*2.2. Microbiological Methodology*

The counts of total mesophilic bacteria, fungi (yeasts and molds), total coliforms, *Escherichia coli*, Enterobacteria, lactic bacteria, *Staphylococcus aureus*, *Salmonella* spp., *Listeria monocytogenes*, *Bacillus cereus*, and sulphite-reducing clostrides were performed. All media were supplied by ThermoFisher Scientific, Oxoid Ltd., Basingstoke, UK. The samples were homogenized with PBS (pH = 7.4) by mixing vigorously. Then serial dilutions in PBS (pH = 7.4) were performed, and the samples were spread aseptically over the media plates. For mesophilic counts, samples were spread over plate count agar (PCA) and incubated at 30 ◦C for 72 h. Malt extract agar with 10% lactic acid (MEA) was utilized to verify the counts of yeasts and molds by incubating at room temperature for 3 days. Lactic acid microorganisms and Enterobacteriacee were investigated using De Man, Rogosa, and Sharpe agar and Violet Red Bile Glucose Agar (MRSA and VRBG) using an incubation period of 72 h at 30 ◦C and 48 h at 37 ◦C, respectively. The determination of *B. cereus* was carried out by using *Bacillus Cereus* Agar Base (PEMBA) after an incubation for 24 h at 37 ◦C. Coliforms and *E. coli* were analyzed using the standard membrane filter technique using Chromogenic Coliform Agar (CCA) and Tryptone Bile X-Gluc agar (TBX) using an incubation period of 24 h at 37 ◦C and of 18/24 h at 44 ◦C. *S. aureus* contamination was detected using the standard membrane filter technique using Baird–Parker Agar (BPA), at 37 ◦C for 48 h. The standard membrane filter technique using Sulfite Polymyxin Sulfadizine agar (SPS) was used for the determination of sulphite-reducing clostrides after an incubation at 37 ◦C for 48 h. The determination of *L. monocytogenes* was carried out according to ISO standard 11290. Briefly, after a two-stage enrichment process, the first

in half Fraser broth for 24 h and then in Fraser broth, the enriched broths were plated on Oxford and BD PALCAM *Lysteria* agars.

The presence of *Salmonella* spp. was investigated according to ISO standard 6579, which includes: pre-enrichment in a non-selective liquid medium (buffered pepton water), followed by selective enrichment (Rappaport–Vessiliadis Soy Broth; ThermoFisher Scientific, Oxoid Ltd., Basingstoke, UK), and then isolation on a selective medium (Hektoen; ThermoFisher Scientific, Oxoid Ltd., Basingstoke, UK). All microbial analyses were carried out in triplicate.
