Parasitology Research

, Volume 73, Issue 4, pp 358–365 | Cite as

Cryopreservation of Dictyocaulus viviparus third-stage larvae and Trichinella spiralis muscle larvae

  • I. Andermatt-Mettler
  • J. Eckert
  • Th. Ramp
  • B. Gottstein
Original Investigations


In cryopreservation studies with third-stage larvae of Dictyocaulus viviparus, best results were achieved by incubating larvae in 0.05% NaOCl at 37‡ C to remove the sheath, followed by cooling at a rate of 1‡ C min per min down to about 0‡ C. After an equilibration time of 10 min at +4‡ C with or without 4% polyethylene glycol-400 as cryoprotectant, samples were frozen at the same cooling rate to an intermediate temperature of −20‡ C, maintained at this temperature for 10 min and finally plunged into liquid nitrogen for storage. Three groups of 3 calves were infected with the following batches of third-stage larvae: (a) fresh, sheated; (b) fresh, exsheathed; (c) exsheathed, cryopreserved for 13 weeks in liquid nitrogen and subsequently thawed. Although 62% of group (c) were regarded as viable in vitro, their infectivity to calves was low and only an average of 0.08% of the inoculated larvae (3000 per animal) developed into adult lungworms (=infectivity rate). Average infectivity rates of fresh, sheathed (a) and fresh, exsheathed (b) larvae were much higher (38.3% and 29.7%) and not significantly different from each other. Two of the calves inoculated with previously frozen larvae and all of the calves infected with fresh larvae excreted first-stage larvae in their faeces, but the latter groups in higher quantities. The results show that cryopreservation of exsheathed third-stage larvae of D. viviparus is possible, but for strain maintenance infection doses greater than 3000 larvae should be used for inoculation of calves. Thawed Trichinella spiralis larvae, which had been cryopreserved in tissue blocks (approx. 2 g) with addition of undiluted ethylene glycol (0.2 ml), revealed in vitro viability rates of 60% after storage at −10‡ C for 2 and 13 weeks and were still infective to rats.


Ethylene Glycol Liquid Nitrogen Cool Rate Infectivity Rate Equilibration Time 
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Copyright information

© Springer-Verlag 1987

Authors and Affiliations

  • I. Andermatt-Mettler
    • 1
  • J. Eckert
    • 1
  • Th. Ramp
    • 1
  • B. Gottstein
    • 1
  1. 1.Institute of ParasitologyUniversity of ZürichZürichSwitzerland

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