The brilliant green dye used in Muller-Kauffmann Tetrathionate Broth has been shown to be critical.
Incubation of Muller-Kauffmann Tetrathionate Broth at 43°C was also shown to be essential.
The technique used for enrichment of the salmonellae is as follows:
Add approximately 10g of sample to 100mL of Muller-Kauffmann Tetrathionate Broth. Shake vigorously and immediately place the flasks of medium in a 45°C water-bath for 15 minutes. Remove the flasks from the water-bath, without drying them, and place in an incubator or another water-bath at 43°C.
Sub-culture the broth after 18 to 24 hours and again after 48 hours. Take one loopful of broth from the edge of
the surface of the fluid and inoculate either two Thermo Scientific Brilliant Green Agar (Part No. PO5033A) without recharging the loop between plates. Incubate the plates at 35°C for 18 to 24 hours.
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References
1. Muller L. (1923) C. R. Soc. Biol. (Paris) 89. 434-443.
2. Kauffmann F. (1930) Z. f. Hyg. 113. 148-157.
3. Kauffmann F. (1935) Z. f. Hyg. 117. 26-32.