Effect of Scalding Temperature on Growth of Staphylococcus aureus and Formation of Staphylococcal Enterotoxin during the Production of Alpine Cheese in a Laboratory Cheesemaking Model.

ABSTRACT: To reduce the number of cheese with potential Staphylococcus aureus contamination reaching consumers, European legislation has stipulated that all cheese must be tested for coagulase-positive staphylococci (CPS) at the point in production when numbers are expected to be highest. When CPS counts exceed 105 CFU/mL, staphylococcal enterotoxin (SE) tests must be conducted. When SE tests are positive, the cheese must be destroyed. Manufacturers of Swiss Alpine cheese are exempt from this legislation because SE formation in hard cheese is expected to be very unlikely because of the high scalding temperatures used for cheeses during production, which inactive CPS in the curd. However, this assumption has not been scientifically tested. A laboratory-scale cheese production experiment was performed in which the conditions corresponded to certain limitations in practical cheesemaking conditions such as temperature and time exposure as for production of Gruyere or Tete de Moine Swiss type cheeses. Raw milk aliquots (200 mL) were inoculated with five strains of CPS, and scalding temperatures of 46 to 56°C were used during cheese production. The temperatures applied after the curd was pressed were meant to reproduce the temperature curve in the peripheral zone of a real cheese wheel. Contrary to expectations, SE formation occurred and differed according to the scalding temperature (52 to 56°C). The differences in SE formation were more associated with strain type rather than temperature. These results indicate that the mechanisms of SE formation in cheese require further study.

Development of a quantitative PCR assay for rapid detection of Lactobacillus plantarum and Lactobacillus fermentum in cocoa bean fermentation.

To monitor dominant species of lactic acid bacteria during cocoa bean fermentation, i.e. Lactobacillus plantarum and Lactobacillus fermentum, a fast and reliable culture-independent qPCR assay was developed. A modified DNA isolation procedure using a commercial kit followed by two species-specific qPCR assays resulted in 100% sensitivity for L. plantarum and L. fermentum. Kruskal-Wallis and post-hoc analyses of data obtained from experiments with cocoa beans that were artificially spiked with decimal concentrations of L. plantarum and L. fermentum strains allowed the calculation of a regression line suitable for the estimation of both species with a detection limit of 3 to 4 Log cells/g cocoa beans. This process was successfully tested for efficacy through the analyses of samples from laboratory-scale cocoa bean fermentations with both the qPCR assay and a culture-dependent method which resulted in comparable results.