Comparison of different culture media and storage temperatures for the long-term preservation of Streptococcus pneumoniae in the tropics

George Siberry, K. N. Brahmadathan, Rajeswar Pandian, M. K. Lalitha, Mark C. Steinhoff, T. John Jacob

Research output: Contribution to journalArticle

Abstract

Objective: The preservation of Streptococcus pneumoniae by standard freezing methods for subsequent tests - such as serotyping and antibiotic susceptibility - is not possible or is difficult in many developing countries because of the high cost of equipment, inadequate equipment maintenance, and irregular power supply. We evaluated alternative low-cost methods, by comparing different culture media and storage temperatures. Methods: Clinical isolates of five capsular types (1, 5, 7, 19, and 23) of S. pneumoniae were preserved in rabbit blood, sheep blood, skimmed milk, or glycerol-chocolate broth, and stored at -20°C or -70°C. The cultures were also preserved by lyophilization or sand desiccation, followed by storage at room temperature and 4°C. The viability of the preserved cultures was determined by making serial colony counts on day 0 and after 1 week, 4 weeks, 4 months and 16 months. The viability of cultures preserved by sand desiccation and storage at 4°C was also determined every 6 months for up to 68 months. Findings: Irrespective of the media used, cultures maintained at -20°C became nonviable by the fourth month, while those maintained at -70°C were still viable at 16 months. Cultures preserved by lyophilization or sand desiccation lost their viability by the fourth month when maintained at local room temperature (30-42°C), but remained viable when stored at 4°C for up to 68 months. Conclusions: Our results confirm that freezing at -70°C, or lyophilization and storage at 4°C are the ideal methods for the preservation of S. pneumoniae. In laboratories where lyophilization is not feasible, sand desiccation and storage at 4°C offers an alternative low-cost method for the long-term preservation of S. pneumoniae.

Original languageEnglish (US)
Pages (from-to)43-47
Number of pages5
JournalBulletin of the World Health Organization
Volume79
Issue number1
StatePublished - 2001

Fingerprint

Streptococcus pneumoniae
Desiccation
Freeze Drying
Culture Media
Temperature
Costs and Cost Analysis
Freezing
Electric Power Supplies
Serotyping
Equipment and Supplies
Glycerol
Developing Countries
Sheep
Milk
Maintenance
Anti-Bacterial Agents
Rabbits

Keywords

  • Cryopreservation, methods
  • Culture media
  • Desiccation, methods
  • Lyophilization, methods
  • Streptococcus pneumoniae, preservation
  • Tropical climate

ASJC Scopus subject areas

  • Public Health, Environmental and Occupational Health

Cite this

Siberry, G., Brahmadathan, K. N., Pandian, R., Lalitha, M. K., Steinhoff, M. C., & Jacob, T. J. (2001). Comparison of different culture media and storage temperatures for the long-term preservation of Streptococcus pneumoniae in the tropics. Bulletin of the World Health Organization, 79(1), 43-47.

Comparison of different culture media and storage temperatures for the long-term preservation of Streptococcus pneumoniae in the tropics. / Siberry, George; Brahmadathan, K. N.; Pandian, Rajeswar; Lalitha, M. K.; Steinhoff, Mark C.; Jacob, T. John.

In: Bulletin of the World Health Organization, Vol. 79, No. 1, 2001, p. 43-47.

Research output: Contribution to journalArticle

Siberry, G, Brahmadathan, KN, Pandian, R, Lalitha, MK, Steinhoff, MC & Jacob, TJ 2001, 'Comparison of different culture media and storage temperatures for the long-term preservation of Streptococcus pneumoniae in the tropics', Bulletin of the World Health Organization, vol. 79, no. 1, pp. 43-47.
Siberry, George ; Brahmadathan, K. N. ; Pandian, Rajeswar ; Lalitha, M. K. ; Steinhoff, Mark C. ; Jacob, T. John. / Comparison of different culture media and storage temperatures for the long-term preservation of Streptococcus pneumoniae in the tropics. In: Bulletin of the World Health Organization. 2001 ; Vol. 79, No. 1. pp. 43-47.
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AB - Objective: The preservation of Streptococcus pneumoniae by standard freezing methods for subsequent tests - such as serotyping and antibiotic susceptibility - is not possible or is difficult in many developing countries because of the high cost of equipment, inadequate equipment maintenance, and irregular power supply. We evaluated alternative low-cost methods, by comparing different culture media and storage temperatures. Methods: Clinical isolates of five capsular types (1, 5, 7, 19, and 23) of S. pneumoniae were preserved in rabbit blood, sheep blood, skimmed milk, or glycerol-chocolate broth, and stored at -20°C or -70°C. The cultures were also preserved by lyophilization or sand desiccation, followed by storage at room temperature and 4°C. The viability of the preserved cultures was determined by making serial colony counts on day 0 and after 1 week, 4 weeks, 4 months and 16 months. The viability of cultures preserved by sand desiccation and storage at 4°C was also determined every 6 months for up to 68 months. Findings: Irrespective of the media used, cultures maintained at -20°C became nonviable by the fourth month, while those maintained at -70°C were still viable at 16 months. Cultures preserved by lyophilization or sand desiccation lost their viability by the fourth month when maintained at local room temperature (30-42°C), but remained viable when stored at 4°C for up to 68 months. Conclusions: Our results confirm that freezing at -70°C, or lyophilization and storage at 4°C are the ideal methods for the preservation of S. pneumoniae. In laboratories where lyophilization is not feasible, sand desiccation and storage at 4°C offers an alternative low-cost method for the long-term preservation of S. pneumoniae.

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