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Appl Environ Microbiol. 1975 August; 30(2): 167-171
Copyright © 1975 American Society for Microbiology. All Rights Reserved.

Survival of Frozen Mycoplasmas

Moshe Raccach, Shlomo Rottem and Shmuel Razin

Division of Food Technology, Agricultural Research Organization, The Volcani Center Bet Dagan, and Biomembrane Research Laboratory, Department of Clinical Microbiology,* The Hebrew University-Hadassah Medical School, Jerusalem, Israel

ABSTRACT

Cooling to -70 C killed a higher percentage of Acholeplasma laidlawii and Mycoplasma mycoides var. capri cells than cooling to -20 C. However, to preserve cell viability for prolonged periods storage at -70 C was much more preferable. The percentage of cells surviving freezing could be increased by increasing the initial cell concentration or by the addition of dimethyl sulfoxide or glycerol as cryoprotective agents. In the presence of 1.5 M of any one of these agents survival rates of up to 100% could be obtained. The optimal cooling rates for maximal survival of A. laidlawii under the experimental conditions tested were 11 C/min for cooling to -20 C and about 15 C/min for cooling to -70 C. Increasing the warming rate during thawing from 0.6 to 67 C/min increased survival by 3 log. Oleic acid enrichment of A. laidlawii membrane lipids, or reduction in the cholesterol content of M. mycoides var. capri membranes, increased the percentage of organisms surviving freezing. Hence, the composition of membrane lipids appears to have a marked influence on the susceptibility of mycoplasmas to freezing injury.

Appl Environ Microbiol. 1975 August; 30(2): 167-171
Copyright © 1975 American Society for Microbiology. All Rights Reserved.





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Copyright © 1975 by the American Society for Microbiology. All rights reserved.