Principle
P. aeruginosa are classified as Gram-negative microorganisms that grow on selective media containing
cetrimide and produces pyocyanin, or as microorganisms that grow on selective media containing
cetrimide, are oxidase positive, fluoresce under UV light (360 ± 20 nm), and can produce ammonia
from acetamide. Colonies producing pyocyanin on CN Agar medium confirmed for their ability to
synthesize pyoverdine using KING B medium. The medium contains high concentrations of magnesium
sulphate that provide the cations necessary for activation of pyoverdine giving the culture medium a
fluorescent greenish yellow color. Most strains (98%) produce a water-soluble fluorescent pigment.
Theoretical Composition
| Bacteriological peptone “B” | 20 g |
| Di-potassium hydrogen phosphate K2SO2 (anhydrous) | 1.5 g |
| Magnesium sulphate heptahydrate MgSO4, 7H2O | 1.5 g |
| Glycerol | 10 ml |
| Agar | 12 g |
| Distilled water | 1,000 ml |
| Final pH at 25 ⁰C = 7.2 ± 0.2 |
Reconstitution Ratio
Ready to use media, 7 ml x 25 tubes
Storage
Store ready-to-use medium at 2-8 ⁰C
References
– ISO 16266:2006
