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Background: Lactic Acid Bacteria (LAB) are an important group of microorganisms responsible for the fermentation dairy products. This study was done to identify the dominant lactic acid bacteria in naturally fermented milks from Messinese goat’s breed. Methods: Eighteen individual raw milk samples collected from Messinese goat’s breed were acidified at pH 5.20 and left to spontaneously ferment at 37 °C for 4 days. All samples were analyzed for rod- and coccus-shaped LAB. Also, all presumptive LAB were isolated and differentiated according to their phenotypic properties and genetic polymorphisms and then identified by sequencing the 16S rRNA gene. Data were statistically analyzed using SAS 9.2 software. Results: Levels of coccus- and rod-shaped LAB were in the range of 1.78-7.10 log and 1.00-7.09 log colony forming unit/ml, respectively. The microbiological counts on the two different growth media were significantly (p

Fresh ovine “primosale” cheese was processed with the addition of grape pomace powder (GPP). Cheese making was performed using pasteurized ewes’ milk and four selected Lactococcus lactis strains (Mise36, Mise94, Mise169 and Mise190) inoculated individually. For each strain the control cheese (CCP) was not added with GPP, while the experimental cheese (ECP) was enriched with 1% (w/w) GPP. GPP did not influence the starter development that reached levels of 109 CFU/g in all final cheeses. The comparison of the bacterial isolates by randomly amplified polymorphic DNA (RAPD)-PCR showed the dominance of the added strains over indigenous milk bacteria resistant to pasteurization. GPP addition reduced fat content and determined an increase of protein and of secondary lipid oxidation. Sensory tests indicated that cheeses CCP94 and ECP94, produced with the strain Mise94, reached the best appreciation scores. Following in vitro simulated human digestion, bioaccessible fraction of ECP94 showed antioxidant capacity, evaluated as radical scavenging activity and inhibition of membrane lipid oxidation, significantly higher than that from CCP94, with promising increase in functional properties. Thus, the main hypothesis was accepted since the functional aspects of the final cheeses improved, confirming that GPP is relevant for sustainable nutrition by using winemaking by-products.

Red grape Nero d’Avola cultivar grape pomace powder (GPP) was applied during fresh ovine cheese production in order to increase polyphenol content. Before cheeses were produced, the bacteria of a freeze-dried commercial starter culture were isolated and tested in vitro against GPP. Two dominant strains, both resistant to GPP, were identified. Thestarter culture was inoculated in pasteurized ewe’s milk and the curd was divided into two bulks, one added with 1% (w/w) GPP and another one GPP-free. GPP did not influence the starter culture development, since lactic acid bacteria (LAB) counts were 109 CFU/g in both cheeses at 30 d. To exclude the interference of indigenous LAB, the pasteurized milk was analyzed, and several colonies of presumptive LAB were isolated, purified and typed. Four strains were allotted into Enterococcus and Lacticaseibacillus genera. The direct comparison of the polymorphic profiles of cheese bacteria evidenced the dominance of the starter culture over milk LAB. The addition of GPP increased cheese total phenolic compounds by 0.42 g GAE/kg. Sensory evaluation indicated that GPP-enriched cheese was well appreciated by the judges, providing evidence that GPP is a suitable substrate to increase the availability of total phenolic content in fresh ovine cheese.

The present work was developed to select lactic acid bacteria (LAB) to be used as starter cultures in functional cheese production. The indigenous milk LAB populations were isolated from fermented raw ewes’ milks (four bulks) added with 0.5 mg/mL of nine polyphenols commonly found in winery by-products. After 48 h of fermentation, all milks were characterized by an increase of LAB levels of about 3–4 Log cycles. All different colonies were purified and characterized for the main physiological and biochemical traits and then differentiated genetically at strain level and identified. Ten species belonging to the LAB genera Lactobacillus, Streptococcus, Enterococcus, Leuconostoc and Lactococcus were identified. Only Lactococcus lactis and Leuconostoc mesenteroides strains were evaluated for the technological traits including acidification and autolytic kinetics, diacetyl formation, exopolysaccharide production and generation of antimicrobial compounds. A total of four strains (Mise36, Mise94 Mise169 and Mise190) belonging to Lc. lactis displayed potential for production of cheeses containing grape polyphenols.

The present work was carried out to evaluate the microbiological and physicochemical composition of salamis produced with the meat of beef, horse, wild boar and pork. Salami productions occurred under controlled laboratory conditions to exclude butchery environmental contaminations, without the addition of nitrate and nitrite. All trials were monitored during the ripening (13 °C and 90% relative humidity) extended until 45 d. The evolution of physicochemical parameters showed that beef and pork salamis were characterized by a higher content of branched chain fatty acids (FA) and rumenic acid than horse and wild boar salamis, whereas the last two productions showed higher values of secondary lipid oxidation. Plate counts showed that lactic acid bacteria (LAB), yeasts and coagulase-negative staphylococci (CNS) populations dominated the microbial community of all productions with Lactobacillus and Staphylococcus as most frequently isolated bacteria. The microbial diversity evaluated by MiSeq Illumina showed the presence of members of Gammaproteobacteria phylum, Moraxellaceae family, Acinetobacter, Pseudomonas, Carnobacterium and Enterococcus in all salamis. This study showed the natural evolution of indigenous fermented meat starter cultures and confirmed a higher suitability of horse and beef meat for nitrate/nitrite free salami production due to their hygienic quality at 30 d.

The present work was carried out to investigate the Antimicrobial Resistance (AMR) of enterococci isolated from raw ewes’ and cows’ milk. The samples were collected from eighteen semi-extensive dairy sheep and cow farms throughout western Sicily. Plate counts, carried out on Rapid Enterococcus Agar commonly used to detect food enterococci, revealed a maximal enterococcal concentration of approximately 4.58 Log Colony Forming Unit (CFU)/mL. Colonies were isolated and differentiated based on genetic analysis by Randomly Amplified Polymorphic DNA (RAPD)-PCR. Thirty-eight different strains were identified. Analysis by a species-specific multiplex PCR assay grouped the strains into three Enterococcus species such as Enterococcus durans, Enterococcus faecalis and Enterococcus faecium. The 38 strains were also investigated for their antimicrobial resistance by a phenotypic approach. All 38 Enterococcus displayed resistance to at least one or more of the antimicrobials tested confirmed that the dairy enterococci could be a vector for the dissemination of antimicrobial resistance. This work showed that enterococci with AMR traits are commonly present in semiextensive dairy sheep and cow farms of western Sicily pointed out the relevance of informing dairy makers and veterinary regarding the antimicrobial use in order to mitigate problems of public health and veterinary medicine.

An innovative ovine cheese enriched with red grape pomace powder (GPP) was produced to improve the functional properties of Vastedda cheese typology. Vastedda cheese making was performed adding GPP and four selected Lactococcus lactis strains (Mise36, Mise94, Mise169 and Mise190). For each strain, 40 L of pasteurized ewe’s milk was divided into two aliquots representing control and experimental trials. Control cheese (CC) production did not contain GPP, while the experimental cheese (EC) production was enriched with 1% (w/w) GPP. GPP did not slow down starter development and acid generation. Plate counts and randomly amplified polymorphic DNA (RAPD)-PCR analysis confirmed the dominance of the starters in all trials. The evolution of the physicochemical parameters showed that EC productions were characterized by lower fat content, higher protein content, and higher values of secondary lipid oxidation. Sensory evaluation indicated that the cheeses produced with the strain Mise94 were those more appreciated by the judges. Thus, the last cheeses were investigated for some functional aspects: GPP enrichment significantly increased antioxidant activity and lipoperoxyl radical scavenger capacity, confirming that grape polyphenol inclusion in cheese represents an optimal strategy for the valorization of ovine cheeses as well as winemaking industry by-products.

Background: The continuous worldwide increase of consumption of fresh mushrooms has registered in the recent years. The major goal of this study was to determine the microbiological characteristics of wild edible mushrooms and effect of temperature during storage of Morchella conica. Methods: Wild mushrooms of the species Boletus edulis, Cantharellus cibarius, and Leccinum aurantiacum were collected in a mixed forest of Picea abies, Betula pendula, and Pinus sylvestris located in Tartu territory, Estonia. Faecal indicators, potential pathogens, spoilage bacteria, and microfungi (yeasts and moulds) were evaluated. M. conica was microbiologically investigated for 24 days under different thermal regimes, including 4, 8, 12, 15, 20, and 28 °C. The statistical analysis was conducted with SAS 9.2 software. Results: The microbial counts of wild mushrooms, ranging from 6.81 to 7.68 log10 CFU/g for total mesophilic count, were generally higher (p