Enterococcus faecium MMRA is an enterocin A producer isolated from 'Rayeb', a Tunisian fermented milk drink. In this work, safety aspects and its behaviour in raw milk were investigated to assess its suitability as a protective adjunct culture. E. faecium MMRA showed interesting features such as the absence of several virulence traits, susceptibility to vancomycin and other clinically relevant antibiotics, and lack of haemolytic activity. To evaluate its performance as an adjunct culture for Rayeb, changes in the overall composition of control (non-inoculated) and experimental Rayeb (inoculated with 1%, v/v, E. faecium MMRA) were determined throughout duplicate fermentations of raw milk using microbiological, chemical, HPLC and headspace GC-MS analyses. E. faecium MMRA could multiply in raw milk and produced enterocin A. Interestingly, a higher content of volatile compounds including ethanol, diacetyl and 2-propanol was observed in the presence of this bacteriocin producer. Furthermore, this strain was capable of inhibiting the growth of Listeria monocytogenes CECT 4032 in pasteurised milk, although total killing was not achieved. Further experiments confirmed the development of resistant variants to enterocin A. On similar challenge assays, L. monocytogenes CECT 5672 growth was halted by the presence of the enterocin producer, but viability was only slightly reduced during cold storage. According to our results, E. faecium MMRA meets the criteria for an autochthonous protective adjunct culture to enhance both the hygienic and the sensory attributes of Rayeb.
