Aim of the present study was the evaluation of the growth potential of Listeria spp. inoculated in the typical North Italian dry fermented sausage “Bergamasco” salami during its production. As it was necessary to carry out the challenge test in the production line of the industry, according to the guidelines of the European Reference Laboratory for Listeria monocytogenes, a non-pathogenic “surrogate” microorganism was used: for the inoculum, two strains of Listeria innocua (1 ATCC, 1 strain isolated from a similar substrate) were used. The inoculation of the samples occurred during grinding and mixing of the sausage mass, before the filling. To avoid cross-contamination, the control samples were produced before the contaminated ones. After the dripping, salamis were subjected to the normal production process (drying and maturation in five steps at specific temperatures and humidity rates). The inoculated products were subjected to the enumeration of Listeria spp. at T0 (day of inoculation) and at T4 (post-drying), and every 10 days during curing (T10, T20, T30, T40, T50, T60, T70, T80 and T90), as this salami is generally sold as whole piece with varying levels of curing (from T20 to T90). Since the product may be cut in half and vacuumpacked, at each of the times starting from T20, half salami was vacuum-packed and stored for 30 days at 12°C, at the end of the which Listeria spp. enumeration was performed again. At all times and for each type of samples of each of the three batches, the enumeration of the natural microflora (Total Viable Count, lactic acid bacteria, Pseudomonas spp., Enterobacteriaceae) and the determination of water activity and pH were performed on control samples. The product was characterized by a high concentration of microflora (8-8.5 Log UFC/g), consisting mainly of lactic acid bacteria, added to the mixture at the beginning of the production process. The pH showed a decrease over time, expected for this type of products, due to the development of lactic acid bacteria (final pH: 5.42-5.55). The water activity reached values able to inhibit the development of Listeria spp. (final aw: 0.826-0.863). Listeria counts in the tested batches of “Bergamasco” salami showed the absence of significant growth in the product with a reduction of loads if compared to T0, between -0.59 and -1.04 Log CFU/g. Even in the samples subjected to vacuum packaging and storage at 12°C, the absence of significant increase of lactic acid bacteria in the product was highlighted with further decrease of bacterial loads (-0.70/-0.79 Log CFU/g if compared to T20). Considering the worst case scenario (thus the batch with the highest growth potential), in the products stored in the curing room at 14-16°C, at humidity of 80% and in the samples stored at 12°C and vacuum packaged, the threshold indicated by the EURL Lm guidelines (+0.5 Log CFU/g) for the growth of Listeria spp. was not reached, allowing to classify “Bergamasco” salami in the category 1.3 of the EC Reg. 2073/2005 as “Ready-to-eat food unable to support the growth of Listeria monocytogenes”.
Prediction of population behavior of Listeria monocytogenes in food using machine learning and a microbial growth and survival database