Search Results (12)

Autochthonous starter and adjunct cultures have gained increasing practical research interest for use in traditional cheese-making in recent years. This study evaluated the performance of a native starter comprising Streptococcus thermophilus ST1 and two wild Lactococcus lactis strains, including the M78 nisin A-producer, during the semi-industrial production of Kefalotyri cooked hard cheese from thermized sheep milk (TM; 65 °C, 30 s) in the absence (C-cheese) or presence of the native adjunct strains Lactiplantibacillus plantarum H25 and Leuconostoc mesenteroides KFM7 + KFM9 (N-cheese). The growth of the native starter was optimal in all three cheese trials within the first 24 h of processing, but afterward, ST1 failed to exceed 8 log CFU/g in favor of total mesophilic LAB, comprising both L. lactis starter strains, and the prevalent (ca. 8.5 log CFU/g) H25 adjunct in the ripening N-cheeses. Instead, in the C-cheeses, indigenous non-Enterococcus NSLAB survivors from TM prevailed, whereas enterococci failed to increase above 6 log CFU/g in all cheeses. Although the mature (90-day-old) N-cheeses presented no statistically significant differences regarding the pH value, gross composition, and hydrophobic (HO)/hydrophilic (HI) peptide ratios from the mature C-cheeses, they had lower total LAB counts and contained less residual lactose, more acetate, and an overall less diversified volatilome (VOC) profile. The most abundant VOCs in both cheeses were acetone, butyric acid, methyl butyrate, ethyl ether, and ethanol. All mature cheeses were safe and graded of ‘excellent quality’ (i.e., moisture < 35%; fat-in-dry-matter > 47%). Full article

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

In the Alpine region of Austria, Italy, and Switzerland, transhumance is widespread and the production of local traditional dairy products during summer is important. Raw milk cheeses are produced according to traditional recipes, using hurdles as a technique to guarantee food safety. In the present study, we aim to provide an overview of S. aureus and its enterotoxins in raw milk cheeses, identify the key parameters responsible for the enterotoxin production, and identify ways to improve food safety. The results demonstrate that safe artisanal raw milk cheese production is achievable under elementary conditions by applying effective hurdles, including high scalding temperatures or thermization, quality starter cultures, and robust milk quality management. The hurdle index (HI), which we introduce in this paper, is a promising tool for assessing and improving safety in raw milk cheese production. 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 investigated the production of rennet gels from β-casein-depleted retentates obtained through cold microfiltration (MF) of skim milk (SM) that was treated beforehand to ensure microbial safety. The treatments included thermization (65 °C, 20 s), pasteurization (72 °C, 15 s), and microfiltration (50 °C; 1.4 μm pore size). The reduction in β-casein content was 0.98, 0.51 and 0.90%, respectively. All treatments resulted in the partial aggregation of serum proteins, which were slightly concentrated in the retentates obtained post cold MF process. This aggregation, along with concentration effect, likely inhibited β-casein dissociation from casein micelles and permeation, particularly in pasteurized milk. Renneting and coagulation properties of the retentates were comparable to those of the respective SM samples, with no significant differences in syneresis, water-holding capacity, or protein hydration. Notably, the retentate from thermized SM, which showed the best performance with the highest β-casein reduction (0.98%), demonstrated shorter coagulation time compared to retentate from pasteurized milk or the corresponding unfiltered SM. Textural analysis revealed greater firmness, cohesiveness, and viscosity of retentate-based rennet gels compared to gels made from unfiltered SM, attributed to protein concentration during cold MF. Overall, this study successfully produced rennet gels from cold MF retentates without compromising their physicochemical properties. Full article

Reverse osmosis (RO) is known for the economic dewatering of dairy streams without any change in phase. At the household level, surplus milk is fermented and churned to obtain butter, which is subsequently heated to obtain clarified milk fat (ghee). The production of 1 kg ghee generates 15–20 kg sour buttermilk (SBM) as a by-product that is mostly drained. This causes a loss of milk solids and environmental pollution. The processing, preservation and valorization of SBM are quite challenging because of its low total solids (TS) and pH, poor heat stability and limited shelf life. This investigation aimed to transform SBM into a novel dried dairy ingredient. SBM was thermized, filtered, defatted and concentrated at 35 ± 1 °C, employing RO up to 3.62× (12.86%). The RO concentrate was subsequently converted into sour buttermilk powder (SBMP) by employing spray drying. SBMP was further characterized for its physicochemical, reconstitution and functional properties; rheological and morphological characteristics; and amino acid and fatty acid profiling, along with FTIR and XRD spectra. SBMP was “instant soluble-3 s” and exhibited excellent emulsion stability (80.70%), water binding capacity (4.34 g/g of protein), flowability (28.36°) and antioxidant properties. In nutshell, a process was developed for the valorization of sour buttermilk to a novel dairy ingredient by employing reverse osmosis and a spray-drying process. Full article

The use of raw milk is compulsory in the manufacturing process of most of the European protected designation of origin (PDO) cheeses but, for ovine products, it is often responsible for faulty productions. Since pasteurization is hardly compatible with the PDO concept, a milder treatment (thermization) is allowed in some cases. An investigation was undertaken to assess the effect of thermization on the overall quality of Canestrato Pugliese, a PDO ovine hard cheese of Southern Italy that can be manufactured exclusively from raw milk. Three types of cheese were produced using raw, mild-thermized and high-thermized milk inoculated with a thermophilic commercial starter. The results demonstrated that the heat treatment did not cause remarkable differences in the gross composition, but the microbiological profiles had some differences despite the use of the selected starter. The raw milk cheese contained higher levels (0.5–1 log units) of mesophilic lactobacilli, total viables, total coliforms and enterococci with respect to the thermized counterparts, with the high-thermized cheese showing the lowest levels; these microbiological differences fitted well with the higher content and the different High Performance Liquid Chromatography (HPLC) pattern of soluble nitrogen. The sensory analysis revealed that the thermized cheeses lost some typical sensory characteristics, probably as a consequence of the reduced indigenous microbiota populations. It was concluded that milk thermization could be applied to Canestrato Pugliese manufacturing only together with the development and use of an autochthonous starter. Full article

The oxidative stability of milk and dairy products is a very interesting topic for the dairy industry due to the growing demand for foods containing bioactive compounds with positive health effects. The aim was to evaluate the oxidative stability of milk intended for cheese production. The effect of storage time, heat pre-treatment, and milk pasteurization temperature on the characteristics of milk and cheese was investigated. The cheese samples were produced with pasteurized milk at both 72 and 77 °C for a time of 15 s using three types of milk: raw fresh milk processed within 48 h of milking, raw stored milk processed within 96 h, and thermized milk that was heat-treated upon arrival at the dairy and processed within 96 h of milking. In total, three repetitions were carried out for each type of milk and pasteurization. Samples of milk before and after pasteurization and cheese at 14 days of storage were analyzed. Antioxidant activity decreased from starting milk to milk after pasteurization to final cheese. The longer storage time of the milk had significant effects on the antioxidant stability of the cheese (64.95 vs. 59.05% of antioxidant activity). Thermization of the milk further reduced the stability of the cheese (54.05% of antioxidant activity). The greater antioxidant stability of fresh milk and cheeses produced with fresh milk is the first result that encourages the production of cheese from a milk that best preserves its original characteristics. 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

Milk pre-processing steps-storage at 4 °C (with durations of 48, 72 or 96 h) and methods for microbiological stabilization of milk (1.4 μm microfiltration, thermization, thermization + bactofugation, pasteurization) are performed industrially before 0.1 µm-microfiltration (MF) of skimmed milk to ensure the microbiological quality of final fractions. The objective of this study was to better understand the influence of these pre-processing steps and their cumulative effects on MF performances (i.e., transmembrane pressure, and transmission and recovery of serum proteins (SP) in the permeate). Results showed that heat treatment of skimmed milk decreased ceramic MF performances, especially after a long 4 °C storage duration (96 h) of raw milk: when milk was heat treated by pasteurization after 96 h of storage at 4 °C, the transmembrane pressure increased by 25% over a MF run of 330 min with a permeation flux of 75 L·h⁻¹·m⁻² and a volume reduction ratio of 3.0. After 48 h of storage at 4 °C, all other operating conditions being similar, the transmembrane pressure increased by only 6%. When milk was 1.4 µm microfiltered, the transmembrane pressure also increased by only 6%, regardless of the duration of 4 °C storage. The choice of microbiological stabilization method also influenced SP transmission and recovery: the higher the initial heat treatment of milk, the lower the transmission of SP and the lower their recovery in permeate. Moreover, the decline of SP transmission was all the higher that 4 °C storage of raw milk was long. These results were explained by MF membrane fouling, which depends on the load of microorganisms in the skimmed milks to be microfiltered as well as the rate of SP denaturation and/or aggregation resulting from pre-processing steps. Full article