This article details a method for culturing Streptococcus mitis, a pathogenic bacterium, under microaerophilic conditions to study its virulence. The process involves using THY agar plates and a candle jar to create an environment that promotes bacterial growth and virulence phenotype development.
Begin with a sterile Todd-Hewitt Yeast Extract, or THY agar plate, a nutrient-rich medium that supports the growth of Streptococcus mitis, a pathogenic bacterium.
Using a sterile loop, streak the plate with the bacterial strain.
Place the inoculated plate in a candle jar, light a candle, seal the jar air-tight, and incubate.
The burning candle consumes oxygen and increases carbon dioxide levels, creating a microaerophilic environment that mimics host-like conditions.
The nutrients in the medium and the jar's microaerophilic environment enhance bacterial proliferation and promote the development of a virulence phenotype.
Post incubation, remove the jar lid and retrieve the plate.
Using a sterile loop, pick a single isolated colony and transfer it into a sterile conical tube containing THY broth.
Incubate the culture under static conditions to support bacterial growth while preserving the virulence phenotype.
This culture can be used as a pathogenic model for investigating host-pathogen interactions.
Before the preparation of Mitis Group Streptococci, warm the THY agar plates in an incubator to 37 degrees Celsius.
After that, use a sterile loop to streak out desired strains of Streptococci on the plates.
Then, put the plates in a candle jar, and incubate the jar at 37 degrees Celsius overnight for around 18 hours to provide a microaerophilic environment.
The next day, remove the plates from the candle jar and use a sterile loop to pick isolated colonies.
Inoculate 15 milliliter sterile conical tubes containing 2 milliliters of THY broth.
Close the caps tight and incubate the tubes at 37 degrees Celsius under static conditions.
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