Search Results (5)

Autochthonous enterococci surviving mild thermization of raw milk (RM) before traditional Greek cheese processing may simultaneously comprise safe and virulent thermoduric strains with multiple antibiotic resistances (ARs). Therefore, this study biotyped and then compared the ARs of 60 Enterococcus isolates from two antilisterial sheep milks of native Epirus breeds before (RM) and after (TM) thermization at 65 °C for 30 s; the RM isolates were previously genotyped and evaluated for primary safety traits, namely, hemolytic activity, vanA/vanB, cytolysin, and virulence genes, by molecular methods. Biochemically typical and atypical strains of Enterococcus faecium (six biotypes), E. durans (five biotypes), E. faecalis (two biotypes), and E. hirae (one biotype), which were subdominant to other LAB species in RM (19 isolates), prevailed in TM (41 isolates). E. faecium biotypes 1A, 1D, and 1H included multiple-Ent+ (entA/entB/entP or entA/entB) strains with strong antilisterial CFS activity, whereas E. faecium 1X (entA), E. durans 2A, 2B, 2C, and 2X (entA/entP or entP), E. faecalis 3B, and E. hirae 4A (entA) biotypes displayed direct in vitro antilisterial activity only. Biotypes 1D, 1X, and 2A were selected in TM. All E. faecium/durans isolates were susceptible to vancomycin, but the m-Ent + E. faecium biotype 1A and 1D strains were resistant to penicillin, erythromycin, ciprofloxacin, and ampicillin. In contrast, all biotype 1X isolates were susceptible to all antibiotics tested. All E. faecalis and most E. durans isolates were resistant to penicillin but susceptible to erythromycin and ciprofloxacin. Biotype 2X isolates and one virulent (ace; gelE) E. faecalis isolate from RM were tetracycline-resistant. A sporadic RM isolate of E. hirae that was resistant to penicillin and vancomycin was not retrieved from the counterpart TM, but the inclusion of three vancomycin-resistant isolates from TM in the primary biotype 3B of E. faecalis was a cause for concern. In conclusion, based on the results, antibiotic-susceptible representatives of all strain biotypes of the E. faecium/durans group, as well as antagonistic m-Ent+ E. faecium strains from sheep milk that were susceptible to vancomycin and ampicillin and lacking virulence genes, can be included in safe complex natural starters to be developed for onsite use in traditional Greek hard cheese technologies. Full article

The production of Fiore Sardo cheese is regulated by the specification of the Protected Designation of Origin (PDO), which aims to guarantee the specific area of production, the know-how of local producers, and the specific use of raw milk from Sarda sheep. The thermization of milk is a sub-pasteurization process that is commonly used in cheese-making to lower the bacterial load and increase the shelf life of the product; it is therefore a cause of non-compliance with the PDO specification of Fiore Sardo cheese, allowing producers to gain practical and economic advantages. In this work, NIR spectroscopy coupled with multivariate discriminant analysis was used to identify the thermal treatment of milk in Fiore Sardo cheese samples. Cheeses were produced using raw milk (38 °C), low-thermized milk (57 °C for 30 s), and high-thermized milk (68 °C for 30 s). The NIR spectra of the cheeses were used to build discriminant models for individuating the thermal treatment of the processed milk. The obtained discriminant models were able to correctly classify about 90% of the Fiore Sardo cheese samples. This method could be suitable as a screening technique to authenticate Fiore Sardo PDO cheese. Full article

Thermization is a sub-pasteurization heat treatment widely applied in traditional Greek hard cheese technologies. In this study, five bulk milk batches from two native Epirus sheep breeds were analyzed microbiologically before (raw milk; RM) and after thermization at 65 °C for 30 s (TM) followed by characterization of 125 presumptive LAB isolates from each of the counterpart RM and TM samples. Psychrotrophic Pseudomonas-like spoilage bacteria and mesophilic LAB, primarily of the genera Leuconostoc (48.4%) and Lactococcus/Streptococcus (32.8%), co-dominated in RM at mean levels 5.7–6.3 log CFU/mL, whereas thermophilic LAB, Enterococcus, Staphylococcus, coliforms, and yeasts were subdominant at mean levels 4.1 to 5.2 log CFU/mL. Coagulase-positive staphylococci were abundant (3–4 log CFU/mL) in all RM batches. Listeria monocytogenes was found in one batch. Both pathogens were diminished by thermization, which reduced all non-LAB contaminants below 100 CFU/mL. Enterococci (68.6%) were highly selected in all TM batches, followed by thermophilic streptococci (8.6%). Only 7.4% of the total RM microbiota survived in the resultant five TM batches. Leuconostoc showed the lowest (1.3%) survival. Thus, thermization improved the quality and safety of raw sheep milk, but reduced mesophilic LAB by ca. 2 log units in favor of enterococci in TM. Full article

This study was conducted to assess, for the first time, the survival of the pathogenic bacteria Listeria monocytogenes, Salmonella spp., Escherichia coli O157:H7, and Staphylococcus aureus during the ripening of protected designation of origin (PDO) Pecorino Romano cheese. A total of twenty-four cheese-making trials (twelve from raw milk and twelve from thermized milk) were performed under the protocol specified by PDO requirements. Sheep cheese milk was first inoculated before processing with approximately 10⁶ colony-forming unit (CFU) mL⁻¹ of each considered pathogen and the experiment was repeated six times for each selected pathogen. Cheese composition and pathogens count were then evaluated in inoculated raw milk, thermized milk, and cheese after 1, 90, and 150 days of ripening. pH, moisture, water activity, and salt content of cheese were within the range of the commercial PDO Pecorino Romano cheese. All the cheeses made from raw and thermized milk were microbiologically safe after 90 days and 1 day from their production, respectively. In conclusion, when Pecorino Romano cheese is produced under PDO specifications, from raw or thermized milk, a combination of factors including the speed and extent of curd acidification in the first phase of the production, together with an intense salting and a long ripening time, preclude the possibility of growth and survival of L. monocytogenes, Salmonella spp., and E. coli O157:H7. Only S. aureus can be still detectable at such low levels that it does not pose a risk to consumers. Full article

The activity of α-l-fucosidase (FSC) has been measured for the first time in Fiore Sardo PDO (Protected Designation of Origin) raw milk cheese. To do this, a RP-HPLC method has been developed, validated and tested on a reliable sampling of cheese experimentally produced in laboratory batches. Three experimental factors have been considered in this work: the thermal treatment undergone by the milk, the lactation period and the ripening time of cheese. Results obtained have evidenced: (i) a meaningful reduction in the activity of FSC from cheeses produced using raw milk to those obtained by thermized milk; (ii) an increase in the activity of FSC during the first months of lactation period (from December to February), followed by a substantial constancy in the central and final months of lactation (from February to May); (iii) the enzyme activity is independent of the ripening time. This method might be useful in revealing frauds related to the use of mild thermal treatments of the milk when this is not allowed as for Fiore Sardo PDO cheese but also for several PDO cheeses for which the specifications establish that raw milk only must be used for their production. Full article