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Studies on the cryopreservation of eggs of Angiostrongylus cantonensis

Published online by Cambridge University Press:  05 June 2009

Shoji Uga ,
Kazuyoshi Araki ,
Takeo Matsumura  and
Noboru Iwamura
Shoji Uga
Affiliation:
Department of Medical Zoology, Kobe University School of Medicine, Kobe 650, Japan
Kazuyoshi Araki
Affiliation:
Department of Medical Zoology, Kobe University School of Medicine, Kobe 650, Japan
Takeo Matsumura
Affiliation:
Department of Medical Zoology, Kobe University School of Medicine, Kobe 650, Japan
Noboru Iwamura
Affiliation:
International Center for Medical Research, Kobe 650, Japan
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Abstract

The possibility of cryopreserving the eggs of Angiostrongylus cantonensis collected from the uterus of female worms was investigated. Eggs were cultured in NCTC 109 medium containing 50% rat serum, and various growth stages, from one-cell eggs to embryonated eggs, were used in this study.

As a cryoprotective agent, dimethylsulphoxide (Me2SO) was added to the medium at a final concentration of 1 M. Eggs suspended in 0·2 ml of the medium at 37°C were cooled to 0°C at a rate of l°C min−1, then an equal volume of 2M-Me2SO solution was added. After equilibration for 15min, the freezing procedures were started. In the freezing procedures, the effectiveness of (i) a seeding process, (ii) different cooling and warming rates and (iii) the relationship between the growth stages of the eggs and their tolerance to freezing at −20°C were investigated.

It was found the highest level of survival could be obtained with 32-cell eggs cooled at a rate of 0·3° C min−1 or more slowly with seeding at −4°C and warming at a rate of 5°C min−1. Survival was influenced more by cooling rate than by warming rate.

Using these optimum conditions, the survival of eggs was then investigated following cooling to various temperatures. While more than 50% of eggs were found to survive cooling to −30–C, extremely low survival was noted from lower temperatures.

Type
Research Papers
Information
Journal of Helminthology , Volume 57 , Issue 4 , December 1983 , pp. 297 - 303
Copyright
Copyright © Cambridge University Press 1983

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