Abstract
The cryopreservation of rabies virus has been described in detail in the literature. To date, little information is available on the use of cryoprotective agents for cold preservation of this virus, and the available data focus only on short-term virus preservation. In this study, we investigated the medium-term cryopreservation of samples of rabies virus using different cryopreservation protocols. The cryopreservation protocols for the rabies virus samples were performed at -20°C and were divided according to the variables of time and cryoprotectant type used. The laboratory tests (intracerebral inoculation of mice, viral titration and direct immunofluorescence) were performed at regular intervals (360 and 720 days) to assess the viability of the viral samples according to the different preservation techniques used. After 1 year of cryopreservation, the fluorescence intensity of intracellular corpuscles of the rabies virus and the median survival time of the mice differed between the positive controls and the treatments with the cryoprotectants. After 2 years, most of the samples subjected to the cryopreservation protocols (including the controls) did not produce fluorescence. However, the virus samples exposed to the cryoprotectant sucrose (68% solution) responded positively in the direct immunofluorescence assay and in the intracerebral inoculation of the mice. Medium-term cryopreservation of the rabies virus inactivates the viral sample. However, the cryoprotectant agent sucrose (68%) produces a preservative effect in cryopreserved rabies virus samples.
Highlights
The cryopreservation of rabies virus has been described in detail in the literature
After cryopreservation for 1 year (360 days; T360), the viral titer of the sample pool was reduced from 4.69 to 1.50 Log10 50% LD50 compared with the sample preserved for 30 days (T30)
At T360, significant differences were observed in the fluorescence intensity of the rabies virus intracellular corpuscles among the PC-CVS, PBS-PC and cryoprotectant treatments
Summary
The cryopreservation of rabies virus has been described in detail in the literature. To date, little information is available on the use of cryoprotective agents for cold preservation of this virus, and the available data focus only on short-term virus preservation. We investigated the medium-term cryopreservation of samples of rabies virus using different cryopreservation protocols. Methods: The cryopreservation protocols for the rabies virus samples were performed at -20oC and were divided according to the variables of time and cryoprotectant type used. Results: After 1 year of cryopreservation, the fluorescence intensity of intracellular corpuscles of the rabies virus and the median survival time of the mice differed between the positive controls and the treatments with the cryoprotectants. The virus samples exposed to the cryoprotectant sucrose (68% solution) responded positively in the direct immunofluorescence assay and in the intracerebral inoculation of the mice. Conclusions: Medium-term cryopreservation of the rabies virus inactivates the viral sample. The cryoprotectant agent sucrose (68%) produces a preservative effect in cryopreserved rabies virus samples. Because cooling significantly delays the deterioration of perishable goods, the use of low temperatures favors the storage
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