survival of free-living stages of Trichostrongylus colubriformis. Journal of Parasitology, 54, 117–128.Google Scholar
Campbell, W. C., Blair, L. S. & Egerton, J. R. (1972) Motility and Infectivity of Haemonchus Contortus Larvae After Freezing. Veterinary Record, 91, 13.Google Scholar
Campbell, W. C., Blair, L. S. & Egerton, J. R. (1973) Unimpaired infectivity of the nematode Haemonchus contortus after freezing for 44 weeks in the presence of liquid nitrogen. Journal of Parasitology, 59, 425–427.CrossRefGoogle ScholarPubMed
Campbell, W. C. & Thompson, B. M. (1973) Survival of Nematode Larvae after freezing over liquid nitrogen. Australian Veterinary Journal, 49, 111.Google Scholar
Farrant, J. (1965) Mechanisms of cell damage during freezing and thawing and its prevention. Nature, 205, 1284–1287.CrossRefGoogle Scholar
Fuller, B. J. & James, E. R. (1985) Cryopreservation of isolated rat hepatocytes in pellet form. Cryo-letters, 6, 49–56.Google Scholar
Isenstein, R. S. & Herlich, H. (1972) Cryopreservation of infective third-stage larvae of Trichostrongylus axei and T. colubriformis. Proceedings of the Helminthological society of Washington, 39, 140–141.Google Scholar
James, E. R. (1981) Schistosoma Mansoni: Cryopreservation of Schistosomula by two-step addition of ethanediol and rapid cooling. Experimental Parasitology, 52, 105–116.CrossRefGoogle ScholarPubMed
Kelly, J. D. & Campbell, W. C. (1974) Survival of Nippostrongylus Brasiliensis larvae after freezing over liquid nitrogen. International Journal for Parasitology, 4, 173–176.CrossRefGoogle ScholarPubMed
Kelly, J. D., Campbell, W. C. & Whitlock, H. V. (1976) Infectivity of Ancylostoma caninum larvae after freezing over liquid nitrogen. Australian Veterinary Journal, 52, 141–143.CrossRefGoogle ScholarPubMed
Miller, T. A. & Cunningham, M. P. (1965) Freezing of infective larvae of the dog hookworm. East African Trypanosomiasis Research Organization Annual Report, 1965, 19–22.Google Scholar
Parfitt, J. W. (1971) Deep freeze preservation of nematode larvae. Research in Veterinary Science, 12, 488–489.CrossRefGoogle ScholarPubMed
Poynter, D., Peacock, R. & Menear, N. C. (1979) The Prevention and Treatment of Husk. Veterinary Record, 86, 148–160.CrossRefGoogle Scholar
Vetter, J. C. M. & Klaver-Wesseling, J. C. M. (1977) Unimpaired Infectivity of Ancylostoma ceylanicum after storage in liquid nitrogen for one year. Journal of Parasitology, 63, 700.Google Scholar