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Long-term (35 years) cryopreservation of Echinococcus multilocularis metacestodes


Laurimäe, Teivi; Kronenberg, Philipp A; Alvarez Rojas, Cristian A; Ramp, Theodor W; Eckert, Johannes; Deplazes, Peter (2020). Long-term (35 years) cryopreservation of Echinococcus multilocularis metacestodes. Parasitology, 147(9):1048-1054.

Abstract

The metacestode of <jats:italic>Echinococcus multilocularis</jats:italic> is the etiological agent of alveolar echinococcosis. The metacestode stage used for research is maintained in rodents by serial passages. In order to determine whether cryopreservation of <jats:italic>E. multilocularis</jats:italic> metacestodes would be suitable for long-term maintenance and replace serial passages, isolates of different geographic origin were cryopreserved in 1984–1986. The aim of the current study was to test the viability of cryopreserved isolates following long-term cryopreservation (up to 35 years) and to determine the phylogenetic clades these isolates belonged to. Cryopreserved isolates were tested for viability <jats:italic>in vitro</jats:italic> and <jats:italic>in vivo</jats:italic> in gerbils. <jats:italic>In vitro</jats:italic> results of 5 isolates indicated protoscolex survival in 13 of 17 experiments (76%) and metacestode survival in 5 of 12 (42%) <jats:italic>in vivo</jats:italic> experiments. <jats:italic>In vivo</jats:italic> results showed ‘abortive lesions’ in 13 of the 36 animals, 15 were negative and 8 harboured proliferating metacestode tissue containing protoscoleces. Genetic analysis confirmed the isolates belonged to European, Asian and North-American clades. In conclusion, the results of the current study indicate that metacestodes of <jats:italic>E. multilocularis</jats:italic> are able to survive long-term cryopreservation. Therefore, cryopreservation is a suitable method for long-term storage of <jats:italic>E. multilocularis</jats:italic> metacestode isolates and reduces the number of experimental animals.

Abstract

The metacestode of <jats:italic>Echinococcus multilocularis</jats:italic> is the etiological agent of alveolar echinococcosis. The metacestode stage used for research is maintained in rodents by serial passages. In order to determine whether cryopreservation of <jats:italic>E. multilocularis</jats:italic> metacestodes would be suitable for long-term maintenance and replace serial passages, isolates of different geographic origin were cryopreserved in 1984–1986. The aim of the current study was to test the viability of cryopreserved isolates following long-term cryopreservation (up to 35 years) and to determine the phylogenetic clades these isolates belonged to. Cryopreserved isolates were tested for viability <jats:italic>in vitro</jats:italic> and <jats:italic>in vivo</jats:italic> in gerbils. <jats:italic>In vitro</jats:italic> results of 5 isolates indicated protoscolex survival in 13 of 17 experiments (76%) and metacestode survival in 5 of 12 (42%) <jats:italic>in vivo</jats:italic> experiments. <jats:italic>In vivo</jats:italic> results showed ‘abortive lesions’ in 13 of the 36 animals, 15 were negative and 8 harboured proliferating metacestode tissue containing protoscoleces. Genetic analysis confirmed the isolates belonged to European, Asian and North-American clades. In conclusion, the results of the current study indicate that metacestodes of <jats:italic>E. multilocularis</jats:italic> are able to survive long-term cryopreservation. Therefore, cryopreservation is a suitable method for long-term storage of <jats:italic>E. multilocularis</jats:italic> metacestode isolates and reduces the number of experimental animals.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:05 Vetsuisse Faculty > Institute of Parasitology
04 Faculty of Medicine > Institute of Parasitology
Dewey Decimal Classification:570 Life sciences; biology
610 Medicine & health
600 Technology
Scopus Subject Areas:Life Sciences > Parasitology
Life Sciences > Animal Science and Zoology
Health Sciences > Infectious Diseases
Uncontrolled Keywords:Animal Science and Zoology, Parasitology, Infectious Diseases
Language:English
Date:April 2020
Deposited On:01 Sep 2020 14:48
Last Modified:02 Sep 2020 20:00
Publisher:Cambridge University Press
ISSN:0031-1820
OA Status:Closed
Publisher DOI:https://doi.org/10.1017/s003118202000075x
PubMed ID:32364108

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