Abstract
A novel, simple, strictly anaerobic method was developed, which dramatically enhanced the recovery of severely heat-injured Listeria monocytogenes from pasteurized milk, compared to current aerobic methods. Cells of L. monocytogenes were grown in trypticase soy broth supplemented with 0.6% yeast extract (TSB-YE) at 43°C for 18 h and then severely heat-injured at 62.8°C for 5 min in raw milk in sealed, totally submerged thermal-death-time tubes. Recovery in primary enrichment media was enhanced in all cases by purging the headspace with N2 gas. The following treatments (in order of effectiveness) significantly improved recovery compared to conventional aerobic controls: addition of filter-sterilized cysteine + N2 purging > pre-reduced Hungate media + N2 purging = Oxyrase® + lactate + N2 purging > filter-sterilized cysteine - N2 purging > Oxyrase® - lactate + N2 purging. Addition of filter-sterilized cysteine to a final concentration of 0.5 g/l with subsequent N2 purging increased recovery from 0% to 60% wi...

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