BAM Media M178: Wagatsuma Agar

January 2001

Bacteriological Analytical Manual

Wagatsuma Agar

Yeast extract 3 g
Peptone 10 g
NaCl 70 g
K2HPO4 5 g
Mannitol 10 g
Crystal violet 0.001 g
Agar 15 g
Distilled water 1 liter
Human or rabbit
red blood cells,
fresh (24 h), with anticoagulant

50 ml

Mix fresh (within 24 h of drawing) human or rabbit blood with same or larger volume of physiological saline. Centrifuge cells at about 4000 × g at 4°C for 15 min. Pour off saline and wash 2 more times. After 3rd wash, pour off saline and resuspend cells to original volume with saline.

Suspend ingredients, except blood, in distilled water and boil to dissolve agar. Adjust to pH 8.0 ± 0.2. Steam 30 min. DO NOT AUTOCLAVE. Cool to 45-50°C. Add 50 ml of washed red blood cells to the cooled medium. Mix and pour into sterile petri dishes. Dry plates thoroughly and use promptly. Medium can be made in smaller volumes (requiring less blood) when few plates are needed.

Hypertext Source: Bacteriological Analytical Manual, 8th Edition, Revision A, 1998.

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