Optimizing a low-temperature preservation technique for Bacillus anthracis strains

The use of pure microbial cultures is associated with the following key challenges: storage, transportation and resuscitation after a long-term preservation. The currently used anthrax vaccines are produced using various strains of Bacillus anthracis. According to the storage passport data, anthrax...

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Main Authors: A. P. Rodionov, E. A. Artemeva, L. A. Melnikova, D. M. Sahibullina
Format: Article
Language:English
Published: Da Vinci Media 2023-06-01
Series:Ветеринария сегодня
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Online Access:https://veterinary.arriah.ru/jour/article/view/733
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author A. P. Rodionov
E. A. Artemeva
L. A. Melnikova
D. M. Sahibullina
author_facet A. P. Rodionov
E. A. Artemeva
L. A. Melnikova
D. M. Sahibullina
author_sort A. P. Rodionov
collection DOAJ
description The use of pure microbial cultures is associated with the following key challenges: storage, transportation and resuscitation after a long-term preservation. The currently used anthrax vaccines are produced using various strains of Bacillus anthracis. According to the storage passport data, anthrax strains are now stored in 30–40% glycerin solutions, which helps to preserve a sufficient number of viable cells without losses to their pathogenic properties for three years. It is obviously an urgent task to develop a long-term preservation technique for Bacillus anthracis strains. The aim of this study was to optimize a low-temperature preservation method for Bacillus anthracis strains that ensures viability and no losses to biological properties of the pathogen. Two vaccine strains of Bacillus anthracis were selected for the research: i.e. K-STI-79 and 55-VNIIVViM and two cryoprotective media (No. 1 – 15% glycerin solution with 15% glucose solution and No. 2 – 30% neutral glycerin solution in saline solution). At first biological properties of the strains were studied and the number of viable cells was calculated. Later on, the strains were placed into low-temperature preservation facilities, at the temperature of –40 and –70 °C. Six months later, the effect of three thawing cycles on viability and biological properties of the agent was tested: i.e. at room temperature (22 ± 2) °C, in a water bath at a temperature of (37 ± 1) °C and in a household refrigerator at a temperature of (6 ± 2) °C. As demonstrated, the best option is to preserve the cells at –70 °C and thaw them in a water bath at (37 ± 1) °C. Further research will be focused on duration of the low-temperature preservation that will ensure appropriate viability and biological properties of the pathogen.
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institution Kabale University
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spelling doaj-art-e9a85646ebef4dd183f203bc108949d52025-02-06T09:52:10ZengDa Vinci MediaВетеринария сегодня2304-196X2658-69592023-06-0112217117710.29326/2304-196X-2023-12-2-171-177619Optimizing a low-temperature preservation technique for Bacillus anthracis strainsA. P. Rodionov0E. A. Artemeva1L. A. Melnikova2D. M. Sahibullina3FSBSI “Federal Center for Toxicological, Radiation and Biological Safety” (FSBSI “FCTRBS-ARRVI”)FSBSI “Federal Center for Toxicological, Radiation and Biological Safety” (FSBSI “FCTRBS-ARRVI”)FSBSI “Federal Center for Toxicological, Radiation and Biological Safety” (FSBSI “FCTRBS-ARRVI”)FSBSI “Federal Center for Toxicological, Radiation and Biological Safety” (FSBSI “FCTRBS-ARRVI”)The use of pure microbial cultures is associated with the following key challenges: storage, transportation and resuscitation after a long-term preservation. The currently used anthrax vaccines are produced using various strains of Bacillus anthracis. According to the storage passport data, anthrax strains are now stored in 30–40% glycerin solutions, which helps to preserve a sufficient number of viable cells without losses to their pathogenic properties for three years. It is obviously an urgent task to develop a long-term preservation technique for Bacillus anthracis strains. The aim of this study was to optimize a low-temperature preservation method for Bacillus anthracis strains that ensures viability and no losses to biological properties of the pathogen. Two vaccine strains of Bacillus anthracis were selected for the research: i.e. K-STI-79 and 55-VNIIVViM and two cryoprotective media (No. 1 – 15% glycerin solution with 15% glucose solution and No. 2 – 30% neutral glycerin solution in saline solution). At first biological properties of the strains were studied and the number of viable cells was calculated. Later on, the strains were placed into low-temperature preservation facilities, at the temperature of –40 and –70 °C. Six months later, the effect of three thawing cycles on viability and biological properties of the agent was tested: i.e. at room temperature (22 ± 2) °C, in a water bath at a temperature of (37 ± 1) °C and in a household refrigerator at a temperature of (6 ± 2) °C. As demonstrated, the best option is to preserve the cells at –70 °C and thaw them in a water bath at (37 ± 1) °C. Further research will be focused on duration of the low-temperature preservation that will ensure appropriate viability and biological properties of the pathogen.https://veterinary.arriah.ru/jour/article/view/733anthraxbacillus anthracisstrainslow-temperature preservation
spellingShingle A. P. Rodionov
E. A. Artemeva
L. A. Melnikova
D. M. Sahibullina
Optimizing a low-temperature preservation technique for Bacillus anthracis strains
Ветеринария сегодня
anthrax
bacillus anthracis
strains
low-temperature preservation
title Optimizing a low-temperature preservation technique for Bacillus anthracis strains
title_full Optimizing a low-temperature preservation technique for Bacillus anthracis strains
title_fullStr Optimizing a low-temperature preservation technique for Bacillus anthracis strains
title_full_unstemmed Optimizing a low-temperature preservation technique for Bacillus anthracis strains
title_short Optimizing a low-temperature preservation technique for Bacillus anthracis strains
title_sort optimizing a low temperature preservation technique for bacillus anthracis strains
topic anthrax
bacillus anthracis
strains
low-temperature preservation
url https://veterinary.arriah.ru/jour/article/view/733
work_keys_str_mv AT aprodionov optimizingalowtemperaturepreservationtechniqueforbacillusanthracisstrains
AT eaartemeva optimizingalowtemperaturepreservationtechniqueforbacillusanthracisstrains
AT lamelnikova optimizingalowtemperaturepreservationtechniqueforbacillusanthracisstrains
AT dmsahibullina optimizingalowtemperaturepreservationtechniqueforbacillusanthracisstrains