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For different bacterial preservation techniques, there is no single method applicable for all bacteria. This study aimed to assess the viability of seven species/species groups of clinical bacteria isolates on the long-term storage (more than 5 years) by using Tryptic Soy Broth with 15% glycerol in the deep freezer (-70 to -80°C). A total 10,654 clinical bacteria isolates used as samples in this study. The isolates consisted of seven species/species groups (i.e.  Escherichia coli, Campylobacter  spp,  Shigella  spp,  Vibrio  spp,  Salmonella  spp,  Aeromonas hydrophila , and  Neisseria gonorhoeae).  The isolates were collected from some previous studies and preserved in the Tryptic Soy Broth (TSB) with 15% glycerol and stored in the deep freezer (-70 to -80°C) for more than five years. The samples were revived on the suitable medium to evaluate the viability of bacteria. Identification conducted by microscopic examination, biochemical test, and latex agglutination. The study showed that the viability of  Salmonella  spp,  Shigella  spp,  Aeromonas hydrophila  and  E. coli  was 100%, while  Campylobacter  spp,  Vibrio  spp, and  N.  gonorhoeae  were 66.7%, 66.4%, and 52.5% respectively. We concluded that viability of  Salmonella  spp,  Shigella  spp,  A. hydrophila , and  E. coli  was optimum thus better than  Campylobacter  spp,  Vibrio  spp, and  N. gonorhoeae  for more than 5 years storage by using TSB with 15% glycerol in the deep freezer (-70 to -80 °C).

Long-term Storage of Bacterial Isolates by Using Tryptic Soy Broth with 15% Glycerol in The Deep Freezer (-70 to -80 °C)