Japanese Journal of Freezing and Drying

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The long-term survival of lapinized-avianized (LA) rinderpest virus preserved by freeze-drying was examined in order to aim the storage of rinderpest live virus vaccine. The viral materials of LA rinderpest virus multiplied in Vero cell cultures were freeze-dried in vials after mixing with an equal volume of protective agent consisting of 10% succharose (4% or 8% glucose in some cases), 1% glutamate and 1% gelatin. Then the vials were filled up with pure nitrogen gas and stored in refrigerator at 4℃ during over six years. The freeze-dried virus specimens exhibited a survival of 58% on the average by the process of freeze-drying. An average of 65% of survival rate was obtained through the preservation, although some lots showed 16% of minimum survival rate and the survival rate of the viral specimens which stored at 4℃ for 6 years was about 23%. A slightly increasing tendency was seen on the moisture rate of the specimens during the preservation, however, it was difficult to find a certain relationship between moisture and survival rates. The survival rate of specimens filled up with nitrogen gas was significantly high than that of the sealed one in vacuo after the preservation for over seven years. From the results mentioned above, it was suggested that LA rinderpest virus can be stored stably for at least six years by freeze-drying in vial.

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Seawater nutrient broth enriched with yeast extract and calcium lactate (SWNBYL) was used alone or in combination with sucrose, lactose, glucose, skim-milk, or bovine serum as suspending fluid for the freeze-drying of 3 strains of marine, chemoheterotrophic spirilla. The survival rate of freeze-dried Spirillum lunatum strain NCMB 54 was lower than 1.0% in all of the suspending fluids, and that of freeze-dried Oceanospirillum linum strain NCMB 55 (Spirillum atlanticum NCMB 55) was also lower than 1.0% in many of the suspending fluids, although it was 3.15% in MMG (bovine serum, 3ml; SWN-BYL, 1ml; glucose, 0.3g), 1.25%(average) in 4/5 SWNBYL・L_8 (4 parts of SWNBYL and 1 part of a 40% lactose solution) and 0.99%(average) in 4/5 SWNBYL・S_<12> (4 parts of SWNBYL and 1 part of a 60% sucrose solution) (Table 2). The survival rate of strain M55 was 2.73 to 16. 55%. MMG, 4/5 SWNBYL・S_<12>, 4/5 SWNBYL・S_8 (4 parts of SWNBYL and 1 part of a 40% sucrose solution) and 4/5 SWNBYL・L_8 showed a favourable effect for both S. lunatum strain NCMB 54 and O. linum strain NCMB 55; but the presence of skim-milk was unfavourable. The survival of freeze-dried strain M 55 seems to be little affected by type or concentration of the suspending fluids used.

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