Summary
Entamoeba invadens, amoebae of the type “Hartmanella” and cultures of both forms, all of them isolated from cold-blooded animals, further Entamoeba histolytica, Hartmanella castellanii, Trichomonas foetus and Trichomonas columbae were stored in liquid nitrogen. For the amoebae it was found, that an addition of 10% dimethylsulphoxide (DMSO) and a slow cooling (0.5° C/min) gave the best results. For Trichomonas foetus an addition of 5% DMSO and for Trichomonas columbae one of 10% glycerol showed the best cryoprotective effect. Slow freezing down to −55° C, which proved to be the optimum temperature for transferring in liquid nitrogen was carried out in the same manner as described for the amoebae. All strains were controlled during five weeks in cultures after the freeze-preservation for their growth rate. It was shown that the storage in liquid nitrogen was of no influence on the multiplication rate.
Zusammenfassung
Es wurden Entamoeba invadens, Amoeben vom Typ Hartmanella, Mischkulturen dieser beiden, sämtlich isoliert aus Kaltblütlern, Entamoeba histolytica, Hartmanella castellanii, Trichomonas foetus und Trichomonas columbae in flüssigem Stickstoff konserviert. Bei den Amoeben erwies sich ein Zusatz von 10% DMSO und eine kontinuierliche Abkühlung von 0,5° C/min als optimal. Für Trichomonas foetus waren 5% DMSO, für Trichomonas columbae 10% Glycerin der günstigste Zusatz bei gleicher Kühlgeschwindigkeit. Die günstigste Übertragungstemperatur von der langsamen Abkühlungsphase in flüssigen Stickstoff lag bei −55° C. Alle Stämme wurden nach der Gefrierkonservierung während eines Zeitraums von 5 Wochen in den Subkulturen auf ihre Vermehrungsfähigkeit kontrolliert. Dabei zeigte sich, daß die Kältekonservierung keinen Einfluß auf die Besatzdichten und damit auf die Fähigkeit zur Vermehrung hatte.
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Durchgeführt mit Mitteln der DFG und der Grimminger-Stiftung für Anthropozoonosenforschung.
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Bosch, I., Frank, W. Beitrag zur Gefrierkonservierung pathogener Amoeben und Trichomonaden. Z. Parasitenk. 38, 303–312 (1972). https://doi.org/10.1007/BF00329277
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DOI: https://doi.org/10.1007/BF00329277