Bio-processing and Herbal Division, Mahatma Gandhi Institute for Rural Industrialization, Wardha - 442001, Maharashtra, India
*Corresponding Author E-mail: saumyakatre48@gmail.com
Reliable revival of lyophilized bacterial cultures is essential for routine microbiological applications; however, commonly recommended media such as Brain Heart Infusion (BHI) broth are often costly and not readily available. This article describes a simple, reproducible, and cost-effective method for the revival, cultivation, and short-term maintenance of lyophilized Streptococcus mutans obtained from the Microbial Type Culture Collection (MTCC) using tryptic soy and nutrient media as alternatives. Lyophilized cultures were rehydrated and incubated under microaerophilic conditions, followed by isolation on nutrient agar and tryptic soy agar. Method validation was performed using visual turbidity assessment, optical density measurements at 600 nm (OD600), colony-forming unit (CFU) enumeration, colony morphology, and subculture viability. The method enabled rapid revival in tryptic soy media within 12–14h, yielding higher biomass and CFU counts than nutrient media, which exhibited delayed growth. Cultures revived in tryptic soy media and maintained on tryptic soy agar remained viable for up to 10–12 days at 4°C. This validated protocol provides an economical and practical alternative to BHI for routine revival and short-term maintenance of lyophilized S. mutans.
Validated, low-cost method using tryptic soy and nutrient media
Reliable revival of lyophilized Streptococcus mutans
Suitable for routine microbiological and teaching laboratories
Streptococcus mutans, Lyophilized culture revival, Tryptic Soy Broth, Nutrient Broth, Media validation, Microbiological methods
