Comparison of Enrichment and Plating Media for Recovery of Virulent Strains of Yersinia enterocolitica from Inoculated Beef Stew

Abstract

Five plating agar media were evaluated for their ability to recover pure cultures of virulent strains of Yersinia enterocolitica serotypes O:3, O:8 and O:5,27. Cellobiose-arginine-lysine and bismuth sulfite agars were unproductive at 32°C but gave quantitative recovery with 48 h of incubation at 22°C. Adjustment of the pH of bismuth sulfite agar to 7.4 made the medium inhibitory. MacConkey, DHL and cefsulodin-irgasan-novobiocin agars gave quantitative recovery with 24 h of incubation at 32°C. Four preenrichment media incubated at four different temperatures, three selective enrichment media incubated at 22°C, and the five plating media were evaluated for their ability to recovery Y. enterocolitica from beef stew seeded with a background of ten other gram-negative bacteria. None of the plating media was superior for recovery; however, cefsulodin-irgasan-novobiocin agar showed the highest confirmation rate for presumptive colonies. Buffered-sorbitol-bile broth was inferior to richer media such as trypticase soy broth for preenrichment. Of the three selective enrichment media examined, only bile-oxalate-sorbose broth was found useful, especially for strains of serotype O:8 which could be recovered after 1 d of preenrichment and 3 d of selective enrichment at 22°C. Strains of serotypes O:8 and O:3 were recovered when two cells with 10⁷ cells of ten other gram-negative bacteria were added to 10 g of beef stew following preenrichment in trypticase soy broth at 2°C for 7 d and selective enrichment in bile oxalate-sorbose broth at 22°C for 3 to 5 d. Strains of serotype O:5,27 were more difficult to recover even with longer enrichment times. These studies indicated that the most comprehensive enrichment system for recovery of Y. enterocolitica from foods is preenrichment in trypticase soy broth at 22°C for 1 d and 2 to 4°C for 4 to 7 d followed by selective enrichment in bile-oxalate-sorbose broth at 22°C for 3 to 5 d and isolation on cefsulodin-irgasan-novobiocin agar.