Frozen cultures were activated in three successive passes, first inoculating 100 μl in 9 ml of TSB (for E. coli O157:H7) and
TSB + 0.6% yeast extract (for L. monocytogenes) and incubated at 37 °C for 18 to 24 h. On the second day, 1 ml of each bacterial culture was inoculated into 9 ml of respective broth and incubated at 37 °C for 18 to 24 h. On the third day, 1 ml of each bacterial culture was inoculated into 99 ml TSB + glucose (for E. coli O157:H7) and TSB + 0.6% yeast extract (for L. monocytogenes) and incubated at 37 °C for 18 to 24 h. On the fourth day, the bacterial cocktail was prepared, and the experiment was performed. For the cocktail preparation, 100 ml of each of the three strains was mixed into an empty sterile 500 ml bottle through a sterilized glass funnel. The average initial inoculumlevel of E. coli O157:H7 in all experiments was 8.8 log CFU/ml, while this figure was 8.9 log CFU/ml for L. monocytogenes.
Frozen cultures were activated in three successive passes, first inoculating 100 μl in 9 ml of TSB (for E. coli O157:H7) andTSB + 0.6% yeast extract (for L. monocytogenes) and incubated at 37 °C for 18 to 24 h. On the second day, 1 ml of each bacterial culture was inoculated into 9 ml of respective broth and incubated at 37 °C for 18 to 24 h. On the third day, 1 ml of each bacterial culture was inoculated into 99 ml TSB + glucose (for E. coli O157:H7) and TSB + 0.6% yeast extract (for L. monocytogenes) and incubated at 37 °C for 18 to 24 h. On the fourth day, the bacterial cocktail was prepared, and the experiment was performed. For the cocktail preparation, 100 ml of each of the three strains was mixed into an empty sterile 500 ml bottle through a sterilized glass funnel. The average initial inoculumlevel of E. coli O157:H7 in all experiments was 8.8 log CFU/ml, while this figure was 8.9 log CFU/ml for L. monocytogenes.
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