Abstract
The capacity of growth, survival, and adaptive responses of an artificial contamination of a three-strain L. monocytogenes cocktail in factory-scale thermized (65 °C, 30 s) Graviera cheese milk (TGCM) was evaluated. Bulk TGCM samples for inoculation were sequentially taken from the cheese making vat before process initiation (CN-LM) and after addition of a commercial starter culture (CSC), the CSC plus the nisin A-producing (NisA+) costarter strain Lactococcus lactis subsp. cremoris M78 (CSC + M78), and all ingredients with the rennet last (CSC + M78-RT). Additional treatments included Listeria-inoculated TGCM samples coinoculated with the NisA+ costarter strain M78 in the absence of the CSC or with the CSC in previously sterilized TGCM to inactivate the background microbiota (CSC-SM). All cultures were incubated at 37 to 42 °C for 6 h, followed by additional 66 h at 22 °C, and 48 h at 12 °C after addition of 2% edible salt. L. monocytogenes failed to grow and declined in all CSC-inoculated treatments after 24 h. In contrast, the pathogen increased by 3.34 and 1.46 log units in the CN-LM and the CSC-SM treatments, respectively, indicating that the background microbiota or the CSC alone failed to suppress it, but they did so synergistically. Supplementation of the CSC with the NisA+ strain M78 did not deliver additional antilisterial effects, because the CSC Streptococcus thermophilus reduced the growth prevalence rates and counteracted the in situ NisA+ activity of the costarter. In the absence of the CSC, however, strain M78 predominated and caused the strongest in situ nisin-A mediated effects, which resulted in the highest listerial inactivation rates after 24 to 72 h at 22 °C. In all TGCM treatments, however, L. monocytogenes displayed a "tailing" survival (1.63 to 1.96 log CFU/mL), confirming that this pathogen is exceptionally tolerant to cheese-related stresses, and thus, can't be easily eliminated.