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The preservation of bacteriophage H1 of Corynebacterium ulcerans U 103 by freeze-drying

Published online by Cambridge University Press:  15 May 2009

J. D. Davies  and
M. J. Kelly
J. D. Davies
Affiliation:
Department of Pathology, University of Cambridge, Cambridge
M. J. Kelly
Affiliation:
Department of Pathology, University of Cambridge, Cambridge
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Summary

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This paper describes an investigation into the successful preservation at room temperature of the bacteriophage H1 of Corynebacterium ulcerans U 103 which was extremely labile when in suspension at 4° C.

Cooling at a rate of 1° C./min. showed that the survival decreased logarithmically at temperatures between – 14 and – 45° C. Survival of broth suspensions of the corynebacteriophage were found to increase proportionally with an increase in the rate of cooling though there was a marked drop in survival at rates of approximately 900° C./min. The addition of peptone solutions was found to increase the survival over the range studied, whereas the addition of sucrose solutions had only a slight effect.

By avoiding freezing damage by cooling at rates of 450° C./min. in (a) 20% peptone solution, (b) 20% peptone and 10% sucrose, and (c) 20% peptone, 10% sucrose and 2% sodium glutamate, a study was made of the drying stage of the freeze-drying process. On drying at controlled temperatures it was found that there was no damage on rewarming to temperatures below – 21° C. after cooling to – 196° C., but that the survival immediately after drying in the absence of glutamate, showed a logarithmic relationship with the temperature of drying, lower temperatures giving better survival.

On storage for a period of 3 months at room temperature in vacuo and darkness, there was no appreciable loss in survival in the mixtures though suspensions in peptone alone showed a slight decrease. At higher temperatures this decrease in survival could be differentiated into two types of damage, each of which could be influenced by the presence of sucrose or glutamate.

We are grateful to Professor H. R. Carne for providing the initial sample of bacteriophage H 1 of Corynebacterium ulcerans U 103, and to Professor R. I. N. Greaves for his interest and encouragement during the course of this investigation.

Type
Research Article
Information
Journal of Hygiene , Volume 67 , Issue 4 , December 1969 , pp. 573 - 583
Copyright
Copyright © Cambridge University Press 1969

References

REFERENCES

Adams, M. (1959). Bacteriophages. New York: Wiley.CrossRefGoogle Scholar
Davies, J. D. (1966). Thermal analysis in freezing and freeze-drying. In Advances in Freeze-Drying, pp. 9–20. Ed. Rey, L.. Paris: Hermann.Google Scholar
Davies, J. D. (1967). The effects of freezing, thawing and drying on living cells. Ph.D. thesis, University of Cambridge.Google Scholar
Davies, J. D. (1969). The role of peptides in preventing freeze-thaw injury. In Ciba Symposium, The Frozen Cell. London: Churchill. (In the Press.)Google Scholar
Fry, R. M. & Greaves, R. I. N. (1951). The survival of bacteria during and after drying. J. Hyg., Camb. 49, 220–46.Google ScholarPubMed
Greaves, R. I. N. & Davies, J. D. (1965). Separate effects of freezing, thawing and drying living cells. Ann. N.Y. Acad. Sci. 125, 548–58.CrossRefGoogle ScholarPubMed
Leibo, S. P. & Mazur, P. (1967). Freezing of bacteriophage T 4B. Cryobiology 3, 366–7.Google Scholar
Muggleton, R. W. (1958). Freeze-drying of bacteria with special reference to B.C.G. In Recent Research in Freezing and Drying, pp. 229–37. Eds. Parkes, A. S. and Smith, A. U.. Oxford: Blackwell.Google Scholar
Nagington, J. & Greaves, R. I. N. (1962). Preservation of tissue culture cells in liquid nitrogen. Nature, Lond. 194, 993–4.CrossRefGoogle ScholarPubMed
Steele, P. R. M., Davies, J. D. & Greaves, R. I. N. (1969). Some factors affecting the viability of freeze-thawed T 4 bacteriophage. J. Hyg., Camb. 67, 107–14.CrossRefGoogle Scholar