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Evaluation of Two Supplemented Culture Media for Long-Term, Room-Temperature Preservation of Streptococcus pneumoniae

Beatriz Quintero Moreno1, María Araque2, Evelyn Mendoza1

  • 1Departamento de Microbiología y Parasitología Clínicas, Facultad de Medicina, Universidad de Los Andes, Mérida 5101, Venezuela.

Biomed Research International
|January 24, 2018
PubMed
Abstract

Insights

Two new agar formulations, TH-HYC and TH-SYC, successfully preserved pneumococci viability for six months at room temperature. These methods offer a valuable alternative for bacterial storage and transport when other techniques are unavailable.

Area of Science:

  • Microbiology
  • Bacterial Preservation

Background:

  • Maintaining bacterial viability for extended periods is crucial for research and diagnostics.
  • Traditional methods like cryopreservation and lyophilization require specialized equipment and can be costly.
  • Developing accessible storage methods at room temperature is essential for resource-limited settings.

Purpose of the Study:

  • To develop and evaluate two novel supplemented agar media for long-term room temperature storage of *Streptococcus pneumoniae*.
  • To assess the viability and phenotypic stability of pneumococcal isolates after prolonged storage.

Main Methods:

  • Two agar types, Todd-Hewitt/Hemoglobin/Yeast/Charcoal/Agar (TH-HYC) and Todd-Hewitt/Skim-Milk/Yeast/Charcoal/Agar (TH-SYC), were prepared.
  • Nineteen *Streptococcus pneumoniae* isolates with diverse serotypes and multilocus sequence types (MLST) were subcultured and stored at room temperature for 4-6 months.
  • Viability was assessed by colony counts (CFU) and phenotypic characteristics.

Main Results:

  • All non-contaminated subcultures remained viable and retained their phenotypic characteristics after 6 months of storage.
  • A slow decrease in viable CFU was observed, but satisfactory growth levels were maintained.
  • Contamination rates were 6.25% at 2 months and 14.58% at 6 months.

Conclusions:

  • TH-HYC and TH-SYC agar formulations effectively support the viability of *Streptococcus pneumoniae* with diverse genetic profiles for up to 6 months at room temperature.
  • These agar types represent a practical and accessible alternative for pneumococcal preservation and inter-laboratory transport.
  • The study validates these methods as a valuable option when cryopreservation or lyophilization are not feasible.

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