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Frozen stocks of the bacterial isolates were obtained from the Cornell University Food Safety Laboratory (CU-FSL) isolate collection. Cultures were streaked from −80¡ãC frozen stock onto brain heart infusion (Difco, Sparks, MD) agar and incubated for 18 h at 32¡ãC. A colony from each strain was used to inoculate 5 mL of sterile brain heart infusion broth in 16-mm glass test tubes, which were then incubated for 48 h with aeration (shaking at 220 rpm) at 32¡ãC. A 1-mL aliquot of the culture was then centrifuged at 14,000 ¡Á gand the supernatant was removed. The resulting cell pellet was resuspended in 1 mL of phosphate dilution buffer (Weber Scientific, Trenton, NJ), diluted to 1,000 to 10,000 cfu/mL, and inoculated into commercially purchased UHT pasteurized milk (1% milk; Organic Valley, La Farge, WI) for an approximate inoculum of 2 to 3 log cfu/mL. Inocula were enumerated by spiral plating the appropriate dilution on SPC agar (EMD, Gibbstown, NJ), followed by incubation at 32¡ãC for 24 to 48 h. Inoculated UHT milk was incubated at 6¡ãC for 48 h to simulate refrigerated storage conditions. After a 48-h incubation at 6¡ãC, appropriate dilutions of inoculated UHT samples prepared in phosphate dilution buffer were plated in duplicate on (1) SPC pour plates (SPCP); (2) SPC plates inoculated by spiral plating (SPCS); (3) CVTA [plate count agar (EMD, Gibbstown, NJ) with 2,3,5-triphenyltetrazolium chloride (Sigma-Aldrich, St. Louis, MO)] pour plates (CVTAP); (4) CVTA plates inoculated by spiral plating (CVTAS); (5) VRBA plates inoculated by pour plating (Difco, Sparks, MD); (6) VRBA with an overlay of approximately 5 mL of VRBA agar (VRBAO); (7) CPETRI (3M); or (8) MAC (Difco) inoculated by spiral plating. All media were incubated at 32¡ãC for 48 h, with enumeration after 24 and 48 h. Three biological replications of the experiment were completed.


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