The composition of a minimal medium suitable for the demands of modern genetical work is described. The nutritional requirements of some 15 strains of L. monocytogenes were studied on this medium. DL-6,8-thioctic acid was not essential. The organisms also grew without thiamine but for a convenient colony size this vitamin had to be used. The presence of Fe++, Mg++, and Ca++ ions as well as a low redox potential supplied by thioglycollate was necessary. There was some difference in the amino acid requirements of the strains. The most common demand was for leucine, iso-leucine, valine, and cysteine. The serotype, agglutinability, hemolytic effect, virulence, and nutritional requirements of the Listeria were compared. It seems that these properties are not consistently linked. Two 'EMS' mutants were also studied. The arginine, glutamine, and methionine metabolism of the auxotrophic mutant 025/4/4 was damaged and its virulence was lost. Data connected with the virulence markers of Listeria are discussed.