A selective and diagnostic medium for use in the enumeration of Listeria spp. in foods

A new medium, called RAPAMY agar, has been elaborated for the isolation from and the enumeration of Listeria spp. in foods. It is based on Ralovich's nalidixic acid-trypaflavin-agar with the following modifications: (i) the slight inhibitory properties of that medium were overcome by the use of Columbia Blood agar base instead of tryptose agar and the addition of 0.05% ferric ammonium citrate and 2.5% egg yolk emulsion; (ii) selectivity was improved by the addition of 0.25% 2-phenyl ethanol and incubation under microaerobic conditions; (iii) the medium was provided with two diagnostic traits by the addition of (a) aesculin + ferric ammonium citrate; and (b) D-mannitol and phenol red. The growth of Enterococcus spp., the only organisms other than Listeria spp. which grow on RAPAMY agar, was not inhibited by the addition of 20 microgram.ml-1 Cefoxitin (Moxolactam). Higher levels inhibited some Listeria spp., but not the enterococci. The medium recovered Listeria spp. quantitatively and allowed recovery from foods colonized by Enterococcus spp. at levels upto 10(2) per g.