Tetrathionate broth base
|Bile salts||1 g|
|Calcium carbonate||10 g|
|Sodium thiosulfate·5H2O||30 g|
|Distilled water||1 liter|
Suspend ingredients in 1 liter distilled water, mix, and heat to boiling. DO NOT AUTOCLAVE. (Precipitate will not dissolve completely.) Cool to less than 45°C. Store at 5-8°C. Final pH, 8.4 ± 0.2.
Iodine-Potassium Iodide (I2-KI) solution
|Potassium iodide||5 g|
|Iodine, resublimed||6 g|
|Distilled water, sterile||20 ml|
Dissolve potassium iodide in 5 ml sterile distilled water. Add iodine and stir to dissolve. Dilute to 20 ml.
Brilliant green solution
|Brilliant green dye, sterile||0.1 g|
|Distilled water, sterile||100 m1|
On day of use, add 20 ml I2-KI solution and 10 ml brilliant green solution to 1 liter base. Resuspend precipitate by gentle agitation and aseptically dispense 10 ml portions into 20 × 150 or 16 × 150 mm sterile test tubes. Do not heat medium after addition of I2-KI and dye solutions.
Hypertext Source: Bacteriological Analytical Manual, 8th Edition, Revision A, 1998.