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. 1969 Mar;17(3):427–434. doi: 10.1128/am.17.3.427-434.1969

Interacting Factors That Influence Long-term Storage of Live Pasteurella tularensis Vaccine and Rift Valley Fever Virus

Frederick Klein a, Jerry S Walker a,1, Bill G Mahlandt a, Richard C Carter a, Michael D Orlando a, Francis J Weirether a, Ralph E Lincoln a
PMCID: PMC377706  PMID: 5780399

Abstract

Studies were conducted on the interaction of various parameters which affect the storage stability and growth potential of liquid cultures of Pasteurella tularensis live vaccine strain (LVS) and Rift Valley fever virus Van Wyk strain (RVFV). Storage variables studied with LVS included four storage temperatures (4, -20, -65, -175 C), single and multiple freeze-thaw cycles, two freezing and two thawing rates (slow and fast), various inoculum levels (1, 3, 5, and 10%) for the determination of growth potential, and the retention of immunizing potential (mice and guinea pig) after storage. Neither the freezing rate nor the number of freeze-thaw cycles seriously affected the growth of LVS after storage at -175C; however, the slow rate of thaw proved deleterious as were all temperatures of storage except -175 C after 1 year of storage, as shown by both criteria of evaluation. RVFV produced in two combinations of cell lines and media (LM cell line-199 peptone medium and LDR cell line-Eagle's minimum essential medium) was stored at three serum levels (10, 20, 40%), three pH values (6.2., 7.0, 7.8), and three temperatures (-20, -65, -175 C). These studies indicated: (i) virus produced in the LDR cell line and Eagle's medium was more stable than that produced in the LM cell line and 199 peptone medium for either short- or long-term storage; (ii) serum levels did not affect stability; and (iii) low pH resulted in losses during long-term storage under all conditions tested. Thus, cryogenic storage is advantageous for stock culture maintenance of bacteria and viruses and for other similar applications.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Downs C. M., Coriell L. L., Chapman S. S., Klauber A. The Cultivation of Bacterium tularense in Embryonated Eggs. J Bacteriol. 1947 Jan;53(1):89–100. [PMC free article] [PubMed] [Google Scholar]
  2. EIGELSBACH H. T., DOWNS C. M. Prophylactic effectiveness of live and killed tularemia vaccines. I. Production of vaccine and evaluation in the white mouse and guinea pig. J Immunol. 1961 Oct;87:415–425. [PubMed] [Google Scholar]
  3. GREIFF D., MYERS M. OXIDATIVE PHOSPHORYLATION BY SUSPENSIONS OF MITOCHONDRIA FOLLOWING FREEZING AND DRYING BY SUBLIMATION IN VACUO. Biochim Biophys Acta. 1963 Oct 8;78:45–51. doi: 10.1016/0006-3002(63)91607-x. [DOI] [PubMed] [Google Scholar]
  4. Greiff D., Kelly R. T. Cryotolerance of enzymes. I. Freezing of lactic dehydrogenase. Cryobiology. 1966 May-Jun;2(6):335–341. doi: 10.1016/s0011-2240(66)80122-0. [DOI] [PubMed] [Google Scholar]
  5. KUCHLER R. J., MERCHANT D. J. Propagation of strain L (Earle) cells in agitated fluid suspension cultures. Proc Soc Exp Biol Med. 1956 Aug-Sep;92(4):803–806. doi: 10.3181/00379727-92-22620. [DOI] [PubMed] [Google Scholar]
  6. MAZUR P., RHIAN M. A., MAHLANDT B. G. Survival of Pasteurella tularensis in sugar solutions after cooling and warming at sub-zero temperatures. J Bacteriol. 1957 Mar;73(3):394–397. doi: 10.1128/jb.73.3.394-397.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. MERYMAN H. T. Preservation of living cells. Fed Proc. 1963 Jan-Feb;22:81–89. [PubMed] [Google Scholar]
  8. PRIVITERA C. A., GREIFF D., STRENGTH D. R., ANGLIN M., PINKERTON H. Oxidative phosphorylation by mitochondrial suspensions after freezing and storage at low temperatures. J Biol Chem. 1958 Aug;233(2):524–527. [PubMed] [Google Scholar]
  9. Scharer J. M., Klein F., Lincoln R. E. Growth and metabolism of live vaccine strain of Pasteurella tularensis. Appl Microbiol. 1968 Jun;16(6):855–861. doi: 10.1128/am.16.6.855-861.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Weirether F. J., Walker J. S., Lincoln R. E. A precise method for replicating suspension cultures of mammalian cells. Appl Microbiol. 1968 Jun;16(6):841–844. doi: 10.1128/am.16.6.841-844.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]

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