Selective and differential medium for the primary isolation of members of the Proteeae

A new differential and selective medium for the isolation of members of the Proteeae, PIM (Proteeae isolation medium) agar, was developed and evaluated. The medium relies on the ability of all members of the Proteeae (with the exception of a very few Morganella morganii strains) to produce a dark brown pigment in medium containing DL-tryptophan. An additional differential property, tyrosine degradation, was also demonstrated by the medium. Members of the Proteeae appeared as dark brown colonies with a halo of clearing of fine tyrosine crystals when cultured on PIM agar. Occasional strains of Citrobacter sp. and Pseudomonas aeruginosa may degrade tyrosine, but none has the ability to produce dark brown pigmentation on PIM agar. Quantitative recovery studies showed that the addition of 5 mg of clindamycin per liter suppressed gram-positive bacteria without inhibiting any strains of the Proteeae. The addition of 100 mg of colistin per liter made the medium highly selective for strains of the Proteeae, but approximately 10% of the strains were not isolated, thus making this formulation unsuitable for general surveys of the occurrence of members of the Proteeae. PIM agar should aid the investigation of episodes of cross infection caused by members of the Proteeae and the isolation of the new species of the Proteeae recently described.