No abstract

The aim of this study was to compare the microbial communities of different cheeses where Listeria monocytogenes either grew or did not grow. For this purpose, (i) isolates from the most inhibitory cheese ecosystem were identified and their ability to produce anti-Listeria substances was determined, (ii) bacterial communities of cheeses with and without L. monocytogenes growth were compared using the Single Strand Conformation Polymorphism method. The study showed SSCP to be an effective tool for differentiating between the bacterial communities of different cheeses manufactured with the same technology. All the cheeses with the lowest L. monocytogenes counts on day 8 were distinguished by the dominance in their SSCP profiles, after amplification of the V2 region of the 16S rRNA gene, of 3 peaks whose nucleotide sequences comigrated with Enterococcus faecium and Enterococcus saccharominimus, Chryseobacterium sp and Corynebacterium flavescens, Lactococcus garvieae and Lactococcus lactis respectively. However, no anti-Listeria compounds were produced under our experimental conditions. These six bacterial species were inoculated, separately or together, into pasteurised milk and their anti-listerial activity in cheese was evaluated. The area of inhibition between the control and trial curves confirmed that L. monocytogenes is inhibited by E. saccharominimus, C. flavescens, L. lactis, L. garvieae and the mixture of all six bacterial strains. Further studies should be performed to determine the metabolites involved in L. monocytogenes inhibition.

A complex microbial consortium derived from raw milk and composed of populations classified in 4 groups (lactic acid bacteria (A), Gram positive catalase positive bacteria (B), Gram negative bacteria (C) and yeasts (D)) can contribute to the inhibition of Listeria monocytogenes in the core of an uncooked pressed cheese. To identify what groups may be involved in the inhibition, the consortium was simplified by successively omitting one group at a time. Pasteurized milk was inoculated with these more or less complex consortia and their effects on L. monocytogenes count, pH, acids and volatile compounds in the core of uncooked pressed cheese were evaluated. The growth of L. monocytogenes was the highest in cheeses prepared with pasteurized milk and only St. thermophilus. Inhibition in other cheeses was expressed by comparison with growth in these ones. All the consortia containing both lactic acid bacteria (group A) and Gram positive catalase positive bacteria (group B)--ABCD, ABD, ABC, AB--were more inhibitory than those containing lactic acid bacteria on its own (A) or associated only with yeasts (AD) or/and Gram negative (ADC). Consortia without lactic acid bacteria were weakly inhibitory or had no effect. Gram positive catalase positive bacteria alone were not inhibitory although most of the species became established in the cheeses. The Lactobacillus population (Lb. casei, Lb. plantarum, Lb. curvatus and Lb. farciminis) was predominant in cheeses (9 log CFU/g) with a higher count than Leuconostoc (7 log CFU/g) and Enterococcus (7 log CFU/g). Lactobacillus counts were negatively correlated with those of L. monocytogenes (r=-0.84 at 18 days) and with the level of D-lactic acid. There was no correlation between L. monocytogenes and Leuconostoc or Enterococcus counts. Complex consortium ABCD and AB not only had a stronger inhibitory power in cheeses than consortium AD, they were also associated with the highest levels of L-lactic and acetic acids. All cheeses inoculated with lactic acid bacteria differed from those without by higher levels of ethyl formiate, pentane and alcohols (2-butanol, 2-pentanol), and lower levels of ketones (2-hexanone, 2,3-butanedione) and aldehydes (2-methyl-butanal). Levels of 2-methyl-butanal, 2-butanol and 2-pentanol were higher in ABCD and AB cheeses than in AD cheeses. Beside their contribution to the inhibition, their effect on cheese flavour must be evaluated.